Identification of ω-aminotransferase from Caulobacter crescentus and site-directed mutagenesis to broaden substrate specificity.
J Microbiol Biotechnol
; 18(1): 48-54, 2008 Jan.
Article
em En
| MEDLINE
| ID: mdl-18239415
ABSTRACT
A putative aminotransferase gene, cc3143 (aptA), from Caulobacter crescentus was screened by bioinformatical tools and overexpressed in E. coli, and the substrate specificity of the aminotransferase was investigated. AptA showed high activity for short-chain beta-amino acids. It showed the highest activity for 3-amino-n-butyric acid. It showed higher activity toward aromatic amines than aliphatic amines. The 3D model of the aminotransferase was constructed by homology modeling using a dialkylglycine decarboxylase PDB ID 1DGE) as a template. Then, the aminotransferase was rationally redesigned to increase the activity for 3-amino- 3-phenylpropionic acid. The mutants N285A and V227G increased the relative activity for 3-amino-3-phenylpropionic acid to 3-amino-n-butyric acid by 11-fold and 3-fold, respectively, over that of wild type.
Buscar no Google
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Mutagênese Sítio-Dirigida
/
Caulobacter crescentus
/
Transaminases
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Revista:
J Microbiol Biotechnol
Ano de publicação:
2008
Tipo de documento:
Article