Alcohol Selectivity in a Synthetic Thermophilic n-Butanol Pathway Is Driven by Biocatalytic and Thermostability Characteristics of Constituent Enzymes

Loder, Andrew J.; Zeldes, Benjamin M.; Garrison, G. Dale; Lipscomb, Gina L.; Adams, Michael W.  W.; Kelly, Robert M.

doi:10.1128/AEM.02028-15

Title: Alcohol Selectivity in a Synthetic Thermophilic n-Butanol Pathway Is Driven by Biocatalytic and Thermostability Characteristics of Constituent Enzymes

Journal Article · Fri Aug 07 00:00:00 EDT 2015 · Applied and Environmental Microbiology

DOI:https://doi.org/10.1128/AEM.02028-15· OSTI ID:1470144

Loder, Andrew J. ^[1]; Zeldes, Benjamin M. ^[2]; Garrison, G. Dale ^[2]; Lipscomb, Gina L. ^[3]; Adams, Michael W. W. ^[3]; Kelly, Robert M. ^[2]

North Carolina State Univ., Raleigh, NC (United States). Dept. of Chemical and Biomolecular Engineering
North Carolina State Univ., Raleigh, NC (United States). Dept. of Chemical and Biomolecular Engineering
Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology

n-Butanol is generated as a natural product of metabolism by several microorganisms, but almost all grow at mesophilic temperatures. In this paper, a synthetic pathway for n-butanol production from acetyl coenzyme A (acetyl-CoA) that functioned at 70°C was assembled in vitro from enzymes recruited from thermophilic bacteria to inform efforts for engineering butanol production into thermophilic hosts. Recombinant versions of eight thermophilic enzymes (β-ketothiolase [Thl], 3-hydroxybutyryl-CoA dehydrogenase [Hbd], and 3-hydroxybutyryl-CoA dehydratase [Crt] from Caldanaerobacter subterraneus subsp. tengcongensis; trans-2-enoyl-CoA reductase [Ter] from Spirochaeta thermophila; bifunctional acetaldehyde dehydrogenase/alcohol dehydrogenase [AdhE] from Clostridium thermocellum; and AdhE, aldehyde dehydrogenase [Bad], and butanol dehydrogenase [Bdh] from Thermoanaerobacter sp. strain X514) were utilized to examine three possible pathways for n-butanol. These pathways differed in the two steps required to convert butyryl-CoA to n-butanol: Thl-Hbd-Crt-Ter-AdhE (C. thermocellum), Thl-Hbd-Crt-Ter-AdhE (Thermoanaerobacter X514), and Thl-Hbd-Crt-Ter-Bad-Bdh. n-Butanol was produced at 70°C, but with different amounts of ethanol as a coproduct, because of the broad substrate specificities of AdhE, Bad, and Bdh. A reaction kinetics model, validated via comparison to in vitro experiments, was used to determine relative enzyme ratios needed to maximize n-butanol production. By using large relative amounts of Thl and Hbd and small amounts of Bad and Bdh, >70% conversion to n-butanol was observed in vitro, but with a 60% decrease in the predicted pathway flux. With more-selective hypothetical versions of Bad and Bdh, >70% conversion to n-butanol is predicted, with a 19% increase in pathway flux. Finally and thus, more-selective thermophilic versions of Bad, Bdh, and AdhE are needed to fully exploit biocatalytic n-butanol production at elevated temperatures.

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Research Organization:: North Carolina State University, Raleigh, NC (United States); Univ. of Georgia, Athens, GA (United States)

Sponsoring Organization:: USDOE Advanced Research Projects Agency - Energy (ARPA-E); National Science Foundation (NSF); National Inst. of Health (NIH) (United States)

Grant/Contract Number:: AR0000081; CBET-1264052; CBET-1264053; 2T32GM008776

OSTI ID:: 1470144

Journal Information:: Applied and Environmental Microbiology, Vol. 81, Issue 20; ISSN 0099-2240

Publisher:: American Society for MicrobiologyCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 17 works

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References (70)

Systematic Construction of Kinetic Models from Genome-Scale Metabolic Networks Stanford, Natalie J.; Lubitz, Timo; Smallbone, Kieran PLoS ONE, Vol. 8, Issue 11 https://doi.org/10.1371/journal.pone.0079195	journal	November 2013
Biochemical and Structural Characterization of the trans -Enoyl-CoA Reductase from Treponema denticola Bond-Watts, Brooks B.; Weeks, Amy M.; Chang, Michelle C. Y. Biochemistry, Vol. 51, Issue 34 https://doi.org/10.1021/bi300879n	journal	August 2012
The kinetic mechanism and properties of the cytoplasmic acetoacetyl-coenzyme A thiolase from rat liver Middleton, B. Biochemical Journal, Vol. 139, Issue 1 https://doi.org/10.1042/bj1390109	journal	April 1974
New types of experimental data shape the use of enzyme kinetics for dynamic network modeling Tummler, Katja; Lubitz, Timo; Schelker, Max FEBS Journal, Vol. 281, Issue 2 https://doi.org/10.1111/febs.12525	journal	November 2013
Metabolic Engineering of Thermoanaerobacterium thermosaccharolyticum for Increased n-Butanol Production Bhandiwad, Ashwini; Guseva, Anna; Lynd, Lee Advances in Microbiology, Vol. 03, Issue 01 https://doi.org/10.4236/aim.2013.31007	journal	January 2013
Engineering alternative butanol production platforms in heterologous bacteria Nielsen, David R.; Leonard, Effendi; Yoon, Sang-Hwal Metabolic Engineering, Vol. 11, Issue 4-5, p. 262-273 https://doi.org/10.1016/j.ymben.2009.05.003	journal	July 2009
Pyruvate-formate-lyase-deactivase and acetyl-CoA reductase activities of Escherichia coli reside on a polymeric protein particle encoded by adhE Kessler, Dorothea; Leibrecht, Iris; Knappe, Joachim FEBS Letters, Vol. 281, Issue 1-2 https://doi.org/10.1016/0014-5793(91)80358-A	journal	April 1991
Extremely Thermophilic Routes to Microbial Electrofuels Hawkins, Aaron S.; Han, Yejun; Lian, Hong ACS Catalysis, Vol. 1, Issue 9 https://doi.org/10.1021/cs2003017	journal	July 2011
Modular pathway engineering of Bacillus subtilis for improved N-acetylglucosamine production Liu, Yanfeng; Zhu, Yanqiu; Li, Jianghua Metabolic Engineering, Vol. 23 https://doi.org/10.1016/j.ymben.2014.02.005	journal	May 2014
Molar absorptivities of beta-NADH and beta-NADPH. Ziegenhorn, J.; Senn, M.; Bücher, T. Clinical Chemistry, Vol. 22, Issue 2 https://doi.org/10.1093/clinchem/22.2.151	journal	February 1976
Butanol formation byClostridium thermosaccharolyticum at neutral pH Freier-Schr�der, Doris; Wiegel, J�rgen; Gottschalk, Gerhard Biotechnology Letters, Vol. 11, Issue 11 https://doi.org/10.1007/BF01026107	journal	November 1989
Reconstructing the clostridial n-butanol metabolic pathway in Lactobacillus brevis Berezina, Oksana V.; Zakharova, Natalia V.; Brandt, Agnieszka Applied Microbiology and Biotechnology, Vol. 87, Issue 2 https://doi.org/10.1007/s00253-010-2480-z	journal	March 2010
Simulating Plant Metabolic Pathways with Enzyme-Kinetic Models Schallau, Kai; Junker, Björn H. Plant Physiology, Vol. 152, Issue 4 https://doi.org/10.1104/pp.109.149237	journal	January 2010
Pyrodictium abyssi sp. nov. Represents a Novel Heterotrophic Marine Archaeal Hyperthermophile Growing at 110°C Pley, Ursula; Schipka, Jutta; Gambacorta, Agata Systematic and Applied Microbiology, Vol. 14, Issue 3 https://doi.org/10.1016/S0723-2020(11)80376-0	journal	July 1991
A Whole-Cell Computational Model Predicts Phenotype from Genotype Karr, Jonathan R.; Sanghvi, Jayodita C.; Macklin, Derek N. Cell, Vol. 150, Issue 2 https://doi.org/10.1016/j.cell.2012.05.044	journal	July 2012
Screening of a novel strong promoter by RNA sequencing and its application to H2 production in a hyperthermophilic archaeon Lee, Seong Hyuk; Kim, Min-Sik; Jung, Hae Chang Applied Microbiology and Biotechnology, Vol. 99, Issue 9 https://doi.org/10.1007/s00253-015-6444-1	journal	February 2015
Bio-based production of C2-C6 platform chemicals Jang, Yu-Sin; Kim, Byoungjin; Shin, Jae Ho Biotechnology and Bioengineering, Vol. 109, Issue 10 https://doi.org/10.1002/bit.24599	journal	July 2012
Tuning genetic control through promoter engineering Alper, H.; Fischer, C.; Nevoigt, E. Proceedings of the National Academy of Sciences, Vol. 102, Issue 36, p. 12678-12683 https://doi.org/10.1073/pnas.0504604102	journal	August 2005
Utilizing an endogenous pathway for 1-butanol production in Saccharomyces cerevisiae Si, Tong; Luo, Yunzi; Xiao, Han Metabolic Engineering, Vol. 22 https://doi.org/10.1016/j.ymben.2014.01.002	journal	March 2014
High-temperature fermentation: how can processes for ethanol production at high temperatures become superior to the traditional process using mesophilic yeast? Abdel-Banat, Babiker M. A.; Hoshida, Hisashi; Ano, Akihiko Applied Microbiology and Biotechnology, Vol. 85, Issue 4 https://doi.org/10.1007/s00253-009-2248-5	journal	October 2009
ATP drives direct photosynthetic production of 1-butanol in cyanobacteria Lan, E. I.; Liao, J. C. Proceedings of the National Academy of Sciences, Vol. 109, Issue 16 https://doi.org/10.1073/pnas.1200074109	journal	April 2012
Systems metabolic engineering of microorganisms for natural and non-natural chemicals Lee, Jeong Wook; Na, Dokyun; Park, Jong Myoung Nature Chemical Biology, Vol. 8, Issue 6 https://doi.org/10.1038/nchembio.970	journal	May 2012
Enzyme mechanism as a kinetic control element for designing synthetic biofuel pathways Bond-Watts, Brooks B.; Bellerose, Robert J.; Chang, Michelle C. Y. Nature Chemical Biology, Vol. 7, Issue 4 https://doi.org/10.1038/nchembio.537	journal	February 2011
Enzymatic investigations on butanol dehydrogenase and butyraldehyde dehydrogenase in extracts of Clostridium acetobutylicum D�rre, Peter; Kuhn, Anita; Gottwald, Matthias Applied Microbiology and Biotechnology, Vol. 26, Issue 3 https://doi.org/10.1007/BF00286322	journal	June 1987
Mathematical models of ABE fermentation: review and analysis Mayank, Rahul; Ranjan, Amrita; Moholkar, Vijayanand S. Critical Reviews in Biotechnology, Vol. 33, Issue 4 https://doi.org/10.3109/07388551.2012.726208	journal	September 2012
A hybrid synthetic pathway for butanol production by a hyperthermophilic microbe Keller, Matthew W.; Lipscomb, Gina L.; Loder, Andrew J. Metabolic Engineering, Vol. 27 https://doi.org/10.1016/j.ymben.2014.11.004	journal	January 2015
Coupled Ferredoxin and Crotonyl Coenzyme A (CoA) Reduction with NADH Catalyzed by the Butyryl-CoA Dehydrogenase/Etf Complex from Clostridium kluyveri Li, F.; Hinderberger, J.; Seedorf, H. Journal of Bacteriology, Vol. 190, Issue 3 https://doi.org/10.1128/JB.01417-07	journal	November 2007
Why measure enzyme activities in the era of systems biology? Stitt, Mark; Gibon, Yves Trends in Plant Science, Vol. 19, Issue 4 https://doi.org/10.1016/j.tplants.2013.11.003	journal	April 2014
Redirecting carbon flux through exogenous pyruvate kinase to achieve high ethanol yields in Clostridium thermocellum Deng, Yu; Olson, Daniel G.; Zhou, Jilai Metabolic Engineering, Vol. 15, p. 151-158 https://doi.org/10.1016/j.ymben.2012.11.006	journal	January 2013
Multivariate modular metabolic engineering for pathway and strain optimization Biggs, Bradley Walters; De Paepe, Brecht; Santos, Christine Nicole S. Current Opinion in Biotechnology, Vol. 29 https://doi.org/10.1016/j.copbio.2014.05.005	journal	October 2014
The aldehyde/alcohol dehydrogenase (AdhE) in relation to the ethanol formation in Thermoanaerobacter ethanolicus JW200 Peng, Hui; Wu, Guogan; Shao, Weilan Anaerobe, Vol. 14, Issue 2 https://doi.org/10.1016/j.anaerobe.2007.09.004	journal	April 2008
Molecular Characterization and Transcriptional Analysis of adhE2, the Gene Encoding the NADH-Dependent Aldehyde/Alcohol Dehydrogenase Responsible for Butanol Production in Alcohologenic Cultures of Clostridium acetobutylicum ATCC 824 Fontaine, L.; Meynial-Salles, I.; Girbal, L. Journal of Bacteriology, Vol. 184, Issue 3 https://doi.org/10.1128/JB.184.3.821-830.2002	journal	February 2002
Overexpression of groESL in Clostridium acetobutylicum Results in Increased Solvent Production and Tolerance, Prolonged Metabolism, and Changes in the Cell's Transcriptional Program Tomas, C. A.; Welker, N. E.; Papoutsakis, E. T. Applied and Environmental Microbiology, Vol. 69, Issue 8 https://doi.org/10.1128/AEM.69.8.4951-4965.2003	journal	August 2003
Engineering Considerations for the Application of Extremophiles in Biotechnology Ludlow, Jan M.; Clark, Douglas S. Critical Reviews in Biotechnology, Vol. 10, Issue 4 https://doi.org/10.3109/07388559109038214	journal	January 1991
Purification and molecular characterization of the NAD+-dependent acetaldehyde/alcohol dehydrogenase from Entamoeba histolytica Bruchhaus, I.; Tannich, E. Biochemical Journal, Vol. 303, Issue 3 https://doi.org/10.1042/bj3030743	journal	November 1994
Kinetic study of butanol production from various sugars by Clostridium acetobutylicum using a dynamic model Raganati, Francesca; Procentese, Alessandra; Olivieri, Giuseppe Biochemical Engineering Journal, Vol. 99 https://doi.org/10.1016/j.bej.2015.03.001	journal	July 2015
The zero-sum game of pathway optimization: Emerging paradigms for tuning gene expression Solomon, Kevin V.; Prather, Kristala L. J. Biotechnology Journal, Vol. 6, Issue 9 https://doi.org/10.1002/biot.201100086	journal	June 2011
Pathway Thermodynamics Highlights Kinetic Obstacles in Central Metabolism Noor, Elad; Bar-Even, Arren; Flamholz, Avi PLoS Computational Biology, Vol. 10, Issue 2 https://doi.org/10.1371/journal.pcbi.1003483	journal	February 2014
eQuilibrator--the biochemical thermodynamics calculator Flamholz, A.; Noor, E.; Bar-Even, A. Nucleic Acids Research, Vol. 40, Issue D1 https://doi.org/10.1093/nar/gkr874	journal	November 2011
The Bifunctional Alcohol and Aldehyde Dehydrogenase Gene, adhE , Is Necessary for Ethanol Production in Clostridium thermocellum and Thermoanaerobacterium saccharolyticum Lo, Jonathan; Zheng, Tianyong; Hon, Shuen Journal of Bacteriology, Vol. 197, Issue 8 https://doi.org/10.1128/JB.02450-14	journal	February 2015
Metabolic engineering of Saccharomyces cerevisiae for the production of n-butanol Steen, Eric J.; Chan, Rossana; Prasad, Nilu Microbial Cell Factories, Vol. 7, Issue 1, Article No. 36 https://doi.org/10.1186/1475-2859-7-36	journal	January 2008
Direct conversion of plant biomass to ethanol by engineered Caldicellulosiruptor bescii Chung, D.; Cha, M.; Guss, A. M. Proceedings of the National Academy of Sciences, Vol. 111, Issue 24, p. 8931-8936 https://doi.org/10.1073/pnas.1402210111	journal	June 2014
Protein production by auto-induction in high-density shaking cultures Studier, F. William Protein Expression and Purification, Vol. 41, Issue 1, p. 207-234 https://doi.org/10.1016/j.pep.2005.01.016	journal	May 2005
Butanol-Ethanol Dehydrogenase and Butanol-Ethanol-Isopropanol Dehydrogenase: Different Alcohol Dehydrogenases in Two Strains of Clostridium beijerinckii (Clostridium butylicum) Hiu, Stephen F.; Zhu, Chang-Xi; Yan, Run-Tao Applied and Environmental Microbiology, Vol. 53, Issue 4 https://doi.org/10.1128/AEM.53.4.697-703.1987	journal	January 1987
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Bradford, Marion M. Analytical Biochemistry, Vol. 72, Issue 1-2 https://doi.org/10.1016/0003-2697(76)90527-3	journal	May 1976
Bioprocessing analysis of Pyrococcus furiosus strains engineered for CO ₂ -based 3-hydroxypropionate production : 3-Hydroxypropionate Production by Hawkins, Aaron B.; Lian, Hong; Zeldes, Benjamin M. Biotechnology and Bioengineering, Vol. 112, Issue 8 https://doi.org/10.1002/bit.25584	journal	June 2015
Bioengineering of microorganisms for C3 to C5 alcohols production Mainguet, Samuel E.; Liao, James C. Biotechnology Journal, Vol. 5, Issue 12 https://doi.org/10.1002/biot.201000276	journal	December 2010
Deletion of acetyl-CoA synthetases I and II increases production of 3-hydroxypropionate by the metabolically-engineered hyperthermophile Pyrococcus furiosus Thorgersen, Michael P.; Lipscomb, Gina L.; Schut, Gerrit J. Metabolic Engineering, Vol. 22 https://doi.org/10.1016/j.ymben.2013.12.006	journal	March 2014
Influence of divalent cations on the structural thermostability and thermal inactivation kinetics of class II xylose isomerases: Thermostability of class II xylose isomerases Epting, Kevin L.; Vieille, Claire; Zeikus, J. Gregory FEBS Journal, Vol. 272, Issue 6, p. 1454-1464 https://doi.org/10.1111/j.1742-4658.2005.04577.x	journal	March 2005
Enzymatic Assembly of Overlapping DNA Fragments Gibson, Daniel G. Methods in Enzymology https://doi.org/10.1016/B978-0-12-385120-8.00015-2	book	May 2011
Purification and characterization of the NADH-dependent butanol dehydrogenase from Clostridium acetobutylicum (ATCC 824) Welch, Richard W.; Rudolph, Frederick B.; Papoutsakis, E. Terry Archives of Biochemistry and Biophysics, Vol. 273, Issue 2 https://doi.org/10.1016/0003-9861(89)90489-X	journal	September 1989
Exploiting microbial hyperthermophilicity to produce an industrial chemical, using hydrogen and carbon dioxide Keller, M. W.; Schut, G. J.; Lipscomb, G. L. Proceedings of the National Academy of Sciences, Vol. 110, Issue 15, p. 5840-5845 https://doi.org/10.1073/pnas.1222607110	journal	March 2013
Solventogenic enzymes of Clostridium acetobutylicum: catalytic properties, genetic organization, and transcriptional regulation Dürre, Peter; Fischer, Ralf-Jörg; Kuhn, Anita FEMS Microbiology Reviews, Vol. 17, Issue 3, p. 251-262 https://doi.org/10.1111/j.1574-6976.1995.tb00209.x	journal	October 1995
Elucidating and reprogramming Escherichia coli metabolisms for obligate anaerobic n-butanol and isobutanol production Trinh, Cong T. Applied Microbiology and Biotechnology, Vol. 95, Issue 4 https://doi.org/10.1007/s00253-012-4197-7	journal	June 2012
Towards kinetic modeling of genome-scale metabolic networks without sacrificing stoichiometric, thermodynamic and physiological constraints Chakrabarti, Anirikh; Miskovic, Ljubisa; Soh, Keng Cher Biotechnology Journal, Vol. 8, Issue 9 https://doi.org/10.1002/biot.201300091	journal	August 2013
Separation technologies for the recovery and dehydration of alcohols from fermentation broths Vane, Leland M. Biofuels, Bioproducts and Biorefining, Vol. 2, Issue 6 https://doi.org/10.1002/bbb.108	journal	November 2008
Structure of a bifunctional alcohol dehydrogenase involved in bioethanol generation in Geobacillus thermoglucosidasius Extance, Jonathan; Crennell, Susan J.; Eley, Kirstin Acta Crystallographica Section D Biological Crystallography, Vol. 69, Issue 10 https://doi.org/10.1107/S0907444913020349	journal	September 2013
Advanced biofuel production in microbes Peralta-Yahya, Pamela P.; Keasling, Jay D. Biotechnology Journal, Vol. 5, Issue 2, p. 147-162 https://doi.org/10.1002/biot.200900220	journal	January 2010
Enhanced Butanol Production Obtained by Reinforcing the Direct Butanol-Forming Route in Clostridium acetobutylicum Jang, Yu-Sin; Lee, Jin Young; Lee, Joungmin mBio, Vol. 3, Issue 5 https://doi.org/10.1128/mBio.00314-12	journal	October 2012
Isoprenoid Pathway Optimization for Taxol Precursor Overproduction in Escherichia coli Ajikumar, P. K.; Xiao, W.-H.; Tyo, K. E. J. Science, Vol. 330, Issue 6000, p. 70-74 https://doi.org/10.1126/science.1191652	journal	September 2010
Isobutanol production at elevated temperatures in thermophilic Geobacillus thermoglucosidasius Lin, Paul P.; Rabe, Kersten S.; Takasumi, Jennifer L. Metabolic Engineering, Vol. 24 https://doi.org/10.1016/j.ymben.2014.03.006	journal	July 2014
Complete Proteomic-Based Enzyme Reaction and Inhibition Kinetics Reveal How Monolignol Biosynthetic Enzyme Families Affect Metabolic Flux and Lignin in Populus trichocarpa Wang, Jack P.; Naik, Punith P.; Chen, Hsi-Chuan The Plant Cell, Vol. 26, Issue 3 https://doi.org/10.1105/tpc.113.120881	journal	March 2014
Metabolic engineering of Thermoanaerobacterium saccharolyticum for n-butanol production Bhandiwad, Ashwini; Shaw, A. Joe; Guss, Adam Metabolic Engineering, Vol. 21 https://doi.org/10.1016/j.ymben.2013.10.012	journal	January 2014
Fermentative butanol production by clostridia Lee, Sang Yup; Park, Jin Hwan; Jang, Seh Hee Biotechnology and Bioengineering, Vol. 101, Issue 2, p. 209-228 https://doi.org/10.1002/bit.22003	journal	October 2008
Applications of computational modeling in metabolic engineering of yeast Kerkhoven, Eduard J.; Lahtvee, Petri-Jaan; Nielsen, Jens FEMS Yeast Research https://doi.org/10.1111/1567-1364.12199	journal	September 2014
Mutant alcohol dehydrogenase leads to improved ethanol tolerance in Clostridium thermocellum Brown, S. D.; Guss, A. M.; Karpinets, T. V. Proceedings of the National Academy of Sciences, Vol. 108, Issue 33 https://doi.org/10.1073/pnas.1102444108	journal	August 2011
A Toolbox for Nonlinear Regression in R : The Package nlstools Baty, Florent; Ritz, Christian; Charles, Sandrine Journal of Statistical Software, Vol. 66, Issue 5 https://doi.org/10.18637/jss.v066.i05	journal	January 2015
Uptake and activation of acetate and butyrate in Clostridium acetobutylicum Hartmanis, MarisG. N.; Klason, Tomas; Gatenbeck, Sten Applied Microbiology and Biotechnology, Vol. 20, Issue 1 https://doi.org/10.1007/BF00254648	journal	July 1984
Coenzyme A-acylating aldehyde dehydrogenase from Clostridium beijerinckii NRRL B592. Yan, R. T.; Chen, J. S. Applied and Environmental Microbiology, Vol. 56, Issue 9 https://doi.org/10.1128/aem.56.9.2591-2599.1990	journal	January 1990
Purification and Characterization of Crotonase from Clostridium acetobutylicum Waterson, Robert M.; Castellino, Francis J.; Hass, G. Michael Journal of Biological Chemistry, Vol. 247, Issue 16 https://doi.org/10.1016/S0021-9258(19)44966-1	journal	August 1972

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