spinosa trans1

compared with 100 (± 7.7) mg L−1 in the parental strain. Quantitative real time polymerase chain reaction analysis of three selected genes (spnH, spnI, and spnK) confirmed the positive effect of the overexpression of these genes on the spinosyn production. This study provides a simple avenue for enhancing spinosyn Lapatinib solubility dmso production. The strategies could also be used to improve the yield of other secondary metabolites. Saccharopolyspora spinosa was originally isolated in 1982 from a soil sample collected in a Caribbean island (Mertz & Yao, 1990). Fermentation broth extracts from this strain contain a series of spinosyn factors that are highly efficient against a broad range of pests, and appear to Selumetinib purchase have little or no effect on non-target insects and mammals (Sparks et al., 1998). Previous studies showed that spinosyns are derived from nine acetate and two propionate units, which produce a cyclized polyketide molecule; three carbon–carbon bonds are soon formed to obtain the tetracyclic aglycone (AGL). The rhamnose is subsequently attached and is tri-O-methylated to yield the intermediate pseudoaglycone (PSA), followed by the incorporation of forosamine sugar, giving the final spinosyns product. The most active and abundant spinosyns from S. spinosa fermentation broth are spinosyn A and spinosyn D. They differ from each

other by a single methyl substituent at position 6 of the polyketide. Other factors of the spinosyn family, produced as minor components, exhibit different methylation patterns and are significantly less active (Crouse et al., 2001). A naturally occurring mixture of spinosyn A (c. 85% of spinosad) and spinosyn D (c. 15%

of spinosad) is called spinosad (Waldron et al., 2001). The c. 74-kb spinosyn biosynthetic Adenosine triphosphate gene cluster contains 23 open reading frames (ORF) including five genes encoding a type I polyketide synthase (PKS) (spnA, B, C, D, and E); four genes involved in intramolecular C–C bond formation (spnF, J, L, and M); four genes responsible for rhamnose attachment and methylation (spnG, I, K, and H); six genes participating in forosamine biosynthesis (spnP, O, N, Q, R, and S) and four genes (ORF-L15, ORF-L16, ORF-R1, and ORF-R2) with no proven role in spinosyn biosynthesis (Waldron et al., 2001). The genes involved in rhamnose biosynthesis (gtt, gdh, epi, and kre) are not linked to this cluster (Madduri et al., 2001b). Traditionally, improvement of secondary metabolite-producing strains is achieved by random mutagenesis and selection techniques (Parekh et al., 2000). Although these techniques have succeeded in generating many industrial strains, they are time-consuming and costly. Rational strain improvement strategies overlap with classical approaches in generating a mutant population (Adrio & Demain, 2006).