Iwahashi H, Nwaka S, Obuchi K
National Institute of Bioscience and Human Technology, Tsukuba, Ibaraki 305, Japan.
Appl Environ Microbiol. 2000 Dec;66(12):5182-5. doi: 10.1128/AEM.66.12.5182-5185.2000.
In yeast, trehalose accumulation and its hydrolysis, which is catalyzed by neutral trehalase, are believed to be important for thermotolerance. We have shown that trehalose is one of the important factors for barotolerance (resistance to hydrostatic pressure); however, nothing is known about the role of neutral trehalase in barotolerance. To estimate the contribution of neutral trehalase in resisting high hydrostatic pressure, we measured the barotolerance of neutral trehalase I and/or neutral trehalase II deletion strains. Under 180 MPa of pressure for 2 h, the neutral trehalase I deletion strain showed higher barotolerance in logarithmic-phase cells and lower barotolerance in stationary-phase cells than the wild-type strain. Introduction of the neutral trehalase I gene (NTH1) into the deletion mutant restored barotolerance defects in stationary-phase cells. Furthermore, we assessed the contribution of neutral trehalase during pressure and recovery conditions by varying the expression of NTH1 or neutral trehalase activity with a galactose-inducible GAL1 promoter with either glucose or galactose. The low barotolerance observed with glucose repression of neutral trehalase from the GAL1 promoter was restored during recovery with galactose induction. Our results suggest that neutral trehalase contributes to barotolerance, especially during recovery.
在酵母中,海藻糖的积累及其由中性海藻糖酶催化的水解被认为对耐热性很重要。我们已经表明,海藻糖是耐压性(对静水压力的抗性)的重要因素之一;然而,关于中性海藻糖酶在耐压性中的作用却一无所知。为了评估中性海藻糖酶在抵抗高静水压力中的作用,我们测量了中性海藻糖酶I和/或中性海藻糖酶II缺失菌株的耐压性。在180MPa压力下处理2小时,中性海藻糖酶I缺失菌株在对数期细胞中显示出比野生型菌株更高的耐压性,而在稳定期细胞中则显示出较低的耐压性。将中性海藻糖酶I基因(NTH1)导入缺失突变体可恢复稳定期细胞中的耐压性缺陷。此外,我们通过用葡萄糖或半乳糖的半乳糖诱导型GAL1启动子改变NTH1的表达或中性海藻糖酶活性,评估了中性海藻糖酶在压力和恢复条件下的作用。在半乳糖诱导恢复期间,观察到的由GAL1启动子对中性海藻糖酶的葡萄糖抑制导致的低耐压性得以恢复。我们的结果表明,中性海藻糖酶有助于耐压性,尤其是在恢复过程中。