A modified pathway for the production of acetone in Escherichia coli |
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| Institution: | 1. Abteilung Mikrobiologie, Institut für Biowissenschaften, Universität Rostock, Albert-Einstein-Str. 3, D-18051 Rostock, Germany;2. Institut für Mikrobiologie und Biotechnologie, Universität Ulm, Albert-Einstein-Allee 11, D-89081 Ulm, Germany;3. Evonik Industries AG, Creavis Technologies & Innovation, Paul-Baumann-Strasse 1, D-45772 Marl, Germany;4. Evonik Industries AG, Service Center Biocatalysis, Rodenbacher Chaussee 4, D-63457 Hanau-Wolfgang, Germany;1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China;2. China Shijiazhuang Pharmaceutical Group Co., Ltd., Shijiazhuang 050038, China;3. School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, China;4. School of Life Science and Food Engineering, Huaiyin Institute of Technology, Huaian 223003, China;1. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, No. 30, Puzhu South Road, Nanjing 211816, PR China;2. Jiangsu National Synergetic Innovation Center for Advance Material (SICAM), No. 30, Puzhu South Road, Nanjing 211816, PR China;3. State Key Laboratory of Bioreactor Engineering, Biomedical Nanotechnology Center, East China University of Science and Technology, Shanghai 200237, PR China;1. School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea;2. School of Energy and Chemical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919, Republic of Korea;1. Laboratory for Bioinformatics, Graduate School of Systems Biosciences, Kyushu University, 804 Westwing, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan;2. Laboratory of Plant Physiology and Metabolism, Graduate School of Life and Environmental Sciences, University of Tsukuba, Tennodai, Tsukuba 305-8572, Japan;1. Department of Chemical Engineering, Feng Chia University, 100 Wenhwa Road, Taichung 40724, Taiwan;2. Department of Medical Laboratory Science and Biotechnology, China Medical University, No. 91, Hsueh-Shih Road, Taichung 40402, Taiwan;3. Department of Health and Nutrition Biotechnology, Asia University, Taichung 41354, Taiwan;4. Department of Medical Research, China Medical University Hospital, Taichung 40447, Taiwan |
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| Abstract: | A modified synthetic acetone operon was constructed. It consists of two genes from Clostridium acetobutylicum (thlA coding for thiolase and adc coding for acetoacetate decarboxylase) and one from Bacillus subtilis or Haemophilus influenzae (teIIsrf or ybgC, respectively, for thioesterase). Expression of this operon in Escherichia coli resulted in the production of acetone starting from the common metabolite acetyl-CoA via acetoacetyl-CoA and acetoacetate. The thioesterases do not need a CoA acceptor for acetoacetyl-CoA hydrolysis. Thus, in contrast to the classic acetone pathway of Clostridium acetobutylicum and related microorganisms which employ a CoA transferase, the new pathway is acetate independent. The genetic background of the host strains was crucial. Only E. coli strains HB101 and WL3 were able to produce acetone via the modified plasmid based pathway, up to 64 mM and 42 mM in 5-ml cultures, respectively. Using glucose fed-batch cultures the concentration could be increased up to 122 mM acetone with HB101 carrying the recombinant plasmid pUC19ayt (thioesterase from H. influenzae). The formation of acetone led to a decreased acetate production by E. coli. |
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