Development of a novel directed evolution method to improve recombinant protein productivity in insect factories

2023 Fiscal Year Final Research Report

• Project/Area Number: 22K20581
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Multi-year Fund
• Review Section: 0602:Agricultural and environmental biology and related fields
• Research Institution: Kyushu University
• Principal Investigator: Masuda Akitsu 九州大学, 農学研究院, 助教 (10967079)
• Project Period (FY): 2022-08-31 – 2024-03-31
• Keywords: カイコ / バキュロウイルス / 変異導入 / 組換えタンパク質 / 指向性進化

Outline of Final Research Achievements

In the silkworm-baculovirus expression system, which can efficiently produce recombinant proteins, it is known that there are some difficult-to-express proteins. Therefore, insect directed evolutionary tools that enable mutagenesis and selection of favourable mutants are needed to solve this problem. In this study, we attempted to develop a tool for introducing mutations into specific regions of the baculovirus genome and to identify baculovirus genes that affect viral proliferation, which is necessary for selecting desired mutants. The mutagenic enzyme was found to be expressed by the baculovirus in cultured insect cells, but no mutations in the target sequence were identified. In the identification of genes affecting viral proliferability, eight candidate genes were examined and several genes that could manipulate proliferability were identified.

Free Research Field

タンパク質工学

Academic Significance and Societal Importance of the Research Achievements

高いタンパク質生産能力を持つカイコの発現系には、その工程の煩雑さからタンパク質の変異体の発現をスループット性高く検証するツールが存在しなかった。今回の知見を元に昆虫における独自の指向性進化ツールが実現できれば、他の発現系では作れないようなタンパク質でも機能改変や生産性改善が可能となり、バイオ医薬品やタンパク質材料の分野で大きく貢献できる可能性がある。