Open AccessExpression and processing of bacterial beta-lactamase in the yeast Saccharomyces cerevisiae.
Author(s) -
Rainer Roggenkamp,
B. Kustermann-Kuhn,
Cornelis P. Hollenberg
Publication year - 1981
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.78.7.4466
Subject(s) - saccharomyces cerevisiae , yeast , escherichia coli , biology , plasmid , biochemistry , enzyme , microbiology and biotechnology , gene
The mode of expression in Saccharomyces cerevisiae of the bacterial antibiotic resistance gene coding for beta-lactamase (EC 3.5.2.6) is described. Yeast transformants, containing hybrid plasmid pMP78-1 consisting of pBR325 in a 2-micrometers DNA vector, synthesize an active beta-lactamase protein. The enzyme was purified about 100-fold over crude extracts. With regard to activity, molecular weight, and binding to specific antibodies the yeast beta-lactamase was indistinguishable from the purified enzyme from Escherichia coli. Because the bacterial enzyme is synthesized as a preprotein with subsequent maturation, the results suggest that S. cerevisiae is able to convert the preprotein to the mature beta-lactamase. This was confirmed by in vitro experiments showing that the bacterial preprotein can be processed by crude extracts of S. cerevisiae.