Alkaline pH enhances farnesol production by Saccharomyces cerevisiae

Muramatsu, Masayoshi; Ohto, Chikara; Obata, Seiji; Sakuradani, Eiji; Shimizu, Sakayu

doi:10.1016/j.jbiosc.2009.02.012

Cited by 25 publications

(14 citation statements)

References 20 publications

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“…2e). This is consistent with previous reports describing production of farnesol in strains over-expressing the mevalonate pathway with or without a sesquiterpene synthase (Muramatsu et al, 2009;Scalcinati et al, 2012a;Song, 2003). This activity is attributed to the presense of endogeneous phosphatase(s) (Farhi et al, 2011;Scalcinati et al, 2012a) The improved performance of the E. faecalis genes (strain N6) might be due to either higher catalytic efficiency of the enzymes encoded by EfmvaS and EfmvaE relative to the cognate yeast genes (ERG10/ERG13 and tHMG1, respectively), higher transcription/translation rate, or higher protein/mRNA stability.…”

Section: Increasing Trans-nerolidol Production By Enhancing the Mva Psupporting

confidence: 94%

A squalene synthase protein degradation method for improved sesquiterpene production in Saccharomyces cerevisiae

Peng

Plan

Chrysanthopoulos

et al. 2017

Metabolic Engineering

101

128

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A squalene synthase protein degradation method for improved sesquiterpene production in Saccharomyces cerevisiae, Metabolic Engineering, http://dx.doi.org/10. 1016/j.ymben.2016.12.003 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting galley proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. AbstractSesquiterpenes are C15 isoprenoids with utility as fragrances, flavours, pharmaceuticals, and potential biofuels. Microbial fermentation is being examined as a competitive approach for bulk production of these compounds. Competition for carbon allocation between synthesis of endogenous sterols and production of the introduced sesquiterpene limits yields. Achieving balance between endogenous sterols and heterologous sesquiterpenes is therefore required to achieve economical yields. In the current study, the yeast Saccharomyces cerevisiae was used to produce the acyclic sesquiterpene alcohol, trans-nerolidol. Nerolidol production was first improved by enhancing the upstream mevalonate pathway for the synthesis of the precursor farnesyl pyrophosphate (FPP). However, excess FPP was partially directed towards squalene by squalene synthase (Erg9p), resulting in squalene accumulation to 1 % biomass; moreover, the specific growth rate declined. In order to redirect carbon away from sterol production and towards the desired heterologous sesquiterpene, a novel protein destabilisation approach was developed for Erg9p. It was shown that Erg9p is located on endoplasmic reticulum and lipid droplets through a C-terminal ER-targeted transmembrane peptide. 2A PEST (rich in Pro, Glu/Asp, Ser, and Thr) sequence-dependent endoplasmic reticulum-associated protein degradation (ERAD) mechanism was established to decrease cellular levels of Erg9p without relying on inducers, repressors or specific repressing conditions. This improved nerolidol titre by 86 % to ~100 mg L -1 . In this strain, squalene levels were similar to the wild-type control strain, and downstream ergosterol levels were slightly decreased relative to the control, indicating redirection of carbon away from sterols and towards sesquiterpene production. There was no negative effect on cell growth under these conditions. Protein degradation is an efficient mechanism to control carbon allocation at flux-competing nodes in metabolic engineering applications. This study demonstrates that an engineered ERAD mechanism can be used to balance flux competition between the endogenous sterol pathway and an introduced bio-product pathways at the FPP node. The approach of protein degradation in general might be more widely applied to improve metabolic engineering outcomes.

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Section: Increasing Trans-nerolidol Production By Enhancing the Mva Psupporting

confidence: 94%

A squalene synthase protein degradation method for improved sesquiterpene production in Saccharomyces cerevisiae

Peng

Plan

Chrysanthopoulos

et al. 2017

Metabolic Engineering

101

128

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“…Farnesol and its isomer nerolidol arise from farnesyl diphosphate instability at low pH, like that in the yeast vacuole or in the exocellular medium [38]. Therefore, it is possible that the transporter Qdr2p is responsible of the export of either farnesyl diphosphate or of nerolidol.…”

Section: Discussionmentioning

confidence: 99%

QTL mapping of the production of wine aroma compounds by yeast

et al. 2012

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BackgroundWine aroma results from the combination of numerous volatile compounds, some produced by yeast and others produced in the grapes and further metabolized by yeast. However, little is known about the consequences of the genetic variation of yeast on the production of these volatile metabolites, or on the metabolic pathways involved in the metabolism of grape compounds. As a tool to decipher how wine aroma develops, we analyzed, under two experimental conditions, the production of 44 compounds by a population of 30 segregants from a cross between a laboratory strain and an industrial strain genotyped at high density.ResultsWe detected eight genomic regions explaining the diversity concerning 15 compounds, some produced de novo by yeast, such as nerolidol, ethyl esters and phenyl ethanol, and others derived from grape compounds such as citronellol, and cis-rose oxide. In three of these eight regions, we identified genes involved in the phenotype. Hemizygote comparison allowed the attribution of differences in the production of nerolidol and 2-phenyl ethanol to the PDR8 and ABZ1 genes, respectively. Deletion of a PLB2 gene confirmed its involvement in the production of ethyl esters. A comparison of allelic variants of PDR8 and ABZ1 in a set of available sequences revealed that both genes present a higher than expected number of non-synonymous mutations indicating possible balancing selection.ConclusionsThis study illustrates the value of QTL analysis for the analysis of metabolic traits, and in particular the production of wine aromas. It also identifies the particular role of the PDR8 gene in the production of farnesyldiphosphate derivatives, of ABZ1 in the production of numerous compounds and of PLB2 in ethyl ester synthesis. This work also provides a basis for elucidating the metabolism of various grape compounds, such as citronellol and cis-rose oxide.

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“…The resulting FPP could be directed to the formation of FOH in the yeast. Optimization of culture condition such as alkaline pH (Muramatsu et al, 2009) and supplementation with oils and detergents (Muramatsu et al, 2008b) could increase FOH production in yeast. It has been reported that FOH was produced at a oncerntration of approximately 80 mg/L in a mevalonate (MVA) pathway engineered yeast strain after 8 days culture (Takahashi et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

Farnesol production from Escherichia coli by harnessing the exogenous mevalonate pathway

Wang

Yoon

Shah

et al. 2010

Biotech & Bioengineering

104

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Farnesol (FOH) production has been carried out in metabolically engineered Escherichia coli. FOH is formed through the depyrophosphorylation of farnesyl pyrophosphate (FPP), which is synthesized from isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) by FPP synthase. In order to increase FPP synthesis, E. coli was metabolically engineered to overexpress ispA and to utilize the foreign mevalonate (MVA) pathway for the efficient synthesis of IPP and DMAPP. Two-phase culture using a decane overlay of the culture broth was applied to reduce volatile loss of FOH produced during culture and to extract FOH from the culture broth. A FOH production of 135.5 mg/L was obtained from the recombinant E. coli harboring the pTispA and pSNA plasmids for ispA overexpression and MVA pathway utilization, respectively. It is interesting to observe that a large amount of FOH could be produced from E. coli without FOH synthase by the augmentation of FPP synthesis. Introduction of the exogenous MVA pathway enabled the dramatic production of FOH by E. coli while no detectable FOH production was observed in the endogenous MEP pathway-only control.

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Alkaline pH enhances farnesol production by Saccharomyces cerevisiae

Cited by 25 publications

References 20 publications

A squalene synthase protein degradation method for improved sesquiterpene production in Saccharomyces cerevisiae

A squalene synthase protein degradation method for improved sesquiterpene production in Saccharomyces cerevisiae

QTL mapping of the production of wine aroma compounds by yeast

Farnesol production from Escherichia coli by harnessing the exogenous mevalonate pathway

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