Journal
JOURNAL OF APPLIED MICROBIOLOGY
Volume 133, Issue 4, Pages 2390-2402Publisher
OXFORD UNIV PRESS
DOI: 10.1111/jam.15700
Keywords
AGT1 overexpression; baker's yeast; freeze-thaw stress; intracellular trehalose; trehalose transporter
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Funding
- Cornell University
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By controlling and regulating intracellular trehalose levels, the study aims to improve the survival rate of yeast spores in frozen state. Trehalose plays a crucial role in enhancing the freeze-thaw tolerance of cells, which can aid in the better application of baker's yeast in the frozen baked goods industry.
Aim This study is to investigate the use of a constitutively expressed trehalose transport protein to directly control intracellular trehalose levels and protect baker's yeast (Saccharomyces cerevisiae) cells against freeze-thaw stress in vivo. Methods and Results We used a constitutively overexpressed Agt1 transporter to investigate the role of trehalose in the freeze-thaw tolerance of yeast cells by regulating intracellular trehalose concentrations independently of intracellular biosynthesis. Using this method, we found that increasing intracellular trehalose in yeast cells improved cell survival rate after 8 days of freezing at -80 and -20 degrees C. We also observed that freeze-thaw tolerance promoted by intracellular trehalose only occurs in highly concentrated cell pellets rather than cells in liquid suspension. Conclusions Trehalose is sufficient to provide freeze-thaw tolerance using our Agt1 overexpression system. Freeze-thaw tolerance can be further enhanced by deletion of genes encoding intracellular trehalose degradation enzymes. Significance and Impact of Study These findings are relevant to improving the freeze-thaw tolerance of baker's yeast in the frozen baked goods industry through engineering strains that can accumulate intracellular trehalose via a constitutively expressed trehalose transporter and inclusion of trehalose into the growth medium.
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