Cloning, sequencing and high expression in Escherichia coli of D-hydantoinase gene from Burkholderia pickettii / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 149-154, 2002.
Artículo
en Chino
| WPRIM
| ID: wpr-231359
ABSTRACT
A strain, MMR003, used for D-p-HPG production in industry was classified as Burkholderia pickettii by morphological observation and biochemical characterization. The gene encoding the D-hydantoinase enzyme was cloned, sequenced and expressed in Escherichia coli. The nucleotide sequence of the 5.0 kb insert of subclone pXZ-total was determined. One open reading frame of 1374 bp was found and predicted to encode a polypeptide consisting of 458 amino acids in size of 50 kD. The amino acid sequence alignment of D-hydantoinase from Burkholderia pickettii shows the 85% homologous with the corresponding enzyme from Agrobacterium radiobacter NRRL B11291. The D-hydantoinase gene (dha) harboured in the plasmid pXZPH2 in E. coli BL21(DE3) was highly expressed by IPTG induction. The D-hydantoinase activity for D, L-p-hydroxyphenylhydantion is 0.66 u/mL broth, which is 2-fold increase compared to the parent strain Burkholderia pickettii.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Recombinación Genética
/
Datos de Secuencia Molecular
/
Sondas de ADN
/
Expresión Génica
/
Secuencia de Aminoácidos
/
Clasificación
/
Clonación Molecular
/
Homología de Secuencia de Aminoácido
/
Análisis de Secuencia de ADN
/
Burkholderia
Límite:
Animales
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2002
Tipo del documento:
Artículo
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