添加化的转录形状分析,以改善树突类型的总化物生产
Xu Qian1, Yitong Qin1, Surendra Sarasiya2
1College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, 211800, Jiangsu, China.
Applied microbiology and biotechnology
|January 3, 2024
概括
优化Trichoderma longibrachiatum UN32介质与葡萄糖,牛肉提取物和化显著提高了生物质和树突类型总化物 (DTTA) 产量. 转录组分析显示增强了乙-CoA的流入,并提出了一种用于DTTA积累的ROS介导信号通路.
科学领域:
- 微生物学 微生物学
- 生物技术是生物技术.
- 生物化学 生物化学
背景情况:
- 特里科德尔玛长甲基 (Trichoderma longibrachiatum UN32) 是一个著名的,有价值的树突蛋白型总化物 (DTTAs) 的生产者.
- 优化发酵条件对于最大限度地提高微生物菌株的二次代谢物产量至关重要.
- 响应表面方法 (RSM) 是优化复杂生物过程的有效统计工具.
研究的目的:
- 为了确定Trichoderma longibrachiatum增强的树突蛋白类型总化物 (DTTA) 生产的最佳介质组成,UN32.
- 调查关键介质成分对生物质和DTTA产量的影响.
- 探索底层DTTA生物合成调节的分子机制.
主要方法:
- 普莱克特-伯曼设计和最的登方法用于初始因素选和优化.
- 盒子-Behnken设计 (一种响应表面方法) 用于微调介质组成.
- 发酵规模扩大到5升生物反应器.
- 转录组分析 (RNA-Seq) 来识别基因表达变化.
主要成果:
- 最佳介质:29.4 g/L 葡萄糖,17.3 g/L 牛肉提取物,0.28 mmol/L CoCl2.
- 菌根干重量增加了80.8% (12.303 g/L) 和DTTA产量增加了76.4% (541.63 μg).
- 在5L生物反应器中扩大规模的产量是对照组的1.95倍多的DTTA.
- 补充CoCl2改变了TCA循环和氨基酸生物合成基因,增加了用于DTTA生产的乙-CoA流入.
- 抗氧化酶基因表达的调节表明反应性氧物种 (ROS) 在DTTA生物合成调节中的作用.
结论:
- 优化的介质显著提高了T. longibrachiatum UN32.32中的生物质和DTTA生产.
- 化通过影响中央碳代谢,并可能作为信号分子,在调节DTTA生物合成方面发挥关键作用.
- 提出了一种涉及ROS信号在DTTA积累中的新机制,为代谢工程提供了新的途径.
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