Delineating the earliest steps of gilvocarcin biosynthesis: role of GilP and GilQ in starter unit specificity

Abstract

In vivo and in vitro investigations of GilP and GilQ, two acyltransferases encoded by the gilvocarcin gene cluster, show that GilQ confers unique starter unit specificity when catalyzing an early as well as rate limiting step of gilvocarcin biosynthesis.

Enzymatic Total Synthesis of Defucogilvocarcin M and Its Implications for Gilvocarcin Biosynthesis

“…[1a] The function of biosynthetic enzymes was then confirmed by both in vitro and in vivo studies. [7a, 9, 11–13] The cluster encodes a set of typical type II polyketide synthase (PKS) enzymes, which includes a ketosynthase (GilA), a chain-length factor (GilB), an acyl carrier protein (GilC), two malonyl-CoA:acyl carrier protein transacylase (MCAT) homologues (GilP, GilQ), a PKS-associated keto reductase (GilF), and two cyclases (GilK, GilG). Together, this group of enzymes was able to catalyze the formation of the angucy-clinone UWM6 ( 10 ).…”

“…JadF has previously been shown to complement the GilOIV-minus mutant S. lividans TK24 (cosG9B3-ΔOIV), which proves the functional identity of the two enzymes. [11] Considering that GilOI and GilOIV are oxygenases that contain flavin adenine dinucleotide (FAD, Figure S3 in the Supporting Information) and require their cofactor to be recycled, a reductase was needed to generate FADH 2 . The putative nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) flavin reductase GilH [7a] showed only very poor activity.…”

Delineation of gilvocarcin, jadomycin, and landomycin pathways through combinatorial biosynthetic enzymology

“…The gilvocarcin biosynthetic gene cluster ( gil ) was cloned and heterologously expressed [30], and the clusters of chrysomycin ( chry ) and ravidomycin ( rav ) were cloned and analyzed (Figure 3) [74 • ,75 • ,76,77 • ,78 • ]. The functions of the post-PKS gene products were assigned after gene inactivation, complementation, cross-feeding experiments along with few in vitro and in vivo studies of activity of individual enzymes or enzyme mixtures [75 • ,79–82,83 • ].…”

Engineered Biosynthesis of Gilvocarcin Analogues with Altered Deoxyhexopyranose Moieties

“…As before, structural elucidation of structure 20 was carried out through NMR (Table 3) (structures 1 to 3) are naturally produced as three congeners differing only in their side chains attached at C-8 (39). This is also illustrated in the host strains' ability to produce compounds 4 to 6, as seen in Fig.…”