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Cloning, Expressing and Characterizing a Novel Cold-Adapted Laminarinase from Marine Bacterium Wenyingzhuangia aestuarii OF219

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Abstract

Laminarinases reveal potential application in the field of food and biotechnology. In this research, a novel GH16 family laminarinase, designated as Lam16A_Wa, was cloned from the genome of marine bacterium Wenyingzhuangia aestuarii OF219 and expressed in Escherichia coli. Lam16A_Wa demonstrates a relatively low optimal reaction temperature (35°C) and a cold-adapted feature. Its optimal pH value is 6.0 and is stable in a broad pH range from 3.0 to 11.0. A glycomics strategy was employed to investigate the hydrolytic pattern of Lam16A_Wa. The enzyme was confirmed as a random endo-acting glycoside hydrolase. Its minimum substrate was laminarin pentasaccharide, and the major final products are oligosaccharides, including disaccharide to pentasaccharide. The Lam16A_Wa provides a novel and well-defined tool for the molecular tailoring of laminarin.

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References

  • Altschul, S., Madden, T., Schaffer, A., Zhang, J. H., Zhang, Z., Miller, W., et al., 1998. Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Faseb Journal, 12(8, S): A1326.

    Google Scholar 

  • Balla, S., Laurent, L., Sujata, S. O., Aruna, V., Richard, D., Hsei-Wei, W., et al., 2014. Oligo-β-(1→3)-glucans: Impact of thiobridges on immunostimulating activities and the development of cancer stem cells. Journal of Medicinal Chemistry, 57(20): 8280–8292.

    Article  Google Scholar 

  • Bl Ttel, V., Larisika, M., Pfeiffer, P., Nowak, C., Eich, A., Eckelt, J., et al., 2011. Beta-1,3-glucanase from Delftia tsuruhatensis strain MV01 and its potential application in vinification. Applied & Environmental Microbiology, 77(3): 983–990.

    Article  Google Scholar 

  • Burkhardt, C., Schaefers, C., Claren, J., Schirrmacher, G., and Antranikian, G., 2019. Comparative analysis and biochemical characterization of two endo-beta-1,3-glucanases from the thermophilic bacterium Fervidobacterium sp.. CATALYSTS, 9: 830.

    Article  Google Scholar 

  • Chang, Y., Hu, Y., and McClements, D. J., 2016. Competitive adsorption and displacement of anionic polysaccharides (fucoidan and gum arabic) on the surface of protein-coated lipid droplets. Food Hydrocolloid, 52: 820–826.

    Article  Google Scholar 

  • Cheng, R., Chen, J., Yu, X., Wang, Y., Wang, S., and Zhang, J., 2013. Recombinant production and characterization of full-length and truncated beta-1,3-glucanase PglA from Paenibacillus sp S09. BMC Biotechnology, 13: 105.

    Article  Google Scholar 

  • Cheng, Y., Hong, T., Liu, C., and Meng, M., 2009. Cloning and functional characterization of a complex endo-beta-1,3-glucanase from Paenibacillus sp.. Applied Microbiology and Biotechnology, 81(6): 1051–1061.

    Article  Google Scholar 

  • Hartl, L., Gastebois, A., Aimanianda, V., and Latge, J., 2011. Characterization of the GPI-anchored endo beta-1,3-glucanase Eng2 of Aspergillus fumigatus. Fungal Genetics and Biology, 48(2): 185–191.

    Article  Google Scholar 

  • Hong, T. Y., and Meng, M., 2003. Biochemical characterization and antifungal activity of an endo-1,3-beta-glucanase of Paenibacillus sp. isolated from garden soil. Applied Microbiology and Biotechnology, 61(5–6): 472–478.

    Article  Google Scholar 

  • Hong, T. Y., Cheng, C. W., Huang, J. W., and Meng, M. S., 2002. Isolation and biochemical characterization of an endo-1,3-beta-glucanase from Streptomyces sioyaensis containing a C-terminal family 6 carbohydrate-binding module that binds to 1,3-beta-glucan. Microbiology-SGM, 148(Pt 4): 1151.

    Article  Google Scholar 

  • Jaafar, N. R., Khoiri, N. M., Ismail, N. F., Mahmood, N. A. N., Abdul Murad, A. M., Abu Bakar, F. D., et al., 2020. Functional characterisation and product specificity of Endo-beta-1,3-glucanase from alkalophilic bacterium, Bacillus lehensis G1. Enzyme and Microbial Technology, 140: 109625.

    Article  Google Scholar 

  • Kadam, S. U., O’Donnell, C. P., Rai, D. K., Hossain, M. B., Burgess, C. M., Walsh, D., et al., 2015. Laminarin from Irish brown seaweeds Ascophyllum nodosum and laminaria hyperborea: Ultrasound assisted extraction, characterization and bioactivity. Marine Drugs, 13(7): 4270–4280.

    Article  Google Scholar 

  • Kim, K. H., Kim, Y. W., Han, B. K., Lee, B. J., and Dong, S. L., 2006. Anti-apoptotic activity of laminarin polysaccharides and their enzymatically hydrolyzed oligosaccharides from Laminaria japonica. Biotechnology Letters, 28(6): 439–446.

    Article  Google Scholar 

  • Klarzynski, O., Plesse, B., Joubert, J. M., Yvin, J. C., Kopp, M., Kloareg, B., et al., 2000. Linear beta-1,3 glucans are elicitors of defense responses in tobacco. Plant Physiology, 124(3): 1027–1038.

    Article  Google Scholar 

  • Krah, M., Misselwitz, R., Politz, O., Thomsen, K., Welfle, H., and Borriss, R., 2010. The laminarinase from thermophilic eubacterium Rhodothermus marinus–Conformation, stability, and identification of active site carboxylic residues by site-directed mutagenesis. European Journal of Biochemistry, 257(1): 101–111.

    Article  Google Scholar 

  • Kumar, K., Correia, M. A. S., Pires, V. M. R., Dhillon, A., Sharma, K., Rajulapati, V., et al., 2018. Novel insights into the degradation of beta-1,3-glucans by the cellulosome of Clostridium thermocellum revealed by structure and function studies of a family 81 glycoside hydrolase. International Journal of Biological Macromolecules, 117: 890–901.

    Article  Google Scholar 

  • Kusaykin, M. I., Belik, A. A., Kovalchuk, S. N., Dmitrenok, P. S., Rasskazov, V. A., Isakov, V. V., et al., 2017. A new recombinant endo-1,3-beta-D-glucanase from the marine bacterium Formosa algae KMM 3553: Enzyme characteristics and transglycosylation products analysis. World Journal of Microbiology & Biotechnology, 33(2): 40.

    Article  Google Scholar 

  • Labourel, A., Jam, M., Jeudy, A., Hehemann, J., Czjzek, M., and Michel, G., 2014. The beta-glucanase ZgLamA from Zobellia galactanivorans evolved a bent active site adapted for efficient degradation of algal laminarin. Journal of Biological Chemistry, 289(4): 2027–2042.

    Article  Google Scholar 

  • Labourel, A., Jam, M., Legentil, L., Sylla, B., Hehemann, J., Ferrieres, V., et al., 2015. Structural and biochemical characterization of the laminarinase ZgLamC (GH16) from Zobellia galactanivorans suggests preferred recognition of branched laminarin. Acta Crystallographica Section D–Structural Biology, 71(2): 173–184.

    Article  Google Scholar 

  • Lee, K. C., Arai, T., Ibrahim, D., Kosugi, A., Prawitwong, P., Lan, D., et al., 2014. Purification and characterization of a thermostable laminarinase from Penicillium rolfsii c3-2 (1) IBRL. BioResources, 9(1): 1072–1084.

    Article  Google Scholar 

  • Lever, M., 1972. A new reaction for colorimetric determination of carbohydrates. Analytical Biochemistry, 47(1): 273–279.

    Article  Google Scholar 

  • Liu, Z., Xiong, Y., Yi, L., Dai, R., Wang, Y., Sun, M., et al., 2018. Endo-β-1,3-glucanase digestion combined with the HPAEC-PAD-MS/MS analysis reveals the structural differences between two laminarins with different bioactivities. Carbohydrate Polymers, 194: 339.

    Article  Google Scholar 

  • Masuda, S., Endo, K., Koizumi, N., Hayami, T., Fukazawa, T., Yatsunami, R., et al., 2006. Molecular identification of a novel beta-1,3-glucanase from alkaliphilic Nocardiopsis sp strain F96. Extremophiles, 10(3): 251–255.

    Article  Google Scholar 

  • Mitsuya, D., Sugiyama, T., Zhang, S., Takeuchi, Y., Okai, M., Urano, N., et al., 2018. Enzymatic properties and the gene structure of a cold-adapted laminarinase from Pseudoalteromonas species LA. Journal of Bioscience and Bioengineering, 126(2): 169–175.

    Article  Google Scholar 

  • Miyanishi, N., Iwamoto, Y., Watanabe, E., and Odaz, T., 2003. Induction of TNF-α production from human peripheral blood monocytes with β-1,3-glucan oligomer prepared from laminarin with β-1,3-glucanase from Bacillus clausii NM-1. Journal of Bioscience & Bioengineering, 95(2): 192–195.

    Article  Google Scholar 

  • Petersen, T. N., Brunak, S., von Heijne, G., and Nielsen, H., 2011. SignalP 4.0: Discriminating signal peptides from transmembrane regions. Nature Methods, 8(10): 785–786.

    Article  Google Scholar 

  • Shi, P. J., Yao, G. Y., Yang, P. L., Li, N., Luo, H. Y., Bai, Y. G., et al., 2010. Cloning, characterization, and antifungal activity of an endo-1,3-beta-D-glucanase from Streptomyces sp. S27. Applied Microbiology & Biotechnology, 85(5): 1483–1490.

    Article  Google Scholar 

  • Tamura, K., Stecher, G., Peterson, D., Filipski, A., and Kumar, S., 2013. MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0. Molecular Biology and Evolution, 30(12): 2725–2729.

    Article  Google Scholar 

  • Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F., and Higgins, D. G., 1997. The CLUSTAL_X windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 25(24): 4876–4882.

    Article  Google Scholar 

  • Wilkins, M. R., Gasteiger, E., Bairoch, A., Sanchez, J. C., Williams, K. L., Appel, R. D., et al., 1999. Protein identification and analysis tools in the ExPASy server. Methods in Molecular Biology, 112(112): 531.

    Google Scholar 

  • Yi, P., Yan, Q., Jiang, Z., and Wang, L., 2018. A first glycoside hydrolase family 50 endo-beta-1,3-D-glucanase from Pseudomonas aeruginosa. Enzyme and Microbial Technology, 108: 34–41.

    Article  Google Scholar 

  • Yin, G., Li, W., Lin, Q., Lin, X., Lin, J., Zhu, Q., et al., 2014. Dietary administration of laminarin improves the growth performance and immune responses in Epinephelus coioides. Fish & Shellfish Immunology, 41(2): 402–406.

    Article  Google Scholar 

  • Yin, Y., Mao, X., Yang, J., Chen, X., Mao, F., and Xu, Y., 2012. dbCAN: A web resource for automated carbohydrate-active enzyme annotation. Nucleic Acids Research, 40(W1): W445–W451.

    Article  Google Scholar 

  • Zakharenko, A. M., Kusaykin, M. I., Kovalchuk, S. N., Sova, V. V., Silchenko, A. S., Belik, A. A., et al., 2012. Catalytic properties and amino acid sequence of endo-1→3-beta-D-glucanase from the marine mollusk Tapes literata. Biochemistry–Moscow, 77(8): 878–888.

    Article  Google Scholar 

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Acknowledgements

This work was supported by the National Key R&D Program of China (No. 2018YFC0311203), and the Fundamental Research Funds for the Central Universities (No. 201941005).

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Authors and Affiliations

  1. College of Food Science and Engineering, Ocean University of China, Qingdao, 266003, China

    Jiajing Li, Changhu Xue, Yaoguang Chang, Jingjing Shen & Yuying Zhang

  2. Laboratory for Marine Drugs and Bioproducts, Qingdao Marine Science and Technology Center, Qingdao, 266237, China

    Changhu Xue & Yaoguang Chang

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  1. Jiajing Li
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  2. Changhu Xue
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  3. Yaoguang Chang
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  4. Jingjing Shen
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  5. Yuying Zhang
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Correspondence to Yaoguang Chang.

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Li, J., Xue, C., Chang, Y. et al. Cloning, Expressing and Characterizing a Novel Cold-Adapted Laminarinase from Marine Bacterium Wenyingzhuangia aestuarii OF219. J. Ocean Univ. China 22, 1034–1040 (2023). https://doi.org/10.1007/s11802-023-5394-y

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  • DOI: https://doi.org/10.1007/s11802-023-5394-y

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