Abstract
An alginate lyase gene aly644 encoding a member of polysaccharide lyase family 6 was obtained from a metagenome of Antarctic macroalgae-associated microbes. The gene was expressed heterologously in Escherichia coli, and the recombinant protein was purified using a Ni-NTA His Tag Kit. With sodium alginate as the substrate, recombinant Aly644 exhibited an optimum reaction temperature of 50°C and an optimum reaction pH of 7.0. The Vmax and Km values of Aly644 toward sodium alginate were 112.36 mg/mL·min and 16.75 mg/mL, respectively. Substrate specificity analysis showed that Aly644 was a bifunctional alginate lyase that hydrolyzed both polyguluronic acid and polymannuronic acid. The hydrolysis products of Aly644 with sodium alginate as the substrate were detected by thin-layer chromatography, and were mainly di- and trisaccharides. The oligosaccharides produced by degradation of sodium alginate by Aly644 inhibited the mycelial growth of the plant pathogens Phytophthora capsici and Fulvia fulva; the 50% maximal effective concentration (EC50) values were 297.45 and 452.89 mg/L, and the 90% maximal effective concentration (EC90) values were 1341.45 and 2693.83 mg/L, respectively. This highlights that Aly644 is a potential candidate enzyme for the industrial production of alginate oligosaccharides with low degree of polymerization. Enzyme-hydrolyzed alginate oligosaccharides could support the development of green agriculture as natural antimicrobial agents.
Key points
• An alginate lyase was obtained from a metagenome of Antarctic macroalgae-associated microbes.
• Aly644 is a bifunctional alginate lyase with excellent thermostability and pH stability.
• The enzymatic hydrolysates of Aly644 directly inhibited Phytophthora capsici and Fulvia fulva.
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Acknowledgements
This work was supported by Impact and Response of Antarctic Seas to Climate Change (IRFSOCC2023-2025). We thank Liwen Bianji (Edanz) (http://www.liwenbianji.cn/) for editing the language of a draft of this manuscript.
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JL, TL, and AZ conceived and designed research. AZ and ZC conducted experiments. AZ wrote the manuscript. LYZ contributed analytical tools. QZ and LPF analyzed data. All authors read and approved the manuscript.
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Zhang, A., Cao, Z., Zhao, L. et al. Characterization of bifunctional alginate lyase Aly644 and antimicrobial activity of enzymatic hydrolysates. Appl Microbiol Biotechnol 107, 6845–6857 (2023). https://doi.org/10.1007/s00253-023-12745-4
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DOI: https://doi.org/10.1007/s00253-023-12745-4