Expression and Processing of Bacterial Beta-lactamase in the Yeast Saccharomyces Cerevisiae

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1981 Jul 1

PMID 7027263

Citations 26

Authors

R Roggenkamp

B Kustermann-Kuhn

C P Hollenberg

Affiliations

Soon will be listed here.

Abstract

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.

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