Enhanced production of β-carotene in recombinant Saccharomyces cerevisiae by inverse metabolic engineering with supplementation of unsaturated fatty acids

We, and others, have identified a novel Saccharomyces cerevisiae peroxisomal protein that belongs to the isomerase/hydratase family. The protein, named Dci1p, shares 50% identity with Eci1p, a delta(3)-cis-delta(2)-trans-enoyl-CoA isomerase that acts as an auxiliary enzyme in the beta-oxidation of unsaturated fatty acids. Both of these proteins are localized to peroxisomes, and both contain motifs at their amino- and carboxyl termini that resemble peroxisome targeting signals (PTS) 1 and 2. However, we demonstrate that the putative type 1 signaling motif is not required for the peroxisomal localization of either of these proteins. Furthermore, the correct targeting of Eci1p and Dci1p occurs in the absence of the receptors for the type 1 or type 2 peroxisome targeting pathway. Together, these data suggest a novel mechanism for the intracellular targeting of these peroxisomal proteins.

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DETERMINATION OF CHEMICAL PROPERTIES, COMPOSITION OF FATTY ACID, CAROTENOIDS AND TOCOPHEROLS OF DEGUMMED AND NEUTRALIZED RED FRUIT (Pandanus conoideus) OIL

Jurnal Teknologi ◽

10.11113/jurnalteknologi.v82.14820 ◽

2020 ◽

Vol 82 (6) ◽

pp. 71-78

Author(s):

Zita Letviany Sarungallo ◽

Budi Santoso ◽

Risma Uli Situngkir ◽

Mathelda Kurniaty Roreng ◽

Meike Meilan Lisangan

Keyword(s):

Fatty Acids ◽

Fatty Acid Composition ◽

Fatty Acid ◽

Water Content ◽

Acid Composition ◽

Unsaturated Fatty Acids ◽

Chemical Properties ◽

Carotenoid Content ◽

Fruit Oil ◽

Β Carotene

Refining of crude red fruit oil (CRFO) through the degumming and neutralization steps intended to produce oil free of impurities (non triglycerides) such as phospholipids, proteins, residues and carbohydrates, and also reducing the amount of free fatty acids (FFA). This study aims to determine the effect of red fruit oil purification through degumming and neutralization stages on chemical properties, fatty acid composition, carotenoid content and tocopherol of red fruit oil (RFO). The results showed that degumming of CRFO did not affect the decrease in water content, FFA levels, peroxide numbers, iodine values, carotenoids and tocopherols content; but decrease in levels of phosphorus, β-carotene and α-tocopherol. Neutralization of degummed-RFO (DRFO) did not affect the decrease in water content, iodine value, carotenoid, tocopherol and α-tocopherol; but the FFA levels, peroxide number, phosphorus and β-carotene levels decreased significantly. The fatty acid composition of RFO was dominated by unsaturated fatty acids (± 75%), which increases through degumming and neutralization stages. β-carotene is more sensitive than α-tocopherol during refining process of crude oil, but in general, this process can improve the RFO quality.

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Effect of aeration and unsaturated fatty acids on expression of the Saccharomyces cerevisiae alcohol acetyltransferase gene.

Applied and Environmental Microbiology ◽

10.1128/aem.63.3.910-915.1997 ◽

1997 ◽

Vol 63 (3) ◽

pp. 910-915 ◽

Cited By ~ 102

Author(s):

T Fujii ◽

O Kobayashi ◽

H Yoshimoto ◽

S Furukawa ◽

Y Tamai

Keyword(s):

Saccharomyces Cerevisiae ◽

Fatty Acids ◽

Unsaturated Fatty Acids ◽

Alcohol Acetyltransferase

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Build Your Bioprocess on a Solid Strain—β-Carotene Production in Recombinant Saccharomyces cerevisiae

Frontiers in Bioengineering and Biotechnology ◽

10.3389/fbioe.2019.00171 ◽

2019 ◽

Vol 7 ◽

Cited By ~ 6

Author(s):

Javiera López ◽

Vicente F. Cataldo ◽

Manuel Peña ◽

Pedro A. Saa ◽

Francisco Saitua ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Recombinant Saccharomyces Cerevisiae ◽

Β Carotene

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The effect of pantothenate on sulfate metabolism and sulfur isotope fractionation by Saccharomyces cerevisiae

Canadian Journal of Microbiology ◽

10.1139/m79-177 ◽

1979 ◽

Vol 25 (10) ◽

pp. 1139-1144 ◽

Cited By ~ 1

Author(s):

R. G. L. McCready ◽

G. A. Din ◽

H. R. Krouse

Keyword(s):

Saccharomyces Cerevisiae ◽

Fatty Acids ◽

Sulfate Reduction ◽

Lipid Content ◽

Isotope Fractionation ◽

Sulfur Isotope ◽

Unsaturated Fatty Acids ◽

Cellular Lipid ◽

Sulfur Isotope Fractionation ◽

Sulfate Metabolism

Growth of Saccharomyces cerevisiae in minimal salts – glucose – SO42− medium with varying concentrations of pantothenate (0–1000 μg/L) produced changes in the cellular lipid content and in the ratio of saturated to unsaturated fatty acids. Substantial differences in SO42−diffusion were also observed with changes in pantothenate concentration. During sulfate reduction, the δ34S value of the evolved sulfide varied with the pantothenate concentration ranging from −31‰ in the absence of pantothenate to 0‰ at 400−1000 μg/L pantothenate. The isotope selectivity is related to the effect of pantothenate concentration on cellular metabolism.

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Combinatorial Metabolic Engineering in Saccharomyces cerevisiae for the Enhanced Production of the FPP-Derived Sesquiterpene Germacrene

Bioengineering ◽

10.3390/bioengineering7040135 ◽

2020 ◽

Vol 7 (4) ◽

pp. 135

Author(s):

Jan Niklas Bröker ◽

Boje Müller ◽

Dirk Prüfer ◽

Christian Schulze Gronover

Keyword(s):

Saccharomyces Cerevisiae ◽

Metabolic Engineering ◽

Metabolic Flux ◽

Mevalonate Pathway ◽

Plant Secondary Metabolites ◽

Product Formation ◽

Culture Conditions ◽

Efficient Production ◽

Enhanced Production ◽

Engineering Strategy

Farnesyl diphosphate (FPP)-derived isoprenoids represent a diverse group of plant secondary metabolites with great economic potential. To enable their efficient production in the heterologous host Saccharomyces cerevisiae, we refined a metabolic engineering strategy using the CRISPR/Cas9 system with the aim of increasing the availability of FPP for downstream reactions. The strategy included the overexpression of mevalonate pathway (MVA) genes, the redirection of metabolic flux towards desired product formation and the knockout of genes responsible for competitive reactions. Following the optimisation of culture conditions, the availability of the improved FPP biosynthesis for downstream reactions was demonstrated by the expression of a germacrene synthase from dandelion. Subsequently, biosynthesis of significant amounts of germacrene-A was observed in the most productive strain compared to the wild type. Thus, the presented strategy is an excellent tool to increase FPP-derived isoprenoid biosynthesis in yeast.

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Expressing a cytosolic pyruvate dehydrogenase complex to increase free fatty acid production in Saccharomyces cerevisiae

Microbial Cell Factories ◽

10.1186/s12934-020-01493-z ◽

2020 ◽

Vol 19 (1) ◽

Author(s):

Yiming Zhang ◽

Mo Su ◽

Ning Qin ◽

Jens Nielsen ◽

Zihe Liu

Keyword(s):

Saccharomyces Cerevisiae ◽

Fatty Acids ◽

Fatty Acid ◽

Free Fatty Acid ◽

Pyruvate Dehydrogenase ◽

Unsaturated Fatty Acids ◽

Pyruvate Dehydrogenase Complex ◽

Cell Factory ◽

Acetyl Coa ◽

Fatty Acid Production

Abstract Background Saccharomyces cerevisiae is being exploited as a cell factory to produce fatty acids and their derivatives as biofuels. Previous studies found that both precursor supply and fatty acid metabolism deregulation are essential for enhanced fatty acid synthesis. A bacterial pyruvate dehydrogenase (PDH) complex expressed in the yeast cytosol was reported to enable production of cytosolic acetyl-CoA with lower energy cost and no toxic intermediate. Results Overexpression of the PDH complex significantly increased cell growth, ethanol consumption and reduced glycerol accumulation. Furthermore, to optimize the redox imbalance in production of fatty acids from glucose, two endogenous NAD+-dependent glycerol-3-phosphate dehydrogenases were deleted, and a heterologous NADP+-dependent glyceraldehyde-3-phosphate dehydrogenase was introduced. The best fatty acid producing strain PDH7 with engineering of precursor and co-factor metabolism could produce 840.5 mg/L free fatty acids (FFAs) in shake flask, which was 83.2% higher than the control strain YJZ08. Profile analysis of free fatty acid suggested the cytosolic PDH complex mainly resulted in the increases of unsaturated fatty acids (C16:1 and C18:1). Conclusions We demonstrated that cytosolic PDH pathway enabled more efficient acetyl-CoA provision with the lower ATP cost, and improved FFA production. Together with engineering of the redox factor rebalance, the cytosolic PDH pathway could achieve high level of FFA production at similar levels of other best acetyl-CoA producing pathways.

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