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木糖操纵子在 和 中的转录变化及其在发酵优化中的应用。

Transcriptional Changes in the Xylose Operon in and Their Use in Fermentation Optimization.

机构信息

Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China.

National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China.

出版信息

Int J Mol Sci. 2019 Sep 18;20(18):4615. doi: 10.3390/ijms20184615.

Abstract

The xylose operon is an efficient biological element used for the regulation of gene expression in . Although the mechanism underlying the xylose-mediated regulation of this operon has been elucidated, the transcriptional changes that occur under various fermentation conditions remain unclear. In this study, the effects of different conditions on xylose operon expression were investigated. Significant upregulation was observed during the transition from the logarithmic phase to the stationary phase (2.5-fold, = 3, < 0.01). Glucose suppressed transcription over 168-fold ( = 3, < 0.01). Meanwhile, the inhibitory effect of glucose hardly strengthened at concentrations from 20 to 180 g/L. Furthermore, the transcription of the xylose operon increased at elevated temperatures (25-42 °C) and was optimal at a neutral pH (pH 6.5-7.0). Based on these findings, relevant fermentation strategies (delaying the induction time, using dextrin as a carbon source, increasing the fermentation temperature, and maintaining a neutral pH) were proposed. Subsequently, these strategies were validated through the use of maltogenic amylase as a reporter protein, as an 8-fold ( = 3, < 0.01) increase in recombinant enzyme activity compared to that under unoptimized conditions was observed. This work contributes to the development of fermentation optimization and furthers the use of the xylose operon as an efficient expression element.

摘要

木糖操纵子是一种高效的生物学元件,用于调控 中的基因表达。尽管已经阐明了木糖介导该操纵子调控的机制,但在各种发酵条件下发生的转录变化仍不清楚。在本研究中,研究了不同条件对木糖操纵子表达的影响。在从对数期到静止期的转变过程中观察到显著的上调(2.5 倍, = 3, < 0.01)。葡萄糖的转录抑制作用超过 168 倍( = 3, < 0.01)。同时,葡萄糖的抑制作用在 20 到 180 g/L 的浓度下几乎没有加强。此外,木糖操纵子的转录在较高温度(25-42°C)下增加,在中性 pH(6.5-7.0)下最佳。基于这些发现,提出了相关的发酵策略(延迟诱导时间、使用糊精作为碳源、提高发酵温度和保持中性 pH)。随后,通过使用麦芽糖淀粉酶作为报告蛋白来验证这些策略,与未优化条件下相比,重组酶活性增加了 8 倍( = 3, < 0.01)。这项工作有助于发酵优化的发展,并进一步利用木糖操纵子作为高效的表达元件。

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