Biosynthesis of 2-phenylethanol from styrene using engineered Escherichia coli whole cells.

IF 3.4 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Enzyme and Microbial Technology Pub Date : 2025-01-08 DOI:10.1016/j.enzmictec.2025.110582

Tianzhen Xiong, Qiuyue Gao, Wei Liu, Wei Li, Guangyan Fan

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Abstract

2-Phenylethanol, an aromatic alcohol with a rose scent, is widely used in the cosmetics, food, and pharmaceutical industries. We designed an efficient multi-enzyme cascade pathway for production of 2-phenylethanol from styrene as the substrate. Initially, 2-phenylethanol was produced by overexpression of styrene monooxygenase A (styA), styrene monooxygenase B (styB), styrene oxide isomerase (SOI), alcohol dehydrogenase (yahK), and glucose dehydrogenase (gdh) in Escherichia coli to give 6.28 mM 2-phenylethanol. Subsequently, plasmids with different copy numbers were employed to balance the expression of pathway enzymes to produce 10.28 mM 2-phenylethanol, resulting in a 63.7 % increase in the final yield. Furthermore, the pH and temperature of the whole-cell conversion reaction were optimized, the optimum pH and temperature are 7.5 and 35℃, respectively. Finally, whole-cell conversion experiment was conducted, and the production of 2-phenylethanol reached 48.17 mM within 10 h. This study provides a theoretical and practical foundation for production of 2-phenylethanol.

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利用工程大肠杆菌全细胞从苯乙烯生物合成2-苯乙醇。

2- 苯乙醇是一种具有玫瑰香味的芳香醇，广泛应用于化妆品、食品和制药行业。我们设计了一种以苯乙烯为底物生产 2-苯乙醇的高效多酶级联途径。最初，通过在大肠杆菌中过表达苯乙烯单加氧酶 A（styA）、苯乙烯单加氧酶 B（styB）、苯乙烯氧化异构酶（SOI）、醇脱氢酶（yahK）和葡萄糖脱氢酶（ghdh）来生产 2-苯乙醇，得到 6.28 mM 的 2-苯乙醇。随后，利用不同拷贝数的质粒平衡途径酶的表达，生产出 10.28 毫摩尔的 2-苯乙醇，最终产量增加了 63.7%。此外，还对全细胞转化反应的 pH 值和温度进行了优化，最佳 pH 值和温度分别为 7.5 和 35℃。最后，进行了全细胞转化实验，10 h 内 2-苯基乙醇的产量达到 48.17 mM。这项研究为生产 2-苯基乙醇提供了理论和实践基础。

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来源期刊

Enzyme and Microbial Technology 生物-生物工程与应用微生物

CiteScore

7.60

自引率

5.90%

发文量

142

审稿时长

38 days

期刊介绍： Enzyme and Microbial Technology is an international, peer-reviewed journal publishing original research and reviews, of biotechnological significance and novelty, on basic and applied aspects of the science and technology of processes involving the use of enzymes, micro-organisms, animal cells and plant cells. We especially encourage submissions on: Biocatalysis and the use of Directed Evolution in Synthetic Biology and Biotechnology Biotechnological Production of New Bioactive Molecules, Biomaterials, Biopharmaceuticals, and Biofuels New Imaging Techniques and Biosensors, especially as applicable to Healthcare and Systems Biology New Biotechnological Approaches in Genomics, Proteomics and Metabolomics Metabolic Engineering, Biomolecular Engineering and Nanobiotechnology Manuscripts which report isolation, purification, immobilization or utilization of organisms or enzymes which are already well-described in the literature are not suitable for publication in EMT, unless their primary purpose is to report significant new findings or approaches which are of broad biotechnological importance. Similarly, manuscripts which report optimization studies on well-established processes are inappropriate. EMT does not accept papers dealing with mathematical modeling unless they report significant, new experimental data.