Cargando…

Combined Biosynthetic Pathway Engineering and Storage Pool Expansion for High-Level Production of Ergosterol in Industrial Saccharomyces cerevisiae

Ergosterol, a terpenoid compound produced by fungi, is an economically important metabolite serving as the direct precursor of steroid drugs. Herein, ergsosterol biosynthetic pathway modification combined with storage capacity enhancement was proposed to synergistically improve the production of erg...

Descripción completa

Detalles Bibliográficos
Autores principales: Sun, Zhi-Jiao, Lian, Jia-Zhang, Zhu, Li, Jiang, Yi-Qi, Li, Guo-Si, Xue, Hai-Long, Wu, Mian-Bin, Yang, Li-Rong, Lin, Jian-Ping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8275991/
https://www.ncbi.nlm.nih.gov/pubmed/34268298
http://dx.doi.org/10.3389/fbioe.2021.681666
_version_ 1783721824365838336
author Sun, Zhi-Jiao
Lian, Jia-Zhang
Zhu, Li
Jiang, Yi-Qi
Li, Guo-Si
Xue, Hai-Long
Wu, Mian-Bin
Yang, Li-Rong
Lin, Jian-Ping
author_facet Sun, Zhi-Jiao
Lian, Jia-Zhang
Zhu, Li
Jiang, Yi-Qi
Li, Guo-Si
Xue, Hai-Long
Wu, Mian-Bin
Yang, Li-Rong
Lin, Jian-Ping
author_sort Sun, Zhi-Jiao
collection PubMed
description Ergosterol, a terpenoid compound produced by fungi, is an economically important metabolite serving as the direct precursor of steroid drugs. Herein, ergsosterol biosynthetic pathway modification combined with storage capacity enhancement was proposed to synergistically improve the production of ergosterol in Saccharomyces cerevisiae. S. cerevisiae strain S1 accumulated the highest amount of ergosterol [7.8 mg/g dry cell weight (DCW)] among the wild-type yeast strains tested and was first selected as the host for subsequent metabolic engineering studies. Then, the push and pull of ergosterol biosynthesis were engineered to increase the metabolic flux, overexpression of the sterol acyltransferase gene ARE2 increased ergosterol content to 10 mg/g DCW and additional overexpression of a global regulatory factor allele (UPC2-1) increased the ergosterol content to 16.7 mg/g DCW. Furthermore, considering the hydrophobicity sterol esters and accumulation in lipid droplets, the fatty acid biosynthetic pathway was enhanced to expand the storage pool for ergosterol. Overexpression of ACC1 coding for the acetyl-CoA carboxylase increased ergosterol content from 16.7 to 20.7 mg/g DCW. To address growth inhibition resulted from premature accumulation of ergosterol, auto-inducible promoters were employed to dynamically control the expression of ARE2, UPC2-1, and ACC1. Consequently, better cell growth led to an increase of ergosterol content to 40.6 mg/g DCW, which is 4.2-fold higher than that of the starting strain. Finally, a two-stage feeding strategy was employed for high-density cell fermentation, with an ergosterol yield of 2986.7 mg/L and content of 29.5 mg/g DCW. This study provided an effective approach for the production of ergosterol and other related terpenoid molecules.
format Online
Article
Text
id pubmed-8275991
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-82759912021-07-14 Combined Biosynthetic Pathway Engineering and Storage Pool Expansion for High-Level Production of Ergosterol in Industrial Saccharomyces cerevisiae Sun, Zhi-Jiao Lian, Jia-Zhang Zhu, Li Jiang, Yi-Qi Li, Guo-Si Xue, Hai-Long Wu, Mian-Bin Yang, Li-Rong Lin, Jian-Ping Front Bioeng Biotechnol Bioengineering and Biotechnology Ergosterol, a terpenoid compound produced by fungi, is an economically important metabolite serving as the direct precursor of steroid drugs. Herein, ergsosterol biosynthetic pathway modification combined with storage capacity enhancement was proposed to synergistically improve the production of ergosterol in Saccharomyces cerevisiae. S. cerevisiae strain S1 accumulated the highest amount of ergosterol [7.8 mg/g dry cell weight (DCW)] among the wild-type yeast strains tested and was first selected as the host for subsequent metabolic engineering studies. Then, the push and pull of ergosterol biosynthesis were engineered to increase the metabolic flux, overexpression of the sterol acyltransferase gene ARE2 increased ergosterol content to 10 mg/g DCW and additional overexpression of a global regulatory factor allele (UPC2-1) increased the ergosterol content to 16.7 mg/g DCW. Furthermore, considering the hydrophobicity sterol esters and accumulation in lipid droplets, the fatty acid biosynthetic pathway was enhanced to expand the storage pool for ergosterol. Overexpression of ACC1 coding for the acetyl-CoA carboxylase increased ergosterol content from 16.7 to 20.7 mg/g DCW. To address growth inhibition resulted from premature accumulation of ergosterol, auto-inducible promoters were employed to dynamically control the expression of ARE2, UPC2-1, and ACC1. Consequently, better cell growth led to an increase of ergosterol content to 40.6 mg/g DCW, which is 4.2-fold higher than that of the starting strain. Finally, a two-stage feeding strategy was employed for high-density cell fermentation, with an ergosterol yield of 2986.7 mg/L and content of 29.5 mg/g DCW. This study provided an effective approach for the production of ergosterol and other related terpenoid molecules. Frontiers Media S.A. 2021-06-29 /pmc/articles/PMC8275991/ /pubmed/34268298 http://dx.doi.org/10.3389/fbioe.2021.681666 Text en Copyright © 2021 Sun, Lian, Zhu, Jiang, Li, Xue, Wu, Yang and Lin. https://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 Bioengineering and Biotechnology
Sun, Zhi-Jiao
Lian, Jia-Zhang
Zhu, Li
Jiang, Yi-Qi
Li, Guo-Si
Xue, Hai-Long
Wu, Mian-Bin
Yang, Li-Rong
Lin, Jian-Ping
Combined Biosynthetic Pathway Engineering and Storage Pool Expansion for High-Level Production of Ergosterol in Industrial Saccharomyces cerevisiae
title Combined Biosynthetic Pathway Engineering and Storage Pool Expansion for High-Level Production of Ergosterol in Industrial Saccharomyces cerevisiae
title_full Combined Biosynthetic Pathway Engineering and Storage Pool Expansion for High-Level Production of Ergosterol in Industrial Saccharomyces cerevisiae
title_fullStr Combined Biosynthetic Pathway Engineering and Storage Pool Expansion for High-Level Production of Ergosterol in Industrial Saccharomyces cerevisiae
title_full_unstemmed Combined Biosynthetic Pathway Engineering and Storage Pool Expansion for High-Level Production of Ergosterol in Industrial Saccharomyces cerevisiae
title_short Combined Biosynthetic Pathway Engineering and Storage Pool Expansion for High-Level Production of Ergosterol in Industrial Saccharomyces cerevisiae
title_sort combined biosynthetic pathway engineering and storage pool expansion for high-level production of ergosterol in industrial saccharomyces cerevisiae
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8275991/
https://www.ncbi.nlm.nih.gov/pubmed/34268298
http://dx.doi.org/10.3389/fbioe.2021.681666
work_keys_str_mv AT sunzhijiao combinedbiosyntheticpathwayengineeringandstoragepoolexpansionforhighlevelproductionofergosterolinindustrialsaccharomycescerevisiae
AT lianjiazhang combinedbiosyntheticpathwayengineeringandstoragepoolexpansionforhighlevelproductionofergosterolinindustrialsaccharomycescerevisiae
AT zhuli combinedbiosyntheticpathwayengineeringandstoragepoolexpansionforhighlevelproductionofergosterolinindustrialsaccharomycescerevisiae
AT jiangyiqi combinedbiosyntheticpathwayengineeringandstoragepoolexpansionforhighlevelproductionofergosterolinindustrialsaccharomycescerevisiae
AT liguosi combinedbiosyntheticpathwayengineeringandstoragepoolexpansionforhighlevelproductionofergosterolinindustrialsaccharomycescerevisiae
AT xuehailong combinedbiosyntheticpathwayengineeringandstoragepoolexpansionforhighlevelproductionofergosterolinindustrialsaccharomycescerevisiae
AT wumianbin combinedbiosyntheticpathwayengineeringandstoragepoolexpansionforhighlevelproductionofergosterolinindustrialsaccharomycescerevisiae
AT yanglirong combinedbiosyntheticpathwayengineeringandstoragepoolexpansionforhighlevelproductionofergosterolinindustrialsaccharomycescerevisiae
AT linjianping combinedbiosyntheticpathwayengineeringandstoragepoolexpansionforhighlevelproductionofergosterolinindustrialsaccharomycescerevisiae