Cloning and characterization of a gene of Streptomyces griseus that increases production of extracellular enzymes in several species of Streptomyces.
A 7.2 kb Bg/II restriction fragment, which increases the production of several extracellular enzymes, including alkaline phosphatase, amylase, protease, lipase and beta-galactosidase, was cloned in Streptomyces lividans from the DNA of S. griseus ATCC 10137. This gene (named saf) showed a positive gene dosage effect on production of extracellular enzymes. When the saf gene was introduced into cells in high copy numbers it delayed the formation of pigments and spores in S. lividans and also retarded actinorhodin production in Streptomyces coelicolor. The saf gene hybridized with specific bands in the DNA of several Streptomyces strains tested. A 1 kb fragment containing the saf gene was sequenced and contains an open reading frame (ORF) of 306 nucleotides which encodes a polypeptide of Mr 10,500. This ORF is contained within a fragment of 432 bp which retained activity in Streptomyces. A fragment with promoter activity is present upstream of the saf reading frame. The predicted Saf polypeptide has a strong positive charge, and does not show a typical amino acid composition for a membrane protein, and contains a DNA-binding domain similar to those found in several regulatory proteins.
Daza
A
A
Department of Ecology, Genetics and Microbiology, University of León, Spain.
Gil
J A
JA
Vigal
T
T
Martin
J F
JF
D000469:Alkaline Phosphatase / Q000096:biosynthesis; D000595:Amino Acid Sequence; D000681:Amylases / Q000096:biosynthesis; D001426:Bacterial Proteins; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D010450:Endopeptidases / Q000096:biosynthesis; D004790:Enzyme Induction / Q000235:genetics; D005798:Genes, Bacterial; D008049:Lipase / Q000096:biosynthesis; D008969:Molecular Sequence Data; D010860:Pigments, Biological / Q000096:biosynthesis; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D013171:Spores, Bacterial / Q000254:growth & development; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics; D013305:Streptomyces griseus / Q000201:enzymology / Q000235:genetics*; D014170:Transformation, Genetic; D001616:beta-Galactosidase / Q000096:biosynthesis
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D010860:Pigments, Biological; D008049:Lipase; C024404:BglII endonuclease; D015252:Deoxyribonucleases, Type II Site-Specific; D000469:Alkaline Phosphatase; D000681:Amylases; D001616:beta-Galactosidase; D010450:Endopeptidases