利用表达卡塞特的系统工程作为一种强大的工具来增强Trichoderma reesei中的异质基因表达
Wangli Ji1, Li Xu2, Xianhua Sun2
1Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, No. 12 South Zhongguancun Street, Haidian District, Beijing 100081, China.
Journal of agricultural and food chemistry
|March 1, 2024
概括
在Trichoderma reesei表达盒的系统工程显著提高了Aspergillus niger mannanase的产量. 修改促进剂和科扎克序列等关键元素,使酶产量增加了27倍以上.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 微生物工程 微生物工程
背景情况:
- 微生物宿主中的异质基因表达通常使用简单的促进器-终结器结构.
- 为了改善蛋白质生产,对表达盒本身的系统工程尚未得到充分研究.
研究的目的:
- 调查系统表达卡塞特突变发生在异质基因表达上的影响.
- 为了优化Aspergillus niger mannanase在Trichoderma reesei中的生产.
主要方法:
- 对表达盒组件的系统工程:促进剂,信号,科扎克序列和3'-UTR.
- 作为模型基因利用了Aspergillus niger mannanase和Trichoderma reesei作为真核丝状真菌宿主.
- 量化了细胞外曼南酶的产生.
主要成果:
- 对表达卡塞特组件的逐步修改导致了曼南生产的显著改进.
- 细胞外曼南酶的产生从17U/mL增加到471U/mL,增长了27.7倍.
- 在表达卡塞特内证明了向突变发生的有效性.
结论:
- 包括促进体和cis元素在内的表达盒的有针对性的工程是增强异质蛋白质生产的强大策略.
- 开发的策略广泛适用于改善真核微生物系统中的重组蛋白产量.
- 优化表达卡塞特可以显著提高微生物发酵的工业应用.
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