title: Biochemical investigations of two 6-DMATS enzymes from Streptomyces reveal new features of L-tryptophan prenyltransferases.
abstract: Two putative prenyltransferase genes, SAML0654 and Strvi8510, were identified in Streptomyces ambofaciens and Streptomyces violaceusniger, respectively. Their deduced products share 63% sequence identity. Biochemical investigations with recombinant proteins demonstrated that L-tryptophan and derivatives, including D-tryptophan, 4-, 5-, 6- and 7-methyl-dl-tryptophan, were well accepted by both enzymes in the presence of DMAPP. Structural elucidation of the isolated products revealed regiospecific prenylation at C-6 of the indole ring and proved unequivocally the identification of two very similar 6-dimethylallyltryptophan synthases (6-DMATS). Detailed biochemical investigations with SAML0654 proved L-tryptophan to be the best substrate (K(m) 18 μm, turnover 0.3 s(-1)). Incubation with different prenyl donors showed that they also accepted GPP and catalyzed the same specific prenylation. Utilizing GPP as a prenyl donor has not been reported for tryptophan prenyltransferases previously. Both enzymes also catalyzed prenylation of some hydroxynaphthalenes; this has not previously been described for bacterial indole prenyltransferases. Interestingly, SAML0654 transferred prenyl moieties onto the unsubstituted ring of hydroxynaphthalenes.
authors.affiliation: Philipps-Universität Marburg, Institut für Pharmazeutische Biologie und Biotechnologie, Deutschhausstrasse 17A, 35037 Marburg (Germany); Zentrum für Synthetische Mikrobiologie, Philipps-Universität Marburg, Hans-Meerwein-Strasse, 35032 Marburg (Germany).
authors.forename: Julia
authors.initials: J
authors.lastname: Winkelblech
authors.forename: Shu-Ming
authors.initials: SM
authors.lastname: Li
mesh_terms: D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004122:Dimethylallyltranstransferase / Q000235:genetics / Q000378:metabolism*; D045782:Hemiterpenes / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D009284:Naphthols / Q000737:chemistry / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000737:chemistry / Q000378:metabolism; D011106:Polyisoprenyl Phosphates / Q000737:chemistry / Q000378:metabolism; D054876:Prenylation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012717:Sesquiterpenes / Q000737:chemistry / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics; D014364:Tryptophan / Q000378:metabolism
chemical_list: D045782:Hemiterpenes; D009284:Naphthols; D009943:Organophosphorus Compounds; D011106:Polyisoprenyl Phosphates; D011993:Recombinant Fusion Proteins; D012717:Sesquiterpenes; C043060:3,3-dimethylallyl pyrophosphate; C004808:farnesyl pyrophosphate; D014364:Tryptophan; D004122:Dimethylallyltranstransferase
keywords: Streptomyces; dimethylallyltryptophan synthases; enzymes; hydroxynaphthalenes; indole prenyltransferases; natural products