Cloning and comparative sequence analysis of the gene coding for isopenicillin N synthase in Streptomyces.
The genes coding for isopenicillin N synthase (IPNS) in Streptomyces jumonjinensis and S. lipmanii were isolated from recombinant phage lambda libraries using the S. clavuligerus IPNS gene as a heterologous probe. The S. jumonjinensis IPNS gene has an open reading frame coding for 329 amino acids, identical in size to that of the previously cloned S. clavuligerus IPNS gene. A partial nucleotide sequence was also determined for the S. lipmanii IPNS gene. Comparison of the predicted amino acid sequences of all three streptomycete IPNS proteins shows that they exhibit more than 70% similarity, close to that found in comparisons among fungal IPNS proteins and significantly greater than that found, approximately 60%, between Streptomyces and fungal IPNS proteins. We conclude that procaryotic and eucaryotic IPNS genes are subgroups of a single family of microbial IPNS genes. Hybridization probes prepared from IPNS genes of the above streptomycete species were used to detect analogous genes in eight other strains that included both penicillin and cephalosporin producers and non-producers. Each producer strain responded with all three probes implying the presence of an IPNS gene. Surprisingly, several non-producer strains also responded with one or two of the probes. Our results suggest that IPNS-related genes may be more prevalent in Streptomyces than previously believed.
Shiffman
D
D
Tel Aviv University, George S. Wise Faculty of Life Sciences, Department of Microbiology, Ramat Aviv, Israel.
Mevarech
M
M
Jensen
S E
SE
Cohen
G
G
Aharonowitz
Y
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003062:Codon; D015342:DNA Probes; D004798:Enzymes / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010088:Oxidoreductases; D015183:Restriction Mapping; D013045:Species Specificity; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*
D003062:Codon; D015342:DNA Probes; D004798:Enzymes; D005656:Fungal Proteins; D010088:Oxidoreductases; C038370:isopenicillin N synthetase