相关实验视频

Updated: Sep 15, 2025

Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris

Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris

Published on: February 25, 2010

在Pichia pastoris中构建CRISPR-Cas9系统用于代谢重编程和cordycepin生物制造

Bingjie Zhao¹, Jinlei Shi¹, Ruoyu Zhao¹

¹College of Resources and Environmental Sciences, China Agricultural University, Beijing 100193, China; Beijing Key Laboratory of Biodiversity and Organic Farming, China Agricultural University, Beijing 100193, China.

Bioresource technology

|July 12, 2025

概括

我们使用CRISPR-Cas9.9开发了一种无抗生素标记物的方法,用于在Pichia pastoris中进行cordycepin生物合成. 与传统方法相比,这种工程菌株实现了高产量,并显著减少了碳排放.

科学领域:

生物技术是生物技术.
合成生物学合成生物学
代谢工程是代谢工程.

背景情况:

科迪塞宾是一种有价值的核酸类比物,主要由Cordyceps militaris 生产.
传统的微生物工程方法通常依赖于抗生素耐药性基因,造成环境问题.

研究的目的:

为了设计Pichia pastoris用于无抗生素标记物的cordycepin生物合成.
为了优化cordycepin的生产和减少对环境的影响.

主要方法:

使用了优化的CRISPR-Cas9系统与gRNA-tRNA阵列和Brex27增强的同源重组.
采用模块化代谢工程策略来增强促进剂组合,基因拷贝数,甲醇同化,前体供应和ATP/NADPH平衡.

主要成果:

在摇瓶发酵中获得2509.7 mg/L的cordycepin,使用工程菌株PC19.
在料批发发酵 (3.05 g/L/d,122.2 mg/g DCW) 中达到18.3 g/L的创纪录产量.
与Cordyceps militaris和其他微生物工厂相比,碳排放量低3.3-57.6倍.

结论:

开发的CRISPR-Cas9系统可以在Pichia pastoris中高效和低碳生物合成cordycepin.
该平台有助于可持续生产cordycepin和其他核酸类类似物.

关键词:

Brex27 域名域名温室气体排放量排放量温室气体排放量甲醇的同化方式多重基因组编辑多重基因组编辑这是一个gRNA-tRNA数组.

更多相关视频

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins

Published on: October 18, 2022

Methanol Independent Expression by Pichia Pastoris Employing De-repression Technologies

Methanol Independent Expression by Pichia Pastoris Employing De-repression Technologies

Published on: January 23, 2019

相关概念视频

CRISPR/Cas9 Genome Editing

CRISPR/Cas9 Genome Editing

The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...

CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...