Chloroacetaldehyde dehydrogenase from Ancylobacter aquaticus UV5: Cloning, expression, characterization and molecular modeling.
Int J Biol Macromol
; 114: 1117-1126, 2018 Jul 15.
Article
en En
| MEDLINE
| ID: mdl-29605256
ABSTRACT
1,2-Dichloroethane (1,2-DCE) is oxidatively converted to a carcinogenic intermediate compound, chloroacetaldehyde by chloroacetaldehyde dehydrogenase (CAldA) during its biodegradation by many bacterial strains, including Xanthobacter autotrophicus and Ancylobacter aquaticus. In this study, a 55kDa NAD-dependent CAldA expressed by chromosomally encoded aldA gene, is reported in an indigenous Ancylobacter aquaticus UV5. A. aquaticus UV5 aldA gene was found to be 99% homologous to the plasmid (pXAU1) encoded aldA gene reported in X. autotrophicus GJ10. Pulse-field gel electrophoresis (PFGE) and PCR experiments revealed the absence of pXAU1 in A. aquaticus UV5 and that aldA was chromosomal encoded. A 6× His-tag fused CAldA cloned in pET15b, overexpressed and purified on Co-agarose affinity column using AKTA purification system showed Mr of 57,526. CAldA was active optimally at pH9 and 30°C. The Km and vmax for the substrate, acetaldehyde were found to be 115µM and 650mU/mg, respectively. CAldA substrate specificity was found to be low for chloroacetaldehyde, formaldehyde, propionaldehyde, butyraldehyde, benzaldehyde and glutaraldehyde as compared to acetaldehyde. Computational modeling revealed a predicted structure of CAldA consisting of five ß-sheets that comprise seven antiparallel ß-strands and 11 mix strands. The Molecular Dynamics and Docking studies showed that acetaldehyde bind to CaldA more tightly as compared to chloroacetaldehyde.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Proteínas Bacterianas
/
Clonación Molecular
/
Alphaproteobacteria
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Aldehído Oxidorreductasas
/
Simulación de Dinámica Molecular
/
Simulación del Acoplamiento Molecular
Tipo de estudio:
Prognostic_studies
Idioma:
En
Revista:
Int J Biol Macromol
Año:
2018
Tipo del documento:
Article
País de afiliación:
Sudáfrica