Key Deployment Project of Chinese Academy of Sciences (No. KSZD-EW-Z-016-3), National Natural Science Foundation of China (Nos. 21072151, 21102100), National Basic Research Program of China (973 Program) (No. 2011CB710801).
In industrial application of NAD(P)H-dependent dehydrogenases, NAD(H) has the advantages over NADP(H) in higher stability, lower price and wider recycling system. Recently, a meso-2,6-diaminopimelate dehydrogenase from Symbiobacterium thermophilum (StDAPDH) has been found to be a useful biocatalyst for the production of D-amino acids, but it requires NADP(H) as co-enzyme. To switch the co-enzyme specificity from NADP(H) to NAD(H), we studied the effect of Y76 on the co-enzyme specificity of StDAPDH, because the crystal structural analysis indicated that residue Y76 is near the adenine ring. The mutation of Y76 exerted significant effect on the co-enzyme specificity. Furthermore, the double mutant R35S/R36V significantly lowered the specific activity toward NADP+, and the combination of R35S/R36V with some of the Y76 mutants resulted in mutant enzymes favorable NAD+ over NADP+. This study should provide useful guidance for the further development of highly active NAD+-dependent StDAPDH by enzyme engineering.
赵雷明,刘卫东,陈曦,王敏,冯进辉,吴洽庆,朱敦明. 嗜热共生杆菌内消旋-2,6-二氨基庚二酸脱氢酶中Y76对辅酶偏好性影响[J]. Chinese Journal of Biotechnology, 2015, 31(7): 1108-1118
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