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嗜热微生物中α-氨基异丁酸转运的特性

Properties of alpha-aminoisobutyric acid transport in a thermophilic microorganism.

作者信息

Reizer J, Grossowicz N

出版信息

J Bacteriol. 1974 May;118(2):414-24. doi: 10.1128/jb.118.2.414-424.1974.

Abstract

Uptake of alpha-aminoisobutyric acid (AIB) by a leucine-tyrosine auxotroph of a thermophilic microorganism starved for leucine was studied. AIB was taken up by the cells against a substantial concentration gradient (300:1) and was present there in a free and unchanged form. Various energy inhibitors and sulfhydryl reagents strongly inhibited the accumulation of AIB. AIB uptake obeyed saturation kinetics, and the Lineweaver-Burk plot is characterized by a biphasic curve. AIB most probably shares a common transport system(s) with alanine, serine, and glycine. A mutant defective in l-alanine uptake was isolated by using the suicide effect due to accumulation of the tritiated substrate. The mutant also exhibited impaired transport activity towards AIB, glycine, and l-serine, but not to phenylalanine or valine. The transport of AIB, glycine, l-alanine, and l-serine was induced by d-alanine (5 x 10(-3) M) during growth in a succinate- and ammonia-containing medium. De novo protein synthesis was required for the induction of AIB transport; the induction was inhibited when growth occurred in glucose-containing media. The apparent differential rate of synthesis of the AIB transport system was decreased considerably in glucose-grown cells as compared to succinate-grown cells. A common genetic basis of either the regulatory or structural nature for the transport of AIB, alanine, glycine, and serine in a thermophilic microorganism is suggested.

摘要

对一种嗜热微生物的亮氨酸 - 酪氨酸营养缺陷型菌株在缺乏亮氨酸的情况下对α - 氨基异丁酸(AIB)的摄取进行了研究。细胞逆着相当大的浓度梯度(300:1)摄取AIB,且AIB以游离且未改变的形式存在于细胞中。各种能量抑制剂和巯基试剂强烈抑制AIB的积累。AIB摄取遵循饱和动力学,Lineweaver - Burk图的特征是一条双相曲线。AIB很可能与丙氨酸、丝氨酸和甘氨酸共享一个共同的转运系统。利用氚标记底物积累导致的自杀效应分离出了一种l - 丙氨酸摄取缺陷的突变体。该突变体对AIB、甘氨酸和l - 丝氨酸的转运活性也受损,但对苯丙氨酸或缬氨酸的转运活性未受损。在含有琥珀酸盐和氨的培养基中生长期间,d - 丙氨酸(5×10⁻³ M)可诱导AIB、甘氨酸、l - 丙氨酸和l - 丝氨酸的转运。AIB转运的诱导需要从头合成蛋白质;当在含葡萄糖的培养基中生长时,诱导作用受到抑制。与在琥珀酸盐培养基中生长的细胞相比,在葡萄糖培养基中生长的细胞中AIB转运系统的表观合成差异速率显著降低。这表明嗜热微生物中AIB、丙氨酸、甘氨酸和丝氨酸转运在调控或结构性质上存在共同的遗传基础。

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