Open AccessIdentification of putative multifunctional peptide synthetase genes using highly conserved oligonucleotide sequences derived from known synthetases
Author(s) -
Borchert Stefan,
Patil Suresh S.,
Marahiel Mohamed A.
Publication year - 1992
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1992.tb05255.x
Subject(s) - operon , biology , bacillus subtilis , gene , oligonucleotide , amino acid , peptide , biochemistry , peptide sequence , genetics , surfactin , conserved sequence , microbiology and biotechnology , bacteria , escherichia coli
Many peptide antibiotics in prokaryotes and lower eukaryotes are produced non‐ribosomally by multi‐enzyme complexes. Analysis of gene‐derived amino acid sequences of some peptide synthetases of bacterial and fungal origins revealed a high degree of conservation (35–50% identity). The genes encoding those peptide synthetases are clustered into large operons with repetitive domains (about 600 amino acids), in the case of synthetases activating more than one amino acid. We used two 35‐mer oligonucleotides derived from two highly conserved regions of known peptide synthetases to identify the surfactin synthetase operon in Bacillus subtilis ATCC 21332, a strain not accessible to genetic manipulation. We show that the derived oligonucleotides can be used not only for the identification of unknown peptide synthetase genes by hybridization experiments but also in sequencing reactions as primers to identify internal domain sequences. Using this method, a 25.8‐kb chromosomal DNA fragment bearing a part of the surfactin biosynthesis operon was cloned and partial sequences of two internal domains were obtained.