Heterologous expression and large-scale production ofbasidiomycetous lignin-degrading enzyme in heterobasidiomycetousyeast

• Project/Area Number: 22580376
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Boundary agriculture
• Research Institution: Kagawa University
• Principal Investigator: ASADA Ysauhiko 香川大学, 農学部, 教授 (70151032)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2010: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: バイオマス / リグニン分解酵素 / ラッカーゼ / 担子菌キノコ / 異担子菌酵母 / 担子菌

Research Abstract

It was found that Grifola frondosa produces a laccase (one of lignin-degrading enzymes), which is effective for decolorizing of synthetic dyes and degrading of bisphenol A (recalcitrant environmental pollutant having an endocrine-disrupting effect). The transformation system of heterobasidiomycetous yeast Rhodosporidium toruloides was established using electroporation technique. The attempt of heterologous expression of laccase cDNA in R. toruloides as a host was unsuccessful, however, it was found that Pichia pastoris transformed with the laccase cDNA of G. frondosaproduced a recombinant laccase.

Research Products

• [Journal Article] Biodegradation of Bisphenol A and decolorization of synthetic dyes by laccase from white-rot fungus, Trametes polyzona (2013)
  • Author(s): Chairin, T., Nitheranont, T., Watanabe, A., Asada, Y., Khanongnuch, C., and Lumyong, S.
  • Journal Title: Appl. Biochem. Biotechnol.
  • Peer Reviewed
• [Journal Article] Purification and characterization of the extracellular laccase produced 1 from Trametes polyzona WR710-1 under solid state fermentation (2013)
  • Author(s): Chairin, T., Nitheranont, T., Watanabe, A., Asada, Y., Khanongnuch, C., and Lumyong, S.
  • Journal Title: J. Basic Microbiol. Volume: 169 Issue: 2 Pages: 539-545
  • DOI: 10.1007/s12010-012-9990-3
  • Peer Reviewed
• [Journal Article] Purification and characterization of the extracellular laccase produced 1 from Trametes polyzona WR710-1 under solid state fermentation (2013)
  • Author(s): Thanunchanok Chairin, et al.
  • Journal Title: Journal of Basic Microbiology Volume: 未定 Issue: 1 Pages: 35-43
  • DOI: 10.1002/jobm.201200456
  • Peer Reviewed
• [Journal Article] マイタケが生産するラッカーゼの有用機能 (2012)
  • Author(s): 渡邉 彰,麻田恭彦
  • Journal Title: バイオサイエンスとインダストリー Volume: 70 Pages: 376-377
  • NAID: 10031003453
• [Journal Article] Decolorization of Synthetic Dyes and Biodegradation of Bisphenol A by Laccase from the Edible Mushroom, Grifola frondosa (2011)
  • Author(s): Nitheranont, T., Watanabe, A., Suzuki, T., Katayama, T., and Asada, Y.
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 75 Issue: 9 Pages: 1845-1847
  • DOI: 10.1271/bbb.110329
  • NAID: 10029757777
  • ISSN: 0916-8451, 1347-6947
  • Peer Reviewed
• [Journal Article] Extracellular Laccase Produced by an Edible Basidiomycetous Mushroom, Grifola frondosa: Purification and Characterization (2011)
  • Author(s): Nitheranont, T., Watanabe, A., and Asada, Y.
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 75 Issue: 3 Pages: 538-543
  • DOI: 10.1271/bbb.100790
  • NAID: 10028201860
  • ISSN: 0916-8451, 1347-6947
  • Peer Reviewed
• [Journal Article] Extracellular laccase produced by an edible basidiomycetous mushroom, Grifola frondosa : Purification and characterization (2011)
  • Author(s): Thitinard Nitheranont
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 75 Pages: 538-543
  • Peer Reviewed
• [Presentation] 担子菌きのこにおける子実体形成とラッカーゼの関連性について (2012)
  • Author(s): 麻田恭彦、渡邉 彰
  • Organizer: 日本きのこ学会第14回ワークショップ
  • Place of Presentation: 近畿大学会館(大阪府)
  • Year and Date: 2012-03-02
• [Presentation] 担子菌 Flammulina velutipes の子実体形成過程において発現するラッカーゼアイソザイムの誘導条件の解析 (2011)
  • Author(s): 山本亮輔, 渡邉 彰, 馬替由美, 麻田恭彦
  • Organizer: 平成 23 年度日本農芸化学会西日本支部・中四国支部合同大会
  • Place of Presentation: 宮崎大学
  • Year and Date: 2011-09-16
• [Presentation] 担子菌Flammulina velutipesの子実体形成過程において発現するラッカーゼアイソザイムの誘導条件の解析 (2011)
  • Author(s): 山本亮輔, 他
  • Organizer: 平成23年度日本農芸化学会西日本支部・中四国支部合同大会
  • Place of Presentation: 宮崎大学(宮崎県)
  • Year and Date: 2011-09-16
• [Presentation] エノキタケの子実体形成過程において発現するラッカーゼの解析 (2010)
  • Author(s): 山本亮輔, 渡邉 彰, 馬替由美, 麻田恭彦
  • Organizer: 2010 年度日本農芸化学会中四国支部大会
  • Place of Presentation: 香川大学
  • Year and Date: 2010-09-24
• [Presentation] 異担子菌酵母 Rhodosporidium toruloides の形質転換系の確立 (2010)
  • Author(s): 川勝早智, 岡田典子, 渡邉 彰, 麻田恭彦
  • Organizer: 2010 年度日本農芸化学会中四国支部大会
  • Place of Presentation: 香川大学
  • Year and Date: 2010-09-24
• [Presentation] 異担子菌酵母Rhodosporidium toruloidesの形質転換系の確立 (2010)
  • Author(s): 川勝早智
  • Organizer: 2010年度日本農芸化学会中四国支部大会
  • Place of Presentation: 香川大学農学部(香川県)
  • Year and Date: 2010-09-24
• [Presentation] エノキタケの子実体形成過程において発現するラッカーゼの解析 (2010)
  • Author(s): 山本亮輔
  • Organizer: 2010年度日本農芸化学会中四国支部大会
  • Place of Presentation: 香川大学農学部(香川県)
  • Year and Date: 2010-09-24
• [Presentation] Molecular cloning of two laccase complementary DNAs from the cultivated mushroom Grifola frondosa and heterologous expression in Pichia pastoris (2010)
  • Author(s): Thitinard Nitheranont, Akira Watanabe, Yasuhiko Asada
  • Organizer: 日本きのこ学会第14回大会
  • Place of Presentation: 東京大学
  • Year and Date: 2010-09-17
• [Presentation] Molecular cloning of two laccase complementary DNAs from the cultivated mushroom Grlfola frondosa and heterologous expression in Pichia pastoris (2010)
  • Author(s): Thitinard Nitheranont
  • Organizer: 日本きのこ学会第14回大会
  • Place of Presentation: 東京大学農学部(東京都)
  • Year and Date: 2010-09-17