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Characterization and Mechanism Study of a Novel PL7 Family Exolytic Alginate Lyase from Marine Bacteria Vibrio sp. W13

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Abstract

Alginate lyase can degrade alginate into oligosaccharides through β-elimination for various biological, biorefinery, and agricultural purposes. Here, we report a novel PL7 family exolytic alginate lyase VwAlg7A from marine bacteria Vibrio sp. W13 and achieve the heterologous expression in E. coli BL21 (DE3). VwAlg7A is 348aa with a calculated molecular weight of 36 kDa, containing an alginate lyase 2 domain. VwAlg7A exhibits specificity towards poly-guluronate. The optimal temperature and pH of VwAlg7A are 30 °C and 7.0, respectively. The activity of VwAlg7A can be significantly inhibited by the Ni2+, Zn2+, and NaCl. The Km and Vmax of VwAlg7A are 36.9 mg/ml and 395.6 μM/min, respectively. The ESI and HPAEC-PAD results indicate that VwAlg7A cleaves the sugar bond in an exolytic mode. Based on the molecular docking and mutagenesis results, we further confirmed that R98, H169, and Y303 are important catalytic residues.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This work was supported by the Dalian Science and Technology Innovation Fund-Key&Major Subject (2020JJ25CY017); Liaoning Provincial Marine Economic Development Project (2022-47); ANSO Collaborative Research Program (ANSO-CR-KP-2020-14); Natural Science Foundation of Liaoning Province, China (2020-MS-025); and Outstanding Member Fund of CAS Youth Innovation Promotion Association (Y201939).

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Authors and Affiliations

  1. Dalian Engineering Research Center for Carbohydrate Agricultural Preparations, Dalian Technology Innovation Center for Green Agriculture, Liaoning Provincial Key Laboratory of Carbohydrates, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116023, China

    Zhongbin Xiao, Kuikui Li, Tang Li, Fanxing Zhang, Jiayi Xue & Heng Yin

  2. Department of Materials and Chemicals, Dalian Polytechnic University, Dalian, 116023, China

    Zhongbin Xiao, Fanxing Zhang, Jiayi Xue & Miao Zhao

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  1. Zhongbin Xiao
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  2. Kuikui Li
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  3. Tang Li
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  4. Fanxing Zhang
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  5. Jiayi Xue
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  6. Miao Zhao
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  7. Heng Yin
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Contributions

All authors contributed to the study conception and design. Material preparation was performed by ZBX, FXZ, and JYX. Data collection and analysis were performed by ZBX. Method development was performed by KKL, TL, MZ, and HY. The first draft of the manuscript was written by ZBX and all authors commented on previous versions of the manuscript. Funding acquisition and supervision were performed by HY. All authors read and approved the final manuscript.

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Correspondence to Heng Yin.

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Xiao, Z., Li, K., Li, T. et al. Characterization and Mechanism Study of a Novel PL7 Family Exolytic Alginate Lyase from Marine Bacteria Vibrio sp. W13. Appl Biochem Biotechnol 196, 68–84 (2024). https://doi.org/10.1007/s12010-023-04483-0

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