THE ROLE OF TREHALOSE SYNTHESIS FOR THE ACQUISITION OF THERMOTOLERANCE IN YEAST .2. PHYSIOLOGICAL CONCENTRATIONS OF TREHALOSE INCREASE THE THERMAL-STABILITY OF PROTEINS IN-VITRO
THE ROLE OF TREHALOSE SYNTHESIS FOR THE ACQUISITION OF THERMOTOLERANCE IN YEAST .2. PHYSIOLOGICAL CONCENTRATIONS OF TREHALOSE INCREASE THE THERMAL-STABILITY OF PROTEINS IN-VITRO
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DOI：
10.1111/j.1432-1033.1994.tb19929.x
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发表时间：
1994-01-15
期刊：
影响因子：
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通讯作者：
WIEMKEN, A
中科院分区：
文献类型：
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作者：
HOTTIGER, T;DEVIRGILIO, C;WIEMKEN, A
在面包酵母（酿酒酵母）中，非还原性二糖海藻糖的积累是由激活热休克反应的刺激触发的。以前，海藻糖水平已被证明与耐热性密切相关，表明这种物质的保护功能。支持这一观点的遗传学证据在随附的论文中提出[De Pastilio，C.，Hottiger，T.，Dominguez，J.，Boller，T. & Wiemken，A.（1993）Eur. 219，179-186]。在这项研究中，我们研究了海藻糖对蛋白质热稳定性的影响，这一参数被认为是耐热性的主要决定因素。发现生理浓度的海藻糖（高达0.5 M）可有效保护酵母酶（葡萄糖-6P-脱氢酶、磷酸葡萄糖异构酶）以及非酵母来源的酶（牛）。谷氨酸脱氢酶（EcoRI）对体外热失活的抑制作用。海藻糖还减少了热诱导的蛋白质聚集体的形成。二糖被证明是一种相容的溶质，因为即使在非常高的浓度下（高达1 M），它也不会显著干扰试验酶的活性。海藻糖作为蛋白质稳定剂至少与其他相容性溶质（包括糖、多元醇和氨基酸）一样好或更好，而结构相关的海藻糖-6P没有任何保护作用。海藻糖对酶的热保护作用是明显的，即使在含有高浓度酵母蛋白或底物的溶液中也是如此。这些数据表明海藻糖的积累可能通过增强完整细胞中蛋白质的稳定性来提高酵母的耐热性。
In-baker's yeast (Saccharomyces cerevisiae), accumulation of the non-reducing disaccharide, trehalose, is triggered by stimuli that activate the heat-shock response. Previously, trehalose levels have been shown to be closely correlated with thermotolerance, suggesting a protective function of this substance. Genetic evidence in support of this view is presented in an accompanying paper [De Virgilio, C., Hottiger, T., Dominguez, J., Boller, T. & Wiemken, A. (1993) Eur. J. Biochem. 219, 179-186]. In this study, we have examined the effect of trehalose:on the thermal stability of proteins, a parameter thought to be a major determinant of thermotolerance. Physiological concentrations of trehalose (up to 0.5 M) were found to efficiently protect enzymes of yeast (glucose-6P-dehydrogenase, phosphoglucose-isomerase) as well as enzymes of non-yeast origin (bovine. glutamic dehydrogenase, EcoRI) against heat inactivation in vitro. Trehalose also reduced the heat-induced formation of protein aggregates. The disaccharide proved to be a compatible solute, as even at very high concentrations (up to 1 M) it did not significantly interfere with the activity of test enzymes. Trehalose was at least as good of better a protein stabilizer than any of a number of other compatible solutes (including sugars, polyalcohols and amino acids), while the structurally related trehalose-6P was devoid of any protective effect. Thermoprotection of enzymes by trehalose was evident even in solutions containing high concentrations of yeast protein or substrate. The data-indicate that trehalose accumulation may-increase-the thermotolerance of yeast-by enhancing protein stability in intact cells.