细菌β-内酰胺酶在酿酒酵母中的表达与加工。

Roggenkamp R, Kustermann-Kuhn B, Hollenberg C P

Proc Natl Acad Sci U S A. 1981 Jul;78(7):4466-70. doi: 10.1073/pnas.78.7.4466.

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.

本文描述了编码β-内酰胺酶（EC 3.5.2.6）的细菌抗生素抗性基因在酿酒酵母中的表达模式。含有由2μm DNA载体中的pBR325组成的杂交质粒pMP78-1的酵母转化体合成了一种活性β-内酰胺酶蛋白。该酶比粗提物纯化了约100倍。就活性、分子量和与特异性抗体的结合而言，酵母β-内酰胺酶与来自大肠杆菌的纯化酶没有区别。由于细菌酶是以前体蛋白形式合成并随后成熟的，结果表明酿酒酵母能够将前体蛋白转化为成熟的β-内酰胺酶。体外实验证实了这一点，该实验表明细菌前体蛋白可以被酿酒酵母的粗提物加工处理。