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.
Article
in Chinese
| 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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Recombination, Genetic
/
Molecular Sequence Data
/
DNA Probes
/
Gene Expression
/
Amino Acid Sequence
/
Classification
/
Cloning, Molecular
/
Sequence Homology, Amino Acid
/
Sequence Analysis, DNA
/
Burkholderia
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2002
Type:
Article
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