Effects of Mutating the Alanine Residue in the Consensus Pentapeptide on Biochemical and Structural Characteristics of Bacillus licheniformis Lipase

IF 1.8 4区农林科学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Food Biotechnology Pub Date : 2023-07-03 DOI:10.1080/08905436.2023.2236689

Junxin Zhao, Maomao Ma, Zheling Zeng, Dongman Wan, Xianghui Yan, Jiaheng Xia, Ping Yu, D. Gong

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Abstract

ABSTRACT This study aimed to investigate the effect of mutating the consensus pentapeptide on the enzymatic properties and structural characteristics of Bacillus licheniformis lipase. Site-directed mutagenesis was used to construct mutants and then expressed in Escherichia coli BL21. After purification and enzyme activity analysis, only A75C and A75G retained activity. Compared to the wild-type lipase and A75G, the A75C showed higher thermostability at 40°C and better alkali resistance in pH 9.0–10.0 after incubation for 1 h. The kinetic parameters, structural characteristics and surface hydrophobicity of the lipases were determined. The results indicated that the secondary structure was hardly changed, while mutants exhibited higher activity, thermostability and hydrophobicity. Finally, the reasons for the increased enzyme activity were conducted through molecular dynamics simulation. The strategy of mutating the first residue of the lipase consensus pentapeptide may provide new insights into the enzyme engineering for industrial applications.

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共有五肽丙氨酸残基突变对地衣芽孢杆菌脂肪酶生化和结构特征的影响

摘要本研究旨在探讨共识五肽突变对地衣芽孢杆菌脂肪酶酶学性质和结构特征的影响。利用定点诱变技术构建突变体，并在大肠杆菌BL21中表达。经纯化和酶活性分析，仅A75C和A75G保留活性。与野生型脂肪酶和A75G相比，A75C在40℃时表现出更高的热稳定性，在pH 9.0 ~ 10.0条件下培养1 h后表现出更好的耐碱性。测定了脂肪酶的动力学参数、结构特性和表面疏水性。结果表明，突变体的二级结构几乎没有变化，但具有较高的活性、耐热性和疏水性。最后，通过分子动力学模拟分析了酶活性升高的原因。突变脂肪酶共识五肽第一残基的策略可能为酶工程的工业应用提供新的见解。

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来源期刊

Food Biotechnology 工程技术-生物工程与应用微生物

CiteScore

3.80

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： Food Biotechnology is an international, peer-reviewed journal that is focused on current and emerging developments and applications of modern genetics, enzymatic, metabolic and systems-based biochemical processes in food and food-related biological systems. The goal is to help produce and improve foods, food ingredients, and functional foods at the processing stage and beyond agricultural production. Other areas of strong interest are microbial and fermentation-based metabolic processing to improve foods, food microbiomes for health, metabolic basis for food ingredients with health benefits, molecular and metabolic approaches to functional foods, and biochemical processes for food waste remediation. In addition, articles addressing the topics of modern molecular, metabolic and biochemical approaches to improving food safety and quality are also published. Researchers in agriculture, food science and nutrition, including food and biotechnology consultants around the world will benefit from the research published in Food Biotechnology. The published research and reviews can be utilized to further educational and research programs and may also be applied to food quality and value added processing challenges, which are continuously evolving and expanding based upon the peer reviewed research conducted and published in the journal.