Isolation, sequence and expression in Escherichia coli, Bacillus subtilis and Lactococcus lactis of the DNase (streptodornase)-encoding gene from Streptococcus equisimilis H46A.
A partial library of BclI-generated chromosomal DNA fragments from Streptococcus equisimilis H64A (Lancefield Group C) was constructed in Escherichia coli. Clones displaying either streptokinase or deoxyribonuclease (streptodornase; SDC) activities were isolated. The gene (sdc) expressing the SDC activity was allocated on the 1.1-kb AccI DNA subfragment. Sequence analysis of this DNA fragment revealed the presence of one open reading frame, which could encode a protein of 36.8 kDa. The N-terminal portion of the deduced protein exhibited features characteristic of prokaryotic signal peptides. The sdc gene was expressed in E. coli, Bacillus subtilis and Lactococcus lactis. As observed for S. equisimilis, in the heterologous Gram + hosts, at least part of the SDC protein was secreted into the medium.
Department of Pharmaceutical Microbiology, Medical Academy, Warsaw, Poland.
R
R
Wolinowska
P
P
Cegłowski
J
J
Kok
G
G
Venema
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D003851:Deoxyribonucleases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D013294:Lactococcus lactis / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D013291:Streptococcus / Q000201:enzymology / Q000235:genetics*; D013300:Streptokinase / Q000096:biosynthesis / Q000378:metabolism
D004269:DNA, Bacterial; D003851:Deoxyribonucleases; D013300:Streptokinase