Cell surface expression of proteins using Pichia yeast: Development and application of high-throuput construction of expression system

Research Project

Project/Area Number	25450126
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Applied biochemistry
Research Institution	Kyoto University
Principal Investigator	Mizutani Kimihiko 京都大学, 農学研究科, 助教 (40314281)
Project Period (FY)	2013-04-01 – 2017-03-31
Project Status	Completed (Fiscal Year 2016)
Budget Amount *help	¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000) Fiscal Year 2016: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000) Fiscal Year 2015: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000) Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000) Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Keywords	タンパク質工学 / Pichia pastoris / 発現系構築 / X線結晶構造解析
Outline of Final Research Achievements	Using High-throuput method constructing Pichia yeast protein expression system, cell surface expression of green fluorescence protein and the target protein using Pichia yeast has been achieved. Pichia yeast producing alpha-amylase on the cell surface had starch diesting activity. Using Pichia yeast, cellulose digesting enzymes of molluscs were produced. For the enzyme of a snail, molecular structure and catalytic mechanism were analyzed.