海洋弧菌HN897 β-琼脂糖酶基因vas1-1339异源表达及活性分析

doi:10.11978/2021065

Abstract

Abstract:

The marine Vibrio HN897 strain is an agarose-producing Vibrio astriarenae. Studies indicated that the absence of the gene Vas1-1339, encoding a β-agarase, significantly reduced the hydrolysis effect of Vibrio HN897 strain on agarose. Herein, we further analyzed the expression and enzymatic characterization of the β-agarase gene in heterologous bacteria, Escherichia coli (E. coli). Vas1-1339 agarase was successfully expressed in E. coli, and the optimal concentration of inducer, IPTG, was at 10 μmol·L^-1. Immunoblotting analysis showed that the C-terminal of the gene expression product might be cleavage in E. coli, suggesting complex maturation process of this protein. Lugol’s iodine staining analysis revealed that E. coli, expressing Vas1-1339 agarase, was highly effective in degrading agarose, and purified protein also had this similar function, which suggests that the β-agarase Vas1-1339 of the HN897 strain can independently play the role on agarolytic degradation. These results lay a preliminary foundation for the functional study and related technology application of β-agarase derived from marine vibrion.

Key words: β-agarase, marine Vibrio, agarose hydrolysis, protein expression

CLC Number:

P735.51

YU Fei, JIN Xingkun, LEI Tianying, CAO Haihang, CHEN Qianghui, YANG Yaofan, LI Jiahang, ZHAO Zhe. Heterologous expression and enzymatic characterization of marine Vibrio astriarenae-derived β-Agarase gene vas1-1339[J].Journal of Tropical Oceanography, 2022, 41(2): 170-176.

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References 22

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Heterologous expression and enzymatic characterization of marine Vibrio astriarenae-derived β-Agarase gene vas1-1339

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