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Biosynthesis of acetylacetone inspired by its biodegradation.
Biotechnology for Biofuels ( IF 6.3 ) Pub Date : 2020-05-15 , DOI: 10.1186/s13068-020-01725-9
Yifei Zhou 1, 2 , Yamei Ding 3 , Wenjie Gao 1 , Jichao Wang 1 , Xiutao Liu 1, 2 , Mo Xian 1 , Xinjun Feng 1 , Guang Zhao 1, 4
Affiliation  

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. 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. This study presents the first intuitive biosynthetic pathway for acetylacetone inspired by its biodegradation, and shows the potential for large-scale production.

中文翻译:

乙酰丙酮的生物合成受到其生物降解的启发。

乙酰丙酮是一种具有多种应用的商业散装化学品。然而,传统的制造方法存在步骤多、条件苛刻、收率低、环境问题等诸多弊端,阻碍了石化基乙酰丙酮的进一步应用。与传统的化学方法相比,生物合成方法具有环境友好、条件温和、选择性高、潜在成本低等优点。迫切需要开发乙酰丙酮的生物合成路线,以避免目前的问题。通过逆转其生物降解途径构建了乙酰丙酮的生物合成途径,并通过工程大肠杆菌(E.coli)过表达约氏不动杆菌的乙酰丙酮裂解酶(Dke1)成功生产乙酰丙酮。基于序列比对和结构分析,选择了几个有希望的氨基酸残基进行酶改良,并通过 Dke1 的定点诱变提高了乙酰丙酮的产量。双突变株(K15Q/A60D)的乙酰丙酮生产能力最高,是野生型蛋白的3.6倍。最后,该菌株在厌氧条件下的补料分批发酵中积累了 556.3 ± 15.2 mg/L 乙酰丙酮。这项研究提出了第一个受生物降解启发的乙酰丙酮生物合成途径,并展示了大规模生产的潜力。双突变株(K15Q/A60D)的乙酰丙酮生产能力最高,是野生型蛋白的3.6倍。最后,该菌株在厌氧条件下的补料分批发酵中积累了 556.3 ± 15.2 mg/L 乙酰丙酮。这项研究提出了第一个受生物降解启发的乙酰丙酮生物合成途径,并展示了大规模生产的潜力。双突变株(K15Q/A60D)的乙酰丙酮生产能力最高,是野生型蛋白的3.6倍。最后,该菌株在厌氧条件下的补料分批发酵中积累了 556.3 ± 15.2 mg/L 乙酰丙酮。这项研究提出了第一个受生物降解启发的乙酰丙酮生物合成途径,并展示了大规模生产的潜力。
更新日期:2020-05-15
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