期刊
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
卷 98, 期 6, 页码 2819-2828出版社
SPRINGER
DOI: 10.1007/s00253-013-5287-x
关键词
Soil metagenome; Metagenomic library; Chitinolytic enzyme; Phytopathogen control; Antifungal; Heterologous protein expression
资金
- EU Metaexplore project [KBBE-222625]
- Foundation for Baltic and East European Studies
- Ministero Italiano della Ricerca e della Universita
- Consorzio Interuniversitario per le Biotecnologie (CIB)
Plant disease caused by fungal pathogens results in vast crop damage globally. Microbial communities of soil that is suppressive to fungal crop disease provide a source for the identification of novel enzymes functioning as bioshields against plant pathogens. In this study, we targeted chitin-degrading enzymes of the uncultured bacterial community through a functional metagenomics approach, using a fosmid library of a suppressive soil metagenome. We identified a novel bacterial chitinase, Chi18H8, with antifungal activity against several important crop pathogens. Sequence analyses show that the chi18H8 gene encodes a 425-amino acid protein of 46 kDa with an N-terminal signal peptide, a catalytic domain with the conserved active site F(175)DGIDIDWE(183), and a chitinase insertion domain. Chi18H8 was expressed (pGEX-6P-3 vector) in Escherichia coli and purified. Enzyme characterization shows that Chi18H8 has a prevalent chitobiosidase activity with a maximum activity at 35 A degrees C at pH lower than 6, suggesting a role as exochitinase on native chitin. To our knowledge, Chi18H8 is the first chitinase isolated from a metagenome library obtained in pure form and which has the potential to be used as a candidate agent for controlling fungal crop diseases. Furthermore, Chi18H8 may also answer to the demand for novel chitin-degrading enzymes for a broad range of other industrial processes and medical purposes.
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