Metabolic diversity in myxobacteria: identification of the myxalamid and the stigmatellin biosynthetic gene cluster of Stigmatella aurantiaca Sg a15 and a combined polyketide-(poly)peptide gene cluster from the epothilone producing strain Sorangium cellulosum So ce90
S. Beyer et al., Metabolic diversity in myxobacteria: identification of the myxalamid and the stigmatellin biosynthetic gene cluster of Stigmatella aurantiaca Sg a15 and a combined polyketide-(poly)peptide gene cluster from the epothilone producing strain Sorangium cellulosum So ce90, BBA-GENE ST, 1445(2), 1999, pp. 185-195
Citations number
41
Categorie Soggetti
Molecular Biology & Genetics
Journal title
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION
Myxobacterial strains producing polyketides (PKs) assumed to be biosynthesi
zed by a type I polyketide synthase (PKS) were analysed. Myxobacteria also
produce a variety of polypeptides (PP) and PKs with incorporated amino acid
s ('mixed PK-PP'). In order to be able to identify the biosynthetic gene cl
usters for these metabolites a PCR based approach has been developed to clo
ne ketosynthase (KS) domains of PKS genes from these organisms. Conserved r
egions of peptide synthetases of the non-ribosomal type (NRPS) were also am
plified via PCR. KS fragments from Stigmatella aurantiaca Sg a15 were used
for chromosomal gene inactivation experiments resulting in a series of muta
nts including such that were unable to produce stigmatellins and myxalamids
. A NRPS fragment and PKS fragments from Sorangium cellulosum So ce90 were
used to identify cosmids hybridizing with both types of probes from a genom
ic library. Both a NRPS and a PKS fragment were cloned and sequenced from a
relatively short restriction fragment of one of these cosmids. The method
described here should be very useful to clone and identify PKS, NRPS and mi
xed PKS-NRPS from myxobacteria in general and thereby open opportunities to
use the biochemical diversity of these bacteria for genetic engineering an
d combinatorial biosynthesis. (C) 1999 Elsevier Science B.V. All rights res
erved.