BACKGROUND

Surfactants, or surface active agents, are amphiphilic organic molecules used as detergents, emulsifiers, wetting agents, dispersants, foaming agents, antifoam agents, and humectants. The annual global production of surfactants was 13 million metric tons in 2008 [Ashby (2013) J Surfactants Deterg 16:683-691]. Between 40 and 65% of surfactants are derived from petroleum [Deleu (2004) C R Chim 7:641-646; Rust (2008) Surfactants: a market opportunity study update. OmniTech International Ltd, Midland, Mich.]. The remaining feedstocks are primarily plant oils such as palm or coconut oil. Due to environmental and food security concerns, renewable and more environmentally friendly alternatives are being sought. Naturally produced microbial biosurfactants are appealing alternatives because they are produced renewably and sustainably [Makkar (2011) AMB express 1:1-19], have low ecotoxicity [Renkin (2003) Riv Ital Sostanze Gr 80:249-252], are highly biodegradable [Mulligan (2005) Environ Pollut 133:183-198; Renkin, supra], and are active under a broad range of conditions [Khan (2014) Biosurfactants: Production and Utilization—Processes, Technologies, and Economics 159:269; Marchant (2012) Trends Biotechnol 30:558-565]. Microbial biosurfactants display a unique combination of market attractiveness and high demand growth, and market prices range $10-$30 per kg, depending on the type, the market and the application [Microbial biosurfactants market: Global industry analysis, size, share, growth, trends and forecast 2014-2020. Transparency Market Research, City, pp 1-74]. Microbial biosurfactants are used in numerous applications including industrial and household cleaning products, cosmetics, lubricants, adhesives, agro-chemicals, mining, petroleum extraction and cleanup, and in pulp and paper industries.

Yeast glycolipids (G L, seeFIG. 1) are a group of microbial biosurfactants comprising sophorolipids (SL), mannosylerythritol lipid (MEL), cellobiose lipids (CL) and polyol lipids (PL) [Marchant, supra]. PL comprise two subgroups: (1) liamocins (LM), which consist of a single partially acetylated polyol head group with three or four 3,5-dihydroxydecanoic tails polyesterified through the 5-hydroxy group and are currently the best understood (FIG. 1e) [Price (2016) J Antibiot DOI 10.1038/ja.2016.92; Price (2013) Carbohydr Res 370:24-32], and (2) polyol esters of fatty acids (PEFA) produced by yeasts taxonomically close to theRhodotorula glutinis/graminisclade [Cajka (2016) J Nat Prod 79:2580-2589; Tulloch (1964) Can J Chem 42:830-835]. PEFA are amphiphilic molecules composed of an acetylated (R)-3-hydroxy fatty acid esterified through the carboxyl end to a 5 or 6 carbon polyol, typically D-mannitol or D-arabitol, with varying degrees of acetylations (FIG. 1d) [Cajka, supra]. They differ from LM in that the 45 carbon is not hydroxylated and thus, only have a single acetylated (R)-3-hydroxylated acyl chain of varying numbers of carbons, usually C16:0 and C18:0. LM polyol head group is also non-acetylated, while PEFA's polyol head group can present different degrees of acetylation. Commercial production of SL, MEL and CL requires provision of both a hydrophobic carbon source such as vegetable oil, and a hydrophilic carbon source such as glucose. In contrast, PL could be synthesized in commercially relevant amounts without a hydrophobic carbon source. A few ascomycete and basidiomycete yeast species have been reported to produce PL (see Table 1). Interestingly, all the basidiomycetes reported to produce PEFA are taxonomically close to theRhodotorula glutinis/graminisclade [Kurtzman (2011) The Yeasts: A Taxonomic Study. Elsevier, Oxford].Rhodotorula babjevaeUCDFST 04-877 secreted five times more PEFA than its phylogenetic neighbors [Cajka, supra], raising the question on whether this feature could be present in related species as well.

Recommended strategies for improving biosurfactant production include identifying new hyper-producing microorganisms, genetically modifying production strains, optimizing media and growth conditions, using wastes or other low-cost raw materials, improving downstream processing, and capturing multiple co-products [Dhanaraj an (2014) Biosurfactants: Production and Utilization—Processes, Technologies, and Economics 159:153]. Addressing the first strategy to improve SL production, Kurtzman [Kurtzman (2010) FEMS Microbiol Lett 311:140-146] analyzed 26 strains from the USDA-ARS Culture Collection (http://nrrl.ncaur.usda.gov) belonging to 18 yeast species within theStarmerellaclade. This clade containsStarmerellaspecies, several relatedCandidaspecies, and numerous species that currently lack valid species names. They confirmed that clade memberCandida apicolaproduces SL, which had been previously known [Gorin (1961) Can J Chem 39:846-855; Stodola (1967) Bacteriol Rev 31:194; Tulloch (1968) Can J Chem 46:345-348]. They found three additional SL-secreting yeast species within theStarmerellaclade [Kurtzman (2012) Intl J Syst Evol Microbiol 62:2307-2311; Kurtzman (2010) FEMS Microbiol Lett 311:140-146]. Targeting this taxonomic clade was therefore a successful strategy. It is important to point out that SL production was strain-specific. For example, of two strains ofCandida apicolathat were analyzed, one produced very high levels of SL, and one produced no detectable SL.

The main disadvantages of the previous art were that no detailed chemical structures of PEFA from any basidiomycetous yeast was previously described, and that the yields within the basidiomycetous yeast were low, and not suitable for commercialization. In all cases, yeast cultures were grown in media where no iron was supplemented, and the level of aeration was not as high as in cultures and methods described herein.

SUMMARY

In one aspect, a yeast culture is provided. In varying embodiments, the yeast culture comprises a population of basidiomycetous yeast cells, one or more hydrophilic (e.g., non-hydrophobic) carbon sources, and/or at least about 1 g/L polyol lipid (e.g., polyol ester of fatty acid (PEFA)), e.g., at least about 2 g/L, 3 g/L, 4 g/L, 5 g/L, 6 g/L, 7 g/L, 8 g/L, 9 g/L, 10 g/L, 11 g/L, 12 g/L, 13 g/L, 14 g/L, 15 g/L, 20 g/L, 25 g/L, 30 g/L, 35 g/L, 40 g/L, 45 g/L, 50 g/L, 100 g/L, 150 g/L, 200 g/L, 250 g/L, 300 g/L, 350 g/L, 400 g/L polyol lipid, e.g., up to about 450 g/L of polyol lipid. In varying embodiments, the culture does not comprise one or more hydrophobic carbon sources. In varying embodiments, the culture does not comprise one or more hydrophobic carbon sources selected from the group consisting of oils, alkanes, fatty acids, fatty esters and mixtures thereof. In varying embodiments, the yeast culture comprises a population of basidiomycetous yeast cells, one or more hydrophilic (e.g., non-hydrophobic) carbon sources, one or more hydrophobic carbons sources, and at least about 1 g/L polyol lipid (e.g., polyol ester of fatty acid (PEFA)), e.g., at least about 2 g/L, 3 g/L, 4 g/L, 5 g/L, 6 g/L, 7 g/L, 8 g/L, 9 g/L, 10 g/L, 11 g/L, 12 g/L, 13 g/L, 14 g/L, 15 g/L, 20 g/L, 25 g/L, 30 g/L, 35 g/L, 40 g/L, 45 g/L, 50 g/L, 100 g/L, 150 g/L, 200 g/L, 250 g/L, 300 g/L, 350 g/L, 400 g/L polyol lipid, e.g., up to about 450 g/L of polyol lipid. In varying embodiments, total polyol lipids are as measured after 1, 2, 3, 4, 5, 6, 7 or 8 days growth. In varying embodiments, the culture is a liquid culture. In varying embodiments, the volume of the culture is at least about 0.5 L, 1 L, 2 L, 3 L, 4 L, 5 L, 10 L, 25 L, 50 L, 75 L, 100 L, 250 L, 500 L, 1000 L, or more. In varying embodiments, the one or more hydrophilic carbon sources comprise non-hydrophobic or hydrophilic waste stream and/or hydrolysates. In varying embodiments, the culture comprises less than about 2% (w/v) nitrogen, e.g., in the range of about 0.005% (w/v) to about 2% (w/v) nitrogen. In varying embodiments, the nitrogen source is selected from the group consisting of ammonia, ammonium salt, nitrate, nitrite, nucleotides, nucleosides, proteins, peptides, amino acids, urea and its derivatives, and mixtures thereof. In varying embodiments, the culture comprises about 0.05% (w/v) ammonium chloride. In varying embodiments, the yeast culture comprises an iron salt, e.g., an iron salt selected from the group consisting of iron chloride (II) & (III), iron sulfate (II) & (III), iron nitrate (II) & (III), iron phosphate (II) & (III), fumarate, iron edetate, iron citrate, iron malate, iron oxalate, iron tartrate, iron succinate, iron acetate, in their anhydrous or their hydrated states, and organometallic complexes thereof. In some embodiments, the yeast culture comprises the iron salt in a concentration in the range of about 0.001 mg/L to about 10 g/L, e.g., from about 1 mg/L to about 0.5 g/L, e.g., from about 0.01 g/L to about 0.5 g/L. In varying embodiments, the culture comprises one or more hydrophilic carbon sources at a concentration in the range of about 0.2% (w/v) to about 70% (w/v), e.g., in the range of about 0.2% (w/v) to about 10%, 20%, 30%, 40%, 50% or 60% (w/v). In varying embodiments, the yeast culture is supplemented with extra hydrophilic carbon source (e.g., a mono- or di-saccharide, e.g., glucose and/or sucrose) to a concentration of at least about 50 g/L, e.g., at least about 75 g/L, 100 g/L, 125 g/L, 150 g/L, 175 g/L, 200 g/L, 225 g/L, 250 g/L, 275 g/L or 300 g/L. In varying embodiments, the culture comprises a carbon to nitrogen ratio of about 5:1 to about 400:1, e.g., from about 10:1 to about 200:1, from about 20:1 to about 100:1, e.g., from about 25:1 to about 75:1, e.g., from about 30:1 to about 70:1, e.g., from about 40:1 to about 60:1, e.g., from about 30:1 to about 40:1, e.g., using a nitrogen source that can be consumed or utilized by the yeast cells. In some embodiments, the culture comprises dissolved oxygen at a concentration of a least about 10%, e.g., at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, and up to 60%, 70%, 80%, 90%, 100% dissolved oxygen, e.g., from about 15% to about 80%, e.g., from about 20% to about 60% dissolved oxygen. In varying embodiments, the hydrophilic carbon source is selected from the group consisting of monosaccharides, oligosaccharides, polysaccharides, sugar alcohols, polyols, polyol lipids (including polyol esters of fatty acid (PEFAs)), organic acids, esters, aldehydes, ketones, alcohols, waste streams, plant materials, lignocellulosic hydrolysates, industrial co-products and mixtures thereof. In varying embodiments, the hydrophilic carbon source is a monosaccharide, an oligosaccharide or a polysaccharide and comprises one or more sugar moieties selected from the group consisting of glucose, sucrose, xylose, galactose, rhamnose, arabinose, mannose, cellobiose, galacturonic acid, lactose, sophorose, glycerol, and mixtures thereof. In varying embodiments, the population of basidiomycetous yeast cells comprises cells in the taxonomic class Microbotryomycetes. In varying embodiments, the population of basidiomycetous yeast cells comprises cells in the taxonomic order Sporidiobolales. In varying embodiments, the population of basidiomycetous yeast cells is from genera classified within the taxonomic order Sporidiobolales selected from the group consisting ofRhodosporidiobolus, Rhodotorula, andSporobolomyces. In varying embodiments, the population of basidiomycetous yeast cells comprisesRhodotorulacells. In varying embodiments, theRhodotorulacells areRhodotorula babjevae(syn.Rhodosporidium babjevae) cells. In varying embodiments, the population of basidiomycetous yeast cells comprises one or more species selected from the group consisting ofRhodotorula babjevae(syn.Rhodosporidium babjevae),Rhodotorula diobovata(syn.Rhodosporidium diobovatum),Rhodotorula kratochvilovae(syn.Rhodosporidium kratochvilovae),Rhodotorula paludigena(syn.Rhodosporidium paludigenum),Rhodotorulaaff.paludigena(syn.Rhodosporidiumaff.paludigenum), andRhodotorula dairenensis. In varying embodiments, the population of basidiomycetous yeast cells comprises one or more strains selected from the group consisting of:a)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain NRRL Y-67018 (also deposited as UCDFST 04-877),b)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain NRRL Y-67017 (also deposited as UCDFST 05-775),c)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 68-916.1,d)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 67-458,e)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 05-736,f)Rhodotorula diobovata(syn.Rhodosporidium diobovatum) strain UCDFST 04-830,g)Rhodotorula diobovata(syn.Rhodosporidium diobovatum) strain NRRL Y-67015 (also deposited as UCDFST 08-225),h)Rhodotorula kratochvilovae(syn.Rhodosporidium kratochvilovae) strain NRRL Y-67016 (also deposited as UCDFST 05-632),i)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain NRRL Y-67012 (also deposited asUCDFST 09-163),j)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain UCDFST 81-492,k)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain UCDFST 82-646.2,1)Rhodotorulaaff.paludigena(syn.Rhodosporidiumaff.paludigenumstrain NRRL Y-67009 (also deposited as UCDFST 81-84), andm)Rhodotorula dairenensisstrain NRRL Y-67011 (also deposited as UCDFST 68-257). In varying embodiments, the population of basidiomycetous yeast cells comprises one or more strains selected from the group consisting of: a)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 68-916.1, b)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 67-458, c)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 05-736, d)Rhodotorula diobovata(syn.Rhodosporidium diobovatum) strain UCDFST 04-830, e)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain UCDFST 81-492, and f)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain UCDFST 82-646.2. In varying embodiments, the cells do not comprisePseudohyphozyma bogoriensis(syn.Rhodotorula bogoriensis). In varying embodiments, the polyol lipids comprise polyol esters of fatty acids (PEFAs). In varying embodiments, the polyol lipid comprises an (R)-3-hydroxy fatty acid moiety comprising from 10 to 24 carbon atoms in length, e.g., from 14 to 24 carbon atoms in length. In varying embodiments, the polyol lipid comprises a sugar alcohol moiety esterified to the carboxyl end of the (R)-3-hydroxy fatty acyl moiety. In varying embodiments, the (R)-3-hydroxy fatty acyl moiety can be acetylated. In varying embodiments, the polyol lipid sugar alcohol moiety comprises non-esterified hydroxy groups that can be acetylated. In varying embodiments, the polyol lipid sugar alcohol moiety comprises a backbone selected from D-mannitol and D-arabitol. In varying embodiments, the polyol lipid sugar alcohol moiety does not comprise xylitol.

In varying embodiments of the yeast culture, the polyol lipid comprises one or more polyol esters of fatty acids (PEFAs) listed in Table 4.

In varying embodiments, the yeast culture comprises a polyol lipid (e.g. PEFA) profile comprising:i) at least about 20%, e.g., at least about 25%, 30%, 35%, 40%, up to about 45% of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12);ii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12); and/oriii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, up to about 20% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13).

In varying embodiments, the yeast culture comprises a polyol lipid (e.g. PEFA) comprising at least about 25%, e.g., at least about 30%, 35%, 40%, 45%, up to about 50% of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12).

In varying embodiments, the yeast culture comprises a polyol lipid (e.g. PEFA) comprising:i) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, up to about 18%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 3 acetylations (Molecular Formula: C32H56O12);ii) at least about 1%, e.g., at least about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, up to about 20%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C32H54O13);iii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13);iv) at least about 15%, e.g., at least about 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, up to about 50% of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12); and/orv) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, up to about 35% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12).

In varying embodiments, the yeast culture further comprises a polyol lipid (e.g. PEFA) comprising:vi) at least about 2%, e.g., at least about 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C34H56O14); and/orvii) at least about 1%, e.g., at least about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol (Molecular Formula: C36H60O14).

In varying embodiments, the yeast culture comprises a polyol lipid (e.g. PEFA) comprising:i) at least about 1%, e.g., at least about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13);ii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, up to about 45%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C34H56O14);iii) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C36H60O14);iv) at least about 15%, e.g., at least about 20%, 25%, 30%, 35%, 40%, up to about 45%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12); and/orv) at least about 15%, e.g., at least about 20%, 25%, 30%, 35%, 40%, up to about 45% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12).

In varying embodiments, the yeast culture comprises a polyol lipid (e.g. PEFA) comprising:i) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13);ii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, up to about 30%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C34H56O14);iii) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C36H60O14);iv) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12);v) at least about 15%, e.g., at least about 20%, 25%, 30%, 35%, 40%, up to about 45%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12); and/orvi) at least about 0.5%, e.g., at least about 1%, 2%, 3%, 4%, 5% up to about 6% of an acetylated C14:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C29H48O12).

In varying embodiments, the yeast culture comprises a polyol lipid (e.g. PEFA) comprising:i) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13);ii) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, up to about 25%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C34H56O14);iii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C36H60O14);iv) at least about 15%, e.g., at least about 20%, 25%, 30%, 35%, 40%, up to about 45%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12);v) at least about 35%, e.g., at least about 36%, 37%, 38%, 39%, 40%, up to about 45% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12); and/orvi) at least about 0.5%, e.g., at least about 1%, 2%, 3%, 4% up to about 5%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 3 acetylations (Molecular Formula: C30H52O12).

In varying embodiments, the yeast culture comprises a polyol lipid (e.g. PEFA) comprising:i) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, up to about 20%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C32H54O13);ii) at least about 45%, e.g., at least about 46%, 47%, 48%, 49%, up to about 50% of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12); and/oriii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, up to about 12% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12).

In varying embodiments, the one or more hydrophobic carbon sources are selected from the group consisting of oils, alkanes, fatty acids, fatty esters and mixtures thereof. In varying embodiments, the one or more polyol lipids are present in the medium in a form that can be harvested without solvent extraction.

Further provided is a bioreactor comprising a yeast culture, as described above and herein.

In a further aspect, provided are methods of producing one or more polyol lipids. In some embodiments, the methods comprise culturing a population of basidiomycetous yeast cells in a yeast cell culture comprising one or more hydrophilic (e.g., non-hydrophobic) carbon sources, whereby the basidiomycetous yeast cells produce one or more polyol esters of fatty acids (PEFAs). In varying embodiments, the culture does not comprise one or more hydrophobic carbon sources. In some embodiments, the methods comprise culturing a population of basidiomycetous yeast cells in a yeast cell culture comprising one or more hydrophilic (e.g., non-hydrophobic) carbon sources and one or more hydrophobic carbon sources, whereby the basidiomycetous yeast cells produce one or more polyol lipids (e.g., polyol esters of fatty acids (PEFAs)). In varying embodiments, additional hydrophilic carbon source is added, e.g., during any stage of growth including exponential growth, and after the population of basidiomycetous yeast cells reaches stationary phase. In varying embodiments, at least about 1 g/L polyol lipid (e.g., polyol esters of fatty acids (PEFAs)), e.g., at least about 2 g/L, 3 g/L, 4 g/L, 5 g/L, 6 g/L, 7 g/L, 8 g/L, 9 g/L, 10 g/L, 11 g/L, 12 g/L, 13 g/L, 14 g/L, 15 g/L, 20 g/L, 25 g/L, 30 g/L, 35 g/L, 40 g/L, 45 g/L, 50 g/L, 100 g/L, 150 g/L, 200 g/L, 250 g/L, 300 g/L, 350 g/L, 400 g/L polyol lipid, e.g., up to about 450 g/L of polyol lipid, is produced. In some embodiments, a polyol lipid yield in the range of about 0.5 g/L to about 30 g/L can be obtained from yeast cultures comprising in the range of about 10 g/L to about 250 g/L glucose. In some embodiments, a polyol lipid yield of at least about 14 g/L, e.g., from about 14 g/L to about 17 g/L, can be obtained from yeast cultures comprising about 50 g/L glucose. In varying embodiments, total polyol lipids are as measured after 1, 2, 3, 4, 5, 6, 7 or 8 days growth. In varying embodiments, the volume of the culture is at least about 0.5 L, 1 L, 2 L, 3 L, 4 L, 5 L, 10 L, 25 L, 50 L, 75 L, 100 L, 250 L, 500 L, 1000 L, or more. In varying embodiments, the culture comprises less than about 2% (w/v) nitrogen, e.g., in the range of about 0.005% (w/v) to about 2% (w/v) nitrogen. In varying embodiments, the nitrogen source is selected from the group consisting of ammonia, ammonium salt, nitrate, nitrite, nucleotides, nucleosides, proteins, peptides, amino acids, urea and its derivatives, and mixtures thereof. In varying embodiments, the culture comprises about 0.05% (w/v) ammonium chloride. In varying embodiments, the yeast culture comprises an iron salt, e.g., an iron salt selected from the group consisting of iron chloride (II) & (III), iron sulfate (II) & (III), iron nitrate (II) & (III), iron phosphate (II) & (III), fumarate, iron edetate, iron citrate, iron malate, iron oxalate, iron tartrate, iron succinate, iron acetate, in their anhydrous or their hydrated states, and organometallic complexes thereof. In some embodiments, the yeast culture comprises the iron salt in a concentration in the range of about 0.001 mg/L to about 10 g/L, e.g., from about 1 mg/L to about 0.5 g/L, e.g., from about 0.01 g/L to about 0.5 g/L. In varying embodiments, the culture comprises one or more hydrophilic carbon sources at a concentration in the range of about 0.2% (w/v) to about 70% (w/v), e.g., in the range of about 0.2% (w/v) to about 10%, 20%, 30%, 40%, 50% or 60% (w/v). In varying embodiments, the culture comprises a carbon to nitrogen ratio of about 5:1 to about 400:1, e.g., from about 10:1 to about 200:1, from about 20:1 to about 100:1, e.g., from about 25:1 to about 75:1, e.g., from about 30:1 to about 70:1, e.g., from about 40:1 to about 60:1, e.g., from about 30:1 to about 40:1, e.g., using a nitrogen source that can be consumed or utilized by the yeast cells. In some embodiments, the yeast culture is maintained at a temperature in the range of between about 24° C. to about 27° C. In varying embodiments, the hydrophilic carbon source is selected from the group consisting of monosaccharides, oligosaccharides, polysaccharides, sugar alcohols, polyols, polyol lipids (e.g., polyol esters of fatty acids (PEFAs)), organic acids, esters, aldehydes, ketones, alcohols, waste streams, plant materials, lignocellulosic hydrolysates, industrial co-products and mixtures thereof. In varying embodiments, the hydrophilic carbon source is a monosaccharide, an oligosaccharide or a polysaccharide and comprises one or more sugar moieties selected from the group consisting of glucose, sucrose, xylose, galactose, rhamnose, arabinose, mannose, cellobiose, galacturonic acid, lactose, sophorose, glycerol, and mixtures thereof. In varying embodiments, the population of basidiomycetous yeast cells comprises cells in the taxonomic class Microbotryomycetes. In varying embodiments, the population of basidiomycetous yeast cells comprises cells in the taxonomic order Sporidiobolales. In varying embodiments, the population of basidiomycetous yeast cells is from genera classified within the taxonomic order Sporidiobolales selected from the group consisting ofRhodosporidiobolus, Rhodotorula, andSporobolomyces. In varying embodiments, the population of basidiomycetous yeast cells comprisesRhodotorulacells. In varying embodiments, theRhodotorulacells areRhodotorula babjevae(syn.Rhodosporidium babjevae) cells. In varying embodiments, the population of basidiomycetous yeast cells comprises one or more species selected from the group consisting ofRhodotorula babjevae(syn.Rhodosporidium babjevae),Rhodotorula diobovata(syn.Rhodosporidium diobovatum),Rhodotorula kratochvilovae(syn.Rhodosporidium kratochvilovae),Rhodotorula paludigena(syn.Rhodosporidium paludigenum),Rhodotorulaaff.paludigena(syn.Rhodosporidiumaff.paludigenum), andRhodotorula dairenensis. In varying embodiments, the population of basidiomycetous yeast cells comprises one or more strains selected from the group consisting of:a)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain NRRL Y-67018 (also deposited as UCDFST 04-877),b)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain NRRL Y-67017 (also deposited as UCDFST 05-775),c)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 68-916.1,d)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 67-458,e)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 05-736,f)Rhodotorula diobovata(syn.Rhodosporidium diobovatum) strain UCDFST 04-830,g)Rhodotorula diobovata(syn.Rhodosporidium diobovatum) strain NRRL Y-67015 (also deposited as UCDFST 08-225),h)Rhodotorula kratochvilovae(syn.Rhodosporidium kratochvilovae) strain NRRL Y-67016 (also deposited as UCDFST 05-632),i)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain NRRL Y-67012 (also deposited asUCDFST 09-163),j)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain UCDFST 81-492,k)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain UCDFST 82-646.2,l)Rhodotorulaaff.paludigena(syn.Rhodosporidiumaff.paludigenumstrain NRRL Y-67009 (also deposited as UCDFST 81-84), andm)Rhodotorula dairenensisstrain NRRL Y-67011 (also deposited as UCDFST 68-257). In varying embodiments, the population of basidiomycetous yeast cells comprises one or more strains selected from the group consisting of: a)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 68-916.1, b)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 67-458, c)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 05-736, d)Rhodotorula diobovata(syn.Rhodosporidium diobovatum) strain UCDFST 04-830, e)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain UCDFST 81-492, and f)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain UCDFST 82-646.2. In varying embodiments, the cells do not comprisePseudohyphozyma bogoriensis(syn.Rhodotorula bogoriensis). In varying embodiments, the polyol lipid comprises one or more polyol esters of fatty acids (PEFAs). In varying embodiments, the polyol lipid comprises a 3-hydroxy fatty acid comprising from 10 to 24 carbon atoms in length, e.g., from 14 to 24 carbon atoms in length. In varying embodiments, the polyol lipid comprises a sugar alcohol moiety esterified to the carboxyl end of the 3-hydroxy fatty acyl moiety. In varying embodiments, the 3 hydroxyl fatty acyl moiety can be acetylated. In varying embodiments, the polyol lipid sugar alcohol moiety comprises non-esterified hydroxy groups that can be acetylated. In varying embodiments, the polyol lipid sugar alcohol moiety comprises a backbone selected from D-mannitol and D-arabitol. In varying embodiments, the polyol lipid sugar alcohol moiety does not comprise xylitol. In varying embodiments of the yeast culture, the polyol lipid comprises one or more polyol esters of fatty acids (PEFAs) listed in Table 4.

In varying embodiments, the methods produce a yeast culture comprising a polyol lipid (e.g. PEFA) comprising:i) at least about 20%, e.g., at least about 25%, 30%, 35%, 40%, up to about 45% of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12);ii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12); and/oriii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, up to about 20% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13).

In varying embodiments, the methods produce a yeast culture comprising a polyol lipid (e.g. PEFA) comprising at least about 25%, e.g., at least about 30%, 35%, 40%, 45%, up to about 50% of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12).

In varying embodiments, the methods produce a yeast culture comprising a polyol lipid (e.g. PEFA) comprising:i) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, up to about 18%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 3 acetylations (Molecular Formula: C32H56O12);ii) at least about 1%, e.g., at least about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, up to about 20%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C32H54O13);iii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13);iv) at least about 15%, e.g., at least about 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, up to about 50% of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12); and/orv) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, up to about 35% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12).

In varying embodiments, the methods produce a yeast culture comprising a polyol lipid (e.g. PEFA) comprising:vi) at least about 2%, e.g., at least about 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C34H56O14); and/orvii) at least about 1%, e.g., at least about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol (Molecular Formula: C36H60O14).

In varying embodiments, the methods produce a yeast culture comprising a polyol lipid (e.g. PEFA) comprising:i) at least about 1%, e.g., at least about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13);ii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, up to about 45%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C34H56O14);iii) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C36H60O14);iv) at least about 15%, e.g., at least about 20%, 25%, 30%, 35%, 40%, up to about 45%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12); and/orv) at least about 15%, e.g., at least about 20%, 25%, 30%, 35%, 40%, up to about 45% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12).

In varying embodiments, the methods produce a yeast culture comprising a polyol lipid (e.g. PEFA) comprising:i) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13);ii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, up to about 30%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C34H56O14);iii) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C36H60O14);iv) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12);v) at least about 15%, e.g., at least about 20%, 25%, 30%, 35%, 40%, up to about 45%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12); and/orvi) at least about 0.5%, e.g., at least about 1%, 2%, 3%, 4%, 5% up to about 6% of an acetylated C14:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C29H48O12).

In varying embodiments, the methods produce a yeast culture comprising a polyol lipid (e.g. PEFA) comprising:i) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13);ii) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, up to about 25%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C34H56O14);iii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C36H60O14);iv) at least about 15%, e.g., at least about 20%, 25%, 30%, 35%, 40%, up to about 45%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12);v) at least about 35%, e.g., at least about 36%, 37%, 38%, 39%, 40%, up to about 45% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12); and/orvi) at least about 0.5%, e.g., at least about 1%, 2%, 3%, 4% up to about 5%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 3 acetylations (Molecular Formula: C30H52O12).

In varying embodiments, the methods produce a yeast culture comprising a polyol lipid (e.g. PEFA) comprising:i) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, up to about 20%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C32H54O13);ii) at least about 45%, e.g., at least about 46%, 47%, 48%, 49%, up to about 50% of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12); and/oriii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, up to about 12% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12).

In varying embodiments of the methods of producing, the one or more hydrophobic carbon sources are selected from the group consisting of oils, alkanes, fatty acids, fatty esters and mixtures thereof. In varying embodiments, the method does not comprise actively pH-adjusting the culture. In some embodiments, the culture volume to container volume ratio is 1:3 or greater, e.g., 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10 or greater. In some embodiments, the container is a baffled flask, e.g., covered in such a manner to allow air/oxygen exchange, e.g., with a foam stopper. In some embodiments, the culture comprises dissolved oxygen at a concentration of a least about 10%, e.g., at least about 15%, 20%, 25%, 30%, 35%, 40%, 45%, and up to 60%, 70%, 80%, 90%, 100% dissolved oxygen, e.g., from about 15% to about 80%, e.g., from about 20% to about 60% dissolved oxygen. In varying embodiments, the methods further comprise purifying and/or isolating the polyol lipid. In varying embodiments, the polyol lipid does not need to be purified from a hydrophobic carbon source. In varying embodiments, the polyol lipid can be purified and/or isolated without cell lysis, e.g., directly from the media. In some embodiments, the yeast cells secrete the one or more polyol lipids into the medium in a form that can be harvested without solvent extraction.

In a related aspect, provided are polyol lipid compositions produced according to the methods described above and herein. In another aspect, provided are compositions comprising one or more polyol esters of fatty acids (PEFAs) listed in Table 4. In varying embodiments, the polyol lipid compositions are selected from the group consisting of a cleanser, a detergent, a surfactant (e.g., for recovery of oil), a wetting agent, an antifoam agent, an emulsifier, an emollient, a dispersant (e.g., for cleanup of oil including spilled petroleum), a humectant, an antibacterial agent, an antiviral agent, an antifungal agent, a spermicide, an insecticide, a lubricant, an adhesive, a crystal modifier, an instantizer, a viscosity modifier, a mixing/blending aid, a release agent, a cream, a foam, a mousse, a lotion, a balm, and an ointment. In varying embodiments, polyol lipid composition is free of any hydrophobic carbon source. In varying embodiments, the method is performed as a batch, fed batch or continuous-feed process.

In varying embodiments, the composition comprises a polyol lipid (e.g. PEFA) comprising:i) at least about 20%, e.g., at least about 25%, 30%, 35%, 40%, up to about 45% of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12);ii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12); and/oriii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, up to about 20% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13).

In varying embodiments, the composition comprises a polyol lipid (e.g. PEFA) comprising at least about 25%, e.g., at least about 30%, 35%, 40%, 45%, up to about 50% of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12).

In varying embodiments, the composition comprises a polyol lipid (e.g. PEFA) comprising:i) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, up to about 18%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 3 acetylations (Molecular Formula: C32H56O12);ii) at least about 1%, e.g., at least about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, up to about 20%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C32H54O13);iii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13);iv) at least about 15%, e.g., at least about 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, up to about 50% of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12); and/orv) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, up to about 35% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12).

In varying embodiments, the compositions further comprise a polyol lipid (e.g. PEFA) comprising:vi) at least about 2%, e.g., at least about 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C34H56O14); and/orvii) at least about 1%, e.g., at least about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol (Molecular Formula: C36H60O14).

In varying embodiments, the composition comprises a polyol lipid (e.g. PEFA) comprising:i) at least about 1%, e.g., at least about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13);ii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, up to about 45%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C34H56O14);iii) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C36H60O14);iv) at least about 15%, e.g., at least about 20%, 25%, 30%, 35%, 40%, up to about 45%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12); and/orv) at least about 15%, e.g., at least about 20%, 25%, 30%, 35%, 40%, up to about 45% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12).

In varying embodiments, the composition comprises a polyol lipid (e.g. PEFA) comprising:i) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13);ii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, up to about 30%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C34H56O14);iii) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, up to about 25%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C36H60O14);iv) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12);v) at least about 15%, e.g., at least about 20%, 25%, 30%, 35%, 40%, up to about 45%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12); and/orvi) at least about 0.5%, e.g., at least about 1%, 2%, 3%, 4%, 5% up to about 6% of an acetylated C14:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C29H48O12).

In varying embodiments, the composition comprises a polyol lipid (e.g. PEFA) comprising:i) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C34H58O13);ii) at least about 10%, e.g., at least about 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, up to about 25%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C34H56O14);iii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, up to about 12%, of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 5 acetylations (Molecular Formula: C36H60O14);iv) at least about 15%, e.g., at least about 20%, 25%, 30%, 35%, 40%, up to about 45%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12);v) at least about 35%, e.g., at least about 36%, 37%, 38%, 39%, 40%, up to about 45% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12); and/orvi) at least about 0.5%, e.g., at least about 1%, 2%, 3%, 4% up to about 5%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 3 acetylations (Molecular Formula: C30H52O12).

In varying embodiments, the composition comprises a polyol lipid (e.g. PEFA) comprising:i) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, 15%, up to about 20%, of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-mannitol comprising 4 acetylations (Molecular Formula: C32H54O13);ii) at least about 45%, e.g., at least about 46%, 47%, 48%, 49%, up to about 50% of an acetylated C16:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C31H52O12); and/oriii) at least about 5%, e.g., at least about 6%, 7%, 8%, 9%, 10%, up to about 12% of an acetylated C18:0 (R)-3-hydroxy fatty acid esterified to D-arabitol comprising 4 acetylations (Molecular Formula: C33H56O12).

In a further aspect, provided is a yeast cell of a yeast strain selected from the group consisting of:a)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain NRRL Y-67018 (also deposited as UCDFST 04-877),b)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain NRRL Y-67017 (also deposited as UCDFST 05-775),c)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 68-916.1,d)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 67-458,e)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 05-736,f)Rhodotorula diobovata(syn.Rhodosporidium diobovatum) strain UCDFST 04-830,g)Rhodotorula diobovata(syn.Rhodosporidium diobovatum) strain NRRL Y-67015 (also deposited as UCDFST 08-225),h)Rhodotorula kratochvilovae(syn.Rhodosporidium kratochvilovae) strain NRRL Y-67016 (also deposited as UCDFST 05-632),i)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain NRRL Y-67012 (also deposited asUCDFST 09-163),j)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain UCDFST 81-492,k)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain UCDFST 82-646.2,1)Rhodotorulaaff.paludigena(syn.Rhodosporidiumaffpaludigenumstrain NRRL Y-67009 (also deposited as UCDFST 81-84), andm)Rhodotorula dairenensisstrain NRRL Y-67011 (also deposited as UCDFST 68-257). In another aspect, provided is a yeast cell of a yeast strain selected from the group consisting of: a)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 68-916.1; b)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 67-458; c)Rhodotorula babjevae(syn.Rhodosporidium babjevae) strain UCDFST 05-736; d)Rhodotorula diobovata(syn.Rhodosporidium diobovatum) strain UCDFST 04-830; e)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain UCDFST 81-492; and f)Rhodotorula paludigena(syn.Rhodosporidium paludigenum) strain UCDFST 82-646.2.

Further provided is a population of yeast cells comprising one or more yeast strains as described above and herein. Further provided is a yeast culture comprising one or more yeast strains as described above and herein.

Definitions

The term “population of yeast cells” refers to two or more yeast cells.

The term “hydrophobic carbon source” refers to an organic compound that is insoluble in water or has a solubility in water of less than 1 g/L.

The term “hydrophilic carbon source refers to an organic compound that is soluble in water at concentrations greater than 1 g/L.

