Evaluation of 3-hydroxypropionate biosynthesis in vitro by partial introduction of the 3-hydroxypropionate/4-hydroxybutyrate cycle from <Emphasis Type="Italic">Metallosphaera sedula</Emphasis> |
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| Authors: | Ziling Ye Xiaowei Li Yongbo Cheng Zhijie Liu Gaoyi Tan Fayin Zhu Shuai Fu Zixin Deng Tiangang Liu |
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| Institution: | 1.Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education,Wuhan University School of Pharmaceutical Sciences,Wuhan,People’s Republic of China;2.Hubei Engineering Laboratory for Synthetic Microbiology,Wuhan Institute of Biotechnology,Wuhan,People’s Republic of China;3.J1 Biotech, Co., Ltd,Wuhan,People’s Republic of China;4.Hubei Provincial Cooperative Innovation Center of Industrial Fermentation,Wuhan,People’s Republic of China;5.The State Key Laboratory of Microbial Metabolism,Shanghai Jiao Tong University,Shanghai,People’s Republic of China |
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| Abstract: | The chemical 3-hydroxypropionate (3HP) is an important starting reagent for the commercial synthesis of specialty chemicals. In this study, a part of the 3-hydroxypropionate/4-hydroxybutyrate cycle from Metallosphaera sedula was utilized for 3HP production. To study the basic biochemistry of this pathway, an in vitro-reconstituted system was established using acetyl-CoA as the substrate for the kinetic analysis of this system. The results indicated that 3HP formation was sensitive to acetyl-CoA carboxylase and malonyl-CoA reductase, but not malonate semialdehyde reductase. Also, the competition between 3HP formation and fatty acid production was analyzed both in vitro and in vivo. This study has highlighted how metabolic flux is controlled by different catalytic components. We believe that this reconstituted system would be valuable for understanding 3HP biosynthesis pathway and for future engineering studies to enhance 3HP production. |
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