Homologous expression of recombinant manganese peroxidase genes in ligninolytic fungus Pleurotus ostreatus.
Pleurotus ostreatus is a white-rot fungus known as an efficient degrader of lignin and also various organo-pollutants. Using a DNA-mediated transformation system, a molecular breeding approach to isolate over-producers of a manganese peroxidase isozyme, MnP3, was carried out. Recombinant mnp3 constructs under the control of P. ostreatus sdi1 expression signals were introduced into the wild-type P. ostreatus strain by co-transformation with a carboxin-resistant vector plasmid, pTM1. One of the recombinants obtained by a mating between two monokaryotic transformants, TMG9-C1, showed a several times higher level of MnP activity than the wild-type control in the early stage of liquid culture. Predominant transcription of the recombinant mnp3 in the strain was demonstrated by RT-PCR experiments. This is the first report of a genetically modified P. ostreatus strain with an expression system for recombinant genes.
Irie
T
T
Wood Research Institute, Kyoto University, Japan.
Honda
Y
Y
Watanabe
T
T
Kuwahara
M
M
D001483:Base Sequence; D001709:Biotechnology; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D005800:Genes, Fungal; D005822:Genetic Vectors; D008031:Lignin / Q000378:metabolism; D010544:Peroxidases / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D020076:Pleurotus / Q000201:enzymology* / Q000235:genetics*; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014170:Transformation, Genetic
D017931:DNA Primers; D012331:RNA, Fungal; D012333:RNA, Messenger; D011994:Recombinant Proteins; D008031:Lignin; D010544:Peroxidases; C051129:manganese peroxidase