High-level soluble expression of human aldehyde dehydrogenase 2 in Escherichia coli achieved through lactose-mediated induction optimization

IF 1.2 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2025-09-04 DOI:10.1016/j.pep.2025.106810

Hongxia Li, Xiong Wang, Xinye Wang, Zhang Zhang, Linmin Ran

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引用次数: 0

Abstract

Aldehyde dehydrogenase 2 (ALDH2) plays a critical role in ethanol metabolism by converting toxic acetaldehyde to acetate. To investigate its functional mechanisms and potential therapeutic applications for alcohol-related diseases, heterologous expression of ALDH2 is essential. However, ALDH2 often forms inclusion bodies when expressed in Escherichia coli. In this work, the solubility of ALDH2 was enhanced by systematic optimization of expression conditions using IPTG and lactose as respective inducers. Under optimized conditions, the media yield of ALDH2 induced by IPTG and lactose reached 44.5 ± 1.3 and 48.7 ± 1.2 μg/mL respectively, representing 7.1- and 7.8-fold improvements over unoptimized conditions. Enzymatic characterization revealed that purified ALDH2 exhibited optimal activity of 9.7 U/mL at 37 °C and pH 8.0. This research demonstrates that optimizing expression conditions is an effective strategy to enhance the solubility of recombinant enzymes, while providing a practical solution for other enzymes prone to inclusion body formation.

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通过乳糖介导的诱导优化，实现了人醛脱氢酶2在大肠杆菌中的高可溶性表达

醛脱氢酶2 （ALDH2）通过将有毒的乙醛转化为乙酸，在乙醇代谢中起关键作用。为了研究其功能机制及其在酒精相关疾病中的潜在治疗应用，异源表达ALDH2是必不可少的。然而，ALDH2在大肠杆菌中表达时往往形成包涵体。本研究以IPTG和乳糖为诱导剂，通过系统优化表达条件，提高了ALDH2的溶解度。优化条件下，IPTG和乳糖诱导ALDH2的培养基产率分别达到44.5±1.3和48.7±1.2 μg/mL，分别比未优化条件提高7.1和7.8倍。酶学鉴定表明，纯化后的ALDH2在37℃、pH 8.0条件下的酶活性为9.7 U/mL。本研究表明，优化表达条件是提高重组酶溶解度的有效策略，同时也为其他易形成包涵体的酶提供了切实可行的解决方案。

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.