Overexpressing enzymes of the Ehrlich pathway and deleting genes of the competing pathway in Saccharomyces cerevisiae for increasing 2-phenylethanol production from glucose

Shen, Li; Nishimura, Yuya; Mizutani, Fumio; Ishii, Jun; Kondo, Akihiko

doi:10.1016/j.jbiosc.2015.12.022

Cited by 53 publications

(38 citation statements)

References 27 publications

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“…Furthermore, YLG18 can even produce 100 mg/L of 2-PE in synthetic medium without supplementation of L-phe via the de novo pathway (Fig. 1B), which surpassed that using metabolically engineered S. cerevisiae (96 mg/L) via de novo pathway [10]. The 18S rDNA genes amplified from the genomic DNA of culture YLG18 showed 99% identity to Meyerozyma guilliermondii when blasted with the microbio sequences in the GenBank database.…”

Section: Resultsmentioning

confidence: 96%

“…For instance, the newly isolated K. marxianus CCT 7735 could generate 3.44 g/L of 2-PE through Ehrlich pathway under optimized conditions [9]. S. cerevisiae BY4741, which overexpressed ARO10 and contained an aro8Δ deletion could produce 96 mg/L of 2-PE from glucose [10]. S. cerevisiae SPO810, in which ARO8 and ARO10 were co-expressed could finally produce 2.61 g/L of 2-PE with fed batch fermentation [3].…”

Section: Introductionmentioning

confidence: 99%

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Enhanced 2-phenylethanol production by newly isolated Meyerozyma sp. strain YLG18 and characterization of its synthetic pathways

Qian

Zhang

Zhou

et al. 2020

Preprint

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Background2-Phenylethanol (2-PE) is an aromatic alcohol which has been widely used in cosmetics, perfume and food industries owning to its delicate rose scent. The newly isolated yeast Meyerozyma guilliermondii YLG18 was able to tolerate high exogenous 2-PE and produce 2-PE with two different pathways.ResultsA unique Meyerozyma sp. strain YLG18 was obtained in this study, which was capable of tolerating 4.0 g/L exogenous 2-PE. Response surface methodology (RSM) was implemented to improve the maximum 2-PE production. At optimized conditions: temperature, 24.7℃; initial glucose, 63.54 g/L; initial L-phe, 10.70 g/L, 2-PE production was increased to 2.55 g/L, leading to 104% increase compared to the pre-optimized one. In situ product recovery (ISPR) could further help improve the final 2-PE production to 3.20 g/L with fatty acid methyl ester as the extractant, representing the highest 2-PE production by using Meyerozyma sp.. Furthermore, genes involved in 2-PE synthesis were identified and their expression levels between Shikimate pathway and Ehrlich pathway were compared. Based on the genomic and transcriptional analysis, a penta-functional enzyme AroM in Shikimate pathway and an aspartate aminotransferase (AAT) with the potential to convert phenylalanine into phenylpyruvate in Ehrlich pathway were identified.ConclusionsThese findings would help broaden our knowledge and add to the pool of known 2-PE generating microbes and genes. Moreover, this study describes a potential, new 2-PE producer that lays foundation for the industrial-scale production of 2-PE and its derivatives in the future. Key words: 2-phenylethanol; Meyerozyma guilliermondii; RSM; ISPR; gene analysis

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Section: Resultsmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

Enhanced 2-phenylethanol production by newly isolated Meyerozyma sp. strain YLG18 and characterization of its synthetic pathways

Qian

Zhang

Zhou

et al. 2020

Preprint

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“…In recent years, E. coli and yeast have been metabolically engineered to produce 2‐PE, by overexpressing the Ehrlich pathway enzymes 2‐keto acid decarboxylase (KDC) and alcohol dehydrogenase (ADH) (Daisuke, Hayato, Kunihiko, Takashi, & Kiyofumi, ; Kim, Cho, et al., ). Shen, Nishimura, Matsuda, Ishii, & Kondo, ; reported that a 2‐PE biosynthetic S. cerevisiae strain with overexpression of ARO10 and deletion of ARO8 (aromatic aminotransferases) produced 96 mg/L 2‐PE (Shen et al., ). Kang, Zhang, Du, & Chen, 2014; constructed a heterologous pathway to produce 2‐PE in a phenylalanine‐producing E. coli .…”

Section: Discussionmentioning

confidence: 99%

Metabolic engineering of Escherichia coli for production of 2‐Phenylethylacetate from L‐phenylalanine

et al. 2017

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In order to meet the need of consumer preferences for natural flavor compounds, microbial synthesis method has become a very attractive alternative to the chemical production. The 2‐phenylethanol (2‐PE) and its ester 2‐phenylethylacetate (2‐PEAc) are two extremely important flavor compounds with a rose‐like odor. In recent years, Escherichia coli and yeast have been metabolically engineered to produce 2‐PE. However, a metabolic engineering approach for 2‐PEAc production is rare. Here, we designed and expressed a 2‐PEAc biosynthetic pathway in E. coli. This pathway comprised four steps: aminotransferase (ARO8) for transamination of L‐phenylalanine to phenylpyruvate, 2‐keto acid decarboxylase KDC for the decarboxylation of the phenylpyruvate to phenylacetaldehyde, aldehyde reductase YjgB for the reduction of phenylacetaldehyde to 2‐PE, alcohol acetyltransferase ATF1 for the esterification of 2‐PE to 2‐PEAc. Using the engineered E. coli strain for shake flasks cultivation with 1 g/L L‐phenylalanine, we achieved co‐production of 268 mg/L 2‐PEAc and 277 mg/L 2‐PE. Our results suggest that approximately 65% of L‐phenylalanine was utilized toward 2‐PEAc and 2‐PE biosynthesis and thus demonstrate potential industrial applicability of this microbial platform.

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“…Romagnoli et al constructed a deletion library of non-essential genes in S. cerevisiae by Synthetic Genetic Array (SGA) technology and identiied that ARO8 encoding an aromatic amino acid transaminase is a target to improve phenylethanol production from glucose [56]. Recently, Shen et al identiied AAT2 encoding a cytosolic aspartate aminotransferase as another deletion target [57]. Deletion of these two genes in combination with the overexpression of Ehrlich pathway enzymes resulted in a signiicant improvement in 2-PE production from glucose, at a titer of 96 mg/L.…”

Section: -Phenyl Ethanol (2-pe) Is Another Economically Atractive Lamentioning

confidence: 99%

Advances in Metabolic Engineering of Saccharomyces cerevisiae for the Production of Industrially and Clinically Important Chemicals

Turanlı-Yıldız¹,

Hacısalihoğlu²,

Çakar³

2017

Old Yeasts - New Questions

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Sustainable production of chemicals is of increasing importance, due to depletion of petroleum and environmental concerns. In addition to its importance in basic research as a simple, eukaryotic model organism, Saccharomyces cerevisiae has long been exploited in industry because of its physiological properties. And today, the development in genetic engineering toolbox and genome-scale metabolic models of S. cerevisiae has extended its application range to new products and bioprocesses. In addition, evolutionary engineering strategies have been useful in improving cellular properties of S. cerevisiae, such as tolerance to product toxicity and inhibitors. In this chapter, recent metabolic and evolutionary engineering studies that involve S. cerevisiae for the production of bulk chemicals and ine chemicals including lavours and pharmaceuticals are reviewed. It was shown that metabolic engineering particularly allowed the improvement of pharmaceuticals production, which will enable economic and large-scale production of many valuable pharmaceuticals. It is clear that S. cerevisiae will continue to be an important host for future metabolic engineering and metabolic pathway engineering applications to produce a variety of industrially and clinically important chemicals.

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Overexpressing enzymes of the Ehrlich pathway and deleting genes of the competing pathway in Saccharomyces cerevisiae for increasing 2-phenylethanol production from glucose

Cited by 53 publications

References 27 publications

Enhanced 2-phenylethanol production by newly isolated Meyerozyma sp. strain YLG18 and characterization of its synthetic pathways

Enhanced 2-phenylethanol production by newly isolated Meyerozyma sp. strain YLG18 and characterization of its synthetic pathways

Metabolic engineering of Escherichia coli for production of 2‐Phenylethylacetate from L‐phenylalanine

Advances in Metabolic Engineering of Saccharomyces cerevisiae for the Production of Industrially and Clinically Important Chemicals

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