In:
Microbiology, Microbiology Society, Vol. 156, No. 1 ( 2010-01-01), p. 278-286
Abstract:
A putative prenyltransferase gene sirD has been identified in the gene cluster encoding the biosynthesis of the phytotoxin sirodesmin PL in Leptosphaeria maculans . The gene product was found to comprise 449 aa, with a molecular mass of 51 kDa. In this study, the coding region of sirD was amplified by PCR from cDNA, cloned into pQE70, and overexpressed in Escherichia coli . The overproduced protein was purified to apparent homogeneity, and characterized biochemically. The dimeric recombinant SirD was found to catalyse the O -prenylation of l -Tyr in the presence of dimethylallyl diphosphate; this was demonstrated unequivocally by isolation and structural elucidation of the enzymic product. Therefore, SirD catalyses the first pathway-specific step in the biosynthesis of sirodesmin PL. K m values for l -Tyr and dimethylallyl diphosphate were determined as 0.13 and 0.17 mM, respectively. Interestingly, SirD was found to share significant sequence similarity with indole prenyltransferases, which catalyse prenyl transfer reactions onto different positions of indole rings. In contrast to indole prenyltransferases, which accept indole derivatives, but not Tyr or structures derived thereof, as substrates, SirD also prenylated l -Trp, resulting in the formation of 7-dimethylallyltryptophan. A K m value of 0.23 mM was determined for l -Trp. Turnover numbers of 1.0 and 0.06 S −1 were calculated for l -Tyr and l -Trp, respectively.
Type of Medium:
Online Resource
ISSN:
1350-0872
,
1465-2080
DOI:
10.1099/mic.0.033886-0
Language:
English
Publisher:
Microbiology Society
Publication Date:
2010
detail.hit.zdb_id:
2008736-6
SSG:
12
