Evaluation of 3-hydroxypropionate biosynthesis in vitro by partial introduction of the 3-hydroxypropionate/4-hydroxybutyrate cycle from Metallosphaera sedula.
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.
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, Wuhan University School of Pharmaceutical Sciences, Wuhan, 430071, People's Republic of China.
Ziling
Z
Ye
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, Wuhan University School of Pharmaceutical Sciences, Wuhan, 430071, People's Republic of China.
Xiaowei
X
Li
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, Wuhan University School of Pharmaceutical Sciences, Wuhan, 430071, People's Republic of China.
Yongbo
Y
Cheng
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, Wuhan University School of Pharmaceutical Sciences, Wuhan, 430071, People's Republic of China.
Zhijie
Z
Liu
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, Wuhan University School of Pharmaceutical Sciences, Wuhan, 430071, People's Republic of China.
Gaoyi
G
Tan
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, Wuhan University School of Pharmaceutical Sciences, Wuhan, 430071, People's Republic of China.
Fayin
F
Zhu
J1 Biotech, Co., Ltd, Wuhan, 430075, People's Republic of China.
Shuai
S
Fu
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, Wuhan University School of Pharmaceutical Sciences, Wuhan, 430071, People's Republic of China.|Hubei Engineering Laboratory for Synthetic Microbiology, Wuhan Institute of Biotechnology, Wuhan, 430075, People's Republic of China.|The State Key Laboratory of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai, 200030, People's Republic of China.
Zixin
Z
Deng
Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, Wuhan University School of Pharmaceutical Sciences, Wuhan, 430071, People's Republic of China. liutg@whu.edu.cn.|Hubei Engineering Laboratory for Synthetic Microbiology, Wuhan Institute of Biotechnology, Wuhan, 430075, People's Republic of China. liutg@whu.edu.cn.|Hubei Provincial Cooperative Innovation Center of Industrial Fermentation, Wuhan, 430068, People's Republic of China. liutg@whu.edu.cn.
Tiangang
T
Liu
D000105:Acetyl Coenzyme A / Q000378:metabolism; D000103:Acetyl-CoA Carboxylase / Q000378:metabolism; D053898:Biosynthetic Pathways; D057486:Carbon Cycle; D007700:Kinetics; D019344:Lactic Acid / Q000031:analogs & derivatives* / Q000096:biosynthesis; D010088:Oxidoreductases / Q000378:metabolism; D012978:Sodium Oxybate / Q000378:metabolism*; D016943:Sulfolobaceae / Q000201:enzymology / Q000378:metabolism*
D019344:Lactic Acid; D000105:Acetyl Coenzyme A; D012978:Sodium Oxybate; C031601:hydracrylic acid; D010088:Oxidoreductases; C454539:malonyl-Coa reductase; D000103:Acetyl-CoA Carboxylase
3-Hydroxypropionate; 3-Hydroxypropionate/4-hydroxybutyrate cycle; Escherichia coli; Metabolic engineering; Metallosphaera sedula