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- Title
Lamivudine production via enantioselective deamination by thermostable Bacillus caldolyticus cytidine deaminase.
- Authors
Ju-Hyung Woo; Hyun-Jeung Shin; Tae-Ho Kim; Sa-Youl Ghim; Lak-Shin Jeong; Jong-Guk Kim; Bang-Ho Song
- Abstract
To decrease the costs of producing the anti-HIV drug, lamivudine, an enzymatic conversion process was developed instead of the traditional chemical method. Thermostable cytidine deaminase was over-produced by cloning the cdd gene into E. coli JF611/pCJH53 from Bacillus caldolyticus. The purified cytidine deaminase was recovered from the lysate of the recombinant E. coli JF611/pCJH53 by removing heat-denatured proteins and eluting sequential chromatography. When the enzyme was used to deaminate (-)-β-l-(2R,5S)- and (+)-β-d-(2S,5R)-1,3-oxathiolanyl-cytosine, about 68% of the (+)-β-d-(2S,5R)-1,3-oxathiolanyl-cytosine was deaminated into the corresponding (+)-thiauridine maximally.
- Subjects
ESCHERICHIA coli; GENETIC engineering; CHROMATOGRAPHIC analysis; ESCHERICHIA; MICROBIOLOGY; MICROBIAL genetics
- Publication
Biotechnology Letters, 2001, Vol 23, Issue 2, p131
- ISSN
0141-5492
- Publication type
Article
- DOI
10.1023/A:1010353502346