NADP+ production using thermostable NAD+ kinase of corynebacterium flaccumfaciens AHU-1622.
Canadian journal of microbiology
confidence
Key findings
Thermostable NAD+ kinase from C. flaccumfaciens produced 75% NADP+ conversion from NAD+ at 50°C; no clinical/biological endpoints reported.
View source on PubMed (PMID 3017521) ↗
- Sample size
- Not reported
- Population
- Corynebacterium flaccumfaciens AHU-1622 (bacterial enzyme production study)
- Dosing
- 8 µmol NAD+, 8 µmol ATP, 16 µmol MgCl2, 1.6 µmol NaN3, 12 mU NAD+ kinase
- Duration
- 10 h incubation
- Route
- In vitro enzymatic reaction
- Blinding
- not_reported
- Controls
- none
- Drug class
- coenzyme
Full abstract
A sonicate of Corynebacterium flaccumfaciens AHU-1622 had the highest NAD+ kinase activity (1.22 mU/mL culture broth) of the strains of bacteria we investigated. This enzyme was thermostable, with activity maintained at 50 degrees C for 1 h. This treatment inactivated phosphatase activity. Resting cells of the bacterium also had NAD+ kinase activity when treated at 60 degrees C for 30 min with 0.2% Triton X-100. NADP+ production was achieved using 8 mumol NAD+, 8 mumol ATP, 16 mumol MgCl2, 1.6 mumol NaN3, and 12 mU NAD+ kinase (0.1 g of permeabilized wet cells) in 2 mL of 0.1 M phosphate buffer, pH 7.5. The conversion ratio of NADP+ from NAD+ was 75% after 10 h of incubation at 50 degrees C, and the amount of accumulated NADP+ was 3 mumol/mL of reaction mixture. The NAD+ kinase activity of the permeabilized cells was stable and did not decrease after repeated use.