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High-level De novo biosynthesis of arbutin in engineered Escherichia coli

熊果苷 生物合成 运动发酵单胞菌 大肠杆菌 生物化学 代谢工程 化学 葡萄糖基转移酶 莽草酸途径 锥形烧瓶 生物 发酵 基因 色谱法 乙醇燃料
作者
Xiaolin Shen,Jia Wang,Jian Wang,Zhenya Chen,Qipeng Yuan,Yajun Yan
出处
期刊:Metabolic Engineering [Elsevier]
卷期号:42: 52-58 被引量:57
标识
DOI:10.1016/j.ymben.2017.06.001
摘要

Arbutin is a hydroquinone glucoside compound existing in various plants. It is widely used in pharmaceutical and cosmetic industries owing to its well-known skin-lightening property as well as anti-oxidant, anti-microbial, and anti-inflammatory activities. Currently, arbutin is usually produced by plant extraction or enzymatic processes, which suffer from low product yield and expensive processing cost. In this work, we established an artificial pathway in Escherichia coli for high-level production of arbutin from simple carbon sources. First, a 4-hydroxybenzoate 1-hydroxylase from Candida parapsilosis CBS604 and a glucosyltransferase from Rauvolfia serpentina were characterized by in vitro enzyme assays. Introduction of these two genes into E. coli led to the production of 54.71 mg/L of arbutin from glucose. Further redirection of carbon flux into arbutin biosynthesis pathway by enhancing shikimate pathway genes enabled production of 3.29 g/L arbutin, which is a 60-fold increase compared with the initial strain. Final optimization of glucose concentration added in the culture medium was able to further improve the titer of arbutin to 4.19 g/L in shake flasks experiments, which is around 77-fold higher than that of initial strain. This work established de novo biosynthesis of arbutin from simple carbon sources and provided a generalizable strategy for the biosynthesis of shikimate pathway derived chemicals. The high titer achieved in our engineered strain also indicates the potential for industrial scale bio-manufacturing of arbutin.
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