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Two GH16 Endo-1,3-β-D-Glucanases from Formosa agariphila and F. algae Bacteria Have Complete Different Modes of Laminarin Digestion

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Abstract

There is a comparative analysis of primary structures and catalytic properties of two recombinant endo-1,3-β-D-glucanases from marine bacteria Formosa agariphila KMM 3901 and previously reported F. algae KMM 3553. Both enzymes had the same molecular mass 61 kDa, temperature optimum 45 °C, and comparable ranges of thermal stability and Km. While the set of products of laminarin hydrolysis with endo-1,3-β-D-glucanase from F. algae was stable of the reaction with pH 4–9, the pH stability of the products of laminarin hydrolysis with endo-1,3-β-D-glucanase from F. agariphila varied at pH 5–6 for DP 2, at pH 4 and 7–8 for DP 5, and at pH 9 for DP 3. There were differences in modes of action of these enzymes on laminarin and 4-methylumbelliferyl-β-D-glucoside (Umb), indicating the presence of transglycosylating activity of endo-1,3-β-D-glucanase from F. algae and its absence in endo-1,3-β-D-glucanase from F. agariphila. While endo-1,3-β-D-glucanase from F. algae produced transglycosylated laminarioligosaccharides with a degree of polymerization 2–10 (predominately 3–4), endo-1,3-β-D-glucanase from F. agariphila did not catalyze transglycosylation in our lab parameters.

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Abbreviations

CM:

Carboxymethylated

DP:

Degree of polymerization

GH:

Glycoside hydrolase

HPLC:

High-performance liquid chromatography

LOS:

Laminarioligosaccharides

NMR:

Nuclear magnetic resonance

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

TLC:

Thin-layer chromatography

Umb:

4-Methylumbelliferyl-β-D-glucoside

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Funding

The work was supported by the Grant of the Ministry of Education and Science of the Russian Federation, Grant No. 13.1902.21.0012 (Number of Agreement: 075–15-2020–796).

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  1. G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far-Eastern Branch of the Russian Academy of Sciences, 159, Prospect 100-let Vladivostoku 159, 690022, Vladivostok, Russia

    Alexey A. Belik, Anton B. Rasin, Mikhail I. Kusaykin & Svetlana P. Ermakova

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  1. Alexey A. Belik
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Belik, A.A., Rasin, A.B., Kusaykin, M.I. et al. Two GH16 Endo-1,3-β-D-Glucanases from Formosa agariphila and F. algae Bacteria Have Complete Different Modes of Laminarin Digestion. Mol Biotechnol 64, 434–446 (2022). https://doi.org/10.1007/s12033-021-00421-9

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