Home > Archive>Volume 63, Issue 9, 2023 >3667-3678. DOI:10.13343/j.cnki.wsxb.20230052
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Cloning, expression, and characterization of three agarases in Vibrio natriegens WPAGA4
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    Abstract:

    [Objective] To clone and express three β-agarase genes agaW3418, agaW3419, and agaW3472 in Vibrio natriegens WPAGA4 isolated from the deep-sea sediment and study the enzymatic properties of the proteins. [Methods] The three agarase genes were expressed in Escherichia coli BL21(DE3) cells, and the 3,5-dinitrosalicylic acid (DNS) method was employed to measure the activities of the recombinant agarases. [Results] The three agarases, AgaW3418, AgaW3419, and AgaW3472, belonged to the GH50 family, with the highest activities at 50, 60, and 30℃ and pH 6.0, 7.0, and 7.0, respectively. AgaW3472 showed the highest activity and maintained good stability at 20℃. The highest production of AgaW3472 was detected in the super optimal broth (SOB) medium with 1% (W/V) lactose as the carbon source, 20 mmol/L MgCl2, and 0.1 mmol/L isopropyl-β-d-thiogalactoside (IPTG) at the induction temperature of 37℃. [Conclusion] The agarase AgaW3472 has high enzyme activity, low temperature adaptability, and good stability, serving as a potential bio-tool for the development of agar degradation-related industries. The three recombinant agarases provided a foundation for understanding the agarose metabolism in V. natriegens WPAGA4 and the role of this strain in the marine carbon cycling.

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TONG Xiufang, WANG Ning, ZHANG Mengyuan, WANG Wenxin, ZHANG Mouyingke, DENG Shuai, WANG Jianxin, ZENG Runying, QU Wu. Cloning, expression, and characterization of three agarases in Vibrio natriegens WPAGA4. [J]. Acta Microbiologica Sinica, 2023, 63(9): 3667-3678

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  • Received:February 05,2023
  • Revised:June 01,2023
  • Online: August 29,2023
  • Published: September 04,2023
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