Development of methods for the enzymatic synthesis of 18O-phosphate

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K14492
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biofunction/Bioprocess
• Research Institution: Hiroshima University
• Principal Investigator: Kuroda Akio 広島大学, 先端物質科学研究科, 教授 (50205241)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: 酸素の安定同位体リン酸 / 酵素合成法 / 亜リン酸デヒドロゲナーゼ / ピロホスファターゼ / ポリホスファターゼ

Outline of Final Research Achievements

Due to the lack of multiple stable isotopes of phosphorus (P), it is difficult to use isotope pair ratios as a means to examine P dynamics in nature. However, if stable isotope of oxygen (18O) is incorporated into phosphate, 18O-phosphate can be used to trace P dynamics. In order to synthesize 18O-phosphate where the number of 18O substitution over 16O is different, we tested several enzymatic reactions. For example, phosphite dehydrogenase is an enzyme to oxidize phosphite to phosphate with a NAD cofactor. The 18O of 18O-H2O was incorporated into phosphite, generating 18O-phosphate. On the contrary, all four oxygen of phosphate was substituted with 18O of 18O-H2O using pyrophosphatase that converts pyrophosphate to phosphate in a reversible manner.

Free Research Field

応用生物工学