ISSN:
1476-5535
Keywords:
Keywords: creatininase; creatinine amidohydrolase; Pseudomonas putida; cloning; DNA sequencing
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Process Engineering, Biotechnology, Nutrition Technology
Notes:
The gene encoding creatininase from Pseudomonas putida RS65 was cloned, sequenced and expressed in Escherichia coli. One plasmid containing a 7.0-kb HindIII insert was selected by its ability to express creatininase activity. After deletion of the adjacent restriction fragments, a 1.1-kb SphI fragment, which contained the full length of the creatininase gene, was subcloned into a pUC18 vector and the nucleotide sequence of the creatininase gene was determined. The gene consists of 771 base pairs and encodes a protein of 257 amino acids. The constitutive creatininase productivity of E. coli DH5α (pCRN741) cultured in broth was about 8.5-fold higher than that of P. putida RS65 cultured in a creatinine-containing medium. The creatininase gene was expressed efficiently in E. coli from its own promoter. Journal of Industrial Microbiology & Biotechnology (2000) 24, 2–6.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1038/sj.jim.2900753
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