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通过在恒化器中以变化的稀释率进行选择来提高β-半乳糖苷酶的催化速率。

Increase in the catalytic rate of beta-galactosidase by selection in chemostats at changing dilution rates.

作者信息

Tsen S D

机构信息

Department of Metabolic Regulation, Boston Biomedical Research Institute, MA 02114.

出版信息

Biochem Biophys Res Commun. 1990 Feb 14;166(3):1245-50. doi: 10.1016/0006-291x(90)90999-4.

Abstract

The experiments presented in this paper explore whether mutants with catalytically more active beta-galactosidase (E.C. 3.2.1.23) can be selected in lactose-limited chemostats. This experimental system has been chosen because lactose metabolism in Escherichia coli is well understood both from a biochemical and genetic point of view. In a lactose-limited chemostat with constant dilution rates, both beta-galactosidase and the lactose permease increased in quantity. The catalytic rate (kcat) of beta-galactosidase in populations showed no change. On the other hand, in chemostats with periodically changing dilution rates, the catalytic rate of beta-galactosidase increased dramatically. Therefore, the selection for beta-galactosidase with improved catalytic rate occurs in chemostats with fluctuating dilution rates but not in ones with constant dilution. These observations may be of value in the selection of other enzymes with enhanced catalytic rates.

摘要

本文所呈现的实验探讨了在乳糖受限的恒化器中,是否能够筛选出具有催化活性更高的β-半乳糖苷酶(E.C. 3.2.1.23)的突变体。选择这个实验系统是因为从生化和遗传学角度来看,大肠杆菌中的乳糖代谢已被充分了解。在具有恒定稀释率的乳糖受限恒化器中,β-半乳糖苷酶和乳糖通透酶的数量均增加。群体中β-半乳糖苷酶的催化速率(kcat)没有变化。另一方面,在稀释率周期性变化的恒化器中,β-半乳糖苷酶的催化速率急剧增加。因此,对具有更高催化速率的β-半乳糖苷酶的选择发生在稀释率波动的恒化器中,而不是在稀释率恒定的恒化器中。这些观察结果对于筛选其他具有更高催化速率的酶可能具有价值。

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