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Bacterial chitinase with phytopathogen control capacity from suppressive soil revealed by functional metagenomics
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
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2014 (English)In: Applied Microbiology and Biotechnology, ISSN 0175-7598, E-ISSN 1432-0614, Vol. 98, no 6, 2819-2828 p.Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
2014. Vol. 98, no 6, 2819-2828 p.
Keyword [en]
Soil metagenome, Metagenomic library, Chitinolytic enzyme, Phytopathogen control, Antifungal, Heterologous protein expression
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:uu:diva-222742DOI: 10.1007/s00253-013-5287-xISI: 000332108100041OAI: oai:DiVA.org:uu-222742DiVA: diva2:712256
Available from: 2014-04-14 Created: 2014-04-14 Last updated: 2017-12-05Bibliographically approved

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Hjort, Karin
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