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One-Step Biosynthesis of Vitamin C in Saccharomyces cerevisiae

Vitamin C (VC) is comprehensively applied in foods, cosmetics, pharmaceuticals, and especially clinical medicine. Nowadays, the industrial production of VC mainly relies on the classic two-step fermentation route, and researchers have explored the way for one-step fermentation of VC in recent years....

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Autores principales: Zhou, Mengyu, Bi, Yanhui, Ding, Mingzhu, Yuan, Yingjin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7947327/
https://www.ncbi.nlm.nih.gov/pubmed/33717042
http://dx.doi.org/10.3389/fmicb.2021.643472
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author Zhou, Mengyu
Bi, Yanhui
Ding, Mingzhu
Yuan, Yingjin
author_facet Zhou, Mengyu
Bi, Yanhui
Ding, Mingzhu
Yuan, Yingjin
author_sort Zhou, Mengyu
collection PubMed
description Vitamin C (VC) is comprehensively applied in foods, cosmetics, pharmaceuticals, and especially clinical medicine. Nowadays, the industrial production of VC mainly relies on the classic two-step fermentation route, and researchers have explored the way for one-step fermentation of VC in recent years. In this study, a VC biosynthesis pathway that directly produced VC from glucose was reconstructed in Saccharomyces cerevisiae, and the protein engineering and metabolic engineering strategies were adopted to improve it. First, five exogenous modules from Arabidopsis were introduced into the chassis cells by synthetic biology approaches to obtain the strain YLAA harboring VC biosynthesis. In addition, L-galactose dehydrogenase (L-GalDH) and L-galactono-1,4-lactone dehydrogenase (L-GLDH) were fused and expressed in S. cerevisiae cells for the first time, which increased the intracellular VC accumulation by 2.78-fold, reaching 9.97 ± 0.09 mg/L. Through copy number engineering, it was further confirmed that the last step catalyzed by L-GLDH is the rate-limiting step. GDP-L-galactose phosphorylase (GPP) encoded by vtc2 is another rate-limiting enzyme confirmed by GAL1p overexpression results. Finally, by balancing gene expression and cell growth, the highest production strain with overexpressing vtc2 by multicopy plasmids was constructed. The VC accumulation reached 24.94 ± 1.16 mg/L, which was currently the highest production from glucose in S. cerevisiae. The production of the recombinant strain reached nearly 44 mg/L with the exogenous addition of L-galactose or glutathione. The results further emphasized the importance of the step catalyzed by GPP. The investigation provided experience for the efficient biosynthesis of VC and the determination of rate-limiting steps.
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spelling pubmed-79473272021-03-12 One-Step Biosynthesis of Vitamin C in Saccharomyces cerevisiae Zhou, Mengyu Bi, Yanhui Ding, Mingzhu Yuan, Yingjin Front Microbiol Microbiology Vitamin C (VC) is comprehensively applied in foods, cosmetics, pharmaceuticals, and especially clinical medicine. Nowadays, the industrial production of VC mainly relies on the classic two-step fermentation route, and researchers have explored the way for one-step fermentation of VC in recent years. In this study, a VC biosynthesis pathway that directly produced VC from glucose was reconstructed in Saccharomyces cerevisiae, and the protein engineering and metabolic engineering strategies were adopted to improve it. First, five exogenous modules from Arabidopsis were introduced into the chassis cells by synthetic biology approaches to obtain the strain YLAA harboring VC biosynthesis. In addition, L-galactose dehydrogenase (L-GalDH) and L-galactono-1,4-lactone dehydrogenase (L-GLDH) were fused and expressed in S. cerevisiae cells for the first time, which increased the intracellular VC accumulation by 2.78-fold, reaching 9.97 ± 0.09 mg/L. Through copy number engineering, it was further confirmed that the last step catalyzed by L-GLDH is the rate-limiting step. GDP-L-galactose phosphorylase (GPP) encoded by vtc2 is another rate-limiting enzyme confirmed by GAL1p overexpression results. Finally, by balancing gene expression and cell growth, the highest production strain with overexpressing vtc2 by multicopy plasmids was constructed. The VC accumulation reached 24.94 ± 1.16 mg/L, which was currently the highest production from glucose in S. cerevisiae. The production of the recombinant strain reached nearly 44 mg/L with the exogenous addition of L-galactose or glutathione. The results further emphasized the importance of the step catalyzed by GPP. The investigation provided experience for the efficient biosynthesis of VC and the determination of rate-limiting steps. Frontiers Media S.A. 2021-02-25 /pmc/articles/PMC7947327/ /pubmed/33717042 http://dx.doi.org/10.3389/fmicb.2021.643472 Text en Copyright © 2021 Zhou, Bi, Ding and Yuan. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Zhou, Mengyu
Bi, Yanhui
Ding, Mingzhu
Yuan, Yingjin
One-Step Biosynthesis of Vitamin C in Saccharomyces cerevisiae
title One-Step Biosynthesis of Vitamin C in Saccharomyces cerevisiae
title_full One-Step Biosynthesis of Vitamin C in Saccharomyces cerevisiae
title_fullStr One-Step Biosynthesis of Vitamin C in Saccharomyces cerevisiae
title_full_unstemmed One-Step Biosynthesis of Vitamin C in Saccharomyces cerevisiae
title_short One-Step Biosynthesis of Vitamin C in Saccharomyces cerevisiae
title_sort one-step biosynthesis of vitamin c in saccharomyces cerevisiae
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7947327/
https://www.ncbi.nlm.nih.gov/pubmed/33717042
http://dx.doi.org/10.3389/fmicb.2021.643472
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