转录激活剂Xyr1中的单一突变增强了Trichoderma reesei在葡萄糖上的细胞酶和氧基酶的产生
Dongmei Lv1, Weixin Zhang1, Xiangfeng Meng1
1State Key Laboratory of Microbial Technology, Shandong University, No. 72 Binhai Road, Qingdao 266237, P. R. China.
Journal of agricultural and food chemistry
|July 31, 2023
概括
在Trichoderma reesei中的工程转录因子增加了纤维细胞分解酶的生产. 一种新的Xyr1突变 (R434L) 显著增强了细胞酶和西兰酶的产量,以改善生物炼油应用.
科学领域:
- 生物技术是生物技术.
- 酶工程是什么? 酶工程是什么?
- 工业微生物学 工业微生物学
背景情况:
- 对生物炼油厂来说,具有成本效益的细胞解酶生产至关重要.
- 工程转录因子是提高Trichoderma reesei中的酶产量的有希望的策略.
- 氨酶调节剂1 (Xyr1) 是细胞酶和氨酶基因表达的关键调节剂.
研究的目的:
- 为了识别Xyr1中增强细胞酶和西兰酶生产的新突变.
- 描述一个特定的Xyr1突变 (R434L) 对酶生产和活性的影响.
- 为了评估工程Xyr1对林氏纤维素生物质的糖化效率的影响.
主要方法:
- 酵母的一混合查系统用于识别Xyr1突变.
- 野生类型Xyr1和Xyr1 (R434L) 在Trichoderma reesei中的构成表达.
- 测量细胞外酶活性 (pNPCase和解活性).
- 细胞酶和西兰酶基因转录水平的量化.
- 使用酶尾酒评估玉米的糖化效率.
主要成果:
- 在Xyr1中等同源区中发现了一种新型突变R434L.
- 与野生型Xyr1 (OEX) 相比,T. reesei (OEXR434L) 中Xyr1 (R434L) 的构成表达显著增加了葡萄糖上的细胞酶和氧基酶的产生.
- 细胞外酶活性增加了pNPCase的0.8倍,xylanolytic活动的0.7倍.
- 在OEXR434L中观察到主要细胞酶和西兰酶基因的转录升高.
- 来自OEX的酶尾酒R434L与OEX相比,提高了21%的玉米的糖化.
结论:
- 在Xyr1中的R434L突变增强了其交换活化功能,从而导致Trichoderma reesei中纤维细胞分解酶的产量增加.
- 这种工程菌株具有在生物炼油工艺中具有成本效益的酶生产的潜力.
- 改进的酶尾酒证明了在基纤维素生物质糖化中提高了效率.
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