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 fIA and fIB genes of the fluorometabolite gene cluster in S. cattleya, were overexpressed. Also, an isomerase gene product that can convert 5-FDRP 6 to 5-FDRibuIP 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.
Original language | English |
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Pages (from-to) | 1268-1276 |
Number of pages | 9 |
Journal | Chemistry & Biology |
Volume | 15 |
Issue number | 12 |
Early online date | 10 Nov 2008 |
DOIs | |
Publication status | Published - 22 Dec 2008 |
Keywords
- 5-methylthioribose 1-phosphate isomerase
- streptomyces-cattleya
- fluorometabolite biosynthesis
- crystal-structure
- L-ribulose-5-phosphate 4-epimerase
- bacillus-subtilis
- bond formation
- gene-cluster
- amino-acids
- identification