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Expression of a Manganese Peroxidase Gene (mnp5) from White rot fungus Phanerochaete chrysosporium in the Pichia pastoris  

Lee, Jae-Won (Department of Forest Sciences, College of Agriculture & Life Sciences, Seoul National University)
Yang, In (Department of Forest Sciences, College of Agriculture & Life Sciences, Seoul National University)
Igarashi, Kiyohiko (Graduate School of Agriculture and Life Sciences, The University of Tokyo)
Samejima, Masahiro (Graduate School of Agriculture and Life Sciences, The University of Tokyo)
Choi, In-Gyu (Department of Forest Sciences, College of Agriculture & Life Sciences, Seoul National University)
Publication Information
Journal of the Korean Wood Science and Technology / v.33, no.4, 2005 , pp. 45-52 More about this Journal
Abstract
The manganese peroxidase (mnp5) from white-rot fungus Phanerochaete chrysosporium has been heterologously expressed in the methylotrophic yeast Pichia pastoris. The majority of the rMnP5 (recombinant MnP5) produced by P. pastoris exhibited an approximate molecular mass 45 kDa considerably larger than that of the predicting mnp5 due to two glycosylation sites of mnp5. After site direct mutation treatment, the effect of N-linked hyperglycosylation was examined by enzyme activity. Analysis by sodium dodesyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Coomassie Brilliant Blue (CBB) staining revealed a major protein band with a molecular mass of 37 kDa. Enzyme activity of M-rMnP5 (mutant recombinant MnP5) was similar to that of rMnP5, indicating that hyperglycosylation did not affect the active site. In this work, active mnp5 was successfully expressed in P. pastoris, suggesting that P. pastoris has potential capability of producing active heme-containing proteins.
Keywords
Phanerochaete chrysosporium; manganese peroxidase; methylotrophic yeast; Pichia pastoris; glycosylation;
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Times Cited By KSCI : 1  (Citation Analysis)
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