Development of a temperature-responsive yeast cell factory using engineered Gal4 as a protein switch.
Biotechnol Bioeng
; 115(5): 1321-1330, 2018 05.
Article
em En
| MEDLINE
| ID: mdl-29315481
ABSTRACT
Conflict between cell growth and product accumulation is frequently encountered in biosynthesis of secondary metabolites. Herein, a temperature-dependent dynamic control strategy was developed by modifying the GAL regulation system to facilitate two-stage fermentation in yeast. A temperature-sensitive Gal4 mutant Gal4M9 was created by directed evolution, and used as a protein switch in ΔGAL80 yeast. After EGFP-reported validation of its temperature-responsive induction capability, the sensitivity and stringency of this system in multi-gene pathway regulation was tested, using lycopene as an example product. When Gal4M9 was used to control the expression of PGAL -driven pathway genes, growth and production was successfully decoupled upon temperature shift during fermentation, accumulating 44% higher biomass and 177% more lycopene than the control strain with wild-type Gal4. This is the first example of adopting temperature as an input signal for metabolic pathway regulation in yeast cell factories.
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Assunto principal:
Saccharomyces cerevisiae
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Fatores de Transcrição
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Regulação Fúngica da Expressão Gênica
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Proteínas de Saccharomyces cerevisiae
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Proteínas de Ligação a DNA
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Engenharia Metabólica
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Metabolismo
Idioma:
En
Ano de publicação:
2018
Tipo de documento:
Article