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工程大肠杆菌中受厌氧诱导的nirB 启动子控制的 5-氨基戊酸的高效生产。

High-efficiency production of 5-aminovalerate in engineered Escherichia coli controlled by an anaerobically-induced nirB promoter.

机构信息

Key Laboratory of Coarse Cereal Processing, Ministry of Agriculture and Rural Affairs, College of Food and Biological Engineering, Chengdu University, Chengdu, 610106, PR China.

Key Laboratory of Coarse Cereal Processing, Ministry of Agriculture and Rural Affairs, College of Food and Biological Engineering, Chengdu University, Chengdu, 610106, PR China.

出版信息

Biochem Biophys Res Commun. 2021 May 7;552:170-175. doi: 10.1016/j.bbrc.2021.03.053. Epub 2021 Mar 19.

Abstract

Biobased production of 5-aminovalerate (5AVA) from biomass can support a sustainable and economic biorefinery process to produce bio-based nylon 5 for food packaging materials. Cost-competitive production of 5AVA from biomass is a key factor in the successful commercialization of nylon 5. Bioproduction of 5AVA is a promising candidate for the industrial process to the current petrochemical route. In this study, we developed an artificial 2-keto-6-aminocaproate-mediated pathway for cost-competitive and high efficiency production of 5AVA in engineered Escherichia coli. Firstly, the combination of native l-lysine α-oxidase (RaiP) from Scomber japonicas, α-ketoacid decarboxylase (KivD) from Lactococcus lactis and aldehyde dehydrogenase (PadA) from Escherichia coli could efficiently convert l-lysine into 5AVA. Moreover, the engineered strains ML03-P-RKP, ML03-P-RKP, ML03-P-RKP induced by anaerobic condition, temperature-induced, constitutive expression instead of expensive isopropyl β-D-thiogalactoside were constructed, respectively. The use of nirB promoter induced by anaerobic condition not only could attain a higher titer of 5AVA than PL-PR and M1-93 promoters, but omit cost of expensive exogenous inducers. After the replacement of industrial materials, 5AVA titer successfully reached 33.68 g/L in engineered strain ML03-P-RKP via biotransformation. This biotransformation process conduces to the cosmically industrial 5AVA bioproduction.

摘要

从生物质中生物合成 5-氨基戊酸(5AVA)可以支持可持续和经济的生物炼制过程,以生产用于食品包装材料的生物基尼龙 5。从生物质中具有成本竞争力的 5AVA 生产是尼龙 5 成功商业化的关键因素。5AVA 的生物生产是替代当前石化路线的工业过程的有前途的候选者。在这项研究中,我们开发了一种人工 2-酮-6-氨基己酸介导的途径,用于在工程大肠杆菌中进行具有成本竞争力和高效率的 5AVA 生产。首先,来自日本竹荚鱼的天然 l-赖氨酸α-氧化酶(RaiP)、来自乳球菌 lactis 的α-酮酸脱羧酶(KivD)和来自大肠杆菌的醛脱氢酶(PadA)的组合可以将 l-赖氨酸有效地转化为 5AVA。此外,分别构建了在厌氧条件、温度诱导、组成型表达下诱导的工程菌株 ML03-P-RKP、ML03-P-RKP、ML03-P-RKP,而不是昂贵的异丙基 β-D-硫代半乳糖苷。厌氧条件下的nirB 启动子的使用不仅可以达到比 PL-PR 和 M1-93 启动子更高的 5AVA 产量,而且还可以省去昂贵的外源诱导剂的成本。在更换工业材料后,通过生物转化,工程菌株 ML03-P-RKP 中的 5AVA 产量成功达到 33.68 g/L。这种生物转化过程有助于实现宇宙工业规模的 5AVA 生物生产。

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