2021 Fiscal Year Final Research Report

Search for new rubber-degrading enzymes produced by fungi and analysis using organic chemical methods

Research Project

PDF

Project/Area Number	19K12389
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Review Section	Basic Section 64020:Environmental load reduction and remediation-related
Research Institution	Kanto Gakuin University
Principal Investigator	Shimizu Yumi 関東学院大学, 理工学部, 教授 (50725124)
Co-Investigator(Kenkyū-buntansha)	香西博明関東学院大学, 理工学部, 教授 (00272089) 清水公徳東京理科大学, 先進工学部生命システム工学科, 教授 (40345004)
Project Period (FY)	2019-04-01 – 2022-03-31
Keywords	白色腐朽菌 / ゴム分解 / 遺伝子クローニング
Outline of Final Research Achievements	(1) Two rubber assimilation fungi were used. We searched for genes with a particularly high expression level and a signal peptide sequence when these fungi were cultured in a rubber medium. As a result, 27 genes were identified in H. marmoreus and 30 genes were identified in Phanerochaete sp. These genes were introduced into basidiomycete yeast. Protein sample preparation was simplified by having yeast synthesize the protein and secrete it extracellularly. (2) From the structural analysis of the rubber sample, SBR or IR which were treated by these fungi, it was confirmed that a hydroxyl group was added to SBR and IR fragmentation occurred. (3) The amino acid sequence of a part of the extracellular lipase of yeast that produces extracellular lipase was determined. Three genes encoding this amino acid sequence were obtained.
Free Research Field	微生物学
Academic Significance and Societal Importance of the Research Achievements	SBR、IRの合成ゴムを分解する白色腐朽菌を2菌種得た。これらの菌を応用すれば、環境負荷の少ない廃ゴム処理方法を見出せる可能性がある。また、SBRとIRの分解物の同定を現在試みている。分解物の分子量は100以上のものであり、構造式が明らかになれば加工利用する方法も具体化する。低温でも活性があり、細胞外分泌性のリパーゼの同定を試みた。本酵素をコードすると予想される遺伝子を3遺伝子まで絞りこんだ。本酵素は脂質を分解することを確認しているが、ゴム分解への関与が明らかになれば、低い温度でも酵素活性が高いため、油による海洋汚染の浄化はもちろん、海洋プラスチック汚染に対応できることが期待される。