L-苯乳酸是一种天然抑菌物质,对多种病原微生物有光谱抑制作用,有望成为一种新型生物防腐剂。通过构建多酶级联催化反应体系,提高大肠杆菌合成L-苯乳酸的能力。利用共表达L-氨基酸脱氨酶和苯丙酮酸还原酶并偶联葡萄糖脱氢酶进行辅酶再生,建立一种新型L-苯乳酸生物合成方法。各基因成功在大肠杆菌中表达,并对全细胞转化条件进行优化,最适转化条件为反应温度30 ℃,初始pH 值为8.0,底物苯丙氨酸30 g/L,菌体OD600为30,辅底物葡萄糖1倍摩尔当量,转化反应12 h,共生成L-苯乳酸21.39 g/L,摩尔转化率为71.33%。提供了一种简洁、高效的生物催化方法,为实现规模化生产奠定了基础。
L-Phenyllactic acid is a natural antibacterial substance, which has a spectral inhibitory effect on a variety of pathogenic microorganisms. It is expected to become a new type of biological preservative. A multi-enzyme cascade catalytic reaction system was constructed to improve the ability of L-phenyllactic acid synthesis by Escherichia coli whole cell. A new L-phenyllactic acid biosynthesis method was established by co-expressing L-amino acid deaminase and phenylpyruvate reductase coupled with glucose dehydrogenase for coenzyme regeneration. The genes were successfully expressed in E.coli. The whole cell transformation conditions were optimized. The optimal transformation conditions were reaction at 30 ℃, initial pH 8.0, substrate phenylalanine of 30 g/L, OD600 of 30, and auxiliary substrate glucose of one-time molar equivalent, conversion reaction time of 12 h. A total of 21.39 g/L of L-phenyllactic acid was produced, and the molar conversion rate was 71.33%. A simple and efficient biocatalysis method is provided, which lays the foundation for the realization of large-scale production.
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