Document Detail

Overproduction of thymidine by recombinant brevibacterium helvolum amplified with thymidine monophosphate phosphohydrolase gene from bacteriophage PBS2.
MedLine Citation:
PMID:  15055759     Owner:  NLM     Status:  MEDLINE    
A microbial fermentation process could be used to produce thymidine biologically but many of the enzymes related to nucleotide biosynthesis are highly regulated. To overcome the complex regulation steps, an analogue mutant of Brevibacterium helvolum resistant to fluorouracil, hydroxyurea, and trimethoprim was constructed. This mutant accumulated 380 mg thymidine 1(-1) in 16 h in shake-flask culture. However, the accumulation of thymidine monophosphate (TMP) inside the cells suggested a low activity of nucleotidase which degrades TMP to thymidine. This limitation was overcome by cloning the TMP phosphohydrolase (TMPase) gene of the unusual bacteriophage, PBS2. As a result, TMP in recombinant cells decreased from 230 micromol g(-1) cell to 20 micromol g(-1) cell with accumulation of 500 mg thymidine l(-1) in the medium.
Hyeon Cheol Lee; Jung Mi Ahn; Sang Nam Lee; Jung Hoe Kim
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Biotechnology letters     Volume:  26     ISSN:  0141-5492     ISO Abbreviation:  Biotechnol. Lett.     Publication Date:  2004 Feb 
Date Detail:
Created Date:  2004-04-01     Completed Date:  2004-12-03     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8008051     Medline TA:  Biotechnol Lett     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  265-8     Citation Subset:  IM    
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1 Guseong-dong, Yuseong-gu, Daejeon 305-701, Republic of Korea.
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MeSH Terms
Bacterial Proteins / chemistry,  genetics*
Bacteriophages / enzymology*,  genetics
Brevibacterium / metabolism*
Chromatography, High Pressure Liquid
Cloning, Molecular
Drug Resistance, Bacterial
Fluorouracil / pharmacology
Hydroxyurea / pharmacology
Nucleotidases / genetics*,  metabolism
Phosphoric Monoester Hydrolases / genetics*,  metabolism
Plasmids / metabolism
Recombinant Proteins / metabolism
Thymidine / biosynthesis*,  metabolism
Thymidine Monophosphate / metabolism*
Trimethoprim / pharmacology
Reg. No./Substance:
0/Bacterial Proteins; 0/Recombinant Proteins; 127-07-1/Hydroxyurea; 365-07-1/Thymidine Monophosphate; 50-89-5/Thymidine; 51-21-8/Fluorouracil; 738-70-5/Trimethoprim; EC 3.1.3.-/Nucleotidases; EC 3.1.3.-/Phosphoric Monoester Hydrolases; EC 5'-phosphatase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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