Cloning and Homologous Expression of Novel Lignin Peroxidase Genes in the White-Rot FungusPhanerochaete sordidaYK-624

• DOI: 10.1271/bbb.90152
• Publication Date: 2009-08-24T05:59:22Z
• Authors: Tatsuki SUGIURA; Kenji YAMAGISHI; Toshiyuki KIMURA; Tomoaki NISHIDA; Hirokazu KAWAGISHI; Hirofumi HIRAI
• Keywords: 0303 health sciences; DNA, Complementary; Molecular Sequence Data; Gene Expression; Phanerochaete; Polymerase Chain Reaction; Recombinant Proteins; 03 medical and health sciences; Transformation, Genetic; Peroxidases; Cloning, Molecular; Genome, Fungal; DNA, Fungal; Plasmids

Two genes, encoding YK-LiP1 and YK-LiP2, were cloned from the white-rot fungus Phanerochaete sordida YK-624, a homologous expression system for gene was constructed. full-length cDNAs (ylpA ylpB) isolated by degenerate RT-PCR RACE-PCR. The results of N-terminal amino acid sequence analysis native YK-LiP2 showed that ylpA ylpB coded respectively. promoter glyceraldehyde-3-phosphate dehydrogenase P. YK-624 (PsGPD) used to drive ylpA. Expression vector pGPD-g-ylpA transformed into uracil auxotrophic mutant, UV-64. YlpA protein secreted in active form transformants after 4 d growth medium containing an excessive nitrogen source, whereas endogenous not produced. physical catalytic properties purified very similar those YK-LiP2. These suggest recombinant successful.