Biosynthesis of acetylacetone inspired by its biodegradation
Zhou, Yifei1,2; Ding, Yamei3; Gao, Wenjie1; Wang, Jichao1; Liu, Xiutao1,2; Xian, Mo1; Feng, Xinjun1; Zhao, Guang1,4
Corresponding AuthorFeng, Xinjun( ; Zhao, Guang(
AbstractBackground Acetylacetone is a commercially bulk chemical with diverse applications. However, the traditional manufacturing methods suffer from many drawbacks such as multiple steps, harsh conditions, low yield, and environmental problems, which hamper further applications of petrochemical-based acetylacetone. Compared to conventional chemical methods, biosynthetic methods possess advantages such as being eco-friendly, and having mild conditions, high selectivity and low potential costs. It is urgent to develop biosynthetic route for acetylacetone to avoid the present problems. Results The biosynthetic pathway of acetylacetone was constructed by reversing its biodegradation route, and the acetylacetone was successfully produced by engineered Escherichia coli (E. coli) by overexpression of acetylacetone-cleaving enzyme (Dke1) from Acinetobacter johnsonii. Several promising amino acid residues were selected for enzyme improvement based on sequence alignment and structure analysis, and the acetylacetone production was improved by site-directed mutagenesis of Dke1. The double-mutant (K15Q/A60D) strain presented the highest acetylacetone-producing capacity which is 3.6-fold higher than that of the wild-type protein. Finally, the strain accumulated 556.3 +/- 15.2 mg/L acetylacetone in fed-batch fermentation under anaerobic conditions. Conclusions This study presents the first intuitive biosynthetic pathway for acetylacetone inspired by its biodegradation, and shows the potential for large-scale production.
KeywordAcetylacetone biosynthesis Acetylacetone-cleaving enzyme Rational design Site-directed mutagenesis beta-Dicarbonyl compounds
Indexed BySCI
Funding ProjectNational Defense Science and Technology Innovation Zone Foundation of China ; National Natural Science Foundation of China[31722001] ; National Natural Science Foundation of China[31670089] ; National Natural Science Foundation of China[31800081] ; Taishan Scholars Program of Shandong Province[ts201712076] ; Natural Science Foundation of Shandong[JQ201707] ; Natural Science Foundation of Shandong[ZR2019QB015] ; Youth Innovation Promotion Association at the Chinese Academy of Sciences
WOS Research AreaBiotechnology & Applied Microbiology ; Energy & Fuels
WOS SubjectBiotechnology & Applied Microbiology ; Energy & Fuels
WOS IDWOS:000536088100001
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Document Type期刊论文
Corresponding AuthorFeng, Xinjun; Zhao, Guang
Affiliation1.Chinese Acad Sci, CAS Key Lab Biobased Mat, Qingdao Inst Bioenergy & Bioproc Technol, Qingdao 266101, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China
4.Shandong Univ, State Key Lab Microbial Technol, Qingdao 266237, Peoples R China
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GB/T 7714
Zhou, Yifei,Ding, Yamei,Gao, Wenjie,et al. Biosynthesis of acetylacetone inspired by its biodegradation[J]. BIOTECHNOLOGY FOR BIOFUELS,2020,13(1):10.
APA Zhou, Yifei.,Ding, Yamei.,Gao, Wenjie.,Wang, Jichao.,Liu, Xiutao.,...&Zhao, Guang.(2020).Biosynthesis of acetylacetone inspired by its biodegradation.BIOTECHNOLOGY FOR BIOFUELS,13(1),10.
MLA Zhou, Yifei,et al."Biosynthesis of acetylacetone inspired by its biodegradation".BIOTECHNOLOGY FOR BIOFUELS 13.1(2020):10.
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