Specific processing of the bacterial beta-lactamase precursor in Saccharomyces cerevisiae.
Synthesis and processing of the bacterial enzyme beta-lactamase (E.C. 3.5. 2.6) were studied in Saccharomyces cerevisiae. The 2-micron DNA vector pADH040-2 containing the yeast ADH1 promoter fused to the bacterial gene was used in order to obtain enhanced synthesis of the bacterial protein in yeast transformants. Both precursor and mature beta-lactamase were shown to be present in yeast cells, the precursor being the major product. The mature enzyme was purified about 500-fold over crude extracts to apparent homogeneity and thus represents nearly 0.2% of the total yeast protein. No difference in specific activity and molecular weight could be observed when compared with the authentic beta-lactamase from Escherichia coli. Specificity of the processing of beta-lactamase in yeast cells was verified by partial amino acid sequence analysis demonstrating the removal of the signal peptide at the correct position.
D000595:Amino Acid Sequence; D004592:Electrophoresis, Starch Gel; D004792:Enzyme Precursors / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D005786:Gene Expression Regulation; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D001618:beta-Lactamases / Q000235:genetics*
D004792:Enzyme Precursors; D001618:beta-Lactamases