2007 Fiscal Year Final Research Report Summary
Molecular breeding of white rot basidiomycete for efficient saccharification in biomass conversion process
| Project/Area Number |
16580133
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
林産科学・木質工学
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| Research Institution | Kyoto University |
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Principal Investigator |
HONDA Yoichi Kyoto University, RISE, Associate Professor (70252517)
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| Project Period (FY) |
2004 – 2007
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| Keywords | biomass conversion / white rot fungi / genetic engineering / transformation / host-vector system / basidiomycete / mushroom / cloning |
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| Research Abstract |
A versatile peroxidase MnP2, secreted by a white rot fungus Pleurotus ostreatus was characterized for its unique property of high molecular weight compounds oxidizing activity. A series of mutant MnP2 were produced by P. ostreatus transformants containing a recombinant mnp2 with site-directed base substitutions. Each mutant protein was purified to homogeneity and characterized with kinetics and reactivity for low- and high-molecular weight substrates, and spectroscopic methods. All variants assayed in this work showed substantially similar kinetic properties for Mn^<2+> and H_2O_2 to those of wild type MnP2. On the other hand, it was demonstrated that substitution of an exposing tryptophan residue (W170) to alanine lost oxidation activity for veratrylacohol (VA), Poly R-478 and Rnase A. Furthermore, substitution of the surrounding amino add residues affected oxidation of the high-molecular weight compounds but VA. From these results, it was clearly demonstrated that MnP2 oxidizes high-molecular weight compounds directly via long range electron transfer pathway from W170 to heme moiety. An extra modification of carbohydrate drain in a mutant MnP2 was also discussed. This is the first report of mutational analysis of fungal versatile peroxidase produced by a homologous expression system.
For the development of a gene-targeting system in basidiomycete, a gene encoding for Ku70 protein which is involved in non-homologous end joining system was cloned and analyzed.
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[Journal Article] Mechanism for oxidation of high-molecular-weight substrates by a fungal versatile peroxidase, MnP22008
Author(s)
Tsukihara, T., Y., Honda, R., Sakai, T., Watanabe, T., Watanabe
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Journal Title
Appl. Environ. Microbiol 74
Pages: 2873-2881
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Exclusive overproduction of recombinant versatile peroxidase MnP2 by genetically modified white rot fungus, Pleurotus ostreatus2006
Author(s)
Tsukihara, T., Y., Honda, R., Sakai, T., Watanabe, T., Watanabe
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Journal Title
J. Biotechnol 126
Pages: 431-439
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Mechanism for oxidation of high-molecular-weight substrates by a fungal versatile peroxidase, MnP22007
Author(s)
Tsukihara, T., R., Sakai, T., Watanabe, Y., Honda, T., Watanabe
Organizer
Annual meeting of Japan Society of Bioscience, Biochemistry and Agrochemistry 2007
Place of Presentation
Tokyo
Year and Date
2007-03-27
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Exclusive overproduction of recombinant versatile peroxidase MnP2 in, Pleurotus ostreatus2006
Author(s)
Tsukihara, T., Y., Honda, R., Sakai, T., Watanabe, T., Watanabe
Organizer
The 50th anniversary meeting of The Mycological Society of Japan
Place of Presentation
Chiba
Year and Date
2006-06-04
Description
「研究成果報告書概要(欧文)」より
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[Book] きのこ学への誘い2004
Author(s)
本田与一, 他(大賀祥治編)
Total Pages
189
Publisher
海青社
Description
「研究成果報告書概要(和文)」より
