Molecular characterization of a highly efficient and thermostable phosphoribosyl anthranilate isomerase from Geobacillus thermopakistaniensis.
Protein Expr Purif
; 166: 105523, 2020 02.
Article
en En
| MEDLINE
| ID: mdl-31669755
ABSTRACT
Phosphoribosyl anthranilate isomerase is involved in the isomerization of phosphoribosyl anthranilate to 1-(o-carboxyphenylamino)-1-deoxyribulose 5-phosphate. In the present study, trpFGt, a gene encoding phosphoribosyl anthranilate isomerase from Geobacillus thermopakistaniensis, was cloned and expressed in Escherichia coli. The gene product, TrpFGt, was produced in E. coli in soluble and active form. Molecular characterization revealed that recombinant TrpFGt was highly efficient and stable. The apparent Vmax and Km values were 480⯵molâ¯min-1 mg-1 and 1.15⯵M, respectively. The half-life of the enzyme was 90â¯minâ¯at 60⯰C. Apart from thermostability, TrpFGt was highly stable against protein denaturants such as urea. There was no significant change in activity even after treatment with 8â¯M urea. To the best of our knowledge, TrpFGt, is the most active and stable phosphoribosyl anthranilate isomerase characterized to date and this is the first characterization of TrpF from the genus Geobacillus.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Contexto en salud:
3_ND
Problema de salud:
3_neglected_diseases
/
3_zoonosis
Asunto principal:
Geobacillus
/
Ortoaminobenzoatos
/
Isomerasas
Idioma:
En
Revista:
Protein Expr Purif
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2020
Tipo del documento:
Article
País de afiliación:
Pakistán