|
type |
Journal Article |
authors |
Deng H, Cross SM, McGlinchey RP, Hamilton JT, O'Hagan D.
|
title |
In vitro reconstituted biotransformation of 4-fluorothreonine from fluoride ion: application of the fluorinase |
journal |
Chem Biol |
Activity |
2.2.1.8 |
Family |
2.2.1.8 |
sel |
selected |
ui |
19101471 |
year |
(2008) |
volume |
15 |
number |
12 |
pages |
1268-76 |
| |
keywords |
doi: 10.1016/j.chembiol.2008.10.012 |
abstract |
In this paper, we report that fluoride ion is converted to the amino acid/antibiotic 4-fluorothreonine 2 in a biotransformation involving five (steps a-e) overexpressed enzymes. The biotransformation validates the biosynthetic pathway to 4-fluorothreonine in the bacterium Streptomyces cattleya (Schaffrath et al., 2002). To achieve an in vitro biotransformation, the fluorinase and the purine nucleoside phosphorylase (PNP) enzymes (steps a and b), which are coded for by the flA and flB genes of the fluorometabolite gene cluster in S. cattleya, were overexpressed. Also, an isomerase gene product that can convert 5-FDRP 6 to 5-FDRibulP 7 (step c) was identified in S. cattleya, and the enzyme was overexpressed for the biotransformation. A fuculose aldolase gene from S. coelicolor was overexpressed in E. coli and was used as a surrogate aldolase (step d) in these experiments. To complete the complement of enzymes, an ORF coding the PLP-dependent transaldolase, the final enzyme of the fluorometabolite pathway, was identified in genomic DNA by a reverse genetics approach, and the S. cattleya gene/enzyme was then overexpressed in S. lividans. This latter enzyme is an unusual PLP-dependent catalyst with some homology to both bacterial serine hydroxymethyl transferases (SHMT) and C5 sugar isomerases/epimerases. The biotransformation demonstrates the power of the fluorinase to initiate C-F bond formation for organo-fluorine synthesis. |
last changed |
2018/02/13 15:42 |
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