Improvement of 2-phenylethanol production in Saccharomyces cerevisiae by evolutionary and rational metabolic engineering.

2-Phenylethanol (2-PE) is a valuable aromatic compound with favorable flavors and good properties, resulting in its widespread application in the cosmetic, food and medical industries. In this study, a mutant strain, AD032, was first obtained by adaptive evolution under 2-PE stress. Then, a fusion p...

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Autores principales:	Linghuan Zhu, Sha Xu, Youran Li, Guiyang Shi
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1b189f80b53b4654ac14c71c12f0244c
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spelling	oai:doaj.org-article:1b189f80b53b4654ac14c71c12f0244c2021-12-02T20:13:38ZImprovement of 2-phenylethanol production in Saccharomyces cerevisiae by evolutionary and rational metabolic engineering.1932-620310.1371/journal.pone.0258180https://doaj.org/article/1b189f80b53b4654ac14c71c12f0244c2021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0258180https://doaj.org/toc/1932-62032-Phenylethanol (2-PE) is a valuable aromatic compound with favorable flavors and good properties, resulting in its widespread application in the cosmetic, food and medical industries. In this study, a mutant strain, AD032, was first obtained by adaptive evolution under 2-PE stress. Then, a fusion protein from the Ehrlich pathway, composed of tyrB from Escherichia coli, kdcA from Lactococcus lactis and ADH2 from Saccharomyces cerevisiae, was constructed and expressed. As a result, 3.14 g/L 2-PE was achieved using L-phenylalanine as a precursor. To further increase 2-PE production, L-glutamate oxidase from Streptomyces overexpression was applied for the first time in our research to improve the supply of α-ketoglutarate in the transamination of 2-PE synthesis. Furthermore, we found that the disruption of the pyruvate decarboxylase encoding gene PDC5 caused an increase in 2-PE production, which has not yet been reported. Finally, assembly of the efficient metabolic modules and process optimization resulted in the strain RM27, which reached 4.02 g/L 2-PE production from 6.7 g/L L-phenylalanine without in situ product recovery. The strain RM27 produced 2-PE (0.8 mol/mol) with L-phenylalanine as a precursor, which was considerably high, and displayed manufacturing potential regarding food safety and process simplification aspects. This study suggests that innovative strategies regarding metabolic modularization provide improved prospects for 2-PE production in food exploitation.Linghuan ZhuSha XuYouran LiGuiyang ShiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 10, p e0258180 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Linghuan Zhu Sha Xu Youran Li Guiyang Shi Improvement of 2-phenylethanol production in Saccharomyces cerevisiae by evolutionary and rational metabolic engineering.
description	2-Phenylethanol (2-PE) is a valuable aromatic compound with favorable flavors and good properties, resulting in its widespread application in the cosmetic, food and medical industries. In this study, a mutant strain, AD032, was first obtained by adaptive evolution under 2-PE stress. Then, a fusion protein from the Ehrlich pathway, composed of tyrB from Escherichia coli, kdcA from Lactococcus lactis and ADH2 from Saccharomyces cerevisiae, was constructed and expressed. As a result, 3.14 g/L 2-PE was achieved using L-phenylalanine as a precursor. To further increase 2-PE production, L-glutamate oxidase from Streptomyces overexpression was applied for the first time in our research to improve the supply of α-ketoglutarate in the transamination of 2-PE synthesis. Furthermore, we found that the disruption of the pyruvate decarboxylase encoding gene PDC5 caused an increase in 2-PE production, which has not yet been reported. Finally, assembly of the efficient metabolic modules and process optimization resulted in the strain RM27, which reached 4.02 g/L 2-PE production from 6.7 g/L L-phenylalanine without in situ product recovery. The strain RM27 produced 2-PE (0.8 mol/mol) with L-phenylalanine as a precursor, which was considerably high, and displayed manufacturing potential regarding food safety and process simplification aspects. This study suggests that innovative strategies regarding metabolic modularization provide improved prospects for 2-PE production in food exploitation.
format	article
author	Linghuan Zhu Sha Xu Youran Li Guiyang Shi
author_facet	Linghuan Zhu Sha Xu Youran Li Guiyang Shi
author_sort	Linghuan Zhu
title	Improvement of 2-phenylethanol production in Saccharomyces cerevisiae by evolutionary and rational metabolic engineering.
title_short	Improvement of 2-phenylethanol production in Saccharomyces cerevisiae by evolutionary and rational metabolic engineering.
title_full	Improvement of 2-phenylethanol production in Saccharomyces cerevisiae by evolutionary and rational metabolic engineering.
title_fullStr	Improvement of 2-phenylethanol production in Saccharomyces cerevisiae by evolutionary and rational metabolic engineering.
title_full_unstemmed	Improvement of 2-phenylethanol production in Saccharomyces cerevisiae by evolutionary and rational metabolic engineering.
title_sort	improvement of 2-phenylethanol production in saccharomyces cerevisiae by evolutionary and rational metabolic engineering.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/1b189f80b53b4654ac14c71c12f0244c
work_keys_str_mv	AT linghuanzhu improvementof2phenylethanolproductioninsaccharomycescerevisiaebyevolutionaryandrationalmetabolicengineering AT shaxu improvementof2phenylethanolproductioninsaccharomycescerevisiaebyevolutionaryandrationalmetabolicengineering AT youranli improvementof2phenylethanolproductioninsaccharomycescerevisiaebyevolutionaryandrationalmetabolicengineering AT guiyangshi improvementof2phenylethanolproductioninsaccharomycescerevisiaebyevolutionaryandrationalmetabolicengineering
_version_	1718374750921687040

Improvement of 2-phenylethanol production in Saccharomyces cerevisiae by evolutionary and rational metabolic engineering.

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