A specific, highly active malate dehydrogenase by redesign of a lactate dehydrogenase framework.
Science
; 242(4885): 1541-4, 1988 Dec 16.
Article
in En
| MEDLINE
| ID: mdl-3201242
ABSTRACT
Three variations to the structure of the nicotinamide adenine dinucleotide (NAD)-dependent L-lactate dehydrogenase from Bacillus stearothermophilus were made to try to change the substrate specificity from lactate to malate Asp197----Asn, Thr246----Gly, and Gln102----Arg). Each modification shifts the specificity from lactate to malate, although only the last (Gln102----Arg) provides an effective and highly specific catalyst for the new substrate. This synthetic enzyme has a ratio of catalytic rate (kcat) to Michaelis constant (Km) for oxaloacetate of 4.2 x 10(6)M-1 s-1, equal to that of native lactate dehydrogenase for its natural substrate, pyruvate, and a maximum velocity (250 s-1), which is double that reported for a natural malate dehydrogenase from B. stearothermophilus.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Geobacillus stearothermophilus
/
L-Lactate Dehydrogenase
/
Malate Dehydrogenase
Language:
En
Journal:
Science
Year:
1988
Document type:
Article
Affiliation country:
Reino Unido