A study of iterative type II polyketide synthases, using bacterial genes cloned from soil DNA: a means to access and use genes from uncultured microorganisms

• DOI: 10.1128/jb.179.23.7360-7368.1997
• Publication Date: 2016-11-10T16:17:28Z
• Authors: K T Seow; G Meurer; M Gerlitz; E Wendt-Pienkowski; C R Hutchinson; J Davies
• Keywords: Gene cluster

To examine as randomly possible the role of beta-ketoacyl and acyl carrier protein (ACP) components bacterial type II polyketide synthases (PKSs), homologs chain-length-factor (CLF) genes were cloned from environmental community microorganisms. With PCR primers derived conserved regions known ketosynthase (KSalpha) ACP specifying formation 16- to 24-carbon polyketides, two CLF (KSbeta) unclassified streptomycetes isolated soil, soil DNA without prior isolation parent microorganism. The sequence deduced product each gene distinct those KSbeta and, by phylogenetic analysis, belonged antibiotic-producing PKS clusters. Hybrid cassettes constructed with novel substituted for actI-ORF2 or tcmL subunit genes, along respective actI-ORF1 tcmK KSalpha, tcmM ACP, tcmN cyclase found produce an octaketide decaketide characteristic ones be made heterologous KSalpha partner. Since substantially less than 1% microorganisms present in are thought cultivatable standard methods, this work demonstrates a potential way gain access more extensive range microbial molecular diversity biosynthetic pathways whose products can tested biological applications.