Oct. 2005, 発酵と代謝研究奨励金, 細胞内コエンザイムAの動態と代謝調節機構の解明, バイオインダストリー協会

Complete genome sequence of Pseudoalteromonas sp. PS1M3, a psychrotrophic bacterium isolated from deep-sea sediment off the Boso Peninsula, Japan Trench
Yoshihito Nikaidou; Yong Guo; Mahoko Taguchi; Shigeru Chohnan; Tomoyasu Nishizawa; Yasurou Kurusu, Elsevier
Marine Genomics, Jun. 2023, [Reviewed]

〔Major achievements〕Enhanced supply of acetyl-CoA by exogenous pantothenate kinase promotes synthesis of poly(3-hydroxybutyrate)
Hirotaka Kudo; Sho Ono; Kenta Abe; Mami Matsuda; Tomohisa Hasunuma; Tomoyasu Nishizawa; Munehiko Asayama; Hirofumi Nishihara; Shigeru Chohnan, Corresponding, BMC
Microbial Cell Factories, 20 Apr. 2023, [Reviewed]

Complete genome sequence of a psychrophilic bacterium, Pseudoalteromonas sp. strain APM04, isolated from the seafloor of the South Mariana Trough, Pacific Ocean
Mahoko Taguchi; Yong Guo; Tomoyasu Nishizawa; Shigeru Chohnan; Yasurou Kurusu, The complete genome sequence of Pseudoalteromonas sp. strain APM04, which is a psychrophilic bacterium that inhabits the seabed of the South Mariana Trough, Pacific Ocean, was determined to characterize the genetic features associated with evolution in extremophilic and oligotrophic deep seawater., American Society for Microbiology
Microbiology Resource Announcements, 27 Jul. 2022, [Reviewed]

〔Major achievements〕Fatty acid production by enhanced malonyl‑CoA supply in Escherichia coli
Moena Kaku; Mei Ishidaira; Shusaku Satoh; Miho Ozaki; Daisuke Kohari; Shigeru Chohnan, Corresponding, Springer Nature
Current Microbiology, 26 Jul. 2022, [Reviewed]

〔Major achievements〕Enhancement of fatty acid biosynthesis by exogenous acetyl-CoA carboxylase and pantothenate kinase in Escherichia coli
Shusaku Satoh; Miho Ozaki; Saki Matsumoto; Takumi Nabatame; Moena Kaku; Takashi Shudo; Munehiko Asayama; Shigeru Chohnan, Corresponding, Springer Nature
Biotechnology Letters, Dec. 2020, [Reviewed]

Sensing of nutrients by CPT1C controls SAC1 activity to regulate AMPA receptor trafficking
Maria Casas; Rut Fadó; José L. Domínguez; Aina Roig; Moena Kaku; Shigeru Chohnan; Montse Solé; Mercedes Unzeta; Alfredo Jesús Miñano-Molina; José Rodríguez-Álvarez; Eamonn James Dickson; Núria Casals, Rockefeller University Press
Journal of Cell Biology, 05 Oct. 2020, [Reviewed]

〔Major achievements〕Coenzyme A and its thioester pools in obese Zucker and Zucker diabetic fatty rats
Shigeru Chohnan; Shiori Matsuno; Kei Shimizu; Yuka Tokutake; Daisuke Kohari; Atsushi Toyoda, Lead, Feeding behavior is closely related to hypothalamic malonyl-CoA level in the brain and diet-induced obesity affects total CoA pools in liver. Herein, we performed a comprehensive analysis of the CoA pools formed in thirteen tissues of Zucker and Zucker diabetic fatty (ZDF) rats. Hypothalamic malonyl-CoA levels in obese rats remained low and were almost the same as those of lean rats, despite obese rats having much higher content of leptin, insulin, and glucose in their sera. Regardless of the fa-genotypes, larger total CoA pools were formed in the livers of ZDF rats and the size of hepatic total CoA pools in Zucker rats showed almost one tenth of the size of ZDF rats. The decreased total CoA pool sizes in Zucker rats was observed in the brown adipose tissues, while ZDF-fatty rats possessed 6% of total CoA pool in the lean rats in response to fa deficiency. This substantially lower CoA content in the obese rats would be disadvantageous to non-shivering thermogenesis. Thus, comparing the intracellular CoA behaviors between Zucker and ZDF rats, as well as the lean and fatty rats of each strain would help to elucidate features of obesity and type 2 diabetes in combination with result (s) of differential gene expression analysis and/or comparative genomics., MDPI
Nutrients, 06 Feb. 2020, [Reviewed]

Efficient butanol recovery from acetone-butanol-ethanol fermentation cultures grown on sweet sorghum juice by pervaporation using silicalite-1 membrane
Miho Kanemoto; Hideyuki Negishi; Keiji Sakaki; Toru Ikegami; Shigeru Chohnan; Youji Nitta; Yasurou Kurusu; Hiroyuki Ohta, We investigated butanol recovery by pervaporation separation, using a silicalite-1 membrane, from batch cultures of butanol-producing Clostridium beijerincicii SBP2 grown on sweet sorghum juice as a fermentation medium. The pervaporation system yielded 73% (w/v) butanol from intact feed cultures containing 1% (w/v) butanol, and had a butanol permeation flux of 11 g m(-2) h(-1). Upon neutralization and activated charcoal treatment of the feed cultures, butanol yield and total flux increased to 82% (w/v) and 40 g m(-2) h(-1), respectively. This system is applicable to refining processes for practical biobutanol production from a promising energy crop, sweet sorghum. (C) 2015 The Society for Biotechnology, Japan. All rights reserved., SOC BIOSCIENCE BIOENGINEERING JAPAN
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, Jun. 2016, [Reviewed]

Decreased hepatic contents of coenzyme A molecular species in mice after subchronic mild social defeat stress
Yoshifumi Kubota; Tatsuhiko Goto; Yuki Hagiya; Shigeru Chohnan; Atsushi Toyoda, Social stress may precipitate psychiatric disorders such as depression, which is related to the occurrence of the metabolic syndrome, including obesity and type 2 diabetes. We have evaluated the effects of social stress on central and peripheral metabolism using a model of depression in mice. In the present study, we focused on coenzyme A (CoA) molecular species [i.e. non-esterified CoA (CoASH), acetyl-CoA and malonyl-CoA] which play important roles in numerous metabolic pathways, and we analyzed changes in expression of these molecules in the hypothalamus and liver of adult male mice (C57BL/6J) subjected to 10 days of subchronic mild social defeat stress (sCSDS) with ICR mice as aggressors. Mice (n=12) exposed to showed hyperphagia- and polydipsia-like symptoms and increased body weight gain compared with control mice which were not affected by exposure to ICR mice (n=12). To elucidate the underlying metabolic features in the sCSDS model, acetyl-CoA, malonyl-CoA and CoASH tissue levels were analyzed using the acyl-CoA cycling method. The levels of hypothalamic malonyl-CoA, which decreases feeding behavior, were not influenced by sCSDS. However, sCSDS reduced levels of acetyl-CoA, malonyl-CoA and total CoA (sum of the three CoA molecular species) in the liver. Hence, hyperphagia-like symptoms in sCSDS mice evidently occurred independently of hypothalamic malonyl-CoA, but might consequently lead to down-regulation of hepatic CoA via altered expression of nudix hydrolase 7. Future studies should investigate the molecular mechanism(s) underlying the down-regulation of liver CoA pools in sCSDS mice., TAYLOR & FRANCIS LTD
STRESS-THE INTERNATIONAL JOURNAL ON THE BIOLOGY OF STRESS, Mar. 2016, [Reviewed]

Genetic engineering and metabolite profiling for overproduction of polyhydroxybutyrate in cyanobacteria
Sayaka Hondo; Masatoshi Takahashi; Takashi Osanai; Mami Matsuda; Tomohisa Hasunuma; Akio Tazuke; Yoichi Nakahira; Shigeru Chohnan; Morifumi Hasegawa; Munehiko Asayama, Genetic engineering and metabolite profiling for the overproduction of polyhydroxybutyrate (PHB), which is a carbon material in biodegradable plastics, were examined in the unicellular cyanobacterium Synechocystis sp. PCC 6803. Transconjugants harboring cyanobacterial expression vectors that carried the pha genes for PHB biosynthesis were constructed. The overproduction of PHB by the engineering cells was confirmed through microscopic observations using Nile red, transmission electron microscopy (TEM), or nuclear magnetic resonance (NMR). We successfully recovered PHB from transconjugants prepared from nitrogen-depleted medium without sugar supplementation in which PHB reached approximately 7% (w/w) of the dry cell weight, showing a value of 12-fold higher productivity in the transconjugant than that in the control strain. We also measured the intracellular levels of acetyl-CoA, acetoacetyl-CoA, and 3-hydroxybutyryl-CoA (3HB-CoA), which are intermediate products for PHB. The results obtained indicated that these products were absent or at markedly low levels when cells were subjected to the steady-state growth phase of cultivation under nitrogen depletion for the overproduction of bioplastics. Based on these results, efficient factors were discussed for the overproduction of PHB in recombinant cyanobacteria. (C) 2015, The Society for Biotechnology, Japan. All rights reserved., SOC BIOSCIENCE BIOENGINEERING JAPAN
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, Nov. 2015, [Reviewed]

Effects of chronic mild food restriction on behavior and the hypothalamic malonyl-CoA signaling pathway
Wataru Iio; Yuka Tokutake; Hiroaki Koike; Noriko Matsukawa; Takamitsu Tsukahara; Shigeru Chohnan; Atsushi Toyoda, Depression induces anorexia, leading to suppressed feeding behaviors and energy intake. Previously, we revealed that chronic social defeat induced a mild suppression of feeding in rats with elevated levels of hypothalamic malonyl-CoA which regulates feeding. Therefore, we attempted to elucidate the effects of chronic mild food restriction on behavior and on hypothalamic malonyl-CoA. The chronic mild food restricted rats were fed a restricted diet approximately 80% to 90% amount of diet compared to the control for 5 weeks. Ratios of restriction were adjusted with feed consumption in the chronic social defeat stressed rats. Chronic mild food restricted rats exhibited a suppression of body weight gain similar to that of the chronic social defeat stressed rats. Also these rats showed increased time spent in the center area of an open field (OF), prolonged immobility time in forced swim, increased phosphorylation of hypothalamic adenosine monophosphate-activated protein kinase (AMPK) and acetyl-CoA carboxylase and a decreased concentration of hypothalamic malonyl-CoA. Weight of the adrenal glands, locomotion in an OF, mitogen-activated protein kinase cascade and calcium/calmodulin-dependent protein kinases II in the hippocampus were not affected by chronic mild food restriction. Our findings suggest that chronic mild food restriction activates AMPK following a decreased hypothalamic malonyl-CoA., WILEY-BLACKWELL
ANIMAL SCIENCE JOURNAL, Feb. 2015, [Reviewed]

Prokaryotic type III pantothenate kinase enhances coenzyme A biosynthesis in Escherichia coli
Yuta Ogata; Shigeru Chohnan, Corresponding, MICROBIOL RES FOUNDATION
JOURNAL OF GENERAL AND APPLIED MICROBIOLOGY, 2015, [Reviewed]

Antimicrobial Activity of Pantothenol against Staphylococci Possessing a Prokaryotic Type II Pantothenate Kinase
Shigeru Chohnan; Misa Murase; Kota Kurikawa; Kodai Higashi; Yuta Ogata, Lead, Pantothenol is a provitamin of pantothenic acid (vitamin B5) that is widely used in healthcare and cosmetic products. This analog of pantothenate has been shown to markedly inhibit the phosphorylation activity of the prokaryotic type II pantothenate kinase of Staphylococcus aureus, which catalyzes the first step of the coenzyme A biosynthetic pathway. Since type II enzymes are found exclusively in staphylococci, pantothenol suppresses the growth of S. aureus, S. epidermidis, and S. saprophyticus, which inhabit the skin of humans. Therefore, the addition of this provitamin to ointment and skincare products may be highly effective in preventing infections by opportunistic pathogens., JAPANESE SOC MICROBIAL ECOLOGY, DEPT BIORESOURCE SCIENCE
MICROBES AND ENVIRONMENTS, Jul. 2014, [Reviewed]

Effects of chronic social defeat stress on peripheral leptin and its hypothalamic actions
Wataru Iio; Haruyoshi Takagi; Yasuki Ogawa; Takamitsu Tsukahara; Shigeru Chohnan; Atsushi Toyoda, Background: Suppression of body weight and symptom of anorexia are major symptoms of depression. Recently, we reported that chronic social defeat stress (CSDS) induced suppression of body weight gain and anorexic feeding behavior in rats. These abnormalities were the result of disrupted malonyl-coenzyme A (CoA) signaling pathway in the hypothalamus. However, the condition of peripheral leptin and its hypothalamic downstream signal molecules which regulate hypothalamic malonyl-CoA level in the CSDS-exposed rats (CSDS rats) is still unknown.
Results: CSDS rats showed suppressed body weight gain and food intake. The weight of the CSDS rats' epididymal white adipose tissues was decreased when compared to the control rats. The plasma cholesterol concentration was decreased significantly in the CSDS rats compared to the control rats (P < 0.05). The plasma glucose concentration was slightly decreased in the CSDS rats compared to the control rats (P < 0.1). The expression of leptin mRNA in epididymal white adipose tissues and the plasma leptin concentration were decreased in CSDS rats. Furthermore, the phosphorylation of the hypothalamic downstream signals of leptin, including extracellular signal-regulated kinase 1/2 (ERK1/2) and signal transducer and activator of transcription 3 (STAT3), was decreased in CSDS rats.
Conclusions: Our results indicated that decreased peripheral leptin expression in CSDS rats could down-regulate the hypothalamic downstream signaling pathways of leptin while suppressed food intake. These data indicate that CSDS induces the down-regulation of hypothalamic AMPK following the elevation of hypothalamic malonyl-CoA levels and is independent of peripheral leptin and glucose., BIOMED CENTRAL LTD
BMC NEUROSCIENCE, Jun. 2014, [Reviewed]

Long-Term Increased Carnitine Palmitoyltransferase 1A Expression in Ventromedial Hypotalamus Causes Hyperphagia and Alters the Hypothalamic Lipidomic Profile
Paula Mera; Joan Francesc Mir; Gemma Fabrias; Josefina Casas; Ana S. H. Costa; Maria Ida Malandrino; Jose-Antonio Fernandez-Lopez; Xavier Remesar; Su Gao; Shigeru Chohnan; Maria Sol Rodriguez-Pena; Harald Petry; Guillermina Asins; Fausto G. Hegardt; Laura Herrero; Dolors Serra, Lipid metabolism in the ventromedial hypothalamus (VMH) has emerged as a crucial pathway in the regulation of feeding and energy homeostasis. Carnitine palmitoyltransferase (CPT) 1A is the rate-limiting enzyme in mitochondrial fatty acid beta-oxidation and it has been proposed as a crucial mediator of fasting and ghrelin orexigenic signalling. However, the relationship between changes in CPT1A activity and the intracellular downstream effectors in the VMH that contribute to appetite modulation is not fully understood. To this end, we examined the effect of long-term expression of a permanently activated CPT1A isoform by using an adeno-associated viral vector injected into the VMH of rats. Peripherally, this procedure provoked hyperghrelinemia and hyperphagia, which led to overweight, hyperglycemia and insulin resistance. In the mediobasal hypothalamus (MBH), long-term CPT1AM expression in the VMH did not modify acyl-CoA or malonyl-CoA levels. However, it altered the MBH lipidomic profile since ceramides and sphingolipids increased and phospholipids decreased. Furthermore, we detected increased vesicular gamma-aminobutyric acid transporter (VGAT) and reduced vesicular glutamate transporter 2 (VGLUT2) expressions, both transporters involved in this orexigenic signal. Taken together, these observations indicate that CPT1A contributes to the regulation of feeding by modulating the expression of neurotransmitter transporters and lipid components that influence the orexigenic pathways in VMH., PUBLIC LIBRARY SCIENCE
PLOS ONE, May 2014, [Reviewed]

Role of prokaryotic type I and III pantothenate kinases in the coenzyme A biosynthetic pathway of Bacillus subtilis
Yuta Ogata; Hiroki Katoh; Munehiko Asayama; Shigeru Chohnan, Corresponding, kinases (CoaAs) catalyze the phosphorylation of pantothenate in the first step of the coenzyme A (CoA) biosynthetic pathway. These bacterial enzymes have been categorized into 3 types, the prokaryotic type I, II, and III CoaAs. Bacteria typically carry a single CoaA gene on their genome, but Bacillus subtilis possesses 2 proteins homologous to type I and III CoaAs, known as BsCoaA and BsCoaX, respectively. Both recombinant proteins exhibited the expected kinase activity and the characteristic properties of type I and III CoaAs, i.e., regulation by CoASH and acyl-CoAs in BsCoaA and the requirement of a monovalent cation in BsCoaX. Both gene disruptants appeared to grow in a manner similar to the wild-type strain. With the BsCoaX disruptant, the BsCoaA had the ability to completely fill the intracellular CoA pool, whereas the BsCoaA disruptant did not. These findings clearly indicate that these 2 CoaAs are employed together in the CoA biosynthetic pathway in B. subtilis and that the contribution of the type I CoaA (BsCoaA) to the formation of the intracellular CoA pool is larger than that of the type III CoaA (BsCoaX)., CANADIAN SCIENCE PUBLISHING, NRC RESEARCH PRESS
CANADIAN JOURNAL OF MICROBIOLOGY, May 2014, [Reviewed]

The importance of tissue environment surrounding the tumor on the development of cancer cachexia
Fumihiro Chiba; Kuniyasu Soda; Shigeki Yamada; Yuka Tokutake; Shigeru Chohnan; Fumio Konishi; Toshiki Rikiyama, The relationship between host factors and cancer cachexia was investigated. A single cell clone (clone 5 tumor) established from colon 26 adenocarcinoma by limiting dilution cell cloning methods was employed to eliminate the inoculation site-dependent differences in the composition of cell clones. Clone 5 tumor did not provoke manifestations of cancer cachexia when inoculated in subcutaneous tissue. However, when inoculated in the gastrocnemius muscle, the peritoneal cavity or the thoracic cavity of CD2F1 male mice, typical manifestations of cancer cachexia were observed in all groups of mice with intergroup variations. The blood levels of various cytokines, chemokines and hormones were increased but with wide intergroup variations. Analyses by stepwise multiple regression models revealed that serum interleukin-10 was the most significant factor associated with manifestations of cancer cachexia, suggesting the possible involvement of mechanisms similar to cancer patients suffering cancer cachexia. White blood cells, especially neutrophils, seemed to have some roles on the induction of cancer cachexia, because massive infiltrations and an increase in peripheral blood were observed in cachectic mice bearing clone 5 tumors. The amount of malonyl-CoA in liver correlated with manifestations of cancer cachexia, however the mRNA levels of spermidine/spermine N-1 acetyl transferase (SSAT) (of which overexpression has been shown to provoke manifestations similar to cancer cachexia) were not necessarily associated with cancer cachexia. These data suggest that the induction of cancer cachexia depends on the environment in which the tumor grows and that the infiltration of host immune cells into the tumor and the resultant increase in inflammation result in the production of cachectic factors, such as cytokines, leading to SSAT activation. Further, multiple factors likely mediate the mechanisms of cancer cachexia. Finally, this animal model was suitable for the investigation of the mechanisms involved in cachexia of cancer patients., SPANDIDOS PUBL LTD
INTERNATIONAL JOURNAL OF ONCOLOGY, Jan. 2014, [Reviewed]

Marked phenotypic differences of endurance performance and exercise-induced oxygen consumption between AMPK and LKB1 deficiency in mouse skeletal muscle: changes occurring in the diaphragm
Shinji Miura; Yuko Kai; Miki Tadaishi; Yuka Tokutake; Kimitoshi Sakamoto; Clinton R. Bruce; Mark A. Febbraio; Kiyoshi Kita; Shigeru Chohnan; Osamu Ezaki, LKB1 phosphorylates members of the AMP-activated protein kinase (AMPK) family. LKB1 and AMPK in the skeletal muscle are believed to regulate not only fuel oxidation during exercise but also exercise capacity. LKB1 was also required to prevent diaphragm fatigue, which was shown to affect exercise performance. Using mice expressing dominant negative (DN) mutants of LKB1 and AMPK, specifically in the skeletal muscle but not in the heart, we investigated the roles of LKB1 and AMPK activity in exercise performance and the effects of these kinases on the characteristics of respiratory and locomotive muscles. In the diaphragm and gastrocnemius, both AMPK-DN and LKB1-DN mice showed complete loss of AMPK alpha 2 activity, and LKB1-DN mice showed a reduction in LKB1 activity. Exercise capacity was significantly reduced in LKB1-DN mice, with a marked reduction in oxygen consumption and carbon dioxide production during exercise. The diaphragm from LKB1-DN mice showed an increase in myosin heavy chain IIB and glycolytic enzyme expression. Normal respiratory chain function and CPT I activity were shown in the isolated mitochondria from LKB1-DN locomotive muscle, and the expression of genes related to fiber type, mitochondria function, glucose and lipid metabolism, and capillarization in locomotive muscle was not different between LKB1-DN and AMPK-DN mice. We concluded that LKB1 in the skeletal muscle contributes significantly to exercise capacity and oxygen uptake during exercise. LKB1 mediated the change of fiber-type distribution in the diaphragm independently of AMPK and might be responsible for the phenotypes we observed., AMER PHYSIOLOGICAL SOC
AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM, Jul. 2013, [Reviewed]

Identification of pantoate kinase and phosphopantothenate synthetase from Methanospirillum hungatei
Hiroki Katoh; Hideyuki Tamaki; Yuka Tokutake; Satoshi Hanada; Shigeru Chohnan, Corresponding, Pantothenate synthetase (PanC) and pantothenate Kinase which function in the canonical coenzyme A (CoA) biosynthetic pathway cannot be found in most archaea. COG1829 and COG1701 intrinsic to archaea were proposed as the candidate proteins for producing 4'-phosphopantothenate instead, and the COG1701 protein from Methanosarcina mazei was assigned as PanC. Meanwhile, the Thermococcus kodakarensis COG1829 and COG1701 proteins were biochemically identified as novel enzymes, i.e., pantoate kinase (PoK) and phosphopantothenate synthetase (PPS). In this study, the functions of Mhun_0831 (COG1829) and Mhun_0832 (COG1701) from Methanospirillum hungatei were identified, and the recombinant enzymes were partially characterized. Plasmids simultaneously possessing the two genes encoding Mhun_0831 and Mhun_0832 complemented the poor growth of the temperature-sensitive Escherichia coli pantothenate Kinase mutant ts9. The recombinant Mhun_0831 and Mhun_0832 expressed in E. coli cells exhibited PoK and PPS activities, respectively, being in accord with the functions of T. kodakarensis proteins. The PoK activity was most active at pH 8.5 and 40 degrees C, and accepted ATP and UTP as a phosphate donor. Although CoA did not affect the PoK activity, the end product considerably accelerated the PPS activity. The homologs of both proteins are widely conserved in most archaeal genomes. Taken together, our findings indicate that archaea can synthesize CoA through the unique pathway involving PoK and PPS, in addition to the canonical one that the order Thermoplasmatales employs. (C) 2012, The Society for Biotechnology, Japan. All rights reserved., SOC BIOSCIENCE BIOENGINEERING JAPAN
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, Apr. 2013, [Reviewed]

Effect of diet composition on coenzyme A and its thioester pools in various rat tissues
Yuka Tokutake; Wataru Iio; Naoki Onizawa; Yuta Ogata; Daisuke Kohari; Atsushi Toyoda; Shigeru Chohnan, Corresponding, Three coenzyme A (CoA) molecular species, i.e., acetyl-CoA, malonyl-CoA, and nonesterified CoA (CoASH), in 13 types of fasted rat tissue were analyzed. A relatively larger pool size of total CoA, consisting of acetyl-CoA, malonyl-CoA, and CoASH, was observed in the medulla oblongata, liver, heart, and brown adipose tissue. Focusing on changes in the CoA pool size in response to the nutrient composition of the diet given, total CoA pools in rats continuously fed a high-fat diet for 4 weeks were significantly higher in the hypothalamus, cerebellum, and kidney, and significantly lower in the liver and skeletal muscle than those of rats fed a high-carbohydrate or high-protein diet. In particular, reductions in the liver were remarkable and were caused by decreased CoASH levels. Consequently, the total CoA pool size was reduced by approximately one-fifth of the hepatic contents of rats fed the other diets. In the hypothalamus, which monitors energy balance, all three CoA molecular species measured were at higher levels when rats were fed the high-fat diet. Thus, it was of interest that feeding rats a high-fat diet affected the behaviors of CoA pools in the hypothalamus, liver, and skeletal muscle, suggesting a significant relationship between CoA pools, especially malonyl-CoA and/or CoASH pools, and lipid metabolism in vivo. (C) 2012 Elsevier Inc. All rights reserved., ACADEMIC PRESS INC ELSEVIER SCIENCE
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, Jul. 2012, [Reviewed]

Anorexic behavior and elevation of hypothalamic malonyl-CoA in socially defeated rats
Wataru Iio; Yuka Tokutake; Noriko Matsukawa; Takamitsu Tsukahara; Shigeru Chohnan; Atsushi Toyoda, Suppression of body weight and eating disorders, such as anorexia, are one of the major symptoms of psychiatric disorders such as depression. However, the mechanisms of weight loss and reduced appetite in depressive patients and in animal models of depression are largely unknown. In this study, we characterized the mechanism of anorexia resulting from depression using socially defeated rats as an animal model of depression. Socially defeated rats showed suppressed body weight gain, enlarged adrenal glands, decreased home cage activity, decreased food intake, and increased immobility in the forced swim test. These results are representative of some of the core symptoms of depression. Simultaneously, we observed decreased levels of phosphorylated AMP-activated protein kinase (AMPK) and acetyl-coenzyme A (CoA) carboxylase (ACC) and increased levels of malonyl-CoA in the hypothalamus of socially defeated rats. Hypothalamic malonyl-CoA controlled feeding behavior and elevation of malonyl-CoA in the hypothalamus induced inhibition of food intake. Our findings suggest that the suppression of body weight gain caused by social defeat stress is caused by anorexic feeding behavior via an increased concentration of malonyl-CoA in the hypothalamus. (C) 2012 Elsevier Inc. All rights reserved., ACADEMIC PRESS INC ELSEVIER SCIENCE
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, May 2012, [Reviewed]

Fuel ethanol production from sweet sorghum using repeated-batch fermentation
Shigeru Chohnan; Megumi Nakane; M. Habibur Rahman; Youji Nitta; Takanori Yoshiura; Hiroyuki Ohta; Yasurou Kurusu, Lead, Ethanol was efficiently produced from three varieties of sweet sorghum using repeated-batch fermentation without pasteurization or acidification. Saccharomyces cerevisiae cells could be recycled in 16 cycles of the fermentation process with good ethanol yields. This technique would make it possible to use a broader range of sweet sorghum varieties for ethanol production. (C) 2010, The Society for Biotechnology, Japan. All rights reserved., SOC BIOSCIENCE BIOENGINEERING JAPAN
JOURNAL OF BIOSCIENCE AND BIOENGINEERING, Apr. 2011, [Reviewed]

Coenzyme A and its thioester pools in fasted and fed rat tissues
Yuka Tokutake; Naoki Onizawa; Hiroki Katoh; Atsushi Toyoda; Shigeru Chohnan, Corresponding, Levels of three coenzyme A (CoA) molecular species, i.e., nonesterified CoA (CoASH), acetyl-CoA, and malonyl-CoA, in fasted and fed rat tissues were analyzed by the acyl-CoA cycling method which makes detection possible at the pmol level. Malonyl-CoA in brain tissues readily increased with feeding, and inversely, acetyl-CoA decreased. This phenomenon occurred in the cerebral cortex, hippocampus, cerebellum, and medulla oblongata, as well as in the hypothalamus which controls energy balance by monitoring malonyl-CoA. In the non-brain tissues, the sizes of the acetyl-CoA, malonyl-CoA, and CoASH pools depended on the tissues. The total CoA pools consisting of the above three CoA species in the liver, heart, and brown adipose tissue were larger and those of the perirenal, epididymal, and ovarian adipose tissues were much smaller, compared with those of other tissues including brain tissues. In addition, the response of each CoA pool to feeding was not uniform, suggesting that the tissue-specific metabolism individually functions in the non-brain tissues. Thus, a comprehensive analysis of thirteen types of rat tissue revealed that CoA pools have different sizes and showed a different response to fasting and feeding depending on the tissue. (C) 2010 Elsevier Inc. All rights reserved., ACADEMIC PRESS INC ELSEVIER SCIENCE
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, Nov. 2010, [Reviewed]

Pantothenate Kinase from the Thermoacidophilic Archaeon Picrophilus torridus
Masakazu Takagi; Hideyuki Tamaki; Yukiko Miyamoto; Roberta Leonardi; Satoshi Hanada; Suzanne Jackowski; Shigeru Chohnan, Corresponding, Pantothenate kinase (CoaA) catalyzes the first step of the coenzyme A (CoA) biosynthetic pathway and controls the intracellular concentrations of CoA through feedback inhibition in bacteria. An alternative enzyme found in archaea, pantoate kinase, is missing in the order Thermoplasmatales. The PTO0232 gene from Picrophilus torridus, a thermoacidophilic euryarchaeon, is shown to be a distant homologue of the prokaryotic type I CoaA. The cloned gene clearly complements the poor growth of the temperature-sensitive Escherichia coli CoaA mutant strain ts9, and the recombinant protein expressed in E. coli cells transfers phosphate to pantothenate at pH 5 and 55 C. In contrast to E. coli CoaA, the P. torridus enzyme is refractory to feedback regulation by CoA, indicating that in P. torridus cells the CoA levels are not regulated by the CoaA step. These data suggest the existence of two subtypes within the class of prokaryotic type I CoaAs., AMER SOC MICROBIOLOGY
JOURNAL OF BACTERIOLOGY, Jan. 2010, [Reviewed]

Decrease in malonyl-CoA and its background metabolic alterations in murine model of cancer cachexia
Alper Celik; Yoshihiko Kano; Shingo Tsujinaka; Sinichirou Okada; Koichi Takao; Masakazu Takagi; Shigeru Chohnan; Kuniyasu Soda; Masanobu Kawakami; Fumio Konishi, The alterations of enzymatic activities involved in lipid degradation in cancer cachexia have not been fully elucidated. One of the two subclones of colon 26 adenocarcinoma, clone 20, with a potent ability to induce cachexia, or clone 5, without such an activity, was transplanted in to CDF-1 male mice. Murine livers were extirpated for analyses on the 14th day after tumor inoculation. The body weights and food intake of mice bearing clone 20 were all significantly lower than those of non-tumor bearing mice and mice bearing the clone 5 tumor. The decline of body weight was accompanied by a shrinkage of epididymal fat pads. Expression of spermidine/spermine N-1 acetyl transferase (SSAT) assessed by real-time PCR was significantly increased in cachectic mice. Conversely, acetyl-CoA carboxylase (ACC) measured by Western blotting and malonyl-CoA levels determined by malonyl-CoA:acetyl-CoA cycling procedures were decreased in cachectic mice. Indomethacin in drinking water reversed the clone 20 induced decrease in body and fat weight and food intake, and simultaneously negated the clone 20 induced increase of SSAT expressions and decrease of ACC and malonyl-CoA amounts. Because malonyl-CoA inhibits the rate-limiting step in the beta-oxidation of fatty acids, the decreased malonyl-CoA and the background metabolic alterations may contribute to the accelerated lipolysis of cancer cachexia., SPANDIDOS PUBL LTD
Oncology Reports, Apr. 2009, [Reviewed]

〔Major achievements〕Fasting-induced hypothermia and reduced energy production in mice lacking acetyl-CoA synthetase 2
Iori Sakakibara; Takahiro Fujino; Makoto Ishii; Toshiya Tanaka; Tatsuo Shimosawa; Shinji Miura; Wei Zhang; Yuka Tokutake; Joji Yamamoto; Mutsumi Awano; Satoshi Iwasaki; Toshiyuki Motoike; Masashi Okamura; Takeshi Inagaki; Kiyoshi Kita; Osamu Ezaki; Makoto Naito; Tomoyuki Kuwaki; Shigeru Chohnan; Tokuo Yamamoto; Robert E. Hammer; Tatsuhiko Kodama; Masashi Yanagisawa; Juro Sakai, Acetate is activated to acetyl-CoA by acetyl-CoA synthetase 2 (AceCS2), a mitochondrial enzyme. Here, we report that the activation of acetate by AceCS2 has a specific and unique role in thermogenesis during fasting. In the skeletal muscle of fasted AceCS2(-/-) mice, ATP levels were reduced by 50% compared to AceCS2(+/+) mice. Fasted AceCS2(-/-) mice were significantly hypothermic and had reduced exercise capacity. Furthermore, when fed a low-carbohydrate diet, 4-week-old weaned AceCS2(-/-) mice also exhibited hypothermia accompanied by sustained hypoglycemia that led to a 50% mortality. Therefore, AceCS2 plays a significant role in acetate oxidation needed to generate ATP and heat. Furthermore, AceCS2(-/-) mice exhibited increased oxygen consumption and reduced weight gain on a low-carbohydrate diet. Our findings demonstrate that activation of acetate by AceCS2 plays a pivotal role in thermogenesis, especially under low-glucose or ketogenic conditions, and is crucially required for survival., CELL PRESS
Cell Metabolism, 04 Feb. 2009, [Reviewed]

〔Major achievements〕Differential effects of central fructose and glucose on hypothalamic malonyl-CoA and food intake
Seung Hun Cha; Michael Wolfgang; Yuka Tokutake; Shigeru Chohnan; M. Daniel Lane, The American diet, especially that of adolescents, contains highly palatable foods of high-energy content and large amounts of high-fructose sweeteners. These factors are believed to contribute to the obesity epidemic and insulin resistance. Previous investigations revealed that the central metabolism of glucose suppresses food intake mediated by the hypothalamic AMP-kinase/malonyl-CoA signaling system. Unlike glucose, centrally administered fructose increases food intake. Evidence presented herein indicates that the more rapid initial steps of central fructose metabolism deplete hypothalamic ATP level, whereas the slower regulated steps of glucose metabolism elevate hypothalamic ATP level. Consistent with effects on the [ATP]/[AMP] ratio, fructose increases phosphorylation/activation of hypothalamic AMP kinase causing phosphorylation/inactivation of acetyl-CoA carboxylase, whereas glucose has the inverse effects. The changes provoked by central fructose administration reduce hypothalamic malonyl-CoA level and thereby increase food intake. These findings explain the paradoxical fructose effect on food intake and lend credence to the malonyl-CoA hypothesis., NATL ACAD SCIENCES
Proceedings of the National Academy of Science USA, 04 Nov. 2008, [Reviewed]

〔Major achievements〕Regulation of hypothalamic malonyl-CoA by central glucose and leptin
Michael J. Wolfgang; Seung Hun Cha; Aniket Sidhave; Shigeru Chohnan; Gary Cline; Gerald I. Shulman; M. Daniel Lane, Hypothalamic malonyl-CoA has been shown to function in global energy homeostasis by modulating food intake and energy expenditure. Little is known, however, about the regulation of malonyl-CoA concentration in the central nervous system. To address this issue we investigated the response of putative intermediates in the malonyl-CoA pathway to metabolic and endocrine cues, notably those provoked by glucose and leptin. Hypothalamic malonyl-CoA rises in proportion to the carbohydrate content of the diet consumed after food deprivation. Malonyl-CoA concentration peaks 1 h after refeeding or after peripheral glucose administration. This response depends on the dose of glucose administered and is blocked by the i.c.v. administration of an inhibitor of glucose metabolism, 2-deoxyglucose (2-DG). The kinetics of change in hypothalamic malonyl-CoA after glucose administration is coincident with the suppression of phosphorylation of AMP kinase and acetyl-CoA carboxylase. Blockade of glucose utilization in the CNS by i.c.v. 2-DG prevented the effects of glucose on 5'AMP-activated protein kinase, malonyl-CoA, hypothalamic neuropeptide expression, and food intake. Finally, we showed that leptin can increase hypothalamic malonyl-CoA and that the increase is additive with glucose administration. Leptin-deficient ob/ob mice, however, showed no defect in the glucose- or refeeding-induced rise in hypothalamic malonyl-CoA after food deprivation, demonstrating that leptin was not required for this effect. These studies show that hypothalamic malonyl-CoA responds to the level of circulating glucose and leptin, both of which affect energy homeostasis., NATL ACAD SCIENCES
Proceedings of the National Academy of Science USA, 04 Dec. 2007, [Reviewed]

〔Major achievements〕Leptin activates hypothalamic acetyl-CoA carboxylase to inhibit food intake
Su Gao; Kimberly P. Kinzig; Susan Aja; Karen A. Scott; Wendy Keung; Sandra Kelly; Ken Strynadka; Shigeru Chohnan; Wanli W. Smith; Kellie L.K. Tamashiro; Ellen E. Ladenheim; Gabriele V. Ronnett; Yajun Tu; Morris J. Birnbaum; Gary D. Lopaschuk; Timothy H. Moran, Hypothalamic fatty acid metabolism has recently been implicated in the controls of food intake and energy homeostasis. We report that intracerebroventricular (ICV) injection of leptin, concomitant with inhibiting AMP-activated kinase (AMPK), activates acetyl-CoA carboxylase (ACC), the key regulatory enzyme in fatty acid biosynthesis, in the arcuate nucleus (Arc) and paraventricular nucleus (PVN) in the hypothalamus. Arc overexpression of constitutively active AMPK prevents the Arc ACC activation in response to ICV leptin, supporting the hypothesis that AMPK lies upstream of ACC in leptin's Arc intracellular signaling pathway. Inhibiting hypothalamic ACC with 5-tetradecyloxy-2-furoic acid, a specific ACC inhibitor, blocks leptin-mediated decreases in food intake, body weight, and mRNA level of the orexigenic neuropeptide NPY. These results show that hypothalamic ACC activation makes an important contribution to leptin's anorectic effects. Furthermore, we find that ICV leptin up-regulates the level of malonyl-CoA (the intermediate of fatty acid biosynthesis) specifically in the Arc and increases the level of palmitoyl-CoA (a major product of fatty acid biosynthesis) specifically in the PVN. The rises of both levels are blocked by 5-tetradecyloxy-2-furoic acid along with the blockade of leptin-mediated hypophagia. These data suggest malonyl-CoA as a downstream mediator of ACC in leptin's signaling pathway in the Arc and imply that palmitoyl-CoA, instead of malonyl-CoA, could be an effector in relaying ACC signaling in the PVN. Together, these findings highlight site-specific impacts of hypothalamic ACC activation in leptin's anorectic signaling cascade., NATL ACAD SCIENCES
Proceedings of the National Academy of Science USA, 30 Oct. 2007, [Reviewed]

〔Major achievements〕Characterization of N-deoxyribosyltransferase from Lactococcus lactis subsp. lactis
Yukiko Miyamoto; Takeharu Masaki; Shigeru Chohnan, Corresponding, A nucleoside N-deoxyribosyltransferase-homologous gene was detected by homological search in the genomic DNA of Lactococcus lactis subsp. lactis. The gene yejD is composed of 477 nucleotide encoding 159 amino acids with only 25% identity, which is low in comparison to the amino acid sequences of the N-deoxyribosyltransferases from other lactic acid bacteria, i.e. Lactobacillus leichmannii and Lactobacillus helveticus. The residues responsible for catalytic and substrate-binding sites in known enzymes are conserved at Gln49, Asp73, Asp93 (or Asp95), and Glu 10 1, respectively. The recombinant YejD expressed in Escherichia coli shows a 2-deoxyribosyl transfer activity to and from both bases of purine and pyrimidine, showing that YejD should be categorized as a class 11 N-deoxyribosyltransferase. Interestingly, the base-exchange activity as well as the heat stability of YejD was enhanced by the presence of monovalent cations such as K+, NH+, and Rb+, indicating that the Lactococcus enzyme is a K+-activated Type II enzyme. However, divalent cations including Mg2+ and Ca2+ significantly inhibit the activity. Whether or not the yejD gene product actually participates in the nucleoside salvage pathway of Lc. lactis remains unclear, but the lactic acid bacterium possesses the gene coding for the nucleoside N-deoxyribosyltransferase activated by K+ on its genome. (C) 2007 Elsevier B.V. All rights reserved., ELSEVIER SCIENCE BV
Biochimica et Biophysica Acta, Oct. 2007, [Reviewed]

〔Major achievements〕Chemical knockout of pantothenate kinase reveals the metabolic and genetic program responsible for hepatic coenzyme A homeostasis
Yong-Mei Zhang; Shigeru Chohnan; Kristopher G. Virga; Robert D. Stevens; Olga R. Ilkayeva; Brett R. Wenner; James R. Bain; Christopher B. Newgard; Richard E. Lee; Charles O. Rock; Suzanne Jackowski, Coenzyme A (CoA) is the major acyl group carrier in intermediary metabolism. Hopantenate (HoPan), a competitive inhibitor of the pantothenate kinases, was used to chemically antagonize CoA biosynthesis. HoPan dramatically reduced liver CoA and mice developed severe hypoglycemia. Insulin was reduced, glucagon and corticosterone were elevated, and fasting accelerated hypoglycemia. Metabolic profiling revealed a large increase in acylcarnitines, illustrating the role of carnitine in buffering acyl groups to maintain the nonesterified CoASH level. HoPan triggered significant changes in hepatic gene expression that substantially increased the thioesterases, which liberate CoASH from acyl-CoA, and increased pyruvate dehydrogenase kinase 1, which prevents the conversion of CoASH to acetyl-CoA. These results identify the metabolic rearrangements that maintain the CoASH pool which is critical to mitochondrial functions, including gluconeogenesis, fatty acid oxidation, and the tricarboxylic acid and urea cycles., CELL PRESS
Chemistry & Biology, Mar. 2007, [Reviewed]

〔Major achievements〕Hypothalamic malonyl-CoA triggers mitochondrial biogenesis and oxidative gene expression in skeletal muscle: role of PGC-1a
Seung-Hun Cha; Joseph T. Rodgers; Pere Puigserver; Shigeru Chohnan; M.Daniel Lane, Previous investigations show that intracerebroventricular administration of a potent inhibitor of fatty acid synthase, C75, increases the level of its substrate, malonyl-CoA, in the hypothalamus. The "malonyl-CoA signal" is rapidly transmitted to skeletal muscle by the sympathetic nervous system, increasing fatty acid oxidation, uncoupling protein-3 (UCP3) expression, and thus, energy expenditure. Here, we show that intracerebroventricular or intraperitoneal administration of C75 increases the number of mitochondria in white and red (soleus) skeletal muscle'. Consistent with signal transmission from the hypothalamus by the sympathetic nervous system, centrally administered C75 rapidly (:52 h) up-regulated the expression (in skeletal muscle) of the beta-adrenergic signaling molecules, i.e., norepinephrine, beta(3)-adrenergic receptor, and cAMP; the transcriptional regulators peroxisomal proliferator activator regulator gamma coactivator 1 alpha (PGC-1 alpha) and estrogen receptor-related receptor alpha (ERR alpha); and the expression of key oxidative mitochondrial enzymes, including pyruvate clehydrogenase kinase, mediumchain length fatty acyl-CoA clehydrogenase, ubiquinone-cytochrome c reductase, cytochrome oxidase, as well as ATP synthase and UCP3. The role of PGC-1 alpha in mediating these responses in muscle was assessed with C2C12 myocytes in cell culture. Consistent with the in vivo response, adenovirus-directed expression of PGC-1 alpha in C2C12 muscle cells provoked the phosphorylation/ inactivation and reduced expression of acetyl-CoA carboxylase 2, causing a reduction of the malonyl-CoA concentration. These effects, coupled with an increased carnitine palmitoyltransferase 1b, led to increased fatty acid oxidation. PGC-1 a also increased the expression of ERR alpha, PPAR alpha, and enzymes that support mitochondrial fatty acid oxidation, ATP synthesis, and thermogenesis, apparently mediated by an increased expression of lUCP3., NATL ACAD SCIENCES
Proceedings of the National Academy of Science USA, 17 Oct. 2006, [Reviewed]

Purification, characterization, and gene analysis of cellulase (Cel8A) from Lysobacter sp. IB-9374
Jiro Ogura; Atsushi Toyoda; Taisuke Kurosawa; Ai Leng Chong; Shigeru Chohnan; Takeharu Masaki, An enzyme that has both beta-1,4-glucanase and chitosanase activities was found in the culture medium of the soil bacterium Lysobacter sp. IB-9374, a high lysyl endopeptidase-producing strain. The enzyme was purified to homogeneity from the culture filtrate using five purification steps and designated Cel8A. The purified Cel8A had a molecular mass of 41 kDa, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A pH optimum of 5.0 was found for the beta-1,4-glucanase activity, and pH optima of 5.0 and 7.0 were found for the chitosanase activity. Nucleotide sequencing of the Ce18A gene yielded a deduced amino acid sequence that comprises a 33-amino acid, N-terminal signal peptide and a mature enzyme consisting of a 381-residue polypeptide with a predicted molecular mass of 41,241 Da. The amino acid sequence of the Cel8A, which contains the catalytic module of glycosyl hydrolase family 8, is homologous to beta-1,3-1,4-D-glucanase from Bacillus circulans WL-12 and endoglucanase N-257 from B. circulans KSM-N257., TAYLOR & FRANCIS LTD
Bioscience, Biotechnology, and Biochemistry, 07 Oct. 2006, [Reviewed]

〔Major achievements〕Prokaryotic type II and type III pantothenate kinases: the same monomer fold creates dimers with distinct catalytic properties
Bum Soo Hong; Mi Kyung Yun; Yong-Mei Zhang; Shigeru Chohnan; Charles O. Rock; Stephen W. White; Suzanne Jackowski; Hee-Won Park; Roberta Leonardi, Three distinct isoforms of pantothenate kinase (CoaA) in bacteria catalyze the first step in coenzyme A biosynthesis. The structures of the type 11 (Staphylococcus aureus, SaCoaA) and type III (Pseudomonas aeruginosa, PaCoaA) enzymes reveal that they assemble nearly identical subunits with actin-like folds into dimers that exhibit distinct biochemical properties. PaCoaA has a fully enclosed pantothenate binding pocket and requires a monovalent cation to weakly bind ATP in an open cavity that does not interact with the adenine nucleotide. Pantothenate binds to an open pocket in SaCoaA that strongly binds ATP by using a classical P loop architecture coupled with specific interactions with the adenine moiety. The PaCoaA(.)Pan binary complex explains the resistance of bacteria possessing this isoform to the pantothenamide antibiotics, and the similarity between SaCoaA and human pantothenate kinase 2 explains the molecular basis for the development of the neurodegenerative phenotype in three mutations in the human protein., CELL PRESS
Structure, Aug. 2006, [Reviewed]

〔Major achievements〕A role for hypothalamic malonyl-CoA in the control of food intake
Zhiyuan Hu; Yun Dai; Marc Prentki; Shigeru Chohnan; M. Daniel Lane, The cellular level of malonyl-CoA, an intermediate in fatty acid biosynthesis, depends on its rate of synthesis catalyzed by acetyl-CoA carboxylase relative to its rate of utilization and degradation catalyzed by fatty acid synthase and malonyl-CoA decarboxylase, respectively. Recent evidence suggests that hypothalamic malonyl-CoA functions in the regulation of feeding behavior by altering the expression of key orexigenic and anorexigenic neuropeptides. Here we report that 5-aminoimidazole-4-carboxamide ribonucleoside ( AICAR), a 5'-AMP kinase activator, rapidly lowers malonyl-CoA both in GT1-7 hypothalamic neurons and in the hypothalami of mice. These effects correlate closely with the phosphorylation of acetyl-CoA carboxylase, an established target of AMP kinase. Intracerebroventricular (i.c.v.)administration of AICAR rapidly lowers hypothalamic [malonyl-CoA] and increases food intake. Expression of an adenoviral cytosolic malonyl-CoA decarboxylase vector ( Ad-cMCD) in hypothalamic GT1-7 cells decreases malonyl-CoA. When delivered by bilateral stereotaxic injection into the ventral hypothalamus ( encompassing the arcuate nucleus) of mice, Ad-cMCD increases food intake and body weight. Ad-MCD delivered into the ventral hypothalamus also reverses the rapid suppression of food intake caused by i.c.v.-administered C75, a fatty acid synthase inhibitor that increases hypothalamic [malonyl-CoA]. Taken together these findings implicate malonyl-CoA in the hypothalamic regulation of feeding behavior., AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
The Journal of Biological Chemistry, 02 Dec. 2005, [Reviewed]

〔Major achievements〕Inhibition of hypothalamic fatty acid synthase trigers rapid activation of fatty acid oxidation in skeletal muscle
Seung Hun Cha; Zhiyuan Hu; Shigeru Chohnan; M. Daniel Lane, Malonyl-CoA functions as a mediator in the hypothalamic sensing of energy balance and regulates the neural physiology that governs feeding behavior and energy expenditure. The central administration of C75, a potent inhibitor of the fatty acid synthase (FAS), increases malonyl-CoA concentration in the hypothalamus and suppresses food intake while activating fatty acid oxidation in skeletal muscle. Closely correlated with the increase in muscle fatty acid oxidation is the phosphorylation/inactivation of acetyl-CoA carboxylase, which leads to reduced malonyl-CoA concentration. Lowering muscle malonyl-CoA, a potent inhibitor of carnitine/ palmitoyl-CoA transferase 1 (CPT1), releases CPT1 from inhibitory constraint, facilitating the entry of fatty acids into mitochondria for 13 oxidation. Also correlated with these events are C75-induced increases in the expression of skeletal muscle peroxisome proliferator-activated receptor alpha (PPAR alpha), a transcriptional activator of fatty acid oxidizing enzymes, and uncoupling protein 3 (UCP3), a thermogenic mitochondrial uncoupling protein. Phentolamine, an a-adrenergic blocking agent, prevents the C75-induced increases of skeletal muscle UCP3 and whole body fatty acid oxidation and C75-induced decrease of skeletal muscle malonyl-CoA. Thus, the sympathetic nervous system is implicated in the transmission of the "malonyl-CoA signal" from brain to skeletal muscle. Consistent with the up-regulation of UCP3 and PPARa is the concomitant increase in the expression of PGC1 alpha, transcriptional coactivator of the UCP3 and PPAR alpha-activated genes. These findings clarify the mechanism by which the hypothalamic malonyl-CoA signal is communicated to metabolic systems in skeletal muscle that regulate fatty acid oxidation and energy expenditure., NATL ACAD SCIENCES
Proceedings of the National Academy of Science USA, 11 Oct. 2005, [Reviewed]

〔Major achievements〕A pantothenate kinase from Staphylococcus aureus refractory to feedback regulation by coenzyme A
Roberta Leonardi; Shigeru Chohnan; Yong-Mei Zhang; Kristopher G. Virga; Richard E.Lee; Charls O. Rock; Suzanne Jackowski, The key regulatory step in CoA biosynthesis in bacteria and mammals is pantothenate kinase (CoaA), which governs the intracellular concentration of CoA through feedback regulation by CoA and its thioesters. CoaA from Staphylococcus aureus (SaCoaA) has a distinct primary sequence that is more similar to the mammalian pantothenate kinases than the prototypical bacterial CoaA of Escherichia coli. In contrast to all known pantothenate kinases, SaCoaA activity is not feedback-regulated by CoA or CoA thioesters. Metabolic labeling of S. aureus confirms that CoA levels are not controlled by CoaA or at steps downstream from CoaA. The pantothenic acid antimetabolite N-heptylpantothenamide (N7-Pan) possesses potent antimicrobial activity against S. aureus and has multiple cellular targets. N7-Pan is a substrate for SaCoaA and is converted to the inactive butyldethia-CoA analog by the downstream pathway enzymes. The analog is also incorporated into acyl carrier protein and D-alanyl carrier protein, the prosthetic groups of which are derived from CoA. The inactivation of acyl carrier protein and the cessation of fatty acid synthesis are the most critical causes of growth inhibition by N7-Pan because the toxicity of the drug is ameliorated by supplementing the growth medium with fatty acids. The absence of feedback regulation at the pantothenate kinase step allows the accumulation of high concentrations of intracellular CoA, consistent with the physiology of S. aureus, which lacks glutathione and relies on the CoA/CoA disulfide reductase redox system for protection from oxidative damage., AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
The Journal of Biological Chemistry, 04 Feb. 2005, [Reviewed]

〔Major achievements〕A second lysine-specific serine protease from Lysobacter sp. strain IB-9374
Shigeru Chohnan; Kentaro Shiraki; Kiyonobu Yokota; Makoto Ohshima; Natsuki Kuroiwa; Kashfia Ahmed; Takeharu Masaki; Fumio Sakiyama, Lead, A second lysyl endopeptidase gene (lepB) was found immediately upstream of the previously isolated lepA gene encoding a highly active lysyl endopeptidase in Lysobacter genomic DNA. The lepB gene consists of 2,034 nucleotides coding for a protein of 678 amino acids. Amino acid sequence alignment between the lepA and lepB gene products (LepA and LepB) revealed that the LepB precursor protein is composed of a prepeptide (20 amino acids [aa]), a propepticle (184 aa), a mature enzyme (274 aa), and a C-terminal extension peptide (200 aa). The mature enzyme region exhibited 72% sequence identity to its LepA counterpart and conserved all essential amino acids constituting the catalytic triad and the primary determining site for lysine specificity. The lepB gene encoding the propeptide and mature-enzyme portions was overexpressed in Escherichia coli, and the inclusion body produced generated active LepB through appropriate refolding and processing. The purified enzyme, a mature 274-aa lysine-specific endopeptidase, was less active and more sensitive to both temperature and denaturation with urea, guanidine hydrochloride, or sodium dodecyl sulfate than LepA. LepA-based modeling implies that LepB can fold into essentially the same three-dimensional structure as LepA by placing a peptide segment, composed of several inserted amino acids found only in LepB, outside the molecule and that the Tyr169 side chain occupies the site in which the indole ring of Trp169, a built-in modulator for unique peptidase functions of LepA, resides. The results suggest that LepB is an isozyme of LepA and probably has a tertiary structure quite similar to it., AMER SOC MICROBIOLOGY
Journal of Bacteriology, Aug. 2004, [Reviewed]

〔Major achievements〕Hypothalamic malonyl-CoA as a mediator of feeding behavior
Zhiyuan Hu; Seung Hun Cha; Shigeru Chohnan; M Daniel Lane, Previous studies showed that i.p. administration of C75, a potent inhibitor of fatty acid synthase (FAS), blocked fasting-induced up-regulation of orexigenic neuropeptides and down-regulation of anorexigenic neuropeptides in the hypothalami of mice. As a result, food intake and body weight were drastically reduced. Here we provide evidence supporting the hypothesis that hypothalamic malonyl-CoA, a substrate of FAS, is an indicator of global energy status and mediates the feeding behavior of mice. We use a sensitive recycling assay to quantify malonyl-CoA to show that the hypothalamic malonyl-CoA level is low in fasted mice and rapidly (less than or equal to2 h) increases (approximate to5-fold) on refeeding. Intracerebroventricular (i.c.v.) administration of C75 to fasted mice rapidly (less than or equal to2 h) increased (by 4-fold) hypothalamic malonyl-CoA and blocked feeding when the mice were presented with food. Moreover, prior i.c.v. administration of an acetyl-CoA carboxylase inhibitor, 5-(tetradecyloxy)-2-furoic acid, rapidly (although only partially) prevented the C75-induced rise of hypothalamic malonyl-CoA and prevented the C75-induced decrease of food intake. These effects correlated closely with the rapid (less than or equal to2 h) and reciprocal effects of i.c.v. C75 on the expression of hypothalamic orexigenic (NPY and AgRP) and anorexigenic (proopiomelanocortin) neuropeptide mRNAs. Previous results showing that C75 administered i.c.v. rapidly activates hypothalamic neurons of the arcuate and paraventricular nuclei are consistent with the results reported in this paper. Together these findings suggest that level of hypothalamic malonyl-CoA, which depends on the relative activities of acetyl-CoA carboxylase and FAS, is an indicator of energy status and mediates feeding behavior., NATL ACAD SCIENCES
Proceedings of the National Academy of Science USA, 28 Oct. 2003, [Reviewed]

Purification, bacteriolytic activity, and specificity of beta-lytic protease from Lysobacter sp. IB-9374
Kashfia Ahmed; Shigeru Chohnan; Hiroyuki Ohashi; Takeshi Hirata; Takeharu Masaki; Fumio Sakiyama, Lysobacter sp. IB-9374, which was isolated from soil as a high lysyl endopeptidase-producing strain (Chohnan et al., FEMS Microbiol. Lett., 213, 13-20, 2002), was found to produce a beta-lytic protease capable of lysing gram-positive bacteria such as Staphylococcus aureus, Microccocus luteus, and Bacillus subtilis. The Lysobacter strain secreted the beta-lytic protease into the culture medium at a 2.4-fold higher level than Achromobacter lyticus. The enzyme was highly purified through a series of six steps with a high yield. The enzyme was strongly inhibited by tetraethylenepentamine and 1,10-phenanthroline. The purified enzyme lysed more efficiently almost all the grain-positive bacteria tested than lysozyme, lysostaphin, and mutanolysin. The enzyme was very similar to Achromobacter beta-lytic protease containing one zinc atom in terms of amino acid composition and N-terminal sequence. The nucleotide sequence revealed that the mature enzyme was composed of 179 amino acid residues with additional 198 amino acids at the amino-terminal end of the enzyme. The deduced amino acid sequence of the mature enzyme coincided with that of the Achromobacter enzyme, although the prepro-region showed a 41% sequence identity with the counterpart. These results indicate that Lysobacter sp. is a useful strain for an efficient large-scale preparation of beta-lytic protease capable of lysing bacteria., SOC BIOSCIENCE BIOENGINEERING JAPAN
Journal of Bioscience and Bioengineering, Jan. 2003, [Reviewed]

〔Major achievements〕Functions of malonate decarboxylase subunits from Pseudomonas putida
Shigeru Chohnan; Kayo Akagi; Yoshichika Takamura, Lead, Malonate decarboxylase from Pseudomonas putida is composed of five subunits, alpha, beta, gamma, delta, and epsilon. Two subunits, delta and epsilon, have been identified as an acyl-carrier protein (ACP) and malonyl-CoA:ACP transacylase, respectively. Functions of the other three subunits have not been identified, because recombinant subunits expressed in Escherichia coli formed inclusion bodies. To resolve this problem, we used a coexpression system with GroEL/ES from E. coli, and obtained active recombinant subunits. Enzymatic analysis of the purified recombinant subunits showed that the alpha subunit was an acetyl-S-ACP:malonate ACP transferase and that the betagamma-subunit complex was a malonyl-S-ACP decarboxylase., TAYLOR & FRANCIS LTD
Bioscience, Biotechnology, and Biochemistry, Jan. 2003, [Reviewed]

〔Major achievements〕Lysobacter strain with high lysyl endopeptidase production
Shigeru Chohnan; Junko Nonaka; Kousei Teramoto; Kouichi Taniguchi; Yuuko Kameda; Hitoshi Tamura; Yasurou Kurusu; Shigemi Norioka; Takeharu Masaki; Fumio Sakiyama, Lead, A new lysyl endopeptidase producing strain, Lysobacter sp. IB-9374, was isolated from soil. This strain secreted the endopeptidase to culture medium at 6-12-fold higher levels relative to Achromobacter lyticus and Lysobacter enzymogenes. The mature Lysobacter sp. enzyme was enzymatically identical to Achromobacter lysyl endopeptidase bearing lysyl bond specificity, a high peptidase activity, a wide pH optimum, and stability against denaturants. Nucleotide sequence analysis of the Lysobacter sp. lysyl endopeptidase gene revealed that the enzyme is synthesized as a precursor protein consisting of signal peptide (20 amino acids (aa)), pro-peptide (185 aa), mature enzyme (268 aa), and C-terminal extension peptide (198 aa). The deduced amino acid sequence of the mature enzyme was totally identical to that of the Achromobacter enzyme. The Lysobacter sp. precursor protein has an 18-aa longer peptide chain following nine consecutive amino acid residues distinct from the Achromobacter counterpart at the C-terminus. Total precursor protein is 671 aa of which only 268 aa are in the finally processed exoenzyme. (C) 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Microbiological Societies., ELSEVIER SCIENCE BV
FEMS Microbiology Letters, 16 Jul. 2002, [Reviewed]

〔Major achievements〕Cloning and characterization of mdc genes encoding malonate decarboxylase from Pseudomonas putida
Shigeru Chohnan; Yasurou Kurusu; Hirofumi Nishihara; Yoshichika Takamura, Lead, The DNA fragment encoding malonate decarboxylase, involved in malonate assimilation, was cloned from Pseudomonas putida. The Il-kb DNA fragment contained nine open reading frames, which were designated mdcABCDEGHLM in the given order. N-terminal protein sequencing established that the mdcA, mdcC, mdcD, mdcE and mdcH genes encoded subunits alpha, delta, beta, gamma and epsilon Of the malonate decarboxylase, respectively. Malonate decarboxylase was functionally expressed in Escherichia coli from plasmid harboring the entire gene cluster or the mdc genes lacking the mdcL and mdcM genes. The mdcL and mdcM genes encode membrane proteins and disruption of the genes of P. putida by the insertion of a kanamycin resistance cassette reduced the malonate uptake activity of the organism. Thus, we conclude that MdcLM is a malonate transporter. (C) 1999 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved., ELSEVIER SCIENCE BV
FEMS Microbiology Letters, 15 May 1999, [Reviewed]

〔Major achievements〕Malonate decarboxylase of Pseudomonas putida is composed of five subunits
Shigeru Chohnan; Tooru Fujio; Toshikazu Takaki; Masami Yonekura; Hirofumi Nishihara; Yoshichika Takamura, Lead, Two different forms of malonate decarboxylase were purified from Pseudomonas putida. The active form was composed of the five different subunits alpha (60 kDa), beta (33 kDa), gamma (28 kDa), delta (13 kDa), and epsilon (30 kDa) and the inactive form was composed of the four subunits lacking the epsilon subunit. The former catalyzed the decarboxylation of malonate to acetate, but the latter could not, although it retained both activities of acetyl-CoA:malonate CoA transferase and malonyl-CoA decarboxylase. The delta subunit of the active form was acylated by the incubation with [2-C-14]malonyl-CoA, but the delta subunit of the inactive form was not labeled. From the above results and the N-terminal amino acid sequence analysis, it was concluded that the epsilon subunit was an essential subunit to function as malonyl-CoA:ACP transacylase, which was an indispensable component of the enzyme for the cyclic decarboxylation of malonate. (C) 1998 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved., ELSEVIER SCIENCE BV
FEMS Microbiology Letters, 01 Dec. 1998, [Reviewed]

〔Major achievements〕Chages in size of intracellular pools of coenzyme A and its thioesters in Escherichia coli K-12 cells to various carbon sources and stresses
Shigeru Chohnan; Hiroaki Izawa; Hirofumi Nishihara; Yoshichika Takamura, Lead, Intracellular pools of three CoA molecular species of coenzyme A, CoASH, acetyl-CoA, and malonyl-CoA, in Escherichia coli K-12 cells were studied by acyl-CoA cycling method in replacement culture. The sizes and compositions of CoA pools starved for a carbon source changed within minutes after the addition of one of various carbon sources. A large acetyl-CoA pool formed after the addition of D-glucose, D-fructose, D-mannose, glycerol, or sorbitol, but there was little change when L-glucose, sucrose, maltose, succinate, or acetate was added, The beta-anomer of D-glucose was assimilated 10 times faster than the alpha-anomer, Intracellular CoA pools also changed with stress: in the pH, incubation temperature, or with osmotic stress. The sizes and compositions of CoA pools were not affected by pH changing between 4 and 8, but the breakdown of acetyl-CoA and CoASH was greater at pH 9 than at pH 4 to 8. Production of acetyl-CoA was greatest at 40 degrees C, and at 50 degrees C, an acetyl-CoA pool did not form at all and the size of the CoASH pool declined. When the organism was stressed by the addition of NaCl at concentrations of more than 0.6 M, little acetyl-CoA was produced. The total CoA pool (the sum of the concentrations of CoASH, acetyl-CoA, and malonyl-CoA) remained within the limits of 0.83-1.40 nmol/mg of dry cell weight (0.30-0.52 mM). Whenever acetyl-CoA increased, CoASH decreased. Therefore, the acetyl-CoA/CoASB ratio is an important index of facultative anaerobes that reflects the state of carbon and energy metabolism in vivo., TAYLOR & FRANCIS LTD
Bioscience, Biotechnology, and Biochemistry, Jun. 1998, [Reviewed]

〔Major achievements〕Changes in the size and composition of intracellular pools of nonesterified coenzyme A and coenzyme A thioesters in aerobic and facultatively anaerobic bacteria
Shigeru Chohnan; Hideki Furukawa; Tooru Fujio; Hirofumi Nishihara; Yoshichika Takamura, Lead, Intracellular levels of three coenzyme A (CoA) molecular species, i.e., nonesterified CoA (CoASH), acetyl-CoA, and malonyl-CoA, in a variety of aerobic and facultatively anaerobic bacteria were analyzed by the acyl-CoA cycling method developed by us, It was demonstrated that there was an intrinsic difference between aerobes and facultative anaerobes in the changes in the size and composition of CoA pools, The CoA pools in the aerobic bacteria hardly changed and were significantly smaller than those of the facultatively anaerobic bacteria, On the other hand, in the facultatively anaerobic bacteria, the size and composition of the CoA pool drastically changed within minutes in response to the carbon and energy source provided, Acetyl-CoA was the major component of the CoA pool in the facultative anaerobes grown on sufficient glucose, although CoASH was dominant in the aerobes, Therefore, the acetyl-CoA/CoASH ratios in facultatively anaerobic bacteria were 10 times higher than those in aerobic bacteria, In Escherichia coli K-12 cells, the addition of reagents to inhibit the respiratory system led to a rapid decrease in the amount of acetyl-CoA with a concomitant increase in the amount of CoASH, whereas the addition of cerulenin, a specific inhibitor of fatty acid synthase, triggered the intracellular accumulation of malonyl-CoA. The acylation and deacylation of the three CoA molecular species coordinated with the energy-yielding systems and the restriction of the fatty acid-synthesizing system of cells, These data suggest that neither the accumulation of acetyl-CoA nor that of malonyl-CoA exerts negative feedback on pyruvate dehydrogenase and acetyl-CoA carboxylase, respectively., AMER SOC MICROBIOLOGY
Applied and Environmental Microbiology, Feb. 1997, [Reviewed]

〔Major achievements〕Investigation of substrate-binding sites of cyclic-ribonucleotide phosphomutase-5'-phosphodiesterase using substrate analogues
Shigeru Chohnan; Yuichirou Midorikawa, Lead, The substrate-binding sites of a bifunctional enzyme, cyclic-ribonucleotide phosphomutase-5'-phosphodiesterase from Aspergillus niger, FS-44, were investigated by kinetic studies. A series of adenine nucleotides' inhibited the mutase and diesterase reactions, but adenosine and adenine did not at all. Adenosine monophosphate derivatives acted as competitive inhibitors with respect to both adenosine 3',5'-cyclic monophosphate in the mutase reaction and adenosine 5'-p-nitrophenyl phosphate in the diesterase reaction with the same decreasing order of inhibition constant. However, the inhibition constants of these compounds in the mutase reaction were 1 order of magnitude lower than those in the diesterase reaction, These results suggest that the active site(s) on the enzyme catalyzing the mutase and diesterase reactions have similar properties, but the substrate-binding sites of both reactions are distinct at least., TAYLOR & FRANCIS LTD
Bioscience, Biotechnology, and Biochemistry, Jun. 1995, [Reviewed]

〔Major achievements〕Purification and characterization of a novel bifunctional enzyme, cyclic-ribonucleotide phosphomutase-5'-phosphodiesterase, from Aspergillus niger
Shigeru Chohnan; Takashi Nagata; Yuichirou Midorikawa, Lead, A 3',5'-cyclic-ribonucleotide was enzymatically converted into a corresponding 2',3'-cyclic-ribonucleotide by a culture extract of 5'-phosphodiesterase (5'-PDase; EC 3.1.4.1) hyperproductive mutant of Aspergillus niger, FS-44. Cyclic-ribonucleotide phosphomutase (cRNPMase) activity catalyzing the preceding reaction was not separated from the 5'-PDase activity during the purification steps. The enzyme was purified to the homogeneity of a single polypeptide with a molecular weight of 62,000. The purified enzyme specifically catalyzed the conversion of 3',5'-cyclic-ribonucleotides to corresponding 2',3'-cyclic-ribonucleotides. The decreasing order of reaction rate was as follows: adenosine 3',5'-cyclic monophosphate > inosine 3',5'-cyclic monophosphate > uridine 3',5'-cyclic monophosphate > guanosine 3',5'-cyclic monophosphate > cytidine 3',5'-cyclic monophosphate. The enzyme did not act on 2'-AMP, 3'-AMP, or 5'-AMP. The optimum pH of cRNPMase activity was pH 3.0, while that of 5'-PDase activity was pH 5.6. Both activities was inhibited by diisopropyl flurophosphate, Fe2+, and Fe3+. While 5'-PDase activity was significantly inhibited by Zn2+ and Cu2+, cRNPMase activity was net at all. The Michaelis constant of cRNPMase was estimated to be 3.72 mM far adenosine 3',5'-cyclic monophosphate and that of 5'-PDase was 0.96 mM for adenosine 5'-p-nitrophenyl phosphate. The enzyme was a glycoprotein containing N-linked sugar chains. The isoelectric point was 4.8. This bifunctional enzyme was named cyclic-ribonucleotide phosphomutase-5'-phosphodiesterase (cRNPMase-5'-PDase)., TAYLOR & FRANCIS LTD
Bioscience, Biotechnology, and Biochemistry, Feb. 1994, [Reviewed]

〔Major achievements〕A Simple micromethod for measurement of CoASH and its use in measuring intracellular levels of CoASH and short chain acyl-CoAs in Escherichia coli K12 cells
Shigeru Chohnan; Yoshichika Takamura, Lead, A simple micromethod for measurment of CoASH was established. CoASH was quantitatively converted to acetyl-CoA with phosphate acetyltransferase (EC 2.3.1.8), then acetyl-CoA originated from CoASH was measuring by the acyl-CoA cycling method using malonate CoA-transferase (EC 2.8.3.3). This method is highly sensitive and can detect 10(-12) mol of CoASH without any loss of CoA-thioesters, and it is specific for CoASH and 3'-dephospho-CoA. The established method was useful to define the rapid changes in the size and composition of in vivo CoA pool (CoASH, acetyl-CoA, and malonyl-CoA) in Escherichia coli K12. It was found that the CoA pool changed rapidly and dramatically in a minute order in ranges of CoASH; 40-300-mu-M (0.11-0.80 nmol/mg dry wt.), acetyl-CoA; 40-350-mu-m (0.12-0.93 nmol/mg dry wt.), and malonyl-CoA; 0-290-mu-M (0-0.79 nmol/mg dry wt.). When glucose in the medium was depleted, CoASH increased to be the major component (82%) of the CoA pool, while when sufficient amount of glucose (28 mM) was provided, CoASH was reduced to a minor component (15%). In contrast with this, acetyl-CoA increased to the maximal level of 350-mu-M (0.93 nmol/mg dry wt.) and to be the major component (66%) 10 min after 28 mM glucose was added. A remarkable demonstration was achieved in confirmation of the reverse relationship between CoASH and short chain acyl-CoAs, particularly acetyl-CoA, of which the in vivo behavior has been mere conjecture until now., JAPAN SOC BIOSCI BIOTECHN AGROCHEM
Agricultural and Biological Chemistry, Jan. 1991, [Reviewed]

日本微生物生態学会第30回大会・第7回JTKシンポジウムの報告(大会開催報告)
太田 寛行; 西澤 智康; 長南 茂
日本微生物生態学会誌, 2016

いばらきBooks 10 茨城大学発 持続可能な世界へ　　　　　　　　　　　　　　　
長南茂, Others
茨城新聞社, 10 Oct. 2010

〔Major achievements〕土壌環境からの新たな水素酸化細菌の探索と分離　　　　　　　　　　　　　　　
寺原 直矢; 今泉 尚志; 入江 敬太; 長南 茂; 西原 宏史
日本農芸化学会2025年度（令和7年度）［札幌］大会, 07 Mar. 2025, 日本農芸化学会
20250304, 20250308

〔Major achievements〕大腸菌でのコエンザイムAとアシルキャリアプロテイン増産が脂肪酸生産に及ぼす影響
吉見 昴、小口 桜、長南 茂
日本農芸化学会 関東支部 2024年度 支部大会, 30 Aug. 2024, 日本農芸化学会 関東支部
20240830, 20240830

〔Major achievements〕Cupriavidus necator H16由来原核Ⅲ型パントテン酸キナーゼの諸性質
入野 晃郁、長南 茂
日本農芸化学会 関東支部 2024年度 支部大会, 30 Aug. 2024, 日本農芸化学会 関東支部
20240830, 20240830

〔Major achievements〕Cupriavidus necator H16由来パントテン酸キナーゼの酵素学的諸性質
入野 晃郁、濱田 美志、長南 茂
第18回日本ゲノム微生物学会年会, 12 Mar. 2024, 日本ゲノム微生物学会
20240312, 20240314

〔Major achievements〕Effects of fabR and fadR on fatty acid accumulation in E. coli biosynthesis　　　　　　　　　　　　　　　
中川満美子、田中真大、長南茂
日本農芸化学会2023年度大会, 15 Mar. 2023, 日本農芸化学会
20230314, 20230317

〔Major achievements〕Free fatty acid production in engineered Escherichia coli with enhancement of malonyl-CoA biosynthesis　　　　　　　　　　　　　　　
濱田美志、中平洋一、西澤智康、長南茂
第74回日本生物工学会大会, 20 Oct. 2022, 日本生物工学会
20221017, 20221020

〔Major achievements〕強化されたCoA生合成による組換え大腸菌でのPHB生産　　　　　　　　　　　　　　　
工藤大嵩、朝山宗彦、西原宏史、長南茂
日本農芸化学会2022年度大会, 17 Mar. 2022, 日本農芸化学会
20220315, 20220318

〔Major achievements〕大腸菌遺伝子破壊株におけるマロニルｰCoA増産が脂肪酸生産に及ぼす影響　　　　　　　　　　　　　　　
加来萌菜; 長南茂
日本農芸化学会 関東支部 2020年度大会, 28 Nov. 2020, 日本農芸化学会 関東支部
20201128, 20201128

〔Major achievements〕大腸菌のコエンザイムA増産株を用いたポリヒドロキシ酪酸生産　　　　　　　　　　　　　　　
小野渉; 阿部健太; 朝山宗彦; 西原宏史; 長南茂
日本微生物生態学会第33回大会, 11 Sep. 2019, 日本微生物生態学会

〔Major achievements〕脂肪酸生産におけるマロニル-CoA供給系強化の有効性　　　　　　　　　　　　　　　
加来萌菜; 佐藤州朔; 石平芽衣; 朝山宗彦; 長南茂
日本微生物生態学会第33回大会, 11 Sep. 2019, 日本微生物生態学会

〔Major achievements〕脂肪酸生産におけるマロニル-CoA増産の有効性　　　　　　　　　　　　　　　
佐藤州朔; 尾崎美帆; 朝山宗彦; 長南茂
第13回日本ゲノム微生物学会年会, 07 Mar. 2019, 日本ゲノム微生物学会

ブタノール生成Clostridium属細菌の比較ゲノム解析　　　　　　　　　　　　　　　
黒坂愛美; 郭永; 上原研人; 長南茂; 久留主泰朗; 太田寛行; 西澤智康
第13回日本ゲノム微生物学会年会, 06 Mar. 2019, 日本ゲノム微生物学会

〔Major achievements〕外来パントテン酸キナーゼ遺伝子の大腸菌ゲノムへの導入によるコエンザイムA生合成経路の強化　　　　　　　　　　　　　　　
根岸剛志; 西澤智康; 三浦隆匡; 長南茂
環境微生物系合同大会2017, 29 Aug. 2017, 日本微生物生態学会・日本土壌微生物学会・環境バイオテクノロジー学会・日本菌学会・日本微生物資源学会

〔Major achievements〕大腸菌由来アセチル-CoAカルボキシラーゼを用いたマロニル-CoAの増産　　　　　　　　　　　　　　　
首藤 誉史; 長南 茂
日本微生物生態学会第31回大会, 23 Oct. 2016, 日本微生物生態学会

ブタノール生産性Clostridium属細菌のゲノム情報に基づく発酵代謝系の分子遺伝学的解析　　　　　　　　　　　　　　　
上原研人; 秋山真成美; 金本美穂; 西澤智康; 長南茂; 新田洋司; 久留主泰朗; 太田寛行
日本微生物生態学会第31回大会, 23 Oct. 2016, 日本微生物生態学会

Genetic engineering for the overproduction of polyhydroxybutyrate in cyanobacteria　　　　　　　　　　　　　　　
Sayaka Hondo; Masatoshi Takahashi; Takashi Osanai; Mami Matsuda; Tomohisa Hasunuma; Akio Tazuke; Yoichi Nakahira; Shigeru Chohnan; Morifumi Hasegawa; Munehiko Asayama
2015 International Chemical Congress of Pacific Basin Societies, 16 Dec. 2015

〔Major achievements〕Effect of the addition of pantothenate and its precursors on coenzyme A content of Escherichia coli expressing exogenous pantothenate kinase　　　　　　　　　　　　　　　
Yuta Ogata; Shigeru Chohnan
日本微生物生態学会第30回大会, 19 Oct. 2015, 日本微生物生態学会

Ammonium acetate increases the production of butanol by Clostridium beijerinckii cultures growing in sweet sorghum juice　　　　　　　　　　　　　　　
Kent Uehara; Manami Akiyama; Miho Kanemoto; Tomoyasu Nishizawa; Shigeru Chohnan; Youji Nitta; Yasurou Kurusu; Hiroyuki Ohta
日本微生物生態学会第30回大会, 19 Oct. 2015, 日本微生物生態学会

Clostridium beijerinckii SBP2-HB株におけるブタノール生成と糖利用性の解析　　　　　　　　　　　　　　　
金本美穂; 秋山真成美; 上原研人; Rahman M. Habibur; 佐藤嘉則; 長南茂; 新田洋司; 久留主康朗; 太田寛行
環境微生物系学会合同大会2014, 22 Oct. 2014, 日本微生物生態学会

スイートソルガム搾汁液を用いたブタノール発酵生産向上化技術の開発　　　　　　　　　　　　　　　
秋山真成美; 金本美穂; Rahman M. Habibur; 佐藤嘉則; 長南茂; 新田洋司; 久留主康朗; 太田寛行
環境微生物系学会合同大会2014, 22 Oct. 2014, 日本微生物生態学会

〔Major achievements〕Enhancement of coenzyme A biosynthetic pathway using pantothenate kinase　　　　　　　　　　　　　　　
Yuta Ogata; Shigeru Chohnan
114th General Meeting, American Society for Microbiology (USA), 18 May 2014, American Society for Microbiology (USA)

〔Major achievements〕Antimicrobial activity of pantothenol against Staphylococci possessing a prokaryotic type II pantothenate kinase　　　　　　　　　　　　　　　
Shigeru Chohnan; Misa Murase; Kota Kurikawa; Kodai Higashi; Yuta Ogata
114th General Meeting, American Society for Microbiology (USA), 18 May 2014, American Society for Microbiology (USA)

慢性社会的敗北ストレスがマウスの増体、摂食量、および肝臓CoAレベルにおよぼす影響　　　　　　　　　　　　　　　
久保田祥史; 後藤達彦; 萩谷友稀; 長南茂; 豊田淳
日本畜産学会 第118回大会, 27 Mar. 2014, 日本畜産学会

混合培養による好気的バイオブタノール生産　　　　　　　　　　　　　　　
秋山真成美; 金本美穂; M; Habibur Rahman; 佐藤嘉則; 長南茂; 新田洋司; 久留主泰朗; 太田寛行
日本土壌微生物学会2013年度大会, 20 Jun. 2013, 日本土壌微生物学会

〔Major achievements〕枯草菌のコエンザイムA生合成　　　　　　　　　　　　　　　
緒形雄太; 朝山宗彦; 長南茂
日本農芸化学会2013年度大会, 26 Mar. 2013

慢性的社会性ストレスによる摂食不振は視床下部マロニル-CoA濃度の 上昇によっておこる　　　　　　　　　　　　　　　
飯尾恒; 小池広明; 徳竹由華; 松川典子; 塚原隆充; 長南茂; 豊田淳
平成24年度関東畜産学会第67回大会, 10 Nov. 2012

慢性的社会性ストレスによる摂食不振は視床下部マロニル-CoA濃度の,上昇によっておこる　　　　　　　　　　　　　　　
飯尾恒、小池広明、徳竹由華、松川典子、塚原隆充、長南茂、豊田淳
平成24年度関東畜産学会第67回大会, 10 Nov. 2012

Anorexia and hypothalamic malonyl-CoA signaling pathway in chronic social defeated rats　　　　　　　　　　　　　　　
Wataru Iio; Yuka Tokutake; Noriko Matsukawa; Takamitsu Tsukahara; Shigeru Chohnan; Atsushi Toyoda
Neuroscience 2012, 13 Oct. 2012

〔Major achievements〕高脂肪食がラット組織内CoAプールに及ぼす影響　　　　　　　　　　　　　　　
徳竹由華; 長南茂
第33回日本肥満学会, 11 Oct. 2012, 日本肥満学会

Anorexic behavior and elevation of hypothalamic malonyl-CoA in socially defeated rats　　　　　　　　　　　　　　　
飯尾恒; 徳竹由華; 松川典子; 塚原隆充; 長南茂; 豊田淳
第35回日本神経科学大会, 21 Sep. 2012

〔Major achievements〕Modification of the acyl-CoA cycling method for analysis of CoA pools　　　　　　　　　　　　　　　
Yuka Tokutake; Arisa Saijoh; Wataru Iio; Yuta Ogata; Atsushi Toyoda; Shigeru Chohnan
112th General Meeting, American Society for Microbiology (USA), 19 Jun. 2012

〔Major achievements〕Characterization of prokaryotic types I and III pantothenate kinases from Bacillus subtilis　　　　　　　　　　　　　　　
Yuta Ogata; Yuka Tokutake; Shigeru Chohnan
112th General Meeting, American Society for Microbiology (USA), 19 Jun. 2012

社会的慢性ストレスモデルにおける増体抑制メカニズムの解明　　　　　　　　　　　　　　　
飯尾恒; 徳竹由華; 松川典子; 塚原隆充; 長南茂; 豊田淳
日本畜産学会第115回大会, 28 Mar. 2012

〔Major achievements〕枯草菌の原核I型およびIII型パントテン酸キナーゼの性質　　　　　　　　　　　　　　　
緒形雄太; 長南茂
日本微生物生態学会第27回大会, 09 Oct. 2011

スイートソルガム搾汁液からブタノールを高生産する細菌株の選抜　　　　　　　　　　　　　　　
金本美穂; Habibur M. Rahman; 佐藤嘉則; 長南茂; 新田洋司; 久留主泰朗; 太田寛行
日本微生物生態学会第27回大会, 09 Oct. 2011

Biomass circulation system through the biobutanol production from a sugar-rich plant, sweet sorghum　　　　　　　　　　　　　　　
Miho Kanemoto; M. Habibur Rahman; Yoshinori Sato; Shigeru Chohnan; Youji Nitta; Yasurou Kurusu; Hiroyuki Ohta
XXth International Symposium on Environmental Biogeochemistry (Turkey), 27 Sep. 2011

〔Major achievements〕食餌の質がラット組織内CoAプールに及ぼす影響　　　　　　　　　　　　　　　
徳竹由華; 鬼澤直樹; 飯尾恒; 豊田淳; 長南茂
第32回日本肥満学会, 23 Sep. 2011

〔Major achievements〕A modified acyl-CoA cycling method accelerates multiple analyses of the in vivo behavior of CoA pools　　　　　　　　　　　　　　　
Yuka Tokutake; Shigeru Chohnan
111th General Meeting, American Society for Microbiology (USA), 24 May 2011

〔Major achievements〕A coenzyme A biosynthetic pathway in archaea　　　　　　　　　　　　　　　
Hiroki Katoh; Hideyuki Tamaki; Yuka Tokutake; Satoshi Hanada; Shigeru Chohnan
111th General Meeting, American Society for Microbiology (USA), 24 May 2011

〔Major achievements〕絶食および飽食ラットにおける組織内CoAプールの動態解析　　　　　　　　　　　　　　　
徳竹由華; 鬼澤直樹; 村田恵; 渡辺央好; 角田梨奈; 豊田淳; 長南茂
日本農芸化学会2011年度大会, 26 Mar. 2011

〔Major achievements〕メタン生成古細菌のコエンザイムA生合成経路　　　　　　　　　　　　　　　
Hiroki Katoh; Shigeru Chohnan
日本微生物生態学会第26回大会, 25 Nov. 2010

Characterization of clostridia able to grow in sweet sorghum juice and produce butanol　　　　　　　　　　　　　　　
Miho Kanemoto; Habibur Rahman; Shigeru Chohnan; Yoji Nitta; Yasurou Kurusu; Hiroyuki Ohta
日本微生物生態学会第26回大会, 25 Nov. 2010

〔Major achievements〕Characterization of clostridia able to grow in sweet sorghum juice and produce butanol　　　　　　　　　　　　　　　
金本 美穂、HABINUR RAHMAN、長南茂、新田洋司、久留主泰朗、太田寛行
日本微生物生態学会第26回大会, 23 Nov. 2010, 日本微生物生態学会
20101123

Isolation of soil bacteria able to grow in sweet sorghum juice and produce butanol　　　　　　　　　　　　　　　
Miho Kanemoto; Habibur Rahman; Shigeru Chohnan; Yoji Nitta; Yasuro Kurusu; Hiroyuki Ohta
13th International Symposium on Microbial Ecology (USA), 23 Aug. 2010

〔Major achievements〕Bioethanol production from sweet sorghum by Saccharomyces cerevisiae　　　　　　　　　　　　　　　
Megumi Nakane; Shigeru Chohnan; Yoji Nitta; Takanori Yoshiura; Hiroyuki Ohta; Yasurou Kurusu
110th General Meeting, American Society for Microbiology (USA), 26 May 2010

〔Major achievements〕Pantothenate kinase from thermoacidophilic archaeon　　　　　　　　　　　　　　　
Masakazu Takagi; Yukiko Miyamoto; Hedeyuki Tamaki; Roberta Leonardi; Satoshi Hanada; Suzanne Jackowski; Shigeru Chohnan
110th General Meeting, American Society for Microbiology (USA), 24 May 2010

〔Major achievements〕メタン生成古細菌のコエンザイムA生合成経路の解析　　　　　　　　　　　　　　　
Hiroki Katoh、Syunta Komatsuzawa、Shigeru Chohnan
日本農芸化学会2010年度大会, 29 Mar. 2010

〔Major achievements〕アシル-CoAサイクリング法によるラット組織内CoAプールの解析　　　　　　　　　　　　　　　
Yuka Tokutake、Naoki Onizawa、Atushi Toyoda、Shigeru Chohnan
日本農芸化学会2010年度大会, 28 Mar. 2010

クレアチンの経口投与が脳内CoA分子種に及ぼす影響　　　　　　　　　　　　　　　
鬼澤直樹; 徳竹由華; 長南茂; 中村豊; 豊田淳
日本畜産学会第112回大会, Mar. 2010, 日本畜産学会

〔Major achievements〕Analysis of pantothenate kinase from thermoacidophilic archaeon　　　　　　　　　　　　　　　
Masakazu Takagi; Yukiko Miyamoto; Koji Mori; Shigeru Chohnan
日本微生物生態学会第24回大会, 26 Nov. 2008, 日本微生物生態学会

高ポリアミン食によるマウスの寿命延長効果　　　　　　　　　　　　　　　
早田邦康; チェリック アルパー; 岡田晋一郎; 加納良彦; 辻仲眞康; 高尾浩一; 長南茂; 小西文雄; 川上正舒
第21回日本バイオセラピィ学会学術集会総会, 18 Nov. 2008, 日本バイオセラピィ学会

Sweet sorghum cultivation as a bio-fuel crop in the suburbs of Tokyo metropolitan area　　　　　　　　　　　　　　　
Akinori Kamiyama; Youji Nitta; Toshiaki Matsuda; Satoshi Nakamura; Yusuke Goto; Eiichi Inoue; Kazuhiko Narisawa; Yasurou Kurusu; Hiroyuki Ohta; Shigeru Chohnan; Atsushi Toyoda; Tasuku Kato; Hisashi Kobayashi; Masakazu Komatsuzaki; Tatuo Sato
5th International Crop Science Congress & Exhibition (Korea), Apr. 2008

〔Major achievements〕Characterization of N-deoxyribosyltransferase from Lactococcus lactis subsp. lactis　　　　　　　　　　　　　　　
Yukiko Miyamoto and Shigeru Chohnan
日本微生物生態学会第23回大会, 16 Sep. 2007, 日本微生物生態学会

〔Major achievements〕乳酸菌由来N-デオキシリボシルトランスフェラーゼの解析　　　　　　　　　　　　　　　
宮本祐規子; 正木武治; 長南茂
日本農芸化学会2007年度大会, 26 Mar. 2007, 日本農芸化学会

Lysobacter sp. IB-9374由来リシルエンドペプチダーゼB変異体の機能解析　　　　　　　　　　　　　　　
武石舞佳; 長南茂; 白木賢太郎; 正木武治; 崎山文夫
日本農芸化学会2007年度大会, 25 Mar. 2007, 日本農芸化学会

Lysobacter sp. IB-9374由来セルラーゼIIの精製と諸性質　　　　　　　　　　　　　　　
Chong AiLeng; 小倉治朗; 長南茂; 正木武治
日本農芸化学会2007年度大会, 25 Mar. 2007

Lysobacter sp. IB-9374由来endoglucanase(Cel8A)遺伝子のクローニング　　　　　　　　　　　　　　　
小倉治朗; 鐘愛玲; 豊田淳; 長南茂; 正木武治
日本農芸化学会2006年度大会講演要旨集, Mar. 2006

Lysobacter sp.由来サブチリシン様プロテアーゼの精製とその特性　　　　　　　　　　　　　　　
牧野裕也; 小倉治朗; 長南茂; 正木武治
日本農芸化学会2006年度大会講演要旨集, Mar. 2006

〔Major achievements〕Pseudomonas属由来パントテン酸キナーゼ遺伝子‐コエンザイムA生合成系での行方不明遺伝子‐　　　　　　　　　　　　　　　
長南茂; Roberta Leonardi; Yong-Mei Zhang,Charles O. Rock; Suzanne Jackowski
日本微生物生態学会第21回大会, Oct. 2005

Lysobacter sp.由来セルラーゼの精製と諸性質　　　　　　　　　　　　　　　
黒澤泰佑; 小倉治朗; Chong AiLeng; 足立吉数; 長南茂; 正木武治
日本農芸化学会2005年度大会, Mar. 2005

Lysobacter sp.由来トリプシン様プロテアーゼの諸性質　　　　　　　　　　　　　　　
上杉亜星; 小倉治朗; 長南茂; 正木武治
日本農芸化学会2005年度大会, Mar. 2005

〔Major achievements〕Staphylococcus aureusのパントテン酸キナーゼはCoAで調節されていない　　　　　　　　　　　　　　　
長南茂; Charles O. Rock; Suzanne Jackowski
日本農芸化学会2004年度大会, Mar. 2004

Lysobacter sp.産生beta-リティックプロテアーゼのペプチドグリカンに対する作用部位について　　　　　　　　　　　　　　　
平田剛士; Ahmed Kashfia; 甲賀さおり; 長南茂; 正木武治
日本農芸化学会2004年度大会, Mar. 2004

A new lysylendopeptidase from Lysobacter sp.　　　　　　　　　　　　　　　
Takeharu Masaki; Shigeru Chohnan; Kentaro Shiraki; Kiyonobu Yokota; Fumio Sakiyama
15th Methods in Protein Structure Analysis (USA), 2004

〔Major achievements〕Chemical knockout of coenzyme A levels in mice induces hepatic steatosis and severe hypoglycemia　　　　　　　　　　　　　　　
Yong-Mei Zhang; Shigeru Chohnan; Richard E. Lee; Charles.O. Rock; Suzanne Jackowski
American Society for Biochemistry and Molecular Biology Annual Meeting and 8th IUBMB Conference (USA), 2004

Lysobacter sp. IB-9374由来トリプシン様セリンプロテアーゼの精製と特性　　　　　　　　　　　　　　　
大島誠; 長南茂; 正木武治
日本農芸化学会2003年度大会, Mar. 2003

Lysobacter sp. IB-9374由来リシルエンドペプチダーゼ類似遺伝子産物の特性解析　　　　　　　　　　　　　　　
黒岩菜月; 長南茂; 白木賢太郎; 横田恭宣; 正木武治; 崎山文夫
日本農芸化学会2003年度大会, Mar. 2003

Lysobacter sp.産生beta-リティックプロテアーゼの溶菌活性　　　　　　　　　　　　　　　
平田剛士; Ahmed Kashfia; 長南茂; 正木武治
日本農芸化学会2003年度大会, Mar. 2003

Brevibacillus sp.由来プロリルオリゴペプチターゼの基質特異性　　　　　　　　　　　　　　　
木村幸男; 矢吹崇吏; 桑原重文; 長南茂; 正木武治
日本農芸化学会2002年度大会, Mar. 2002

大腸菌シャペロニンGroEL/ES共発現系におけるP.putidaマロン酸脱炭酸酵素サブユニットの発現と機能解析　　　　　　　　　　　　　　　
赤木香予; 長南茂; 高村義親; 舟木強
日本農芸化学会2002年度大会, Mar. 2002, 日本農芸化学会

Xanthomonas sp. IB-9374株由来beta-リティックプロテアーゼ遺伝子のクローニング　　　　　　　　　　　　　　　
Kashfia Ahamed; 平田剛士; 大橋広行; 長南茂; 崎山文夫; 正木武治
日本農芸化学会2002年度大会, Mar. 2002

Xanthomonasリジルエンドペプチダーゼに対するリジンクロロメチルケトン誘導体の阻害について　　　　　　　　　　　　　　　
鈴木康史; 稲垣宋彰; 長南茂; 桑原重文; 正木武治
日本農芸化学会2002年度大会, Mar. 2002

大腸菌シャペロニンにおけるGroEL/ES共発現系におけるP.putidaマロン酸脱炭酸酵素サブユニットの発現と機能解析　　　　　　　　　　　　　　　
赤木香予; 長南茂; 高村義親
日本農芸化学会関東支部2001年度支部大会, Oct. 2001

Xanthomonas sp. IB-9374株のリシルエンドペプチダーゼ類似遺伝子の解析　　　　　　　　　　　　　　　
長南茂; 大島誠; 中峯有香; 崎山文夫; 正木武治
日本農芸化学会2001年度大会, Mar. 2001

Brevibacillus sp.産生の新規なプロリルエンドぺプチターゼの単離とその特性　　　　　　　　　　　　　　　
木村幸男; 渡部栄二郎; 近藤礼子; 長南茂; 久留主泰朗; 桑原重文; 正木武治
日本農芸化学会2001年度大会, Mar. 2001

Malonate decarboxylase from Pseudomonas putida　　　　　　　　　　　　　　　
Shigeru Chohnan; Kayo Akagi; Yoshichika Takamura
Pseudomonas 2001 (Belgium), 2001

Xanthomonas sp.産生リジン残基特異的プロテアーゼ遺伝子のクローニング　　　　　　　　　　　　　　　
長南茂; 野中淳子; 谷口浩一; 久留主泰朗; 正木武治
日本農芸化学会2000年度大会, Mar. 2000

Pseudomonas putidaマロン酸脱炭酸酵素のa（MdcA），b（MdcD）およびg（MdcE）サブユニットの機能解析　　　　　　　　　　　　　　　
赤木香予; 長南茂; 高村義親
日本農芸化学会2000年度大会, Mar. 2000

Xanthomonas sp.産生リジン特異的プロテアーゼ遺伝子のクローニング　　　　　　　　　　　　　　　
野中淳子; 谷口浩一; 長南茂; 久留主泰朗; 乗岡茂巳; 崎山文夫; 正木武治
第72回日本生化学会大会, Aug. 1999

Pseudomonas putidaのマロン酸脱炭酸酵素遺伝子のクローニング　　　　　　　　　　　　　　　
長南茂; 久留主泰朗; 西原宏史; 高村義親
日本農芸化学会1999年度大会, Mar. 1999

Pseudomonas putidaのマロン酸脱炭酸酵素の反応機構　　　　　　　　　　　　　　　
長南茂; 高木俊和; 西原宏史; 高村義親
日本農芸化学会1998年度大会, Mar. 1998

Aspergillus niger由来サイクリック-リボヌクレオチドホスホムターゼ‐5'-ホスホジエステラーゼ（cRNPMase-5'-PDase）の精製と性質（第2報）　　　　　　　　　　　　　　　
長南茂; 緑川祐一朗
日本農芸化学会1994年度大会, Mar. 1994

Aspergillus nigerのサイクリック‐リボヌクレオチドホスホムターゼ（cRNPMase）の精製と性質　　　　　　　　　　　　　　　
長南茂; 永田兀; 緑川祐一朗
日本農芸化学会1992年度大会, Mar. 1992

アシルCoA:CoAサイクリング法による大腸菌CoAプールのサイズと組成の解析　　　　　　　　　　　　　　　
長南茂; 高村義親
第63回日本生化学会大会, Jul. 1990, 日本生化学会

Available carbon potential (ACp) -A fundamental parameter of cellular carbon metabolism-　　　　　　　　　　　　　　　
Yoshichika Takamura; Shigeru Chohnan
IUMS Congress: Bacteriology & Mycology-Osaka, Japan-1990, 1990

アシルCoAサイクリング法による遊離CoAの極微量定量法の確立とE.coli K-12における細胞内遊離CoAの変動について　　　　　　　　　　　　　　　
長南茂; 萩原秀美; 圓藤重雄; 高村義親
日本農芸化学会1989年度大会, Mar. 1989, 日本農芸化学会

E.coli K-12の脂肪酸合成に及ぼす細胞内アセチルCoA及びマロニルCoAの影響　　　　　　　　　　　　　　　
圓藤重雄; 高村義親; 長南茂; 萩原秀美
日本農芸化学会1989年度大会, Mar. 1989, 日本農芸化学会

Dec. 2018 - Present, 日本ゲノム微生物学会

Jan. 2013 - Present, The Society for Biotechnology, Japan

Jan. 2010 - Present, American Sciety for Microbiology

Jan. 2004 - Present, The Japanese Society of Microbial Ecology

2000 - Present, バイオインダストリー協会

Jan. 1989 - Present, 日本農芸化学会