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Molecular cloning of the gene encoding β-1,3(4)-glucanase A from a marine bacterium, Pseudomonas sp. PE2, an essential enzyme for the degradation of Pythium porphyrae cell walls

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The β-1,3(4)-glucanase A (GluA)-encoding gene named gluA was cloned from the genomic library of a marine bacterium Pseudomonas sp. PE2 by expression in Escherichia coli, and the complete DNA sequence was determined. The recombinant enzyme from Pseudomonas sp. PE2 was examined to determine the essential enzymes for degrading Pythium porphyrae cell walls, comparatively using other two recombinant enzymes, chitinase A and β-1,3-glucanase B from the same bacterial strain. GluA most degraded the cell walls among these three enzymes, suggesting that GluA seems to be most important to P. porphyrae cell-wall-degrading activity. The deduced GluA is a modular enzyme composed of an N-terminal signal peptide, the tandem-duplicated carbohydrate-binding module family 6 (CBMGluA-1 and CBMGluA-2), and a glycoside hydrolase family 16 catalytic domain. Deletion analysis clearly indicated that GluA lacking CBMGluA-1 and CBMGluA-2 does not bind to Avicel and xylan. These results suggest that the tandem-repeated CBM of GluA may play a key role in the binding of Avicel and xylan as well as β-1,3- and β-1,3;1,4-glucans and is very important to bind to insoluble polysaccharides.

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Acknowledgement

This work was supported in part by grant 13660186 from the Ministry of Education, Culture, Sports, Science and Technology.

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  1. Coastal Bioenvironment Center, Saga University, 152-1 Shonan-cho, Karatsu, Saga, 847-0021, Japan

    Etsushi Kitamura & Yuto Kamei

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  1. Etsushi Kitamura
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  2. Yuto Kamei
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Correspondence to Yuto Kamei.

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Kitamura, E., Kamei, Y. Molecular cloning of the gene encoding β-1,3(4)-glucanase A from a marine bacterium, Pseudomonas sp. PE2, an essential enzyme for the degradation of Pythium porphyrae cell walls. Appl Microbiol Biotechnol 71, 630–637 (2006). https://doi.org/10.1007/s00253-005-0200-x

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  • DOI: https://doi.org/10.1007/s00253-005-0200-x

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