Improving ethanol tolerance of Saccharomyces cerevisiae industrial strain by directed evolution of SPT3.
- Author:
Xinqing ZHAO
1
;
Rujiao JIANG
;
Ning LI
;
Qing YANG
;
Fengwu BAI
Author Information
1. Department of Bioscience and Bioengineering, Dalian University of Technology, Dalian, China. xqzhao@dlut.edu.cn
- Publication Type:Journal Article
- MeSH:
Directed Molecular Evolution;
methods;
Drug Resistance, Fungal;
Drug Tolerance;
Ethanol;
metabolism;
pharmacology;
Industrial Microbiology;
methods;
Saccharomyces cerevisiae;
drug effects;
genetics;
metabolism;
Saccharomyces cerevisiae Proteins;
genetics;
Trans-Activators;
genetics;
Transcription Factors;
genetics
- From:
Chinese Journal of Biotechnology
2010;26(2):159-164
- CountryChina
- Language:Chinese
-
Abstract:
Directed evolution of transcription factors can be employed to effectively improve the phenotypes which are controlled by multiple genetic loci. In this study, we used error-prone PCR for the directed evolution of SPT3, which is the component of yeast Spt-Ada-Gcn5-acetyltransferase (SAGA) complex responsible for the transcription of stress-related genes, and studied its effect on the improvement of ethanol tolerance. Mutant library was constructed by ligating the error-prone PCR products with a modified pYES2.0 plasmid, and the expression plasmids were subsequently transformed to yeast industrial strain Saccharomyces cerevisiae 4126. One mutant strain M25 showing superior growth in presence of 10% ethanol was selected. M25 produced 11.7% more ethanol than the control strain harboring the empty vector when 125 g/L glucose was used as substrate. This study revealed that SPT3 is an important transcription factor for the metabolic engineering of yeast ethanol tolerance.
