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Food Biotechnology Volume 37, 2023 - Issue 3
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Research Article

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

Junxin Zhaoa State Key Laboratory of Food Science and Resources, Nanchang University, Nanchang, China;b Jiangxi Province Key Laboratory of Edible and Medicinal Resources Exploitation, Nanchang University, Nanchang, China;c School of Food Science and Technology, Nanchang University, Nanchang, ChinaView further author information
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Maomao Maa State Key Laboratory of Food Science and Resources, Nanchang University, Nanchang, China;b Jiangxi Province Key Laboratory of Edible and Medicinal Resources Exploitation, Nanchang University, Nanchang, China;c School of Food Science and Technology, Nanchang University, Nanchang, ChinaView further author information
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Zheling Zenga State Key Laboratory of Food Science and Resources, Nanchang University, Nanchang, China;b Jiangxi Province Key Laboratory of Edible and Medicinal Resources Exploitation, Nanchang University, Nanchang, China;d School of Resource and Environmental and Chemical Engineering, Nanchang University, Nanchang, ChinaCorrespondence[email protected] [email protected]
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Dongman Wana State Key Laboratory of Food Science and Resources, Nanchang University, Nanchang, China;b Jiangxi Province Key Laboratory of Edible and Medicinal Resources Exploitation, Nanchang University, Nanchang, China;c School of Food Science and Technology, Nanchang University, Nanchang, ChinaView further author information
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Xianghui Yana State Key Laboratory of Food Science and Resources, Nanchang University, Nanchang, China;b Jiangxi Province Key Laboratory of Edible and Medicinal Resources Exploitation, Nanchang University, Nanchang, China;d School of Resource and Environmental and Chemical Engineering, Nanchang University, Nanchang, ChinaView further author information
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Jiaheng Xiaa State Key Laboratory of Food Science and Resources, Nanchang University, Nanchang, China;b Jiangxi Province Key Laboratory of Edible and Medicinal Resources Exploitation, Nanchang University, Nanchang, China;d School of Resource and Environmental and Chemical Engineering, Nanchang University, Nanchang, ChinaView further author information
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Ping Yua State Key Laboratory of Food Science and Resources, Nanchang University, Nanchang, China;b Jiangxi Province Key Laboratory of Edible and Medicinal Resources Exploitation, Nanchang University, Nanchang, China;d School of Resource and Environmental and Chemical Engineering, Nanchang University, Nanchang, ChinaCorrespondence[email protected]
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Deming Gonga State Key Laboratory of Food Science and Resources, Nanchang University, Nanchang, China;b Jiangxi Province Key Laboratory of Edible and Medicinal Resources Exploitation, Nanchang University, Nanchang, China;e New Zealand Institute of Natural Medicine Research, Auckland, New ZealandView further author information
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Pages 280-300 | Published online: 01 Aug 2023
 
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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.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Supplementary material

Supplemental data for this article can be accessed online at https://doi.org/10.1080/08905436.2023.2236689.

Additional information

Funding

The work was supported by the the International Science and Technology Cooperation Program of China [2011DFA32770]; the National Natural Science Foundation of China [31701651, 32060516]; the Research Program of State Key Laboratory of Food Science and Technology, Nanchang University [SKLF-ZZA-201910, SKLF-ZZB-201916, SKLF-ZZB-202135]; the Jiangxi Provincial Natural Science Foundation [20224BAB215050]; the Central Government Guide Local Special Fund Project for Scientific and Technological Development of Jiangxi Province [20212ZDD02008, 20221ZDD02001]; the Science and Technology Program of Jiangxi Province [20143ACG70015].

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