Expression of the Streptomyces enzyme endoglycosidase H in Escherichia coli.
Endoglycosidase H is one of a large number of enzymes secreted by Streptomyces plicatus and other Streptomyces species. When the structural gene for this enzyme is introduced into Escherichia coli attached to the plasmid pBR-322 or Charon 4 phage, the enzyme is synthesized and is found in the periplasmic space, culture medium, and cells. Attachment of the UV-5 lac promoter to a site in the plasmid adjacent to the Streptomyces insert stimulates enzyme synthesis as much as 100-fold. This result demonstrates that transcription of the Streptomyces gene can be initiated from sequences outside of the Streptomyces insert. Initiation of transcription on a Streptomyces promoter is also a suggested but unproven possibility. In contrast to the situation in Streptomyces, where the enzyme has a molecular weight of 27,000, the enzyme made in E. coli has a molecular weight of approximately 30,000. Possible explanations for this difference in size are lack of cleavage of the Streptomyces secretion "signal sequence" in E. coli or protein "processing" by enzymes secreted into the medium by STreptomyces.
D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005796:Genes; D006026:Glycoside Hydrolases / Q000235:genetics*; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008970:Molecular Weight; D010957:Plasmids; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics; D014158:Transcription, Genetic
D004274:DNA, Recombinant; D004262:DNA Restriction Enzymes; D006026:Glycoside Hydrolases; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase