1887

Abstract

A putative prenyltransferase gene, , was identified in the genome sequence of . EAL92290, encoded by , consists of 472 aa, with a molecular mass of about 53 kDa. The coding sequence of was cloned in pQE60, and overexpressed in The soluble His-fusion protein was purified to apparent homogeneity, and characterized biochemically. The enzyme was found to catalyse the prenylation of Trp at the C-7 position of the indole moiety, in the presence of dimethylallyl diphosphate (DMAPP); therefore, it functions as a 7-dimethylallyltryptophan synthase (7-DMATS). The structure of the enzymic product was elucidated by NMR and MS analysis. values were 67 μM for DMAPP, and 137 μM for -Trp. Geranyl diphosphate was not accepted as prenyl donor, while Trp-containing dipeptides were found to be aromatic substrates of 7-DMATS. 7-DMATS did not need divalent metal ions for its enzymic reaction, although Ca enhanced the reaction velocity slightly. The enzyme is the second dimethylallyltryptophan synthase identified in . Interestingly, it shares a sequence identity of only 31 % at the amino acid level with another known dimethylallyltryptophan synthase, FgaPT2, from the same fungus; FgaPT2 prenylates -Trp at the C-4 position of the indole ring. belongs to a putative biosynthetic gene cluster consisting of eight genes. Orthologous clusters were also identified in the genome sequences of and . The putative roles of the genes in the cluster are discussed.

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2007-10-01
2020-09-26
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