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

• Published in: The journal of microbiology 2012, 50(6), , pp.939-946
• Main Authors: Yu, Gang, Jilin University, Changchun, Jilin, P. R. China, Liu, Jin-Liang, Jilin University, Changchun, Jilin, P. R. China, Xie, Li-Qin, Jilin University, Changchun, Jilin, P. R. China, Wang, Xue-Liang, Jilin University, Changchun, Jilin, P. R. China, Zhang, Shi-Hong, Jilin University, Changchun, Jilin, P. R. China, Pan, Hong-Yu, Jilin University, Changchun, Jilin, P. R. China
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino acids; Base Sequence; Biological control; Biomedical and Life Sciences; characterization; CLONACION; CLONAGE; CLONING; Cloning, Molecular; E coli; Enzyme Activation; Enzymes; Escherichia coli; Fungi; Fusarium moniliforme; Gene Expression; Genetic engineering; Insects; Life Sciences; Microbiology; Molecular Sequence Data; Pathogens; Phylogeny; Protein Stability; Proteins; Sequence Analysis, DNA; serine protease; Serine Proteases - chemistry; Serine Proteases - genetics; Serine Proteases - isolation & purification; Serine Proteases - metabolism; Subtilisin - genetics; Subtilisin - metabolism; Verticillium - classification; Verticillium - genetics; Verticillium - metabolism; VERTICILLIUM LECANII; 생물학
• ISSN: 1225-8873; 1976-3794
• DOI: 10.1007/s12275-012-2199-x

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℃ 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.