Vitamin A Production by Engineered Saccharomyces cerevisiae from Xylose via Two-Phase in Situ Extraction

Sun, Liang; Kwak, Suryang; Jin, Yong-Su

doi:10.1021/acssynbio.9b00217

Title: Vitamin A Production by Engineered Saccharomyces cerevisiae from Xylose via Two-Phase in Situ Extraction

Abstract

Vitamin A is an essential human micronutrient and plays critical roles in vision, reproduction, immune system, and skin health. Current industrial methods for the production of vitamin A rely on chemical synthesis from petroleum-derived substrates, such as acetone and acetylene. In this study, we developed a biotechnological method for production of vitamin A from an abundant and non-edible sugar. Specifically, we engineered Saccharomyces cerevisiae to produce vitamin A from xylose—the second most abundant sugar in plant cell wall hydrolysates—by introducing a β-carotene biosynthetic pathway, and a gene coding for β-carotene 15, 15’-dioxygenase (BCMO) into a xylose-fermenting S. cerevisiae. The resulting yeast strain produced vitamin A from xylose at a titer four-fold higher than from glucose. When a two-phase in situ extraction strategy with dodecane, or olive oil as an extractive agent was employed, vitamin A production improved additional two-fold. Furthermore, a xylose fed-batch fermentation with dodecane in situ extraction achieved a final titer of 3,350 mg/L vitamin A, which consisted of retinal (2,094 mg/L) and retinol (1,256 mg/L). These results suggest that potential limiting factors of vitamin A production in yeast, such as insufficient supply of isoprenoid precursors, and limited intracellular storage capacity, can be effectively addressed by using xylosemore »« less

Authors:

Sun, Liang ^[1]; Kwak, Suryang ^[1];

^[1]

Univ. of Illinois at Urbana-Champaign, Urbana, IL (United States). Dept. of Food Science and Human Nutrition and Carl R. Woese Inst. for Genomic Biology

Publication Date:: Fri Aug 02 00:00:00 EDT 2019

Research Org.:: Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)

Sponsoring Org.:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

OSTI Identifier:: 1547320

Grant/Contract Number:: SC0018420

Resource Type:: Accepted Manuscript

Journal Name:: ACS Synthetic Biology

Additional Journal Information:: Journal Volume: 8; Journal Issue: 9; Journal ID: ISSN 2161-5063

Publisher:: American Chemical Society (ACS)

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; Vitamin A; β-carotene; xylose; Saccharomyces cerevisiae; in situ extraction

Citation Formats


                    Sun, Liang, Kwak, Suryang, and Jin, Yong-Su. Vitamin A Production by Engineered Saccharomyces cerevisiae from Xylose via Two-Phase in Situ Extraction.  United States: N. p., 2019. 
Web.  doi:10.1021/acssynbio.9b00217.

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                    Sun, Liang, Kwak, Suryang, & Jin, Yong-Su. Vitamin A Production by Engineered Saccharomyces cerevisiae from Xylose via Two-Phase in Situ Extraction.  United States.  https://doi.org/10.1021/acssynbio.9b00217

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                    Sun, Liang, Kwak, Suryang, and Jin, Yong-Su. Fri .  
"Vitamin A Production by Engineered Saccharomyces cerevisiae from Xylose via Two-Phase in Situ Extraction".  United States.  https://doi.org/10.1021/acssynbio.9b00217.  https://www.osti.gov/servlets/purl/1547320.

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@article{osti_1547320,

  title        = {Vitamin A Production by Engineered Saccharomyces cerevisiae from Xylose via Two-Phase in Situ Extraction},

  author       = {Sun, Liang and Kwak, Suryang and Jin, Yong-Su},

  abstractNote = {Vitamin A is an essential human micronutrient and plays critical roles in vision, reproduction, immune system, and skin health. Current industrial methods for the production of vitamin A rely on chemical synthesis from petroleum-derived substrates, such as acetone and acetylene. In this study, we developed a biotechnological method for production of vitamin A from an abundant and non-edible sugar. Specifically, we engineered Saccharomyces cerevisiae to produce vitamin A from xylose—the second most abundant sugar in plant cell wall hydrolysates—by introducing a β-carotene biosynthetic pathway, and a gene coding for β-carotene 15, 15’-dioxygenase (BCMO) into a xylose-fermenting S. cerevisiae. The resulting yeast strain produced vitamin A from xylose at a titer four-fold higher than from glucose. When a two-phase in situ extraction strategy with dodecane, or olive oil as an extractive agent was employed, vitamin A production improved additional two-fold. Furthermore, a xylose fed-batch fermentation with dodecane in situ extraction achieved a final titer of 3,350 mg/L vitamin A, which consisted of retinal (2,094 mg/L) and retinol (1,256 mg/L). These results suggest that potential limiting factors of vitamin A production in yeast, such as insufficient supply of isoprenoid precursors, and limited intracellular storage capacity, can be effectively addressed by using xylose as a carbon source, and two-phase in situ extraction. The engineered S. cerevisiae and fermentation strategies described in this study might contribute to sustainable and economic production of vitamin A, and vitamin A-enriched bioproducts from renewable biomass.},

  doi          = {10.1021/acssynbio.9b00217},

  journal      = {ACS Synthetic Biology},

  number       = 9,

  volume       = 8,

  place        = {United States},

  year         = {Fri Aug 02 00:00:00 EDT 2019},

  month        = {Fri Aug 02 00:00:00 EDT 2019}

}

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Journal Article:

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https://doi.org/10.1021/acssynbio.9b00217

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Cited by: 35 works

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Figures / Tables:

Fig. 1: Biosynthetic pathways of vitamin A from glucose and xylose in engineered S. cerevisiae. A heterogolous xylose assimilation pathway contains xylose reductase (XR), xylitol dehydrogenase (XDH), and xylulokinase (XK). A heterologous vitamin A biosynthetic pathway consists of GGPP synthase (CrtE), phytoene desturase (CrtI), bifunctional phytoene synthase, lycopene cyclase (CrtYB),more »

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