2012 Fiscal Year Final Research Report
Development of NADH regeneration system by thermostable phosphite dehydrogenase
| Project/Area Number |
22360346
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Biofunction/Bioprocess
|
|---|
| Research Institution | Hiroshima University |
|---|
Principal Investigator |
KURODA Akio 広島大学, 大学院・先端物質科学研究科, 教授 (50205241)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
HIROTA Ryuichi 広島大学, 大学院・先端物質科学研究科, 助教 (90452614)
|
|---|
| Project Period (FY) |
2010 – 2012
|
| Keywords | 補酵素 / リン酸 / 亜リン酸 / NADH / デヒドロゲナーゼ |
|---|
| Research Abstract |
Phosphite dehydrogenase (PtxD), which catalyzes the nearly irreversible oxidation of phosphite to phosphate with the concomitant reduction of NAD+to NADH, has great potential for NADH regeneration in industrial biocatalysts. In order to obtain a heat-stable PtxD, we isolated a Ralstonia sp. strain 4506 that can grow at 45 oC on a minimal medium containing phosphite as the sole source of phosphorus. NADH regeneration system using PtxD of this strain (RsPtxD) was successfully applied to the production of L-tert-leucine, an important chiral building block used in the pharmaceutical industry. Then RsPtxD was modified by site-specific mutation to increase the substrate specificity for NADP. The resultant enzyme RsPtxD-DM was successfully used for shikimic acid production as a NADPH regeneration system.
|
