Effect of residue Y76 on co-enzyme specificity of meso-diaminopimelate dehydrogenase from Symbiobacterium thermophilum / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 1108-1118, 2015.
Article
in Chinese
| WPRIM
| ID: wpr-240591
ABSTRACT
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Substrate Specificity
/
Chemistry
/
Clostridiales
/
Amino Acid Oxidoreductases
/
Amino Acids
/
Mutation
/
NAD
/
NADP
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2015
Type:
Article
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