U.S. Department of EnergyOffice of Scientific and Technical Information
Data from Enhanced 2′-Fucosyllactose Production by Engineered Saccharomyces cerevisiae using Xylose as a Co-Substrate
Abstract
2′-Fucosyllactose (2′-FL), a human milk oligosaccharide with confirmed benefits for infant health, is a promising infant formula ingredient. Although Escherichia coli, Saccharomyces cerevisiae, Corynebacterium glutamicum, and Bacillus subtilis have been engineered to produce 2′-FL, their titers and productivities need be improved for economic production. Glucose along with lactose have been used as substrates for producing 2′-FL, but accumulation of by-products due to overflow metabolism of glucose hampered efficient production of 2′-FL regardless of a host strain. To circumvent this problem, we used xylose, which is the second most abundant sugar in plant cell wall hydrolysates and is metabolized through oxidative metabolism, for the production of 2′-FL by engineered yeast. Specifically, we modified an engineered S. cerevisiae strain capable of assimilating xylose to produce 2′-FL from a mixture of xylose and lactose. First, a lactose transporter (Lac12) from Kluyveromyces lactis was introduced. Second, a heterologous 2′-FL biosynthetic pathway consisting of enzymes Gmd, WcaG, and WbgL from E. coli was introduced. Third, we adjusted expression levels of the heterologous genes to maximize 2′-FL production. The resulting engineered yeast produced 25.5 g/L of 2′-FL with a volumetric productivity of 0.35 g/L∙h in a fed-batch fermentation with lactose and xylose feeding to mitigate themore »
- Authors:
-
;
;
;
- Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA; Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
- Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA; Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA
- Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
- Department of Biotechnology, Graduate School, Korea University, Seoul, 02841, Republic of Korea
- Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA
- Publication Date:
- DOE Contract Number:
- SC0018420
- Research Org.:
- Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States); University of Illinois Urbana-Champaign
- Sponsoring Org.:
- U.S. Department of Energy (DOE)
- Subject:
- Conversion; Genome Engineering
- OSTI Identifier:
- 3013941
- DOI:
- https://doi.org/10.13012/B2IDB-5204797_V1
Citation Formats
Lee, Jaewon, Kwak, Suryang, Liu, Jing-Jing, Yu, Sora, Yun, Eun Ju, Kim, Dong Hyun, Liu, Cassie, Kim, Kyoung Heon, and Jin, Yong-Su. Data from Enhanced 2′-Fucosyllactose Production by Engineered Saccharomyces cerevisiae using Xylose as a Co-Substrate. United States: N. p., 2020.
Web. doi:10.13012/B2IDB-5204797_V1.
Lee, Jaewon, Kwak, Suryang, Liu, Jing-Jing, Yu, Sora, Yun, Eun Ju, Kim, Dong Hyun, Liu, Cassie, Kim, Kyoung Heon, & Jin, Yong-Su. Data from Enhanced 2′-Fucosyllactose Production by Engineered Saccharomyces cerevisiae using Xylose as a Co-Substrate. United States. doi:https://doi.org/10.13012/B2IDB-5204797_V1
Lee, Jaewon, Kwak, Suryang, Liu, Jing-Jing, Yu, Sora, Yun, Eun Ju, Kim, Dong Hyun, Liu, Cassie, Kim, Kyoung Heon, and Jin, Yong-Su. 2020.
"Data from Enhanced 2′-Fucosyllactose Production by Engineered Saccharomyces cerevisiae using Xylose as a Co-Substrate". United States. doi:https://doi.org/10.13012/B2IDB-5204797_V1. https://www.osti.gov/servlets/purl/3013941. Pub date:Tue Oct 20 00:00:00 UTC 2020
@article{osti_3013941,
title = {Data from Enhanced 2′-Fucosyllactose Production by Engineered Saccharomyces cerevisiae using Xylose as a Co-Substrate},
author = {Lee, Jaewon and Kwak, Suryang and Liu, Jing-Jing and Yu, Sora and Yun, Eun Ju and Kim, Dong Hyun and Liu, Cassie and Kim, Kyoung Heon and Jin, Yong-Su},
abstractNote = {2′-Fucosyllactose (2′-FL), a human milk oligosaccharide with confirmed benefits for infant health, is a promising infant formula ingredient. Although Escherichia coli, Saccharomyces cerevisiae, Corynebacterium glutamicum, and Bacillus subtilis have been engineered to produce 2′-FL, their titers and productivities need be improved for economic production. Glucose along with lactose have been used as substrates for producing 2′-FL, but accumulation of by-products due to overflow metabolism of glucose hampered efficient production of 2′-FL regardless of a host strain. To circumvent this problem, we used xylose, which is the second most abundant sugar in plant cell wall hydrolysates and is metabolized through oxidative metabolism, for the production of 2′-FL by engineered yeast. Specifically, we modified an engineered S. cerevisiae strain capable of assimilating xylose to produce 2′-FL from a mixture of xylose and lactose. First, a lactose transporter (Lac12) from Kluyveromyces lactis was introduced. Second, a heterologous 2′-FL biosynthetic pathway consisting of enzymes Gmd, WcaG, and WbgL from E. coli was introduced. Third, we adjusted expression levels of the heterologous genes to maximize 2′-FL production. The resulting engineered yeast produced 25.5 g/L of 2′-FL with a volumetric productivity of 0.35 g/L∙h in a fed-batch fermentation with lactose and xylose feeding to mitigate the glucose repression. Interestingly, the major location of produced 2′-FL by the engineered yeast can be changed using different culture media. While 72% of the produced 2′-FL was secreted when a complex medium was used, 82% of the produced 2′-FL remained inside the cells when a minimal medium was used. As yeast extract is already used as food and animal feed ingredients, 2′-FL enriched yeast extract can be produced cost-effectively using the 2′-FL-accumulating yeast cells.},
doi = {10.13012/B2IDB-5204797_V1},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Oct 20 00:00:00 UTC 2020},
month = {Tue Oct 20 00:00:00 UTC 2020}
}