相关实验视频

Updated: May 19, 2025

CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art

CRISPR/Cas12a Multiplex Genome Editing of Saccharomyces cerevisiae and the Creation of Yeast Pixel Art

Published on: May 28, 2019

使用CRISPR-Cas9工程酵母生物催化降解氨化

Arne Hagman¹, Olof Stenström², Göran Carlström²

¹Division of Biotechnology and Applied Microbiology, Lund University, Lund, Sweden. nhagman@gmail.com.

Scientific reports

|May 15, 2025

概括

代谢工程酵母通过全细胞生物转化有效地产生奇拉胺. 修改氨酸-酸盐节点并用欧米茄氨酸酶 (cv-ATA) 取代氨酸转移酶 (ALT1) 显著增强了产量.

科学领域:

生物技术是生物技术.
代谢工程是代谢工程.
酶催化酶的催化作用

背景情况:

贝克酵母是一种多功能的宿主,可以通过全细胞生物转化产生的性氨基.
氨酸-酸盐代谢节点在调节代谢流量方面发挥着至关重要的作用,可以调节以提高生物转化效率.

研究的目的:

通过调节氨酸-酸盐代谢节点来改造面包酵母,以增强奇拉胺的产生.
研究将内源性氨酸转移酶 (ALT1) 替换为混杂的氨酸转移酶 (cv-ATA) 对减少性氨基化的影响.
开发一种CRISPR/cas9方法,用于在酵母中快速基因替代,以获得更广泛的应用.

主要方法:

面包酵母的代谢工程是通过多个cv-ATA拷贝的染色体集成来实现的.
使用CRISPR/cas9技术将ALT1基因淘汰并用cv-ATA替换.
在有氧批量培养下生物反应器中的生理特征和代谢中间体的NMR分析.

主要成果:

在乙醇后的双生长过程中,表达cv-ATA的工程酵母显示出活跃的性氨基生产,特别是 (S) -1-甲基-3-烯基胺 (MPPA).
与对照菌株相比,用cv-ATA取代ALT1导致反应产量提高了2.6倍.
在葡萄糖代谢过程中形成的酸盐被确定为氨酸生产的抑制剂,这是NMR研究表明的.

结论:

通过用cv-ATA替换ALT1来调节氨酸-酸盐节点是增强代谢工程酵母中奇拉氨酸生产的有效策略.
开发的CRISPR/cas9系统促进了快速的基因替代,使生物催化剂的有效菌株工程成为可能.
在最佳的生物转化条件下,从基中获得58%的MPPA,突出显示了这种工程酵母系统的潜力.

更多相关视频

CRISPR-mediated Genome Editing of the Human Fungal Pathogen Candida albicans

CRISPR-mediated Genome Editing of the Human Fungal Pathogen Candida albicans

Published on: November 14, 2018

Genetic Engineering of an Unconventional Yeast for Renewable Biofuel and Biochemical Production

Genetic Engineering of an Unconventional Yeast for Renewable Biofuel and Biochemical Production

Published on: September 20, 2016

相关概念视频

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...

CRISPR and crRNAs

CRISPR and crRNAs

Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...

Homologous Recombination

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...