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酿酒酵母中hxt1、hxt5和hxt7转运蛋白表达对细胞外葡萄糖浓度扰动的动态响应。

Dynamic response of the expression of hxt1, hxt5 and hxt7 transport proteins in Saccharomyces cerevisiae to perturbations in the extracellular glucose concentration.

作者信息

Buziol Stefan, Warth Lydia, Magario Ivana, Freund Andreas, Siemann-Herzberg Martin, Reuss Matthias

机构信息

Institute of Biochemical Engineering, University of Stuttgart, Allmandring 31, D-70569 Stuttgart, Germany.

出版信息

J Biotechnol. 2008 Apr 30;134(3-4):203-10. doi: 10.1016/j.jbiotec.2008.02.002. Epub 2008 Feb 13.

Abstract

Glucose transport in Saccharomyces cerevisiae relies on a multi-factorial uptake system. The modulation of its efficiency depends on the differential expression of various sets of hexose transport-related proteins whose glucose affinity differs considerably. The expression of three different glucose transport proteins (HXT1, HXT5 and HXT6/7 with low-, intermediate- and high-affinity, respectively) was monitored as a result of modified extracellular glucose concentrations. Cultivation at glucose-limited (continuous) conditions was instantly replaced by a batch (and thus, non-limited) mode. Further, to mimic concentration gradients in large-scale production bioreactors, multiple and rapid transient glucose pulses were applied to chemostat cultivation. Antibodies against the HXT-proteins were used to monitor the proteins' expression levels prior to and after perturbing the external glucose concentrations. HXT5 and HXT6/7 were either expressed during the starvation-like steady-state phases in the chemostat cultivations, whereas HXT1 could not be detected at all. HXT1, however, is subsequently expressed during the excess of glucose in the batch mode, while the HXT5 and HXT6/7 transporters were at least found to decline. These findings coincide well with the transporters' affinity profiles. As a result of repeated and rapid transient glucose pulses during continuous fermentation, especially HXT6/7 pointed out to alter the protein expression pattern.

摘要

酿酒酵母中的葡萄糖转运依赖于一个多因素摄取系统。其效率的调节取决于多组与己糖转运相关的蛋白质的差异表达,这些蛋白质对葡萄糖的亲和力差异很大。由于细胞外葡萄糖浓度的改变,监测了三种不同葡萄糖转运蛋白(分别为低亲和力的HXT1、中等亲和力的HXT5和高亲和力的HXT6/7)的表达。在葡萄糖受限(连续)条件下的培养立即被分批(即非受限)模式取代。此外,为了模拟大规模生产生物反应器中的浓度梯度,对恒化器培养施加了多次快速的瞬时葡萄糖脉冲。使用针对HXT蛋白的抗体来监测在扰动外部葡萄糖浓度之前和之后蛋白质的表达水平。在恒化器培养中类似饥饿的稳态阶段,HXT5和HXT6/7均有表达,而完全检测不到HXT1。然而,在分批模式下葡萄糖过量时,HXT1随后会表达,而HXT5和HXT6/7转运蛋白至少被发现有所下降。这些发现与转运蛋白的亲和力概况非常吻合。在连续发酵过程中,由于反复快速的瞬时葡萄糖脉冲,尤其是HXT6/7显示出蛋白质表达模式发生了改变。

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