重组 Zymomonas mobilis 锌依赖性醇脱氢酶 I 的强化功能表达。

IF 4.1 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of biotechnology Pub Date : 2024-09-29 DOI:10.1016/j.jbiotec.2024.09.012

Klaudia Žigová , Zuzana Marčeková , Tatiana Petrovičová , Katarína Lorková , František Čacho , Vladimír Krasňan , Martin Rebroš

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引用次数: 0

摘要

莫比莱兹单胞菌（Zymomonas mobilis）的醇脱氢酶 I（zmADH1）是一种锌依赖性氧化还原酶，它以 NAD+/NADH 为辅助因子，催化伯醇或仲醇氧化成相应的醛或酮。在大肠杆菌中以可溶性形式表达zmADH1的努力遇到了溶解性方面的困难。通过在培养基中添加 1mM 的锌，重组的 zmADH1 实现了可溶性。锌的添加有利于重组zmADH1的正常折叠，并显著减少包涵体的形成。重组 zmADH1 的产量约为 30 毫克/1 升 Luria-Bertani 培养基。在发酵罐中的强化生产表明，在不同锌浓度下生产的 zmADH1 的特异活性和总活性之间存在显著差异。zmADH1 对中链醇（尤其是 1-戊醇）具有亲和力，可用于制备醛和醇的新绿色途径。

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Intensified functional expression of recombinant Zymomonas mobilis zinc-dependent alcohol dehydrogenase I

Alcohol dehydrogenase I from Zymomonas mobilis (zmADH1) is a zinc-dependent oxidoreductase that catalyses the oxidation of primary or secondary alcohols to the corresponding aldehydes or ketones using NAD⁺/NADH as a cofactor. Efforts to express zmADH1 in Escherichia coli in a soluble form have been laden with solubility difficulties. A soluble form of recombinant zmADH1 was achieved by the addition of 1 mM zinc into media. Zinc addition facilitates the proper folding of recombinant zmADH1 and significantly reduces the formation of inclusion bodies. The yield of recombinant zmADH1 represents approximately 30 mg/1 L Luria-Bertani media. Intensified production in fermenters showed a striking difference between the specific and total activities of zmADH1 produced at different zinc concentrations. The zmADH1 showed an affinity to medium-chain alcohols, especially 1-pentanol, which could be used in new greener routes for preparation of aldehydes and alcohols.

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来源期刊

Journal of biotechnology 工程技术-生物工程与应用微生物

CiteScore

8.90

自引率

2.40%

发文量

190

审稿时长

45 days

期刊介绍： The Journal of Biotechnology has an open access mirror journal, the Journal of Biotechnology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. The Journal provides a medium for the rapid publication of both full-length articles and short communications on novel and innovative aspects of biotechnology. The Journal will accept papers ranging from genetic or molecular biological positions to those covering biochemical, chemical or bioprocess engineering aspects as well as computer application of new software concepts, provided that in each case the material is directly relevant to biotechnological systems. Papers presenting information of a multidisciplinary nature that would not be suitable for publication in a journal devoted to a single discipline, are particularly welcome.