A novel serine hydroxymethyltransferase from Arthrobacter nicotianae: characterization and improving catalytic efficiency by rational design.
BMC Biotechnol
; 14: 93, 2014 Nov 14.
Article
en En
| MEDLINE
| ID: mdl-25394480
ABSTRACT
BACKGROUND:
Serine hydroxymethyltransferase (SHMT) is the key enzyme in L-serine enzymatic production, suggesting the importance of obtaining a SHMT with high activity.RESULTS:
Here, a novel SHMT gene, glyA, was obtained through degenerate oligonucleotide-primed PCR and encoded a novel SHMT with 54.3% similarity to the known SHMT from Escherichia coli. The obtained protein AnSHMT showed the optimal activity at 40 °C and pH 7.5, and was more stable in weakly alkali conditions (pH 6.5-8.5) than Hyphomicrobium methylovorum's SHMT (pH 6.0-7.5), In order to improve the catalytic efficiency of the wild type, the site-directed mutagenesis based on sequences alignment and bioinformatics prediction, was used and the catalytic efficiency of the mutant I249L was found to be 2.78-fold higher than that of the wild-type, with the replacement of isoleucine by leucine at the 249 position.CONCLUSIONS:
This research provides useful information about the interesting site, and the application of DOP-PCR in cloning a novel glyA gene.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Arthrobacter
/
Glicina Hidroximetiltransferasa
/
Proteínas Bacterianas
Idioma:
En
Revista:
BMC Biotechnol
Asunto de la revista:
BIOTECNOLOGIA
Año:
2014
Tipo del documento:
Article