Development of engineered enzymes for synthesis of highly pure phosphatidylinositol

2015 Fiscal Year Final Research Report

• Project/Area Number: 25420830
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Nagoya University
• Principal Investigator: Iwasaki Yugo 名古屋大学, 生命農学研究科, 准教授 (50273214)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: phospholipase D / phosphatidylinositol

Outline of Final Research Achievements

Protein engineering study on a phospholipase D (PLD) was performed in to synthesize phosphatidylinositol (PI), a functional phospholipid, with high purity. Analysis of many PLD variants with mutations at its substrate-binding pocket successfully identified some variants with excellent positional specificity. In addition, trimming an unstable loop structure from PLD molecule lead to the creation of another variant with both improved stability and high positional specificity. Moreover, during the optimization of the reaction conditions, we found that very high concentration of NaCl enhanced the PI yield. Furthermore, we discovered another enzyme that hydrolyzes phosphatidic acid (a unwanted side product) from a microorganism by screening.

Free Research Field

protein engineering