A metabolic engineering strategy for producing free fatty acids by the <i>Yarrowia lipolytica</i> yeast based on impairment of glycerol metabolism

Yuzbasheva, Evgeniya Y.; Mostova, E. B.; Андреева, Н. И.; Yuzbashev, Tigran V.; Fedorov, A. S.; Konova, I. A.; Sineoky, S. P.

doi:10.1002/bit.26402

Cited by 22 publications

(11 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Various extractive production strategies are known for permeating the cell membrane and continuously stripping the produced oil. These techniques employ a liquid phase biocompatible extractant like long-chained alkanes [29] to increase the miscibility of the extracellular oil in the aqueous broth and thereby promote its efflux. Some also report secretion of bioemulsifiers that aid the extracellular oil.…”

Section: Resultsmentioning

confidence: 99%

Simultaneous lipid biosynthesis and recovery for oleaginous yeast Yarrowia lipolytica

Pawar

Odaneth

Vadgama

et al. 2019

Biotechnol Biofuels

View full text Add to dashboard Cite

Background Recent trends in bioprocessing have underlined the significance of lignocellulosic biomass conversions for biofuel production. These conversions demand at least 90% energy upgradation of cellulosic sugars to generate renewable drop-in biofuel precursors (Heff/C ~ 2). Chemical methods fail to achieve this without substantial loss of carbon; whereas, oleaginous biological systems propose a greener upgradation route by producing oil from sugars with 30% theoretical yields. However, these oleaginous systems cannot compete with the commercial volumes of vegetable oils in terms of overall oil yields and productivities. One of the significant challenges in the commercial exploitation of these microbial oils lies in the inefficient recovery of the produced oil. This issue has been addressed using highly selective oil capturing agents (OCA), which allow a concomitant microbial oil production and in situ oil recovery process. Results Adsorbent-based oil capturing agents were employed for simultaneous in situ oil recovery in the fermentative production broths. Yarrowia lipolytica, a model oleaginous yeast, was milked incessantly for oil production over 380 h in a media comprising of glucose as a sole carbon and nutrient source. This was achieved by continuous online capture of extracellular oil from the aqueous media and also the cell surface, by fluidizing the fermentation broth over an adsorbent bed of oil capturing agents (OCA). A consistent oil yield of 0.33 g per g of glucose consumed, corresponding to theoretical oil yield over glucose, was achieved using this approach. While the incorporation of the OCA increased the oil content up to 89% with complete substrate consumptions, it also caused an overall process integration. Conclusion The nondisruptive oil capture mediated by an OCA helped in accomplishing a trade-off between microbial oil production and its recovery. This strategy helped in realizing theoretically efficient sugar-to-oil bioconversions in a continuous production process. The process, therefore, endorses a sustainable production of molecular drop-in equivalents through oleaginous yeasts, representing as an absolute microbial oil factory.

show abstract

Section: Resultsmentioning

confidence: 99%

Simultaneous lipid biosynthesis and recovery for oleaginous yeast Yarrowia lipolytica

Pawar

Odaneth

Vadgama

et al. 2019

Biotechnol Biofuels

View full text Add to dashboard Cite

show abstract

“…GPD1 encodes glycerol phosphate dehydrogenase. In Y. lipolytica, it is essential for growth in glucose as a carbon source, but non-essential on medium containing glycerol as a carbon source (Yuzbasheva et al, 2017). Cells disrupted for GPD1 were viable when grown on glycerol but failed to grow on glucose.…”

Section: Validation Of Gene Classification Methodsmentioning

confidence: 99%

Functional genomics for the oleaginous yeast Yarrowia lipolytica

Patterson

Landberg

et al. 2018

Metabolic Engineering

View full text Add to dashboard Cite

Oleaginous yeasts are valuable systems for biosustainable production of hydrocarbon-based chemicals. Yarrowia lipolytica is one of the best characterized of these yeast with respect to genome annotation and flux analysis of metabolic processes. Nonetheless, progress is hampered by a dearth of genome-wide tools enabling functional genomics. In order to remedy this deficiency, we developed a library of Y. lipolytica insertion mutants via transposon mutagenesis. The Hermes DNA transposon was expressed to achieve saturation mutagenesis of the genome. Over 534,000 independent insertions were identified by next-generation sequencing. Poisson analysis of insertion density classified ~ 22% of genes as essential. As expected, most essential genes have homologs in Saccharomyces cerevisiae and Schizosaccharomyces pombe, and the majority of those are also essential. As an obligate aerobe, Y. lipolytica has significantly more respiration - related genes that are classified as essential than do S. cerevisiae and S. pombe. Contributions of non-essential genes to growth in glucose and glycerol carbon sources were assessed and used to evaluate two recent genome-scale models of Y. lipolytica metabolism. Fluorescence-activated cell sorting identified mutants in which lipid accumulation is increased. Our findings provide insights into biosynthetic pathways, compartmentalization of enzymes, and distinct functions of paralogs. This functional genomic analysis of the oleaginous yeast Y. lipolytica provides an important resource for modeling, bioengineering, and design of synthetic minimalized strains of respiratory yeasts.

show abstract

“…First, we explored the citrate route-a pathway that has been extensively studied for its capacity to increase lipid production in Y. lipolytica (21)(22)(23). Overexpression of ACC1, which codes for the enzyme that converts acetyl-CoA to malonyl-CoA, has also been shown to promote lipid production (24,25). When the pathway genes (ACL1, ACL2, and AMPD) were concurrently overexpressed, TAL production was significantly reduced and only marginally improved with the addition of ACC1 ( Fig.…”

mentioning

confidence: 99%

Rewiring Yarrowia lipolytica toward triacetic acid lactone for materials generation

Markham

Palmer

Chwatko

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

160

151

View full text Add to dashboard Cite

Polyketides represent an extremely diverse class of secondary metabolites often explored for their bioactive traits. These molecules are also attractive building blocks for chemical catalysis and polymerization. However, the use of polyketides in larger scale chemistry applications is stymied by limited titers and yields from both microbial and chemical production. Here, we demonstrate that an oleaginous organism (specifically, ) can overcome such production limitations owing to a natural propensity for high flux through acetyl-CoA. By exploring three distinct metabolic engineering strategies for acetyl-CoA precursor formation, we demonstrate that a previously uncharacterized pyruvate bypass pathway supports increased production of the polyketide triacetic acid lactone (TAL). Ultimately, we establish a strain capable of producing over 35% of the theoretical conversion yield to TAL in an unoptimized tube culture. This strain also obtained an averaged maximum titer of 35.9 ± 3.9 g/L with an achieved maximum specific productivity of 0.21 ± 0.03 g/L/h in bioreactor fermentation. Additionally, we illustrate that a β-oxidation-related overexpression () can support high TAL production and is capable of achieving over 43% of the theoretical conversion yield under nitrogen starvation in a test tube. Next, through use of this bioproduct, we demonstrate the utility of polyketides like TAL to modify commodity materials such as poly(epichlorohydrin), resulting in an increased molecular weight and shift in glass transition temperature. Collectively, these findings establish an engineering strategy enabling unprecedented production from a type III polyketide synthase as well as establish a route through O-functionalization for converting polyketides into new materials.

show abstract

A metabolic engineering strategy for producing free fatty acids by the Yarrowia lipolytica yeast based on impairment of glycerol metabolism

Cited by 22 publications

References 47 publications

Simultaneous lipid biosynthesis and recovery for oleaginous yeast Yarrowia lipolytica

Simultaneous lipid biosynthesis and recovery for oleaginous yeast Yarrowia lipolytica

Functional genomics for the oleaginous yeast Yarrowia lipolytica

Rewiring Yarrowia lipolytica toward triacetic acid lactone for materials generation

Contact Info

Product

Resources

About