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Molecular characterization of manganese peroxidases from white-rot fungus Polyporus brumalis
Sun-Hwa Ryu1, Boyeong Kim, Myungkil Kim
1Division of Wood Chemistry and Microbiology, Department of Forest Products, Korea Forest Research Institute, Seoul, 130-712, South Korea.
Abstract:
The cDNAs of six manganese-dependent peroxidases (MnPs) were isolated from white-rot fungus Polyporus brumalis. The MnP proteins shared similar properties with each other in terms of size (approximately 360-365 amino acids) and primary structure, showing 62-96 % amino acid sequence identity. RT-PCR analysis indicated that these six genes were predominantly expressed in shallow stationary culture (SSC) in a liquid medium. Gene expression was induced by treatment with dibutyl phthalate (DBP) and wood chips. Expression of pbmnp4 was strongly induced by both treatments, whereas that of pbmnp5 was induced only by DBP, while pbmnp6 was induced by wood chips only. Then, we overexpressed pbmnp4 in P. brumalis under the control of the GPD promoter. Overexpression of pbmnp4 effectively increased MnP activity; the transformant that had the highest MnP activity also demonstrated the most effective decolorization of Remazol Brilliant Blue R dye. Identification of MnP cDNAs can contribute to the efficient production of lignin-degradation enzymes and may lead to utilization of basidiomycetous fungi for degradation of lignin and numerous recalcitrant xenobiotics.
Insights
Researchers isolated six manganese-dependent peroxidases (MnPs) from fungus Polyporus brumalis. Overexpressing one MnP gene enhanced enzyme activity and dye decolorization, showing potential for lignin degradation.
Area of Science:
- Biochemistry
- Mycology
- Enzymology
Background:
- White-rot fungi, such as Polyporus brumalis, produce lignin-degrading enzymes.
- Manganese-dependent peroxidases (MnPs) are key enzymes in lignin biodegradation.
- Understanding MnP gene expression is crucial for biotechnological applications.
Purpose of the Study:
- To isolate and characterize manganese-dependent peroxidases (MnPs) from Polyporus brumalis.
- To investigate the expression patterns and regulation of MnP genes.
- To enhance MnP activity through genetic engineering for improved lignin degradation.
Main Methods:
- Isolation of manganese-dependent peroxidase (MnP) cDNAs from Polyporus brumalis.
- Reverse transcription polymerase chain reaction (RT-PCR) for gene expression analysis.
- Overexpression of the pbmnp4 gene in P. brumalis using the GPD promoter.
- Assay of MnP activity and decolorization of Remazol Brilliant Blue R dye.
Main Results:
- Six MnP cDNAs were isolated, exhibiting high sequence identity (62-96%).
- MnP gene expression was primarily observed in shallow stationary culture (SSC) and induced by dibutyl phthalate (DBP) and wood chips.
- Overexpression of pbmnp4 significantly increased MnP activity and enhanced the decolorization of Remazol Brilliant Blue R dye.
Conclusions:
- The identified MnP genes are predominantly expressed under specific culture conditions and are inducible.
- Overexpression of pbmnp4 in P. brumalis leads to increased enzymatic activity and potential for bioremediation.
- This study contributes to the efficient production of lignin-degrading enzymes and the application of fungi for xenobiotic degradation.
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