Purification, characterization, and cDNA cloning of xylanase from fungus Trichoderma strain SY

Abstract

A xylanase-producing Trichoderma strain was isolated from soil. Xylanase from Trichoderma strain SY was purified 27-fold to an apparent homogeneity, with a 17.4% yield. The optimum pH and temperature were determined to be 5.5 and 50°C, respectively, and its molecular weight was 21-kDa by SDS-PAGE. The corresponding gene, named xyl, was cloned by RT-PCR. DNA blot analysis of xyl showed that this gene is present as a single copy. The amino acid sequence of the Xyl protein showed similarity to those of other xylanases derived from various fungi. mRNA of xyl was highly expressed when this fungus was grown on cellulose or xylan as a sole carbon source, but undetectable when grown on sucrose. Extracts of Escherichia coli cells expressing Xyl were found to have xylanase activity. It was confirmed that xyl from this isolate encodes xylanase.