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Molecular Biology Reagents> | … 0.1 mM PMSF and 1 x protease inhibitor cocktail Complete Mini, Roche Diagnostics, Mannheim, Germany), the homogenate was centrifuged at 30000g for 30 min at 4 °C. The supernatant was combined with 1 mL pre-washed glutathione resin (GenScript, Nanjing, China) for 2 … | Get A Quote |
The flavoprotein l-aspartate oxidase (LASPO) is the first enzyme of the de novo biosynthetic pathway of NAD in plants. Although LASPO is considered pivotal to maintain NAD homeostasis, it has not been hitherto characterized in plants. Here, the cDNA encoding the LASPO from the model plant Arabidopsis thaliana (AtLASPO, At5g14760) has been cloned and expressed in Escherichia coli for subsequent enzyme characterization. The purified AtLASPO enzyme displayed a K of 0.79 mM for l-aspartate and a k of 0.25 s. We could further detect an l-aspartate: fumarate oxidoreductase activity of the recombinant plant enzyme. In addition, results indicated that NADP but not NAD, and even more strongly NADH, inhib... More