Improving ethanol tolerance of Saccharomyces cerevisiae industrial strain by directed evolution of SPT3 / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 159-164, 2010.
Article
in Chinese
| WPRIM
| ID: wpr-336248
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Saccharomyces cerevisiae
/
Transcription Factors
/
Industrial Microbiology
/
Trans-Activators
/
Directed Molecular Evolution
/
Saccharomyces cerevisiae Proteins
/
Drug Resistance, Fungal
/
Ethanol
/
Drug Tolerance
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2010
Type:
Article
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