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Title: Overcoming the thermodynamic equilibrium of an isomerization reaction through oxidoreductive reactions for biotransformation

Isomerases perform biotransformations without cofactors but often cause an undesirable mixture of substrate and product due to unfavorable thermodynamic equilibria. We demonstrate the feasibility of using an engineered yeast strain harboring oxidoreductase reactions to overcome the thermodynamic limit of an isomerization reaction. Specifically, a yeast strain capable of consuming lactose intracellularly is engineered to produce tagatose from lactose through three layers of manipulations. First, GAL1 coding for galactose kinase is deleted to eliminate galactose utilization. Second, heterologous xylose reductase (XR) and galactitol dehydrogenase (GDH) are introduced into the Δgal1 strain. Third, the expression levels of XR and GDH are adjusted to maximize tagatose production. The resulting engineered yeast produces 37.69 g/L of tagatose from lactose with a tagatose and galactose ratio of 9:1 in the reaction broth. These results suggest that in vivo oxidoreaductase reactions can be employed to replace isomerases in vitro for biotransformation.
Authors:
ORCiD logo [1] ; ORCiD logo [2] ;  [2] ;  [2] ;  [3] ;  [4] ; ORCiD logo [5] ;  [2]
  1. Univ. of Illinois, Urbana-Champaign, IL (United States). Carl R. Woese Inst. for Genomic Biology
  2. Univ. of Illinois, Urbana-Champaign, IL (United States). Carl R. Woese Inst. for Genomic Biology, and Dept. of Food Science and Human Nutrition
  3. Univ. of Illinois, Urbana-Champaign, IL (United States). Carl R. Woese Inst. for Genomic Biology; Korea Univ., Seoul (South Korea)
  4. National Center for Genetic Engineering and Biotechnology (BIOTEC), Pathum Thani (Thailand)
  5. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology
Publication Date:
Grant/Contract Number:
SC0018420
Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 10; Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Research Org:
Univ. of Illinois, Champaign, IL (United States). Center for Advanced Bioenergy and Bioproducts Innovation
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1504466

Liu, Jing-Jing, Zhang, Guo-Chang, Kwak, Suryang, Oh, Eun Joong, Yun, Eun Ju, Chomvong, Kulika, Cate, Jamie H. D., and Jin, Yong-Su. Overcoming the thermodynamic equilibrium of an isomerization reaction through oxidoreductive reactions for biotransformation. United States: N. p., Web. doi:10.1038/s41467-019-09288-6.
Liu, Jing-Jing, Zhang, Guo-Chang, Kwak, Suryang, Oh, Eun Joong, Yun, Eun Ju, Chomvong, Kulika, Cate, Jamie H. D., & Jin, Yong-Su. Overcoming the thermodynamic equilibrium of an isomerization reaction through oxidoreductive reactions for biotransformation. United States. doi:10.1038/s41467-019-09288-6.
Liu, Jing-Jing, Zhang, Guo-Chang, Kwak, Suryang, Oh, Eun Joong, Yun, Eun Ju, Chomvong, Kulika, Cate, Jamie H. D., and Jin, Yong-Su. 2019. "Overcoming the thermodynamic equilibrium of an isomerization reaction through oxidoreductive reactions for biotransformation". United States. doi:10.1038/s41467-019-09288-6. https://www.osti.gov/servlets/purl/1504466.
@article{osti_1504466,
title = {Overcoming the thermodynamic equilibrium of an isomerization reaction through oxidoreductive reactions for biotransformation},
author = {Liu, Jing-Jing and Zhang, Guo-Chang and Kwak, Suryang and Oh, Eun Joong and Yun, Eun Ju and Chomvong, Kulika and Cate, Jamie H. D. and Jin, Yong-Su},
abstractNote = {Isomerases perform biotransformations without cofactors but often cause an undesirable mixture of substrate and product due to unfavorable thermodynamic equilibria. We demonstrate the feasibility of using an engineered yeast strain harboring oxidoreductase reactions to overcome the thermodynamic limit of an isomerization reaction. Specifically, a yeast strain capable of consuming lactose intracellularly is engineered to produce tagatose from lactose through three layers of manipulations. First, GAL1 coding for galactose kinase is deleted to eliminate galactose utilization. Second, heterologous xylose reductase (XR) and galactitol dehydrogenase (GDH) are introduced into the Δgal1 strain. Third, the expression levels of XR and GDH are adjusted to maximize tagatose production. The resulting engineered yeast produces 37.69 g/L of tagatose from lactose with a tagatose and galactose ratio of 9:1 in the reaction broth. These results suggest that in vivo oxidoreaductase reactions can be employed to replace isomerases in vitro for biotransformation.},
doi = {10.1038/s41467-019-09288-6},
journal = {Nature Communications},
number = 1,
volume = 10,
place = {United States},
year = {2019},
month = {3}
}

Works referenced in this record:

Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems
journal, March 2013
  • DiCarlo, James E.; Norville, Julie E.; Mali, Prashant
  • Nucleic Acids Research, Vol. 41, Issue 7, p. 4336-4343
  • DOI: 10.1093/nar/gkt135