Characterization, Cloning, and Heterologous Expression of a Subtilisin-Like Serine Protease Gene VlPr1 from Verticillium lecanii

Gang Yu; Jin-Liang Liu; Li-Qin Xie; Xue-Liang Wang; Shi-Hong Zhang; Hong-Yu Pan

doi:10.1007/s12275-012-2199-x

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Characterization, Cloning, and Heterologous Expression of a Subtilisin-Like Serine Protease Gene VlPr1 from Verticillium lecanii: Gang Yu , Jin-Liang Liu , Li-Qin Xie , Xue-Liang Wang , Shi-Hong Zhang , Hong-Yu Pan; Journal of Microbiology 2012;50(6):939-946.
DOI: https://doi.org/10.1007/s12275-012-2199-x
Published online: December 30, 2012

College of Plant Sciences, Jilin University, Changchun, Jilin 130062, P. R. China

Corresponding author: Hong-Yu Pan , Tel: +86-431-87836251,

Received: 16 April 2012 • Accepted: 14 July 2012

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Abstract

The entomopathogenic fungus Verticillium lecanii is a wellknown biocontrol agent. V. lecanii produces subtilisin-like serine protease (Pr1), which is important in the biological control activity of some insect pests by degrading insect cuticles. In this study, a subtilisin-like serine protease gene VlPr1 was cloned from the fungus and the VlPr1 protein was expressed in Escherichia coli. The VlPr1 gene contains an open reading frame (ORF) interrupted by three short introns, and encodes a protein of 379 amino acids. Protein sequence analysis revealed high homology with subtilisin serine proteases. The molecular mass of the protease was 38 kDa, and the serine protease exhibited its maximal activity at 40°C and pH 9.0. Protease activity was also affected by Mg2+ and Ca2+ concentration. The protease showed inhibitory activity against several plant pathogens, especially towards Fusarium moniliforme.

Keywords: serine protease;Verticillium lecanii;cloning;characterization

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Characterization, Cloning, and Heterologous Expression of a Subtilisin-Like Serine Protease Gene VlPr1 from Verticillium lecanii

J. Microbiol. 2012;50(6):939-946. Published online December 30, 2012

DOI: https://doi.org/10.1007/s12275-012-2199-x

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