Hydrophobic microspheres forin situremoval of 2-phenylethanol from yeast fermentation

Achmon, Yigal; Goldshtein, Jenny; Margel, Shlomo; Fishman, Ayelet

doi:10.3109/02652048.2011.599443

Cited by 17 publications

(14 citation statements)

References 32 publications

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“…[46] Meanwhile, various strategies for in situ 2PE removal have also been investigated, including, for example, via its extraction in a biphasic ionic liquid system which gave threeto fivefold increases in 2PE production by S. cerevisiae. [47] Other approaches, meanwhile, including pervaporation [48] and solidphase extraction (i.e., using hydrophobic resins) [49] have shown as high as 10-fold improvements in 2PE production.…”

Section: Discussionmentioning

confidence: 99%

Expanding Upon Styrene Biosynthesis to Engineer a Novel Route to 2‐Phenylethanol

Machas

McKenna

Nielsen

2017

Biotechnology Journal

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2-Phenylethanol (2PE) is a key molecule used in the fragrance and food industries, as well as a potential biofuel. In contrast to its extraction from plant biomass and/or more common chemical synthesis, microbial 2PE production has been demonstrated via both native and heterologous expression of the yeast Ehrlich pathway. Here, a novel alternative to this established pathway is systematically engineered in Escherichia coli and evaluated as a more robust and efficient route. This novel pathway is constructed via the modular extension of a previously engineered styrene biosynthesis pathway, proceeding from endogenous l-phenylalanine in five steps and involving four heterologous enzymes. This "styrene-derived" pathway boasts nearly a 10-fold greater thermodynamic driving force than the Ehrlich pathway, and enables reduced accumulation of acetate byproduct. When directly compared using a host strain engineered for l-phenylalanine over-production, preservation of phosphoenolpyruvate, and reduced formation of byproduct 2-phenylacetic acid, final 2PE titers via the styrene-derived and Ehrlich pathways reached 1817 and 1164 mg L , respectively, at yields of 60.6 and 38.8 mg g . Following optimization of induction timing and initial glucose loading, 2PE titers by the styrene-derived pathway approached 2 g L - nearly a two-fold twofold increase over prior reports for 2PE production by E. coli employing the Ehrlich pathway.

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Section: Discussionmentioning

confidence: 99%

Expanding Upon Styrene Biosynthesis to Engineer a Novel Route to 2‐Phenylethanol

Machas

McKenna

Nielsen

2017

Biotechnology Journal

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“…The microbial production of 2‐phenylethanol has recently received considerable attention as an all‐natural flavour in the food and cosmetics industry. Due to the toxic effect of large amounts of 2‐phenylethanol (>2.5 g L −1 ) in situ removal strategies using organic solvents have been developed to avoid a decrease in growth rate (Stark et al ., ; Eshkol et al ., ; Achmon et al ., ; Hua & Xu, ).…”

Section: Discussionmentioning

confidence: 99%

“…The microbial production of 2-phenylethanol has recently received considerable attention as an all-natural flavour in the food and cosmetics industry. Due to the toxic effect of large amounts of 2-phenylethanol (>2.5 g L À1 ) in situ removal strategies using organic solvents have been developed to avoid a decrease in growth rate (Stark et al, 2003;Eshkol et al, 2009;Achmon et al, 2011;Hua & Xu, 2011). The production of more fruity notes in beverages could benefit from for example co-fermentations using S. cerevisiae and another non-conventional yeast strain.…”

Section: Ag Aro8amentioning

confidence: 99%

Major contribution of the Ehrlich pathway for 2-phenylethanol/rose flavor production inAshbya gossypii

2014

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Aroma alcohols of fermented food and beverages are derived from fungal amino acids catabolism via the Ehrlich pathway. This linear pathway consists of three enzymatic reactions to form fusel alcohols. Regulation of some of the enzymes occurs on the transcriptional level via Aro80. The riboflavin overproducer Ashbya gossypii produces strong fruity flavours in contrast to its much less aromatic relative Eremothecium cymbalariae. Genome comparisons indicated that A. gossypii harbors genes for aromatic amino acid catabolism (ARO8a, ARO8b, ARO10, and ARO80) while E. cymbalariae only encodes ARO8a and thus lacks major components of aromatic amino acid catabolism. Volatile compound (VOC) analysis showed that both Eremothecium species produce large amounts of isoamyl alcohol while A. gossypii also produces high levels of 2-phenylethanol. Deletion of the A. gossypii ARO-genes did not confer any growth deficiencies. However, A. gossypii ARO-mutants (except Agaro8a) were strongly impaired in aroma production, particularly in the production of the rose flavour 2-phenylethanol. Conversely, overexpression of ARO80 via the AgTEF1 promoter resulted in 50% increase in VOC production. Together these data indicate that A. gossypii is a very potent flavour producer and that amongst the non-Saccharomyces biodiversity strains can be identified that could provide positive sensory properties to fermented beverages.

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“…Hence, a significant improvement in 2-PE production can be expected when applying a fed-batch cultivation for the AM1-d strain. However, in the case of AM1-d it would be better to apply the fed-batch culture in conjunction with 1 of the ISPR techniques, since in a batch culture we obtained a boundary concentration for 2-PE of around 4 g/L, which is toxic for yeast and inhibits the production of 2-PE [Achmon et al, 2011;]. …”

Section: Selection Of Diploid S Cerevisiae Am1-d and Testing For 2-pmentioning

confidence: 99%

Mating of 2 Laboratory Saccharomyces cerevisiae Strains Resulted in Enhanced Production of 2-Phenylethanol by Biotransformation of <smlcap>L</smlcap>-Phenylalanine

Mierzejewska

Tymoszewska²,

Chreptowicz³

et al. 2017

Microb Physiol

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2-Phenylethanol (2-PE) is an aromatic alcohol with a rosy scent which is widely used in the food, fragrance, and cosmetic industries. Promising sources of natural 2-PE are microorganisms, especially yeasts, which can produce 2-PE by biosynthesis and biotransformation. Thus, the first challenging goal in the development of biotechnological production of 2-PE is searching for highly productive yeast strains. In the present work, 5 laboratory Saccharomyces cerevisiae strains were tested for the production of 2-PE. Thereafter, 2 of them were hybridized by a mating procedure and, as a result, a new diploid, S. cerevisiae AM1-d, was selected. Within the 72-h batch culture in a medium containing 5 g/L of L-phenylalanine, AM1-d produced 3.83 g/L of 2-PE in a shaking flask. In this way, we managed to select the diploid S. cerevisiae AM1-d strain, showing a 3- and 5-fold increase in 2-PE production in comparison to parental strains. Remarkably, the enhanced production of 2-PE by the hybrid of 2 yeast laboratory strains is demonstrated here for the first time.

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Hydrophobic microspheres forin situremoval of 2-phenylethanol from yeast fermentation

Cited by 17 publications

References 32 publications

Expanding Upon Styrene Biosynthesis to Engineer a Novel Route to 2‐Phenylethanol

Expanding Upon Styrene Biosynthesis to Engineer a Novel Route to 2‐Phenylethanol

Major contribution of the Ehrlich pathway for 2-phenylethanol/rose flavor production inAshbya gossypii

Mating of 2 Laboratory <b><i>Saccharomyces cerevisiae</i></b> Strains Resulted in Enhanced Production of 2-Phenylethanol by Biotransformation of <smlcap>L</smlcap>-Phenylalanine

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