Metabolomics approach used for understanding temperature-related pectinase activity in Bacillus licheniformis DY2
Metabolomics approach used for understanding temperature-related pectinase activity in Bacillus licheniformis DY2
复制标题
用于了解地衣芽孢杆菌 DY2 温度相关果胶酶活性的代谢组学方法
DOI：
10.1093/femsle/fny255
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发表时间：
2018-12-01
影响因子：
2.1
通讯作者：
Ye, Xiuyun
中科院分区：
文献类型：
--
作者：
Guan, Yi;Yin, Di;Ye, Xiuyun
果胶酶是催化果胶降解的酶。由于果胶酶的广泛用途和催化效率，全球对果胶酶的需求很大。优化发酵过程以提高果胶酶活力通常是为了降低工艺成本，但温度是否会影响代谢产物，从而提高果胶酶活力，目前尚不清楚。在这里，我们开发了一种代谢组学方法来探索它。地衣芽孢杆菌DY2在30℃条件下产生的P-DY2果胶酶活力高于37℃条件下生长的细胞，差异代谢组分析表明，在30℃条件下，三羧酸循环基因的转录水平以及丙酮酸脱氢酶和α-酮戊二酸脱氢酶的活性均低于37℃。此外，抑制丙酮酸脱氢酶和琥珀酸脱氢酶提高了P-DY2的活性，支持了果胶酶活性需要丙酮酸代谢失活和TCA循环的结论，并且P-DY2不依赖于TCA循环。总而言之，这些发现表明，发酵温度通过代谢调节影响P-DY2活性，失活的TCA循环是高P-DY2活性的特征。更重要的是，本研究强调了一种通过使用代谢途径抑制剂通过代谢调节来促进果胶酶活性的方法。
Pectinases are enzymes that catalyze pectin degradation. There is a global demand for pectinases because of their wide utility and catalytic efficiency. Optimization of the fermentation process to increase the pectolytic enzyme activity is generally practiced to lower process costs, but whether temperature influences the metabolome, enhancing pectinase activity, is not known. Here, we developed a metabolomics approach to explore it. The activity of P-DY2 pectinase produced by Bacillus licheniformis DY2 was higher in cells grown at 30°C than those grown at 37°C. Differential metabolome analysis revealed fluctuating tricarboxylic acid (TCA) cycle at 30°C. Consistently, the transcripts of TCA cycle genes and activities of pyruvate dehydrogenase and α-Ketoglutaric dehydrogenase were lower at 30°C than 37°C. Furthermore, inhibition of pyruvate dehydrogenase and succinate dehydrogenase enhanced the activity of P-DY2, supporting the conclusion that the inactivated pyruvate metabolism and TCA cycle were required for pectinase activity, and that P-DY2 was TCA cycle-independent. Collectively, these findings indicated that fermentation temperature affected P-DY2 activity by metabolic modulation, with an inactivated TCA cycle as a characteristic feature of high P-DY2 activity. More importantly, the present study highlights an approach of promoting pectinase activity through metabolic modulation by using metabolic pathway inhibitors.