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首页> 外文期刊>Biotechnology Progress >Arabinose-Induction of lac-Derived Promoter Systems for Penicillin Acylase Production in Escherichia coli
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Arabinose-Induction of lac-Derived Promoter Systems for Penicillin Acylase Production in Escherichia coli

机译：阿拉伯糖诱导的lac衍生启动子系统在大肠杆菌中生产青霉素酰化酶

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摘要

Arabinose was shown to serve as an effective inducer for induction of the lac-derived promoters in Escherichia coli using penicillin acylase (PAC)as a model protein.Upon the induction with a conventional inducer,isopropyl-beta-D-thiogalactopyranoside (IPTG),for pac overexpression,which is regulated by the trc or (DE3)/T7 promoter,the production of PAC was limited by the accumulation of PAC precursors (proPAC)as inclusion bodies.Negative cellular responses,such as growth inhibition and cell lysis,were frequently observed,resulting in a low pac expression level and poor culture performance.Interestingly,these technical hurdles can be overcome simply through the use of arabinose as an inducer.The results indicate that arabinose not only induced the lac-derived promoter systems (i.e.,trc and (DE3)/T7)for pac (or LL pac)overexpression but also facilitated the posttranslational processing of proPAC for maturation.However,the arabinose-inducibility appears to be host-dependent and becomes less observable in the strains with a mutation in the ara operon.The arabinose-inducibility was also investigated in the expression system with the coexistence of the trc promoter system regulating pac expression and another arabinose-inducible promoter system of araB regulating degP coexpression.
机译：阿拉伯糖被证明是一种以青霉素酰基转移酶（PAC）为模型蛋白诱导大肠杆菌中lac衍生的启动子的有效诱导剂。在常规诱导剂异丙基-β-D-硫代半乳糖吡喃糖苷（IPTG）诱导下，对于由trc或（DE3）/ T7启动子调节的pac过表达，PAC的产生受到作为包涵体的PAC前体（proPAC）的积累的限制。负面的细胞应答，例如生长抑制和细胞裂解有趣的是，通过使用阿拉伯糖作为诱导剂，可以简单地克服这些技术障碍。结果表明，阿拉伯糖不仅诱导了lac衍生的启动子系统（即， trc和（DE3）/ T7）用于pac（或LL pac）的过表达，但也促进了proPAC的翻译后加工以使其成熟。但是，阿拉伯糖的诱导性似乎与宿主有关，并且变得不易消化。在具有ara操纵子突变的菌株中可以观察到。在表达系统中还研究了阿拉伯糖诱导性，其与调节pac表达的trc启动子系统和调节degP共表达的另一阿拉伯糖诱导型启动子系统共存。

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