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2-脱氧半乳糖,鼠伤寒沙门氏菌半乳糖通透酶的一种特异性底物:其在分离galP突变体中的应用。

2-deoxygalactose, a specific substrate of the Salmonella typhiimurium galactose permease: its use for the isolation of galP mutants.

作者信息

Nagelkerke F, Postma P W

出版信息

J Bacteriol. 1978 Feb;133(2):607-13. doi: 10.1128/jb.133.2.607-613.1978.

Abstract

2-Deoxygalactose is a specific substrate of the galactose permease. The apparent Km is about 500 micron, compared to 45 micron for galactose, whereas the maximal rate of uptake is one-half to one-third of that of galactose. None of the other galactose transport systems, including methyl beta-D-thiogalactosides I and II, the beta-methyl-galactoside permease, and both arabinose systems, is able to catalyze transport of 2-deoxygalactose to a significant extent. 2-Deoxygalactose can also be used to isolate mutants defective in galactose permease, since it is bacteriostatic. Colonies that grow with lactate, malate, or succinate as a carbon source in the presence of 0.5 to 2 mM 2-doexygalactose were found to be mostly galP mutants, lacking galactose permease. Spontaneous 2-deoxygalactose-resistant strains arose with a frequency of about 2 X 10(-6). galP mutants have also been derived from pts deletion mutants that require galactose permease for growth on glucose. Revertants have been obtained that have acquired the parental phenotype.

摘要

2-脱氧半乳糖是半乳糖通透酶的一种特异性底物。其表观米氏常数(Km)约为500微米,而半乳糖的表观米氏常数为45微米,2-脱氧半乳糖的最大摄取速率是半乳糖的最大摄取速率的二分之一到三分之一。其他半乳糖转运系统,包括β-D-硫代半乳糖苷Ⅰ和Ⅱ、β-甲基半乳糖苷通透酶以及两种阿拉伯糖系统,均不能在很大程度上催化2-脱氧半乳糖的转运。2-脱氧半乳糖还可用于分离半乳糖通透酶缺陷型突变体,因为它具有抑菌作用。发现在0.5至2 mM 2-脱氧半乳糖存在的情况下,以乳酸、苹果酸或琥珀酸作为碳源生长的菌落大多是galP突变体,缺乏半乳糖通透酶。自发产生的2-脱氧半乳糖抗性菌株出现的频率约为2×10⁻⁶。galP突变体也源自pts缺失突变体,这些pts缺失突变体在以葡萄糖为碳源生长时需要半乳糖通透酶。现已获得恢复了亲本表型的回复突变体。

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