对CEL3C的功能性表征揭示了它在Trichoderma reesei Rut C30中调节细胞酶基因表达中的关键作用
Lu Wang1, Xiao He1, Tian Tian1
1School of Chemistry and Chemical Engineering, Chongqing University of Science and Technology, Chongqing 401331, China.
Enzyme and microbial technology
|July 5, 2025
概括
在Trichoderma reesei中删除核β-葡萄糖酶CEL3C,通过增强基因转录和酶活性,显著增加细胞酶的产生. 这一发现为优化工业酶产量提供了新的目标.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 酶工程是什么? 酶工程是什么?
背景情况:
- 在Trichoderma reesei中的核β-葡萄糖酶CELC3C与细胞酶调节有关,但其精确的机制尚未完全理解.
- 了解CEL3C的作用对于增强纤维酶生产至关重要,这是生物燃料和生物化学工业的关键过程.
研究的目的:
- 调查核局部β-葡萄糖酶CEL3C在Trichoderma reesei中调节细胞酶生产中的功能.
- 阐明CEL3C影响细胞酶基因表达和酶活性的分子机制.
主要方法:
- 使用CRISPR-Cas9在高细胞解菌株T. reesei Rut C30中创建了一个CEL3C删除突变 (T. reesei ΔCEL3C).
- 细胞质生产在富含糖的MGD诱导下进行了评估.
- 进行了转录组分析和细胞内糖分析,以确定调控变化和代谢转变.
主要成果:
- 删除CEL3C显著增加了总细胞酶活性31.28%,在β-葡萄糖酶 (94.97%),内葡萄糖酶 (19.40%) 和细胞质酶 (28.99%) 活动中显著增强.
- 转录组分析显示了关键细胞酶基因 (例如,CEL7A,CEL6A,CEL12A,CEL5A) 和转录调节器 (XYR1,ACE3,RXE1) 的上调调节,以及抑制抑制剂 (RCE1,RCE2) 的抑制.
- CEL3C的删除导致细胞内葡萄糖和糖水平增加,豆糖和豆糖水平降低,这表明改变了分糖水解和潜在的糖丰富.
结论:
- CEL3C充当双重功能核调节器,通过转录控制和酶活性影响细胞酶合成.
- 干扰CEL3C通过放大索的吸收和诱导来增强细胞酶的产生,为工业酶产量优化提供了一个有前途的战略.
- 这项研究提供了可操作的目标,用于工程 Trichoderma reesei 菌株,以改善细胞酶产量,用于生物技术应用.
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