ISSN:
0730-2312
Keywords:
β-lactamase
;
Saccharomyces cerevisiae
;
heterologous gene expression
;
preprotein
;
specific processing
;
Life and Medical Sciences
;
Cell & Developmental Biology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
Notes:
Synthesis and processing of the bacterial enzyme β-lactamase (E.C. 3.5. 2.6) were studied in Saccharomyces cerevisiae. The 2-μm 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 β-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 β-lactamase from Escherichia coli. Specificity of the processing of β-lactamase in yeast cells was verified by partial amino acid sequence analysis demonstrating the removal of the signal peptide at the correct position.
Additional Material:
3 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/jcb.240220303
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