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利用表达细菌 4-羟基肉桂酰辅酶 A 水合酶/裂解酶的竹细胞生物生产 4-羟基苯甲酸和香草酸的葡萄糖缀合物。

Bioproduction of glucose conjugates of 4-hydroxybenzoic and vanillic acids using bamboo cells transformed to express bacterial 4-hydroxycinnamoyl-CoA hydratase/lyase.

机构信息

Biotechnology Research Center and Department of Biotechnology, Toyama Prefectural University, Imizu, Toyama 939-0398, Japan.

Biotechnology Research Center and Department of Biotechnology, Toyama Prefectural University, Imizu, Toyama 939-0398, Japan.

出版信息

J Biosci Bioeng. 2020 Jul;130(1):89-97. doi: 10.1016/j.jbiosc.2020.02.010. Epub 2020 Mar 16.

Abstract

Rational metabolic-flow switching, which we proposed recently, is an effective strategy to produce an exogenous high-value natural product using transformed plant cells; the proof of this concept was demonstrated using bamboo (Phyllostachys nigra; Pn) cells as a model system. Pn cells were transformed to express 4-hydroxycinnamoyl-CoA hydratase/lyase of Pseudomonas putida KT2440 (PpHCHL), which catalyzes the formation of 4-hydroxybenzaldehyde and vanillin from p-coumaroyl-CoA and feruloyl-CoA, respectively. The PpHCHL-transformed cells accumulated glucose conjugates of 4-hydroxybenzoic acid and vanillic acid, indicating that the PpHCHL products (aldehydes) were further metabolized by inherent enzymes in the Pn cells. The production titers of 4-hydroxybenzoic acid glucose ester, vanillic acid glucose ester, and 4-hydroxybenzoic acid glucoside reached 1.7, 0.17, and 0.14 g/L at the maximum, respectively. These results proved the versatility of Pn cells for producing vanillin-related compounds based on rational metabolic-flow switching.

摘要

我们最近提出的理性代谢流转换是利用转化植物细胞生产外源高价值天然产物的有效策略;这一概念的证明是使用毛竹(Phyllostachys nigra;Pn)细胞作为模型系统来演示的。Pn 细胞被转化为表达恶臭假单胞菌 KT2440 的 4-羟基肉桂酰辅酶 A 水合酶/裂合酶(PpHCHL),该酶分别催化对香豆酰辅酶 A 和阿魏酰辅酶 A 形成 4-羟基苯甲醛和香草醛。PpHCHL 转化细胞积累了 4-羟基苯甲酸和香草酸的葡萄糖缀合物,表明 PpHCHL 产物(醛)被 Pn 细胞中的内源性酶进一步代谢。4-羟基苯甲酸葡萄糖酯、香草酸葡萄糖酯和 4-羟基苯甲酸葡萄糖苷的产量最高分别达到 1.7、0.17 和 0.14 g/L。这些结果证明了 Pn 细胞在基于理性代谢流转换生产香草醛相关化合物方面的多功能性。

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