Challenge for production of therapic glyco-peptides using gene expression system in wood rot fungi

2014 Fiscal Year Final Research Report

• Project/Area Number: 25660137
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Wood science
• Research Institution: Kyoto University
• Principal Investigator: HONDA Yoichi 京都大学, (連合)農学研究科(研究院), 教授 (70252517)
• Co-Investigator(Kenkyū-buntansha): WATANABE Takashi 京都大学, 生存圏研究所, 教授 (80201200)
• Co-Investigator(Renkei-kenkyūsha): NAKAZAWA Takehito 京都大学, 大学院農学研究科, 助教 (80608141) ; YOSHIOKA Koichi 京都府立大学, 大学院生命環境科学研究科, 特任助教 (20540149)
• Research Collaborator: NAGAOKA Jun 京都大学, 大学院農学研究科 ; ITO Keigo 京都大学, 農学部森林科学科
• Project Period (FY): 2013-04-01 – 2015-03-31
• Keywords: 木材腐朽菌 / 糖タンパク質 / 異種発現 / 糖鎖工学

Outline of Final Research Achievements

It was demonstrated that wild type MnP2 enzyme from Pleurotus ostreatus contains a GlcNac2-Hex5 glycan attached to Asp102 using MALDI TOF-MS analysis. This is the first report of glycan structure attached to fungal lignin-degrading peroxidase. We also tried to analyze an additional glycan supposed to its artificially modified mutant, MnP2 R263N, but production and purification of the enzyme was not enough for the carbohydrate analysis. Moreover, a cDNA fragment encoding for cholera toxin B subunit was inserted to an expression vector for Pleurotus ostreatus and tried to introduced into host strain but no protein-producing transformants have been isolated so far.

Free Research Field

森林生化学