Directed Evolution Improves the Enzymatic Synthesis of L-5-Hydroxytryptophan by an Engineered Tryptophan Synthase.

Applied Biochemistry and Biotechnology
Lisheng XuBianling Jiang

Abstract

L-5-Hydroxytryptophan is an important amino acid that is widely used in food and medicine. In this study, L-5-hydroxytryptophan was synthesized by a modified tryptophan synthase. A direct evolution strategy was applied to engineer tryptophan synthase from Escherichia coli to improve the efficiency of L-5-hydroxytryptophan synthesis. Tryptophan synthase was modified by error-prone PCR. A high-activity mutant enzyme (V231A/K382G) was obtained by a high-throughput screening method. The activity of mutant enzyme (V231A/K382G) is 3.79 times higher than that of its parent, and kcat/Km of the mutant enzyme (V231A/K382G) is 4.36 mM-1∙s-1. The mutant enzyme (V231A/K382G) reaction conditions for the production of L-5-hydroxytryptophan were 100 mmol/L L-serine at pH 8.5 and 35°C for 15 h, reaching a yield of L-5-hydroxytryptophan of 86.7%. Directed evolution is an effective strategy to increase the activity of tryptophan synthase.

References

Apr 3, 2001·Current Opinion in Chemical Biology·U T Bornscheuer, M Pohl
Oct 24, 2009·Biochemistry·Adam T DierkersMichael F Dunn
Dec 15, 2010·Journal of the American Chemical Society·Jinfeng LaiLeonard J Mueller
Oct 17, 2012·Journal of Ethnopharmacology·Liang YeJingwei Tian
Jun 22, 2015·Applied Biochemistry and Biotechnology·Zhenming ChenLinshu Jiang
Dec 30, 2015·Applied Biochemistry and Biotechnology·Hui LuoZhongyao Shen
Feb 24, 2016·Applied Microbiology and Biotechnology·Lin ZhuShuang-Yan Tang
Jun 30, 2016·Journal of the American Chemical Society·Michael HergerFrances H Arnold
Jun 2, 2017·Journal of the Neurological Sciences·Gabriella A HorvathBruce Carleton
Sep 28, 2017·Food Chemistry·Huiqi WangXiaoming Zhang
Dec 5, 2017·Bioscience, Biotechnology, and Biochemistry·Dongdong MuZhi Zheng
May 2, 2018·Journal of the American Chemical Society·Andrew R BullerFrances H Arnold
Sep 15, 2018·Angewandte Chemie·Christina E BovilleFrances H Arnold
Feb 14, 2019·Bioresource Technology·Huawen HanXiangkai Li

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