Abstract
Chitinases are digestive enzymes that break down glycosidic bonds in chitin. In the current study, an endochitinase gene Lbchi31 was cloned from Limonium bicolor. The cDNA sequence of Lbchi31 was 1,107 bp in length, encoding 322 amino acid residues with a calculated molecular mass of 31.7 kDa. Clustal analysis showed that there was a highly conserved chitin-binding domains in Lbchi31 protein, containing four sulfide bridges. The Lbchi31 gene was inserted into the pPIC9 vector and transferred into yeast Pichia pastoris GS115 and KM71 for heterologous expression. The transformant harboring the Lbchi31 gene showed a clearly visible protein band with a molecular mass of more than 31 kDa in the SDS-PAGE gel, indicating that it had been translated in P. pastoris. Enzyme characterization showed that the optimal reaction condition for chitinase LbCHI31 activity was: 40°C, pH of 5.0 and 5 mmol l−1 of Mn2+. The maximum enzyme activity was 0.88 U ml−1 following exposure to the cell wall chitin of Valsa sordida. The LbCHI31 enzyme can efficiently degrade cell wall chitin of the phytopathogenic Rhizoctonia solani, Fusarium oxysporum, Sclerotinia sclerotiorum, V. sordida, Septoria tritici and Phytophthora sojae, suggesting that it has the biocontrol function to fungal phytopathogen.
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Acknowledgment
This work has been supported by Heilongjiang Province International Scientific and Technological Cooperation Project (WB07N02) and Natural Science Foundation of Heilongjiang province (QC07C56).
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Liu, Z.H., Wang, Y.C., Qi, X.T. et al. Cloning and characterization of a chitinase gene Lbchi31 from Limonium bicolor and identification of its biological activity. Mol Biol Rep 37, 2447–2453 (2010). https://doi.org/10.1007/s11033-009-9756-3
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DOI: https://doi.org/10.1007/s11033-009-9756-3