Development of yeast strains for ethanol production.

Abstract

Disclosed herein are yeast strains and their derivatives, as well as compositions comprising the yeast strains for use in ethanol production.The disclosure also relates to a method for producing ethanol from biomass using the yeast strains and compositions.In particular, the yeast strains produce less glycerol and more ethanol than strains and products currently used in ethanol production processes, and have high temperature tolerance and high fermentation rates.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This patent application claims priority to U.S. Provisional Application No. 63 / 170,884, filed April 5, 2021, the entire contents of which are incorporated herein by reference.

[0002] Technical Field

[0002] The present disclosure relates to yeast strains and their derivatives, as well as compositions comprising the yeast strains for use in ethanol production. The present disclosure also relates to methods for producing ethanol from biomass using the yeast strains and compositions. In particular, the yeast strains produce less glycerol and more ethanol than strains and products currently used in ethanol production processes, and have high temperature and organic acid tolerance and high fermentation rates. [Background technology]

[0003] Introduction

[0003] Ethanol can be produced from living organisms using various biochemical pathways that are intrinsic to the organism. Ethanol produced from living organisms is called bioethanol, and is therefore distinct from ethanol produced by purely chemical methods. Bioethanol is produced commercially and can be used as a liquid fuel for internal combustion engines (fuel ethanol), as a raw material for industrial products (industrial ethanol), or as a component of alcoholic beverages (potable ethanol).

[0004] Organisms such as yeast produce bioethanol. Selected yeasts are advantageous for ethanol production, allowing optimization of production processes and profitability. Yeasts used for commercial production of ethanol require several characteristics, including adequate ethanol metabolic yield, adequate ethanol tolerance, acceptable by-product yield, adequate fermentation rate, and some degree of thermotolerance during fermentation. Yeasts of the genus Saccharomyces exhibit many of these characteristics required for commercial production of ethanol. Commercially available yeast products containing Saccharomyces include Ethanol Red® (Fermentis®), Thermosacc® (Lallemand®), Angel Super Alcohol® (Angel®), and Fali (AB Mauri®). Although commercially available yeast strains and products have advantageous characteristics for ethanol production, there is an increasing need to improve the efficiency of ethanol production to reduce production costs.

[0005]

[0005] Thus, there is a need for new and improved strains of Saccharomyces for producing new and improved yeast products that can improve the efficiency of commercial ethanol production by providing higher sugar to ethanol conversion, higher temperature tolerance, higher tolerance to fermentation inhibitors, and faster fermentation rates than currently commercially used yeast strains and products. Summary of the Invention

[0006] In one aspect, the disclosure provides: (a) Saccharomyces strain Y1912, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68003; (b) Saccharomyces strain Y1913, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68004; (c) Saccharomyces strain Y1914, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68005; (d) Saccharomyces strain Y1915, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68006; (e) Saccharomyces strain Y1923, which is a representative specimen of the strains deposited under NRRL Patent Deposit Designation No. Y-68007; (f) Saccharomyces strain Y1927, which is a representative specimen of the strains deposited under NRRL Patent Deposit Designation No. Y-68008; and (g) Saccharomyces strain Y1929, which is a representative specimen of the strains deposited under NRRL Patent Deposit Designation No. Y-68009.

[0007] In a further aspect, the disclosure provides a method for the preparation of a Saccharomyces strain, comprising: (a) Saccharomyces strain Y1912, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68003; (b) Saccharomyces strain Y1913, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68004; (c) Saccharomyces strain Y1914, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68005; (d) Saccharomyces strain Y1915, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68006; (e) Saccharomyces strain Y1923, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68007; (f) Saccharomyces strain Y1927, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68008; and (g) Saccharomyces strain Y1929, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68009. In one embodiment, the yeast strain or derivative comprises one or more defining characteristics selected from: (a) a higher ethanol yield than the Saccharomyces Y1609 strain under the same fermentation conditions; (b) increased temperature tolerance compared to the Saccharomyces Y1609 strain at temperatures between 32° C. and 36° C.; (c) a lower glycerol yield than the Saccharomyces Y1609 strain under the same fermentation conditions; (d) increased organic acid tolerance compared to the Saccharomyces Y1609 strain at a pH between about 4.0 and about 5.2 in the presence of an organic acid; and (e) an increased fermentation rate compared to the Saccharomyces Y1609 strain under the same fermentation conditions. In another embodiment, the yeast strain or derivative produces at least about 0.5% more ethanol after 50 hours of fermentation compared to the Saccharomyces cerevisiae Y1609 strain. In another embodiment, the yeast strain or derivative produces at least about 2% less glycerol after 50 hours of fermentation compared to Saccharomyces cerevisiae Y1609 strain. In another embodiment, the yeast strain or derivative has a fermentation rate at least about 2% higher than Saccharomyces cerevisiae Y1609 strain after 24 hours of fermentation. In another embodiment, the temperature is 32°C.In another embodiment, the yeast strain or derivative produces at least about 5% more ethanol after 50 hours of fermentation compared to Saccharomyces cerevisiae Y1609 strain. In another embodiment, the yeast strain or derivative produces at least about 3% less glycerol after 50 hours of fermentation compared to Saccharomyces cerevisiae Y1609 strain. In another embodiment, the yeast strain or derivative has a fermentation rate at least about 10% higher than Saccharomyces cerevisiae Y1609 strain after 24 hours of fermentation. In another embodiment, the temperature is 36° C. In another embodiment, the organic acid comprises lactic acid, acetic acid, succinic acid, citric acid, malic acid, fumaric acid, or a combination thereof.

[0008] Another aspect of the present disclosure provides a method for producing a derivative of a Saccharomyces yeast strain described herein, the method comprising the steps of: (a) providing (i) a first yeast strain selected from Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, and derivatives thereof, and (ii) a second yeast strain belonging to the phylogenetic clade of Saccharomyces sensu stricto; (b) inducing sporulation of the first yeast strain and the second yeast strain; (c) screening and selecting spores from the first yeast strain and spores from the second yeast strain; (d) crossing the selected spores of the first yeast strain with the selected spores of the second yeast strain; and (e) screening or selecting a derivative strain. In one embodiment, step (c) comprises screening or selecting spores exhibiting one or more defining characteristics of Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or derivatives thereof. In another embodiment, step (e) comprises screening or selecting hybrids exhibiting one or more defining characteristics of Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or derivatives thereof.

[0009] Another aspect of the present disclosure provides a method for producing derivatives of the Saccharomyces yeast strains described herein, comprising the steps of: (a) providing (i) a first yeast strain selected from Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, and derivatives thereof, and (ii) one or more additional yeast strains belonging to the phylogenetic clade of Saccharomyces sensu stricto; (b) inducing sporulation of the first yeast strain and the one or more additional yeast strains to produce spores; (c) mixing all the spores from step (b) to allow for spore crossing; and (d) screening or selecting the derivative strains. In one embodiment, step (d) comprises screening or selecting hybrids that exhibit one or more defining characteristics of Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929.

[0010]

[0010] Another aspect of the present disclosure provides a mutant yeast of the yeast strain described herein or a derivative described herein.

[0011] Another aspect of the present disclosure provides a method of producing a mutant yeast as described herein, wherein the mutant yeast is mutated by contacting the yeast strain with a mutagen. In one embodiment, the mutagen is ethyl methanesulfonate (EMS), ultraviolet light (UV), X-rays, methyl methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine mustard, 2-methoxy-6-chloro-9[3-(ethyl-2-chloroethyl)aminopropylamino]acridine·2 (ICR-170), or nitrogen mustard.

[0012] Another aspect of the disclosure provides a method of producing a mutant yeast as described herein, wherein the mutant yeast is mutated by contacting the derivative with a mutagen. In one embodiment, the mutagen is ethyl methanesulfonate (EMS), ultraviolet light (UV), X-rays, methyl methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine mustard, 2-methoxy-6-chloro-9[3-(ethyl-2-chloroethyl)aminopropylamino]acridine·2 (ICR-170), or nitrogen mustard.

[0013]

[0013] Another aspect of the present disclosure provides evolved yeasts of the yeast strains described herein or derivatives described herein.

[0014]

[0014] Another aspect of the present disclosure provides a method of producing an evolved yeast as described herein, wherein the evolution is induced by applying selective pressure to the yeast strain.

[0015]

[0015] Another aspect of the present disclosure provides a method of producing an evolved yeast as described herein, wherein the evolution is induced by applying selective pressure to the derivatives.

[0016]

[0016] Another aspect of the present disclosure provides a genetically modified yeast of the yeast strain described herein or a derivative described herein. In one embodiment, the nucleic acid sequence of the genetically modified yeast is altered using gene editing.

[0017]

[0017] Another aspect of the disclosure provides a recombinant yeast of a yeast strain described herein or a derivative described herein. In one embodiment, the recombinant yeast comprises a modification to suppress expression of a gene, to enhance expression of a gene, to introduce a gene, or to delete a gene.

[0018] Another aspect of the present disclosure is a method for producing ethanol from a substrate by contacting the substrate with a fermenting organism, the fermenting organism being selected from the group consisting of: (a) Saccharomyces strain Y1912, a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68003, or a derivative thereof; (b) Saccharomyces strain Y1913, a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68004, or a derivative thereof; (c) Saccharomyces strain Y1914, a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68005, or a derivative thereof; (d) Saccharomyces strain Y1915, a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68006, or a derivative thereof; (e) Saccharomyces strain Y1923, or a derivative thereof, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68007; (f) Saccharomyces strain Y1927, or a derivative thereof, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68008; and (g) Saccharomyces strain Y1929, or a derivative thereof, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68009. In one embodiment, the substrate comprises or is derived from sugar cane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, cassava, or a combination thereof. In another embodiment, the yeast strain comprises one or more defining characteristics selected from: (a) a higher ethanol yield than the Saccharomyces Y1609 strain under the same fermentation conditions; (b) an increased temperature tolerance compared to the Saccharomyces Y1609 strain at temperatures between 32° C. and 36° C.; (c) a lower glycerol yield than the Saccharomyces Y1609 strain under the same fermentation conditions; (d) an increased organic acid tolerance compared to the Saccharomyces Y1609 strain at a pH between about 4.0 and about 5.2 in the presence of an organic acid; and (e) an increased fermentation rate compared to the Saccharomyces Y1609 strain under the same fermentation conditions. In another embodiment, the ethanol is used for fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof. In another embodiment, the ethanol is produced using starch.In another embodiment, ethanol is produced using simultaneous saccharification and fermentation (SSF) or continuous fermentation. In another embodiment, ethanol is produced using sugars. In another embodiment, ethanol is produced using batch fermentation or continuous fermentation. In another embodiment, ethanol is produced using lignocellulosic sugars. In another embodiment, ethanol is produced using simultaneous saccharification and fermentation (SSF) or separate hydrolysis and fermentation (SHF).

[0019] Another aspect of the disclosure provides a composition comprising a yeast strain as described herein or a derivative as described herein and one or more components selected from surfactants, emulsifiers, thickeners, swelling agents, protectants, and antioxidants. In one embodiment, the composition comprises one or more defining characteristics selected from: (a) a higher ethanol yield than Fali under the same fermentation conditions; (b) an increased temperature tolerance compared to Fali at temperatures between 32° C. and 36° C.; (c) a lower glycerol yield than Fali under the same fermentation conditions; (d) an increased organic acid tolerance compared to Fali at a pH between about 4.0 and about 5.2 in the presence of an organic acid; and (e) an increased fermentation rate compared to Fali under the same fermentation conditions.

[0020] Another aspect of the disclosure provides a method for producing ethanol from biomass by contacting the biomass with a composition described herein. In one embodiment, the ethanol is used for fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof. In another embodiment, the ethanol is produced using starch. In another embodiment, the ethanol is produced using simultaneous saccharification and fermentation (SSF) or continuous fermentation. In another embodiment, the ethanol is produced using sugars. In another embodiment, the ethanol is produced using batch or continuous fermentation. In another embodiment, the ethanol is produced using lignocellulosic sugars. In another embodiment, the ethanol is produced using simultaneous saccharification and fermentation (SSF) or separate hydrolysis and fermentation (SHF).

[0021] Another aspect of the present disclosure is a method for producing a fermented product from a substrate by contacting the substrate with a fermenting organism, the fermenting organism being selected from the group consisting of: (a) Saccharomyces strain Y1912, a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68003, or a derivative thereof; (b) Saccharomyces strain Y1913, a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68004, or a derivative thereof; (c) Saccharomyces strain Y1914, a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68005, or a derivative thereof; (d) Saccharomyces strain Y1915, a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68006, or a derivative thereof; (e) Saccharomyces strain Y1923, or a derivative thereof, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68007; (f) Saccharomyces strain Y1927, or a derivative thereof, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68008; and (g) Saccharomyces strain Y1929, or a derivative thereof, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68009. In one embodiment, the substrate comprises or is derived from sugar cane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, cassava, or a combination thereof. In another embodiment, the fermentation product is ethanol. In another embodiment, the ethanol is used as fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof. In another embodiment, the fermentation product is produced using batch fermentation, continuous fermentation, simultaneous saccharification and fermentation (SSF), or separated hydrolysis and fermentation (SHF).

[0022]

[0022] The present disclosure provides other aspects and embodiments that will become apparent in light of the following detailed description and accompanying drawings. [Brief description of the drawings]

[0023] [Figure 1A]

[0023] Figure 1 shows a method for generating new yeast strains. Figure 1A shows directed mating. [Figure 1B] FIG. 1B shows mass mating. [Figure 1C] FIG. 1C shows the directed evolution of yeast strains. [Figure 1D] FIG. 1D shows the mutagenesis of the yeast strain. [Figure 2A]

[0024] Figure 2 shows the percent difference from the control (Y1609) at the end of 50 hours of fermentation without exogenous addition of organic acids and under normal fermentation temperature (32°C) in proprietary synthetic corn starch medium (SCM) at pH 5.2. The percent difference from the control in ethanol at the end of fermentation (Figure 2A), glycerol at the end of fermentation (Figure 2B), and initial fermentation rate of fermentation (Figure 2C) are shown. [Figure 2B] FIG. 2 shows the percent difference from the control (Y1609) at normal fermentation temperature (32° C.) in a proprietary synthetic corn starch medium (SCM) at pH 5.2 without exogenous addition of organic acids at the end of 50 hours of fermentation. [Figure 2C] FIG. 2 shows the percent difference from the control (Y1609) at normal fermentation temperature (32° C.) in a proprietary synthetic corn starch medium (SCM) at pH 5.2 without exogenous addition of organic acids at the end of 50 hours of fermentation. [Figure 3A]

[0025] Figure 3 shows the percent difference from the control at the end of 50 hours of fermentation under normal fermentation temperature (32°C) in proprietary synthetic corn starch medium (SCM) at pH 4.0 with exogenous addition of 1% w / v lactic acid and 0.05% w / v acetic acid. The percent difference from the control in ethanol at the end of fermentation (Figure 3A), glycerol at the end of fermentation (Figure 3B), and initial fermentation rate of fermentation (Figure 3C) are shown. [Figure 3B]FIG. 3 shows the percent difference from the control at normal fermentation temperature (32° C.) in a proprietary synthetic corn starch medium (SCM) at pH 4.0 with exogenous addition of 1% w / v lactic acid and 0.05% w / v acetic acid at the end of 50 hours of fermentation. [Figure 3C] FIG. 3 shows the percent difference from the control at normal fermentation temperature (32° C.) in a proprietary synthetic corn starch medium (SCM) at pH 4.0 with exogenous addition of 1% w / v lactic acid and 0.05% w / v acetic acid at the end of 50 hours of fermentation. [Figure 4A]

[0026] Figure 4 shows the percent difference compared to the control (Y1609) at the end of 50 hours of fermentation without exogenous addition of organic acids and at high fermentation temperature (36°C) in proprietary synthetic corn starch medium (SCM) at pH 5.2. The percent difference compared to the control in ethanol at the end of fermentation (Figure 4A), glycerol at the end of fermentation (Figure 4B), and initial fermentation rate of fermentation (Figure 4C) are shown. [Figure 4B] FIG. 4 shows the percent difference compared to the control (Y1609) at the end of 50 hours of fermentation without exogenous addition of organic acids in a proprietary synthetic corn starch medium (SCM) at pH 5.2 and under high fermentation temperature (36° C.). [Figure 4C] FIG. 4 shows the percent difference compared to the control (Y1609) at the end of 50 hours of fermentation without exogenous addition of organic acids in a proprietary synthetic corn starch medium (SCM) at pH 5.2 and under high fermentation temperature (36° C.). [Figure 5A]

[0027] Figure 5 shows the percent difference compared to the control (Y1609) under high fermentation temperature (36°C) in proprietary synthetic corn starch medium (SCM) at pH 4.0 with exogenous addition of 1% w / v lactic acid and 0.05% w / v acetic acid at the end of the 50 hour fermentation. The percent difference compared to the control in ethanol at the end of the fermentation (Figure 5A), glycerol at the end of the fermentation (Figure 5B), and initial fermentation rate of the fermentation (Figure 5C) are shown. [Figure 5B] FIG. 5 shows the percent difference compared to the control (Y1609) under high fermentation temperature (36° C.) in proprietary synthetic corn starch medium (SCM) at pH 4.0 with exogenous addition of 1% w / v lactic acid and 0.05% w / v acetic acid at the end of 50 hours of fermentation. [Figure 5C] FIG. 5 shows the percent difference compared to the control (Y1609) under high fermentation temperature (36° C.) in proprietary synthetic corn starch medium (SCM) at pH 4.0 with exogenous addition of 1% w / v lactic acid and 0.05% w / v acetic acid at the end of 50 hours of fermentation. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] Detailed Description

[0028] Described herein are fermenting organisms that include defining characteristics including higher ethanol yield, lower glycerol yield, higher temperature and inhibitor tolerance, and higher fermentation rate, compared to current industry standard yeasts used in yeast products such as Fali under the same fermentation conditions. Also described herein are Saccharomyces yeast strains with improved properties compared to the yeast strains used in Fali. The disclosure relates to methods of producing yeast products from the yeast strains. The disclosure also relates to improved methods of producing ethanol from various fermentable biomass materials using the fermenting organisms described herein.

[0025] 1.Definition

[0029] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. In case of conflict, the present document including definitions shall prevail. Preferred methods and materials are described below, but similar or equivalent methods and materials to those described herein may be used to practice or test the present invention. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are merely illustrative and are not intended to be limiting.

[0026]

[0030] As used herein, the terms "comprise(s)", "include(s)", "having", "has", "can", "contain(s)", and variations thereof are intended to be open-ended transitional phrases, terms, or words that do not exclude the possibility of additional acts or structures. The singular forms "a", "and", and "the" include the plural unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments that "comprise", "consist of", and "consist essentially of" the embodiments or elements presented herein, whether or not expressly stated.

[0027]

[0031] For the recitation of numerical ranges herein, each intervening number is expressly contemplated with the same degree of precision. For example, for the range of 6 to 9, in addition to 6 and 9, 7 and 8 are also contemplated. For the range of 6.0 to 7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are expressly contemplated. For the range of 1 to 5, in addition to 1 and 5, the numbers 2, 3, and 4 are contemplated.

[0028]

[0032] The term "about" or "approximately" as used herein, as applied to one or more values ​​of interest, refers to a value similar to a stated reference value or within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined, such as the limitations of the measurement system. In certain embodiments, the term "about" refers to a range of values ​​that falls within 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater or less) of the stated reference value, unless otherwise specified or clear from the context (except where such value exceeds 100% of the possible value). Alternatively, "about" may mean within or more than 3 standard deviations, as is customary in the art. Alternatively, such as with respect to biological systems or processes, the term "about" can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold of a value.

[0029]

[0033] The term "biomass" as used herein refers to any organic matter of plant origin that can be a carbohydrate source after conversion. Preferably, the biomass can be derived from agricultural or food processing products and / or coproducts. In particular, the biomass is rich in sucrose or starch and is selected from or derived from, for example, sorghum, sugarcane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, cassava, or mixtures thereof.

[0030]

[0034] As used herein, "DNA combination" between yeast strains refers to combining all or part of the genome of yeast strains.The DNA combination between yeast strains can be by any method suitable for combining the DNA of at least two yeast cells, and can include, for example, mating method, including sporulation of yeast strains to generate haploid cells, followed by crossing or mating of compatible haploid cells; cytoduction; or cell fusion, such as protoplast fusion.

[0031]

[0035] As used herein, a "derivative" is a yeast strain derived from a yeast strain disclosed herein (e.g., a phylogenetic clade of the genus Saccharomyces or Saccharomyces sensu stricto), including by crossing between yeast strains, mutagenesis, recombinant DNA techniques, breeding, cell fusion, or cytoplasmic transfer. A derivative strain may be a direct progeny (i.e., the product of a cross between a strain of the invention and another strain or itself).

[0032]

[0036] As used herein, "ethanol yield from glucose" is the yield of ethanol that can be achieved from glucose. In one embodiment, the ethanol yield from glucose is represented by the following description: one molecule of glucose produces two molecules of ethanol and two molecules of carbon dioxide. In another embodiment, the ethanol yield from glucose is represented by the chemical formula: CH 12 O6→2C2H5OH+2CO2, where C6H 12 O6 is the chemical formula for glucose, CHOH is the chemical formula for ethanol, and CO2 is the chemical formula for carbon dioxide. In another embodiment, the ethanol yield from glucose is expressed on a mass basis, with 1.0 gram of glucose producing 0.511 grams of ethanol and 0.489 grams of carbon dioxide. The highest ethanol yield from glucose is 2 molecules of ethanol from 1 molecule of glucose. The highest ethanol yield on a mass basis is 0.511 grams of ethanol from 1 gram of glucose.

[0033]

[0037] The term "control" is used interchangeably with "Y1609" when discussing yeast strains and with "Fali" when discussing yeast products. The Y1609 strain is used in the production of the yeast product Fali. Fali can be produced as an active dry yeast product, a crumble yeast product, and as a liquid yeast product for use in the fermentation of substrates to fuel ethanol, industrial ethanol, and potable ethanol. Fali is particularly suitable for use in the fermentation of sucrose, glucose, and fructose sugar-containing biomass and the fermentation of sugars liberated from starch-containing biomass after liberation of the sugars by the action of enzymes or chemical processes. Fali can be used in batch, continuous, and simultaneous saccharification and fermentation (SSF) of starch substrates. Fali has high tolerance to free glucose, high cell numbers (>20x10 9 It has a pH of 10.5-12.5 (cells / g), moderate ethanol and temperature tolerance, and moderate organic acid tolerance. It rehydrates well in direct pitch applications and can be used in combination with glucoamylase and alpha amylase enzyme systems. Fali performs optimally in the pH range of 4.0-5.0, but can also ferment well in the pH range of 3.5-6.0. The optimum fermentation temperature for Fali depends on the stresses present (e.g. organic acids, ethanol, pH), but generally ferments well in a temperature range of about 32°C. Fali is available commercially from AB Mauri®.

[0034]

[0038] The term "fermentation medium" refers to an environment in which fermentation is carried out using a fermenting organism and contains a fermentable substrate, i.e., a carbohydrate source (e.g., glucose) that can be metabolized by the fermenting organism into a desired fermentation product, such as ethanol. The fermentation medium may contain fermentation nutrients for the fermenting organism. Fermentation nutrients are widely used in the field of fermentation and include nitrogen sources such as ammonia urea, vitamins, minerals, or combinations thereof. A "feed" is a fermentation medium, although a feed may have a different composition than the fermentation medium.

[0035]

[0039] "High yield ethanol production" as used herein means ethanol production by fermentation where the ethanol yield approaches that from glucose. High yield ethanol production requires limiting by-products such as glycerol and yeast growth during fermentation.

[0036]

[0040] The terms "improved," "enhanced," "enhanced," or "better" as used herein refer to an enhancement or improvement of a particular property or trait as compared to other similar organisms, controls, or wild-type organisms. Typically, this is a beneficial fermentation-related trait.

[0037]

[0041] The term "inoculum" is intended to mean the amount of microorganism added to a main fermenter to initiate a fermentation process. In the case of a fermentation process using a seed fermenter, the inoculum is typically a pre-culture in an amount equivalent to 5-20% of the volume of the main fermenter.

[0038]

[0042] The term "isolated" refers to a substance in a form or environment that is not found in nature. Non-limiting examples of isolated substances include: (1) any non-naturally occurring substance; (2) any substance, including but not limited to, any enzyme, mutant, nucleic acid, protein, peptide, or cofactor, that is at least partially removed from one or more or all of the natural components associated with it in nature; (3) any substance that has been modified by the hand of man relative to the substance found in nature; or (4) any substance that has been modified by increasing the amount of the substance relative to other components associated with it in nature (e.g., recombinant production in yeast, multiple copies of the gene encoding the substance; and use of a stronger promoter than the promoter naturally associated with the gene encoding the substance). An isolated substance can be present in a fermentation broth sample. For example, yeast can be genetically modified to express a particular polypeptide. The fermentation broth from the yeast contains the isolated polypeptide.

[0039]

[0043] The term "low pH" as used herein refers to a pH of about 2.5 to about 4.5. A low pH is preferably less than about 4.5. The term "normal pH" as used herein refers to a pH of about 4.0 to about 6.0. A normal pH is preferably about 5.0.

[0040]

[0044] As used herein, the term "main fermentor" is used for the final fermentor used in a fermentation process to produce a fermentation product, the final fermentor in which the fermentation product of interest is produced.

[0041]

[0045] The term "parental" or "parent" strain refers to the yeast strain from which a derivative strain is derived. In some embodiments, a derivative may be a parent.

[0042]

[0046] The term "pre-culture" is understood as an actively growing liquid culture of a microorganism (i.e. yeast) used to inoculate the main fermenter. Actively growing is intended to mean that the culture is at a stage where the microorganism is increasing in cell number. A pre-culture is generally used as an inoculum to avoid or reduce the lag phase in the main fermenter. In fermentation to produce ethanol, the cells in the pre-fermenter are typically habituated. The idea is not to produce yeast biomass, as the carbon in the biomass reduces the carbon going to ethanol. There is fermentative growth, but it is not desirable to promote aerobic growth (e.g., using a seed fermenter). Alternatively, yeast can be added directly to the main fermenter by "direct pitch".

[0043]

[0047] As used herein, the terms "characteristics" and "defining features" of the Saccharomyces cerevisiae strains detailed herein include at least increased ethanol yield as compared to a control (i.e., Fali or Y1609) under the same process conditions. Other "characteristics" and "defining features" include, among others, increased temperature tolerance, increased fermentation rate, increased organic acid tolerance, increased ethanol production, and decreased glycerol production. For example, the fermenting organisms described herein used in the methods described herein can have one or more of the above "characteristics" and "defining features."

[0044]

[0048] The term "pre-fermenter" is intended to mean a fermenter in which a pre-culture is formed by fermenting microorganisms until the yeast is activated and prepared for inoculation of the main fermenter. If a pre-fermenter is not used, a "direct pitch" is used.

[0045]

[0049] As used herein, a "substrate" is a molecule that can be metabolized, directly or indirectly, to ethanol by fermentation by Saccharomyces or any of the yeasts or yeast products described herein.

[0046]

[0050] The term "wild-type" as used herein refers to a typical form of a normally occurring organism or its genetic material, as distinguished from a selected organism.

[0047]

[0051] The terms "yeast product" and "composition" are used interchangeably herein and, as used herein, refer to a composition comprising, inter alia, dry yeast, starch and an emulsifier. The yeast product may also be a liquid composition comprising, inter alia, cream yeast, glycerol and xanthan gum.

[0048]

[0052] Unless otherwise defined herein, scientific and technical terms used in connection with this disclosure shall have the meanings commonly understood by those skilled in the art. For example, the nomenclature used in connection with the techniques of cell biology, molecular biology, microbiology, genetics, and protein and nucleic acid chemistry described herein are well known and commonly used in the art. The meaning and scope of the terms should be clear. However, in the event of potential ambiguity, the definitions provided herein shall take precedence over any dictionary or external definitions. Furthermore, unless otherwise required by context, singular terms shall include the plural and plural terms shall include the singular.

[0049] 2. Yeast strains and yeast strain derivatives

[0053] Yeast strains and yeast strain derivatives, as used herein, can be any yeast useful for ethanol production, including, but not limited to, Saccharomyces, ZygoSaccharomyces, Brettanomyces, and Kluyveromyces. Preferably, the yeast is a Saccharomyces sp., and even more preferably, Saccharomyces cerevisiae.

[0050]

[0054] Additionally, the Saccharomyces yeast strains and derivatives thereof described herein can be readily distinguished from (a) naturally occurring strains of Saccharomyces; (b) contaminating strains of Saccharomyces; and (c) other strains used in the ethanol industry that do not have the ethanol production capabilities and defining characteristics of the strains described herein.

[0051]

[0055] In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly higher ethanol yield than a typical yeast strain used in fermentation (e.g., Saccharomyces Y1609 strain). In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly higher ethanol yield than a typical yeast strain used in fermentation (e.g., Saccharomyces Y1609 strain) at a temperature of 20° C. to 40° C., preferably 32° C. to 36° C. In a particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly higher ethanol yield than a typical yeast strain used in fermentation (e.g., Saccharomyces Y1609 strain) at a temperature of 32° C. In another particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly higher ethanol yield than a typical yeast strain used in fermentation (e.g., Saccharomyces Y1609 strain) at a temperature of 36° C. The inventors have surprisingly found that the yeast strains described herein provide a statistically significantly higher ethanol yield than Y1609 under the same fermentation conditions. The inventors have also surprisingly found that the derivatives described herein generally result in statistically significantly higher ethanol yields than Y1609 under identical conditions (e.g., Y1919 in Figures 2-5).

[0052]

[0056] In some embodiments, the yeast strains and derivatives thereof described herein produce at least about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0%, or 21.0% more ethanol after 50 hours of fermentation compared to a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609). In some embodiments, the yeast strains and derivatives thereof described herein produce up to about 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0%, or 21.0% more ethanol after 50 hours of fermentation compared to a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609). In some embodiments, the yeast strains and derivatives thereof described herein have a fermentation yield of about 0.1% to 21% (i.e., from about 0.1% to about 21%), 0.2% to 21%, 0.3% to 21%, 0.4% to 21%, 0.5% to 21%, 0.6% to 21%, 0.7% to 21%, 0.8% to 21%, 0.9% to 21%, 1% to 21%, 2% to 21%, 3% to 21%, 4% to 21%, 5% to 21%, 6% to 21%, 7% to 21% after 50 hours of fermentation compared to a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609). ,8%~21%,9%~21%,10%~21%,0.2%~20%,0.3%~20%,0.4%~20%,0.5%~20%,0.6%~20%,0.7%~20%,0.8%~20%,0.9%~20%,1%~20%,2%~20%,3%~20%,4%~20%,5%~20%,6%~20%,7%~20%,8%~20%,9%~20%,10%~20%,0.2%~19%,0.3%~19%,0.4%~19%,0.5%~19%,0.6%~19%,0.7%~19%,0.8%~19%,0.9%~19%、1%~19%、2%~19%、3%~19%、4%~19%、5%~19%、6%~19%、7%~19%、8%~19%、9%~19%、10%~19%、0.2%~18%、0.3%~18%、0.4%~18%、0.5%~18%、0.6%~18%、0.7%~18%、0.8%~18%、0.9%~18%、1%~18%、2%~18%、3%~18%、4%~18%、5%~18%、6%~18%、7%~18%、8%~18%、9%~18%、10%~18%、0.2%~17%、0.3%~17%、0.4%~17%、0.5%~17%、0.6%~17%、0.7%~17%、0.8%~17%、0.9%~17%、1%~17%、2%~17%、3%~17%、4%~17%、5%~17%、6%~17%、7%~17%、8%~17%、9%~17%、10%~17%、0.2%~16%、0.3%~16%、0.4%~16%、0.5%~16%、0.6%~16%、0.7%~16%、0.8%~16%、0.9%~16%、1%~16%、2%~16%、3%~16%、4%~16%、5%~16%、6%~16%、7%~16%、8%~16%、9%~16%、10%~16%、0.2%~15%、0.3%~15%、0.4%~15%、0.5%~15%、0.6%~15%、0.7%~15%、0.8%~15%、0.9%~15%、1%~15%、2%~15%、3%~15%、4%~15%、5%~15%、6%~15%、7%~15%、8%~15%、9%~15%、10%~15%、0.1%~14%、0.2%~14%、0.3%~14%、0.4%~14%、0.5%~14%、0.6%~14%、0.7%~14%、0.8%~14%、0.9%~14%、1%~14%、2%~14%、3%~14%、4%~14%、5%~14%、6%~14%、7%~14%、8%~14%、9%~14%、10%~14%、0.1%~13%、0.2%~13%、0.3%~13%、0.4%~13%、0.5%~13%、0.6%~13%、0.7%~13%、0.8%~13%、0.9%~13%、1%~13%、2%~13%、3%~13%、4%~13%、5%~13%、6%~13%、7%~13%、8%~13%、9%~13%、10%~13%、0.1%~12%、0.2%~12%、0.3%~12%、0.4%~12%、0.5%~12%、0.6%~12%, 0.7%~12%, 0.8%~12%, 0.9%~12%, 1%~12%, 2%~12%, 3%~12%, 4%~12%, 5%~12%, 6%~12%, 7%~12%, 8%~12%, 9%~12%, 10%~12%, 0.1%~11%, 0.2%~11%, 0.3%~11%, 0.4%~11%, 0.5%~11%, 0.6%~11%, 0.7%~11%, 0.8%~11%, 0.9%~11%, 1%~11%, 2%~11%, 3%~11%, 4%~11 %, 5%-11%, 6%-11%, 7%-11%, 8%-11%, 9%-11%, 10%-11%, 0.1%-0.2%, 0.1%-0.3%, 0.1%-0.4%, 0.1%-0.5%, 0.1%-0.6%, 0.1%-0.7%, 0.1%-0.8%, 0.1%-0.9%, 0.1%-1%, 0.1%-2%, 0.1%-3%, 0.1%-4%, 0.1%-5%, 0.1%-6%, 0.1%-7%, 0.1%-8%, or 0.1%-9% more ethanol.

[0053]

[0057] In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly lower glycerol yield than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain). In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly lower glycerol yield than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain) at temperatures between 20°C and 40°C, preferably between 32°C and 36°C. In a particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly lower glycerol yield than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain) at a temperature of 32°C. In another particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly lower glycerol yield than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain) at a temperature of 36°C. The inventors have surprisingly found that the yeast strains described herein result in a statistically significantly lower glycerol production compared to Y1609 under identical fermentation conditions. The inventors have also surprisingly found that the derivatives described herein result in statistically significantly lower glycerol production compared to Y1609 under identical conditions (e.g., Y1919 in Figures 2-5).

[0054]

[0058] In some embodiments, the yeast strains and derivatives thereof described herein produce at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, or 22% less glycerol after 50 hours of fermentation compared to a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609). In some embodiments, the yeast strains and derivatives thereof described herein produce up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29% or 30% less glycerol after 50 hours of fermentation compared to a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609). In some embodiments, the yeast strains and derivatives thereof described herein produce up to about 1%-30% (i.e., from about 1% to about 30%), 2%-30%, 3%-30%, 4%-30%, 5%-30%, 6%-30%, 7%-30%, 8%-30%, 9%-30%, 10%-30%, 12%-30%, 14%-30%, 15%-30%, 16%-30%, 17%-30%, 18%-30%, 19%-30%, 20%-30%, 21%-30%, 22%-30%, 23%-30%, 24%-30%, 25%-30%, 26%-30%, 27%-30%, 28%-30%, 29%-30%, 30%-30%, 31%-30%, 32%-30%, 33%-30%, 34%-30%, 35%-30%, 36%-30%, 37%-30%, 38%-30%, 39%-30%, 39%-30%, 39%-30%, 38%-30%, 39%-30%, 39%-30 ~30%, 10%~30%, 11%~30%, 12%~30%, 13%~30%, 14%~30%, 15%~30%, 16%~30%, 17%~30%, 18%~30%, 19%~30%, 20%~30%, 21%~30%, 22%~30%, 23%~30%, 24%~30%, 25%~30%, 26%~30%, 27%~30%, 28%~30%, 2 9%~30%, 1%~29%, 2%~29%, 3%~29%, 4%~29%, 5%~29%, 6%~29%, 7%~29%, 8%~29%, 9%~29%, 10%~29%, 11%~29%, 12%~29%, 13%~29%, 14%~29%, 15%~29%, 16%~29%, 17%~29%, 18%~29%, 19%~29%, 20%~29%, 21%~29%, 22%~29%, 23%~29%, 24%~29%, 25%~29%, 26%~29%, 27%~29%, 28%~29%, 1%~28%, 2%~28%, 3%~28%, 4%~28%, 5%~28%, 6%~28%, 7%~28%, 8%~28%, 9%~28%, 10%~28%, 11%~28%, 12%~28%, 13%~28%,14%~28%、15%~28%、16%~28%、17%~28%、18%~28%、19%~28%、20%~28%、21%~28%、22%~28%、23%~28%、24%~28%、25%~28%、26%~28%、27%~28%、1%~27%、2%~27%、3%~27%、4%~27%、5%~27%、6%~27%、7%~27%、8%~27%、9%~27%、10%~27%、11%~27%、12%~27%、13%~27%、14%~27%、15%~27%、16%~27%、17%~27%、18%~27%、19%~27%、20%~27%、21%~27%、22%~27%、23%~27%、24%~27%、25%~27%、26%~27%、1%~26%、2%~26%、3%~26%、4%~26%、5%~26%、6%~26%、7%~26%、8%~26%、9%~26%、10%~26%、11%~26%、12%~26%、13%~26%、14%~26%、15%~26%、16%~26%、17%~26%、18%~26%、19%~26%、20%~26%、21%~26%、22%~26%、23%~26%、24%~26%、25%~26%、1%~25%、2%~25%、3%~25%、4%~25%、5%~25%、6%~25%、7%~25%、8%~25%、9%~25%、10%~25%、11%~25%、12%~25%、13%~25%、14%~25%、15%~25%、16%~25%、17%~25%、18%~25%、19%~25%、20%~25%、21%~25%、22%~25%、23%~25%、24%~25%、1%~24%、2%~24%、3%~24%、4%~24%、5%~24%、6%~24%、7%~24%、8%~24%、9%~24%、10%~24%、11%~24%、12%~24%、13%~24%、14%~24%、15%~24%、16%~24%、17%~24%、18%~24%、19%~24%、20%~24%、21%~24%、22%~24%、23%~24%、1%~23%、2%~23%、3%~23%、4%~23%、5%~23%、6%~23%、7%~23%、8%~23%、9%~23%、10%~23%、11%~23%、12%~23%、13%~23%、14%~23%、15%~23%、16%~23%、17%~23%、18%~23%、19%~23%、20%~23%、21%~23%、22%~23%、1%~22%、2%~22%、3%~22%、4%~22%、5%~22%、6%~22%、7%~22%、8%~22%、9%~22%、10%~22%、11%~22%、12%~22%、13%~22%、14%~22%、15%~22%、16%~22%、17%~22%、18%~22%、19%~22%、20%~22%、21%~22%、1%~21%、2%~21%、3%~21%、4%~21%、5%~21%、6%~21%、7%~21%、8%~21%、9%~21%、10%~21%、11%~21%、12%~21%、13%~21%、14%~21%、15%~21%、16%~21%、17%~21%、18%~21%、19%~21%、20%~21%、1%~20%、2%~20%、3%~20%、4%~20%、5%~20%、6%~20%、7%~20%、8%~20%、9%~20%、10%~20%、11%~20%、12%~20%、13%~20%、14%~20%、15%~20%、16%~20%、17%~20%、18%~20%、19%~20%、1%~19%、2%~19%、3%~19%、4%~19%、5%~19%、6%~19%、7%~19%、8%~19%、9%~19%、10%~19%、11%~19%、12%~19%、13%~19%、14%~19%、15%~19%、16%~19%、17%~19%、18%~19%、1%~18%、2%~18%、3%~18%、4%~18%、5%~18%、6%~18%、7%~18%、8%~18%、9%~18%、10%~18%、11%~18%、12%~18%、13%~18%、14%~18%、15%~18%、16%~18%、17%~18%、1%~17%、2%~17%、3%~17%、4%~17%、5%~17%、6%~17%、7%~17%、8%~17%、9%~17%、10%~17%、11%~17%、12%~17%、13%~17%、14%~17%、15%~17%、16%~17%、1%~16%、2%~16%、3%~16%、4%~16%、5%~16%、6%~16%、7%~16%、8%~16%、9%~16%、10%~16%、11%~16%、12%~16%、13%~16%、14%~16%、15%~16%、1%~15%、2%~15%、3%~15%、4%~15%, 5%~15%, 6%~15%, 7%~15%, 8%~15%, 9%~15%, 10%~15%, 11%~15%, 12%~15%, 13%~15%, 14%~15%, 1%~14%, 2%~14%, 3%~14%, 4%~14%, 5%~14%, 6%~14%, 7%~14%, 8%~14%, 9%~14%, 10%~14%, 11%~14%, 12%~14%, 13%~14%, 1%~13%, 2%~13%, 3%~13%, 4%~13%, 5%~13%, 6%~13%, 7%~13%, 8%~13%, 9%~13%, 10%~13%, 11%~13%, 12%~13%, 1%~12%, 2%~12%, 3%~12%, 4%~12%, 5%~12%, 6%~12%, 7%~12%, 8%~12%, 9%~12%, 10%~12%, 11%~12%, 1%~11%, 2%~11 %, 3%~11%, 4%~11%, 5%~11%, 6%~11%, 7%~11%, 8%~11%, 9%~11%, 10%~11%, 1%~10%, 2%~10%, 3%~10%, 4%~10%, 5%~10%, 6%~10%, 7%~10%, 8%~10%, 9%~10%, 1%~9%, 2%~9%, 3%~9%, 4%~9%, 5%~9%, 6%~9%, 7%~9%, 8%~9%, 1%~8%, 2 Produces %-8%, 3%-8%, 4%-8%, 5%-8%, 6%-8%, 7%-8%, 1%-7%, 2%-7%, 3%-7%, 4%-7%, 5%-7%, 6%-7%, 1%-6%, 2%-6%, 3%-6%, 4%-6%, 5%-6%, 1%-5%, 2%-5%, 3%-5%, 4%-5%, 1%-4%, 2%-4%, 3%-4%, 1%-3%, 2%-3%, or 1%-2% less glycerol.

[0055]

[0059] In some embodiments, the yeast strains and derivatives thereof described herein have a higher fermentation rate than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain). In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly higher fermentation rate than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain) at temperatures between 20°C and 40°C, preferably between 32°C and 36°C. In a particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly higher fermentation rate than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain) at a temperature of 32°C. In another particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly higher fermentation rate than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain) at a temperature of 36°C. The inventors have surprisingly found that the yeast strains described herein provide a statistically significantly higher fermentation rate compared to Y1609 under the same fermentation conditions. The inventors have also surprisingly found that the derivatives described herein result in statistically significantly higher fermentation rates compared to Y1609 under identical conditions (e.g., Y1919 in Figures 2-5).

[0056]

[0060] In some embodiments, the yeast strains and derivatives thereof described herein have a fermentation rate after 24 hours of fermentation that is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% higher than a typical yeast strain used for fermentation (e.g., Saccharomyces strain Y1609). In some embodiments, the yeast strains and derivatives thereof described herein have a fermentation rate that is up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% higher after 24 hours of fermentation than a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609). In some embodiments, the yeast strains and derivatives thereof described herein, after 24 hours of fermentation, exhibit a fermentation yield of about 1%-50% (i.e., from about 1% to about 50%), 2%-50%, 3%-50%, 4%-50%, 5%-50%, 6%-50%, 7%-50%, 8%-50%, 9%-50%, 10%-50%, 11%-50%, 12%-50%, 13%-50%, 14%-50%, 15%-50%, 16%-50%, 17%-50%, 18%-50%, 19%-50%, 20%-50%, 21%-50%, 22%-50%, 23%-50%, 24%-50%, 25%-50%, 26%-50%, 27%-50%, 28%-50%, 29%-50%, 30%-50%, 31%-50%, 32%-50%, 33%-50%, 34%-50%, 35%-50%, 36%-50%, 37%-50%, 38%-50%, 39%-50%, 40%-50%, 41%-50%, 42%-50%, 43%-50%, 44%-50%, 45%-50%, 46%-50%, 47%-50%, 48%-50%, 49%-50%, 50%-50%, 50%-50%, 51%-50%, 52%-50%, 53%-50%, 54%-50%, 55%-50%, 56%-50%, 57%-50%, 58 50%, 26%~50%, 27%~50%, 28%~50%, 2%~49%, 3%~49%, 4%~49%, 5%~49%, 6%~49%, 7%~49%, 8%~49%, 9%~49%, 10%~49%, 11%~49%, 12%~49%, 13%~49%, 14%~49%, 15%~49%, 16%~49%, 17 %~49%, 18%~49%, 19%~49%, 20%~49%, 21%~49%, 22%~49%, 23%~49%, 24%~49%, 25%~49%, 26%~49%, 27%~49%, 28%~49%, 2%~48%, 3%~48%, 4%~48%, 5%~48%, 6%~48%, 7%~48%, 8%~48%,9%~48%、10%~48%、11%~48%、12%~48%、13%~48%、14%~48%、15%~48%、16%~48%、17%~48%、18%~48%、19%~48%、20%~48%、21%~48%、22%~48%、23%~48%、24%~48%、25%~48%、26%~48%、27%~48%、28%~48%、2%~47%、3%~47%、4%~47%、5%~47%、6%~47%、7%~47%、8%~47%、9%~47%、10%~47%、11%~47%、12%~47%、13%~47%、14%~47%、15%~47%、16%~47%、17%~47%、18%~47%、19%~47%、20%~47%、21%~47%、22%~47%、23%~47%、24%~47%、25%~47%、26%~47%、27%~47%、28%~47%、2%~46%、3%~46%、4%~46%、5%~46%、6%~46%、7%~46%、8%~46%、9%~46%、10%~46%、11%~46%、12%~46%、13%~46%、14%~46%、15%~46%、16%~46%、17%~46%、18%~46%、19%~46%、20%~46%、21%~46%、22%~46%、23%~46%、24%~46%、25%~46%、26%~46%、27%~46%、28%~46%、2%~45%、3%~45%、4%~45%、5%~45%、6%~45%、7%~45%、8%~45%、9%~45%、10%~45%、11%~45%、12%~45%、13%~45%、14%~45%、15%~45%、16%~45%、17%~45%、18%~45%、19%~45%、20%~45%、21%~45%、22%~45%、23%~45%、24%~45%、25%~45%、26%~45%、27%~45%、28%~45%、2%~44%、3%~44%、4%~44%、5%~44%、6%~44%、7%~44%、8%~44%、9%~44%、10%~44%、11%~44%、12%~44%、13%~44%、14%~44%、15%~44%、16%~44%、17%~44%、18%~44%、19%~44%、20%~44%、21%~44%、22%~44%、23%~44%、24%~44%、25%~44%、26%~44%、27%~44%、28%~44%、2%~43%、3%~43%、4%~43%、5%~43%、6%~43%、7%~43%、8%~43%、9%~43%、10%~43%、11%~43%、12%~43%、13%~43%、14%~43%、15%~43%、16%~43%、17%~43%、18%~43%、19%~43%、20%~43%、21%~43%、22%~43%、23%~43%、24%~43%、25%~43%、26%~43%、27%~43%、28%~43%、2%~42%、3%~42%、4%~42%、5%~42%、6%~42%、7%~42%、8%~42%、9%~42%、10%~42%、11%~42%、12%~42%、13%~42%、14%~42%、15%~42%、16%~42%、17%~42%、18%~42%、19%~42%、20%~42%、21%~42%、22%~42%、23%~42%、24%~42%、25%~42%、26%~42%、27%~42%、28%~42%、2%~41%、3%~41%、4%~41%、5%~41%、6%~41%、7%~41%、8%~41%、9%~41%、10%~41%、11%~41%、12%~41%、13%~41%、14%~41%、15%~41%、16%~41%、17%~41%、18%~41%、19%~41%、20%~41%、21%~41%、22%~41%、23%~41%、24%~41%、25%~41%、26%~41%、27%~41%、28%~41%、2%~40%、3%~40%、4%~40%、5%~40%、6%~40%、7%~40%、8%~40%、9%~40%、10%~40%、11%~40%、12%~40%、13%~40%、14%~40%、15%~40%、16%~40%、17%~40%、18%~40%、19%~40%、20%~40%、21%~40%、22%~40%、23%~40%、24%~40%、25%~40%、26%~40%、27%~40%、28%~40%、2%~39%、3%~39%、4%~39%、5%~39%、6%~39%、7%~39%、8%~39%、9%~39%、10%~39%、11%~39%、12%~39%、13%~39%、14%~39%、15%~39%、16%~39%、17%~39%、18%~39%、19%~39%、20%~39%、21%~39%、22%~39%、23%~39%、24%~39%、25%~39%、26%~39%、27%~39%、28%~39%、2%~38%、3%~38%、4%~38%、5%~38%、6%~38%、7%~38%、8%~38%、9%~38%、10%~38%、11%~38%、12%~38%、13%~38%、14%~38%、15%~38%、16%~38%、17%~38%、18%~38%、19%~38%、20%~38%、21%~38%、22%~38%、23%~38%、24%~38%、25%~38%、26%~38%、27%~38%、28%~38%、2%~37%、3%~37%、4%~37%、5%~37%、6%~37%、7%~37%、8%~37%、9%~37%、10%~37%、11%~37%、12%~37%、13%~37%、14%~37%、15%~37%、16%~37%、17%~37%、18%~37%、19%~37%、20%~37%、21%~37%、22%~37%、23%~37%、24%~37%、25%~37%、26%~37%、27%~37%、28%~37%、2%~36%、3%~36%、4%~36%、5%~36%、6%~36%、7%~36%、8%~36%、9%~36%、10%~36%、11%~36%、12%~36%、13%~36%、14%~36%、15%~36%、16%~36%、17%~36%、18%~36%、19%~36%、20%~36%、21%~36%、22%~36%、23%~36%、24%~36%、25%~36%、26%~36%、27%~36%、28%~36%、2%~35%、3%~35%、4%~35%、5%~35%、6%~35%、7%~35%、8%~35%、9%~35%、10%~35%、11%~35%、12%~35%、13%~35%、14%~35%、15%~35%、16%~35%、17%~35%、18%~35%、19%~35%、20%~35%、21%~35%、22%~35%、23%~35%、24%~35%、25%~35%、26%~35%、27%~35%、28%~35%、2%~34%、3%~34%、4%~34%、5%~34%、6%~34%、7%~34%、8%~34%、9%~34%、10%~34%、11%~34%、12%~34%、13%~34%、14%~34%、15%~34%、16%~34%、17%~34%、18%~34%、19%~34%、20%~34%、21%~34%、22%~34%、23%~34%、24%~34%、25%~34%、26%~34%、27%~34%、28%~34%、2%~33%、3%~33%、4%~33%、5%~33%、6%~33%、7%~33%、8%~33%、9%~33%、10%~33%、11%~33%、12%~33%、13%~33%、14%~33%、15%~33%、16%~33%、17%~33%、18%~33%、19%~33%、20%~33%、21%~33%、22%~33%、23%~33%、24%~33%、25%~33%、26%~33%、27%~33%、28%~33%、2%~32%、3%~32%、4%~32%、5%~32%、6%~32%、7%~32%、8%~32%、9%~32%、10%~32%、11%~32%、12%~32%、13%~32%、14%~32%、15%~32%、16%~32%、17%~32%、18%~32%、19%~32%、20%~32%、21%~32%、22%~32%、23%~32%、24%~32%、25%~32%、26%~32%、27%~32%、28%~32%、2%~31%、3%~31%、4%~31%、5%~31%、6%~31%、7%~31%、8%~31%、9%~31%、10%~31%、11%~31%、12%~31%、13%~31%、14%~31%、15%~31%、16%~31%、17%~31%、18%~31%、19%~31%、20%~31%、21%~31%、22%~31%、23%~31%、24%~31%、25%~31%、26%~31%、27%~31%、28%~31%、2%~30%、3%~30%、4%~30%、5%~30%、6%~30%、7%~30%、8%~30%、9%~30%、10%~30%、11%~30%、12%~30%、13%~30%、14%~30%、15%~30%、16%~30%、17%~30%、18%~30%、19%~30%、20%~30%、21%~30%、22%~30%、23%~30%、24%~30%、25%~30%、26%~30%、27%~30%、28%~30%、29%~30%、1%~29%、2%~29%、3%~29%、4%~29%、5%~29%、6%~29%、7%~29%、8%~29%、9%~29%、10%~29%、11%~29%、12%~29%、13%~29%、14%~29%、15%~29%、16%~29%、17%~29%、18%~29%、19%~29%、20%~29%、21%~29%、22%~29%、23%~29%、24%~29%、25%~29%、26%~29%、27%~29%、 28%~29%、1%~28%、2%~28%、3%~28%、4%~28%、5%~28%、6%~28%、7%~28%、8%~28%、9%~28%、10%~28%、11%~28%、12%~28%、13%~28%、14%~28%、15%~28%、16%~28%、17%~28%、18%~28%、19%~28%、20%~28%、21%~28%、22%~28%、23%~28%、24%~28%、25%~28%、26%~28%、27%~28%、1%~27%、2%~27%、3%~27%、4%~27%、5%~27%、6%~27%、7%~27%、8%~27%、9%~27%、10%~27%、11%~27%、12%~27%、13%~27%、14%~27%、15%~27%、16%~27%、17%~27%、18%~27%、19%~27%、20%~27%、21%~27%、22%~27%、23%~27%、24%~27%、25%~27%、26%~27%、1%~26%、2%~26%、3%~26%、4%~26%、5%~26%、6%~26%、7%~26%、8%~26%、9%~26%、10%~26%、11%~26%、12%~26%、13%~26%、14%~26%、15%~26%、16%~26%、17%~26%、18%~26%、19%~26%、20%~26%、21%~26%、22%~26%、23%~26%、24%~26%、25%~26%、1%~25%、2%~25%、3%~25%、4%~25%、5%~25%、6%~25%、7%~25%、8%~25%、9%~25%、10%~25%、11%~25%、12%~25%、13%~25%、14%~25%、15%~25%、16%~25%、17%~25%、18%~25%、19%~25%、20%~25%、21%~25%、22%~25%、23%~25%、24%~25%、1%~24%、2%~24%、3%~24%、4%~24%、5%~24%、6%~24%、7%~24%、8%~24%、9%~24%、10%~24%、11%~24%、12%~24%、13%~24%、14%~24%、15%~24%、16%~24%、17%~24%、18%~24%、19%~24%、20%~24%、21%~24%、22%~24%、23%~24%、1%~23%、2%~23%、3%~23%、4%~23%、5%~23%、6%~23%、7%~23%、8%~23%、9%~23%、10%~23%、11%~23%、12%~23%、13%~23%、14%~23%、15%~23%、16%~23%、17%~23%、18%~23%、19%~23%、20%~23%、21%~23%、22%~23%、1%~22%、2%~22%、3%~22%、4%~22%、5%~22%、6%~22%、7%~22%、8%~22%、9%~22%、10%~22%、11%~22%、12%~22%、13%~22%、14%~22%、15%~22%、16%~22%、17%~22%、18%~22%、19%~22%、20%~22%、21%~22%、1%~21%、2%~21%、3%~21%、4%~21%、5%~21%、6%~21%、7%~21%、8%~21%、9%~21%、10%~21%、11%~21%、12%~21%、13%~21%、14%~21%、15%~21%、16%~21%、17%~21%、18%~21%、19%~21%、20%~21%、1%~20%、2%~20%、3%~20%、4%~20%、5%~20%、6%~20%、7%~20%、8%~20%、9%~20%、10%~20%、11%~20%、12%~20%、13%~20%、14%~20%、15%~20%、16%~20%、17%~20%、18%~20%、19%~20%、1%~19%、2%~19%、3%~19%、4%~19%、5%~19%、6%~19%、7%~19%、8%~19%、9%~19%、10%~19%、11%~19%、12%~19%、13%~19%、14%~19%、15%~19%、16%~19%、17%~19%、18%~19%、1%~18%、2%~18%、3%~18%、4%~18%、5%~18%、6%~18%、7%~18%、8%~18%、9%~18%、10%~18%、11%~18%、12%~18%、13%~18%、14%~18%、15%~18%、16%~18%、17%~18%、1%~17%、2%~17%、3%~17%、4%~17%、5%~17%、6%~17%、7%~17%、8%~17%、9%~17%、10%~17%、11%~17%、12%~17%、13%~17%、14%~17%、15%~17%、16%~17%、1%~16%、2%~16%、3%~16%、4%~16%、5%~16%、6%~16%, 7%~16%, 8%~16%, 9%~16%, 10%~16%, 11%~16%, 12%~16%, 13%~16%, 14%~16%, 15%~16%, 1%~15%, 2%~15%, 3%~15%, 4%~15%, 5%~15%, 6%~15%, 7%~15%, 8%~15%, 9%~15%, 10%~15%, 11%~15%, 12%~15%, 13%~15%, 14%~15%, 1%~14%, 2%~14%, 3%~14%, 4%~14%, 5%~14%, 6%~14%, 7%~14%, 8%~14%, 9%~14%, 10%~14%, 11%~14%, 12%~14%, 13%~14%, 1%~13%, 2%~13%, 3%~13%, 4%~13%, 5%~13%, 6%~13%, 7%~13%, 8%~13%, 9%~13%, 10%~13%, 11%~13%, 12%~13%, 1%~12%, 2%~12%, 3%~12%, 4%~12%, 5%~12%, 6%~12 %, 7%~12%, 8%~12%, 9%~12%, 10%~12%, 11%~12%, 1%~11%, 2%~11%, 3%~11%, 4%~11%, 5%~11%, 6%~11%, 7%~11%, 8%~11%, 9%~11%, 10%~11%, 1%~10%, 2%~10%, 3%~10%, 4%~10%, 5%~10%, 6%~10%, 7%~10%, 8%~10%, 9%~10%, 1%~9%, 2%~9%, 3%~9%, 4%~9%, 5%~9 %, 6%-9%, 7%-9%, 8%-9%, 1%-8%, 2%-8%, 3%-8%, 4%-8%, 5%-8%, 6%-8%, 7%-8%, 1%-7%, 2%-7%, 3%-7%, 4%-7%, 5%-7%, 6%-7%, 1%-6%, 2%-6%, 3%-6%, 4%-6%, 5%-6%, 1%-5%, 2%-5%, 3%-5%, 4%-5%, 1%-4%, 2%-4%, 3%-4%, 1%-3%, 2%-3%, or 1%-2% higher fermentation rate.

[0057]

[0061] In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significant higher temperature tolerance than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain). Temperature tolerance can be indicated by increased ethanol production, decreased glycerol production, and increased fermentation rate. In certain embodiments, the yeast strains and derivatives thereof described herein can tolerate temperatures of about 20°C, about 21°C, about 22°C, about 23°C, about 24°C, about 25°C, about 26°C, about 27°C, about 28°C, about 29°C, about 30°C, about 31°C, about 32°C, about 33°C, about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, and / or about 40°C. The inventors have surprisingly found that the yeast strains and derivatives thereof described herein have a statistically significant higher temperature tolerance compared to Y1609 under the same fermentation conditions.

[0058]

[0062] In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly higher organic acid tolerance at low pH than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain). In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly higher organic acid tolerance at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain). In a particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly higher organic acid tolerance at a temperature of 32° C. than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain). In another particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly higher organic acid tolerance at a temperature of 36° C. than typical yeast strains used in fermentation (e.g., Saccharomyces Y1609 strain). The inventors have surprisingly found that the yeast strains and derivatives thereof described herein have a statistically significantly higher organic acid tolerance compared to Y1609 under the same fermentation conditions. Examples of organic acids include, but are not limited to, lactic acid, acetic acid, succinic acid, citric acid, malic acid, fumaric acid, other carboxylic acids, or combinations thereof.

[0059]

[0063] In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly higher ethanol yield than a typical yeast strain used in fermentation (e.g., Saccharomyces Y1609 strain) at 20°C to 40°C, preferably 32°C to 36°C, and at low pH with organic acids. In a particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly higher ethanol yield than a typical yeast strain used in fermentation (e.g., Saccharomyces Y1609 strain) at a temperature of 32°C and at low pH with organic acids. In another particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly higher ethanol yield than a typical yeast strain used in fermentation (e.g., Saccharomyces Y1609 strain) at a temperature of 36°C and at low pH with organic acids. The inventors have surprisingly found that the yeast strains described herein provide a statistically significantly higher ethanol yield at low pH with organic acids compared to Y1609 under the same fermentation conditions. The inventors have also surprisingly found that the derivatives described herein generally provide statistically significantly higher ethanol yields at low pH with organic acids compared to Y1609 under identical conditions (e.g., Y1919 in Figures 2-5).

[0060]

[0064] In some embodiments, the yeast strains and derivatives thereof described herein produce at least about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0%, or 21.0% more ethanol after 50 hours of fermentation compared to a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609) at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids. In some embodiments, the yeast strains and derivatives thereof described herein produce up to about 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0%, or 21.0% more ethanol after 50 hours of fermentation compared to a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609) at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids. In some embodiments, the yeast strains and derivatives thereof described herein exhibit a fermentation yield of about 0.1% to 21% (i.e., from about 0.1% to about 21%), 0.2% to 21%, 0.3% to 21%, 0.4% to 21%, 0.5% to 21%, 0.6% to 21%, 0.7% to 21%, 0.8% to 21%, 0.9% to 21%, 1% to 1% after 50 hours of fermentation at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids, compared to a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609). ~21%, 2%~21%, 3%~21%, 4%~21%, 5%~21%, 6%~21%, 7%~21%, 8%~21%, 9%~21%, 10%~21%, 0.2%~20%, 0.3%~20%, 0.4%~20%, 0.5%~20%, 0.6%~20%, 0.7%~20%, 0.8%~20%, 0.9%~20%, 1%~20%, 2%~20%, 3%~20%, 4%~20%, 5%~20%, 6%~20%, 7%~20%, 8%~20%, 9%~20%, 10%~20%, 0.2%~19%, 0.3%~19%、0.4%~19%、0.5%~19%、0.6%~19%、0.7%~19%、0.8%~19%、0.9%~19%、1%~19%、2%~19%、3%~19%、4%~19%、5%~19%、6%~19%、7%~19%、8%~19%、9%~19%、10%~19%、0.2%~18%、0.3%~18%、0.4%~18%、0.5%~18%、0.6%~18%、0.7%~18%、0.8%~18%、0.9%~18%、1%~18%、2%~18%、3%~18%、4%~18%、5%~18%、6%~18%、7%~18%、8%~18%、9%~18%、10%~18%、0.2%~17%、0.3%~17%、0.4%~17%、0.5%~17%、0.6%~17%、0.7%~17%、0.8%~17%、0.9%~17%、1%~17%、2%~17%、3%~17%、4%~17%、5%~17%、6%~17%、7%~17%、8%~17%、9%~17%、10%~17%、0.2%~16%、0.3%~16%、0.4%~16%、0.5%~16%、0.6%~16%、0.7%~16%、0.8%~16%、0.9%~16%、1%~16%、2%~16%、3%~16%、4%~16%、5%~16%、6%~16%、7%~16%、8%~16%、9%~16%、10%~16%、0.2%~15%、0.3%~15%、0.4%~15%、0.5%~15%、0.6%~15%、0.7%~15%、0.8%~15%、0.9%~15%、1%~15%、2%~15%、3%~15%、4%~15%、5%~15%、6%~15%、7%~15%、8%~15%、9%~15%、10%~15%、0.1%~14%、0.2%~14%、0.3%~14%、0.4%~14%、0.5%~14%、0.6%~14%、0.7%~14%、0.8%~14%、0.9%~14%、1%~14%、2%~14%、3%~14%、4%~14%、5%~14%、6%~14%、7%~14%、8%~14%、9%~14%、10%~14%、0.1%~13%、0.2%~13%、0.3%~13%、0.4%~13%、0.5%~13%、0.6%~13%、0.7%~13%、0.8%~13%、0.9%~13%, 1%~13%, 2%~13%, 3%~13%, 4%~13%, 5%~13%, 6%~13%, 7%~13%, 8%~13%, 9%~13%, 10%~13%, 0.1%~12%, 0.2%~12%, 0.3%~12%, 0.4%~12%, 0.5%~12%, 0.6%~12%, 0.7%~ 12%, 0.8%~12%, 0.9%~12%, 1%~12%, 2%~12%, 3%~12%, 4%~12%, 5%~12%, 6%~12%, 7%~12%, 8%~12%, 9%~12%, 10%~12%, 0.1%~11%, 0.2%~11%, 0.3%~11%, 0.4%~11%, 0.5%~11% , 0.6%~11%, 0.7%~11%, 0.8%~11%, 0.9%~11%, 1%~11%, 2%~11%, 3%~11%, 4%~11%, 5%~11%, 6%~11%, 7%~11%, 8%~11%, 9%~11%, 10%~11%, 0.1%~0.2%, 0.1%~0.3%, 0.1%~0.4% , 0.1%-0.5%, 0.1%-0.6%, 0.1%-0.7%, 0.1%-0.8%, 0.1%-0.9%, 0.1%-1%, 0.1%-2%, 0.1%-3%, 0.1%-4%, 0.1%-5%, 0.1%-6%, 0.1%-7%, 0.1%-8%, or 0.1%-9% more ethanol.

[0061]

[0065] In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly lower glycerol yield than a typical yeast strain used for fermentation (e.g., Saccharomyces Y1609 strain) at temperatures between 20°C and 40°C, preferably between 32°C and 36°C, and at low pH with organic acid. In a particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly lower glycerol yield than a typical yeast strain used for fermentation (e.g., Saccharomyces Y1609 strain) at temperatures between 32°C and low pH with organic acid. In another particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly lower glycerol yield than a typical yeast strain used for fermentation (e.g., Saccharomyces Y1609 strain) at temperatures between 36°C and low pH with organic acid. The inventors have surprisingly found that the yeast strains described herein provide a statistically significantly lower glycerol production at low pH with organic acid compared to Y1609 under the same fermentation conditions. The inventors have also surprisingly found that the derivatives described herein result in statistically significantly lower glycerol production at low pH with organic acids compared to Y1609 under identical conditions (e.g., Y1919 in Figures 2-5).

[0062]

[0066] In some embodiments, the yeast strains and derivatives thereof described herein produce at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, or 22% less glycerol after 50 hours of fermentation compared to a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609) at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids. In some embodiments, the yeast strains and derivatives thereof described herein produce up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30% less glycerol after 50 hours of fermentation compared to typical yeast strains used in fermentation (e.g., Saccharomyces strain Y1609) at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids. In some embodiments, the yeast strains and derivatives thereof described herein exhibit a fermentation yield of about 1% to 30% (i.e., from about 1% to about 30%), 2% to 30%, 3% to 30%, 4% to 30%, 5% to 30%, 6% to 30%, 7% to 30%, 8% to 30%, 9% to 30%, 10% to 30%, 11% to 30%, 12% to 30%, 13% to 30%, 14% to 30%, 15% to 30%, 16% to 30%, 17% to 30%, 18% to 30%, 19% to 20% and 21% after 50 hours of fermentation at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids, compared to a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609). ~30%, 20%~30%, 21%~30%, 22%~30%, 23%~30%, 24%~30%, 25%~30%, 26%~30%, 27%~30%, 28%~30%, 29%~30%, 1%~29%, 2%~29%, 3%~29%, 4%~29%, 5%~29%, 6%~29%, 7%~29%, 8%~29%, 9%, 9%~29%, 10%~29%, 11%~29%, 12%~29%, 13%~29%, 14%~29%, 15%~29%, 16%~29%, 17%~29%, 18%~29%, 19%~29%, 20%~29%, 21%~29%, 22%~29%, 23%~29%, 24%~29%, 25%~29%,26%~29%、27%~29%、28%~29%、1%~28%、2%~28%、3%~28%、4%~28%、5%~28%、6%~28%、7%~28%、8%~28%、9%~28%、10%~28%、11%~28%、12%~28%、13%~28%、14%~28%、15%~28%、16%~28%、17%~28%、18%~28%、19%~28%、20%~28%、21%~28%、22%~28%、23%~28%、24%~28%、25%~28%、26%~28%、27%~28%、1%~27%、2%~27%、3%~27%、4%~27%、5%~27%、6%~27%、7%~27%、8%~27%、9%~27%、10%~27%、11%~27%、12%~27%、13%~27%、14%~27%、15%~27%、16%~27%、17%~27%、18%~27%、19%~27%、20%~27%、21%~27%、22%~27%、23%~27%、24%~27%、25%~27%、26%~27%、1%~26%、2%~26%、3%~26%、4%~26%、5%~26%、6%~26%、7%~26%、8%~26%、9%~26%、10%~26%、11%~26%、12%~26%、13%~26%、14%~26%、15%~26%、16%~26%、17%~26%、18%~26%、19%~26%、20%~26%、21%~26%、22%~26%、23%~26%、24%~26%、25%~26%、1%~25%、2%~25%、3%~25%、4%~25%、5%~25%、6%~25%、7%~25%、8%~25%、9%~25%、10%~25%、11%~25%、12%~25%、13%~25%、14%~25%、15%~25%、16%~25%、17%~25%、18%~25%、19%~25%、20%~25%、21%~25%、22%~25%、23%~25%、24%~25%、1%~24%、2%~24%、3%~24%、4%~24%、5%~24%、6%~24%、7%~24%、8%~24%、9%~24%、10%~24%、11%~24%、12%~24%、13%~24%、14%~24%、15%~24%、16%~24%、17%~24%、18%~24%、19%~24%、20%~24%、21%~24%、22%~24%、23%~24%、1%~23%、2%~23%、3%~23%、4%~23%、5%~23%、6%~23%、7%~23%、8%~23%、9%~23%、10%~23%、11%~23%、12%~23%、13%~23%、14%~23%、15%~23%、16%~23%、17%~23%、18%~23%、19%~23%、20%~23%、21%~23%、22%~23%、1%~22%、2%~22%、3%~22%、4%~22%、5%~22%、6%~22%、7%~22%、8%~22%、9%~22%、10%~22%、11%~22%、12%~22%、13%~22%、14%~22%、15%~22%、16%~22%、17%~22%、18%~22%、19%~22%、20%~22%、21%~22%、1%~21%、2%~21%、3%~21%、4%~21%、5%~21%、6%~21%、7%~21%、8%~21%、9%~21%、10%~21%、11%~21%、12%~21%、13%~21%、14%~21%、15%~21%、16%~21%、17%~21%、18%~21%、19%~21%、20%~21%、1%~20%、2%~20%、3%~20%、4%~20%、5%~20%、6%~20%、7%~20%、8%~20%、9%~20%、10%~20%、11%~20%、12%~20%、13%~20%、14%~20%、15%~20%、16%~20%、17%~20%、18%~20%、19%~20%、1%~19%、2%~19%、3%~19%、4%~19%、5%~19%、6%~19%、7%~19%、8%~19%、9%~19%、10%~19%、11%~19%、12%~19%、13%~19%、14%~19%、15%~19%、16%~19%、17%~19%、18%~19%、1%~18%、2%~18%、3%~18%、4%~18%、5%~18%、6%~18%、7%~18%、8%~18%、9%~18%、10%~18%、11%~18%、12%~18%、13%~18%、14%~18%、15%~18%、16%~18%、17%~18%、1%~17%、2%~17%、3%~17%、4%~17%、5%~17%、6%~17%、7%~17%、8%~17%、9%~17%、10%~17%、11%~17%、12%~17%、13%~17%、14%~17%、15%~17%、16%~17%、1%~16%、2%~16%、3%~16%、4%~16%, 5%~16%, 6%~16%, 7%~16%, 8%~16%, 9%~16%, 10%~16%, 11%~16%, 12%~16%, 13%~16%, 14%~16%, 15%~16%, 1%~15%, 2%~15%, 3%~15%, 4%~15%, 5%~15%, 6%~15%, 7%~15%, 8%~15%, 9%~15%, 10%~15%, 11%~15%, 12%~15%, 13%~15%, 14%~15%, 1%~14%, 2%~14 %, 3%~14%, 4%~14%, 5%~14%, 6%~14%, 7%~14%, 8%~14%, 9%~14%, 10%~14%, 11%~14%, 12%~14%, 13%~14%, 1%~13%, 2%~13%, 3%~13%, 4%~13%, 5%~13%, 6%~13%, 7%~13%, 8%~13%, 9%~13%, 10%~13%, 11%~13%, 12%~13%, 1%~12%, 2%~12%, 3%~12%, 4%~12%, 5%~12%, 6%~12%, 7%~12%, 8%~12%, 9%~12%, 10%~12%, 11%~12%, 1%~11%, 2%~11%, 3%~11%, 4%~11%, 5%~11%, 6%~11%, 7%~11%, 8%~11%, 9%~11%, 10%~11%, 1%~10%, 2%~10%, 3%~10%, 4%~10%, 5%~10%, 6%~10%, 7%~10%, 8%~10%, 9%~10%, 1%~9%, 2%~9%, 3%~9%, 4%~9%, 5%~ Produces 9%, 6%-9%, 7%-9%, 8%-9%, 1%-8%, 2%-8%, 3%-8%, 4%-8%, 5%-8%, 6%-8%, 7%-8%, 1%-7%, 2%-7%, 3%-7%, 4%-7%, 5%-7%, 6%-7%, 1%-6%, 2%-6%, 3%-6%, 4%-6%, 5%-6%, 1%-5%, 2%-5%, 3%-5%, 4%-5%, 1%-4%, 2%-4%, 3%-4%, 1%-3%, 2%-3%, or 1%-2% less glycerol.

[0063]

[0067] In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly higher fermentation rate than typical yeast strains used for fermentation (e.g., Saccharomyces Y1609) at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids. In a particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly higher fermentation rate than typical yeast strains used for fermentation (e.g., Saccharomyces Y1609) at temperatures between 32° C. and low pH with organic acids. In another particular embodiment, the yeast strains and derivatives thereof described herein have a statistically significantly higher fermentation rate than typical yeast strains used for fermentation (e.g., Saccharomyces Y1609) at temperatures between 36° C. and low pH with organic acids. The inventors have surprisingly found that the yeast strains described herein provide a statistically significantly higher fermentation rate at low pH with organic acids compared to Y1609 under the same fermentation conditions. The inventors have surprisingly found that the derivatives described herein result in statistically significantly higher fermentation rates at low pH with organic acids compared to Y1609 under identical conditions (e.g., Y1919 in Figures 2-5).

[0064]

[0068] In some embodiments, the yeast strains and derivatives thereof described herein have a fermentation rate that is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% higher than a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609) at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids, after 24 hours of fermentation. In some embodiments, the yeast strains and derivatives thereof described herein have a fermentation rate that is up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% higher after 24 hours of fermentation than a typical yeast strain used in fermentation (e.g., Saccharomyces strain Y1609). In some embodiments, the yeast strains and derivatives thereof described herein are about 1%-50% (i.e., from about 1% to about 50%), 2%-50%, 3%-50%, 4%-50%, 5%-50%, 6%-50%, 7%-50%, 8%-50%, 9%-50%, 10%-50%, 11%-50%, 12%-50%, 13%-50%, 14%-50%, 15%-50%, 16%-50%, 17%-50%, 18%-50%, 19%-50%, 20%-50%, 21%-50%, 22%-50%, 23%-50%, 24%-50%, 25%-50%, 26%-50%, 27%-50%, 28%-50%, 29%-50%, 30%-50%, 31%-50%, 32%-50%, 33%-50%, 34%-50%, 35%-50%, 36%-50%, 37%-50%, 38%-50%, 39%-50%, 40%-50%, 41%-50%, 42%-50%, 43%-50%, 44%-50%, 45%-50%, 46%-50%, 47%-50%, 48%-50%, 49%-50%, 50%-50%, 50%-50%, 51%-50%, 52%-50%, 53%-50%, 54%-50%, 55%-50%, 56%-50%, 57%-50%, 58%-50%, 59%-50%, 60%-5 6%~50%, 17%~50%, 18%~50%, 19%~50%, 20%~50%, 21%~50%, 22%~50%, 23%~50%, 24%~50%, 25%~50%, 26%~50%, 27%~50%, 28%~50%, 2%~49%, 3%~49%, 4%~49% , 5%~49%, 6%~49%, 7%~49%, 8%~49%, 9%~49%, 10%~49%, 11%~49%, 12%~49%, 13%~49%, 14%~49%, 15%~49%, 16%~49%, 17%~49%, 18%~49%, 19%~49%, 20%~49%,21%~49%、22%~49%、23%~49%、24%~49%、25%~49%、26%~49%、27%~49%、28%~49%、2%~48%、3%~48%、4%~48%、5%~48%、6%~48%、7%~48%、8%~48%、9%~48%、10%~48%、11%~48%、12%~48%、13%~48%、14%~48%、15%~48%、16%~48%、17%~48%、18%~48%、19%~48%、20%~48%、21%~48%、22%~48%、23%~48%、24%~48%、25%~48%、26%~48%、27%~48%、28%~48%、2%~47%、3%~47%、4%~47%、5%~47%、6%~47%、7%~47%、8%~47%、9%~47%、10%~47%、11%~47%、12%~47%、13%~47%、14%~47%、15%~47%、16%~47%、17%~47%、18%~47%、19%~47%、20%~47%、21%~47%、22%~47%、23%~47%、24%~47%、25%~47%、26%~47%、27%~47%、28%~47%、2%~46%、3%~46%、4%~46%、5%~46%、6%~46%、7%~46%、8%~46%、9%~46%、10%~46%、11%~46%、12%~46%、13%~46%、14%~46%、15%~46%、16%~46%、17%~46%、18%~46%、19%~46%、20%~46%、21%~46%、22%~46%、23%~46%、24%~46%、25%~46%、26%~46%、27%~46%、28%~46%、2%~45%、3%~45%、4%~45%、5%~45%、6%~45%、7%~45%、8%~45%、9%~45%、10%~45%、11%~45%、12%~45%、13%~45%、14%~45%、15%~45%、16%~45%、17%~45%、18%~45%、19%~45%、20%~45%、21%~45%、22%~45%、23%~45%、24%~45%、25%~45%、26%~45%、27%~45%、28%~45%、2%~44%、3%~44%、4%~44%、5%~44%、6%~44%、7%~44%、8%~44%、9%~44%、10%~44%、11%~44%、12%~44%、13%~44%、14%~44%、15%~44%、16%~44%、17%~44%、18%~44%、19%~44%、20%~44%、21%~44%、22%~44%、23%~44%、24%~44%、25%~44%、26%~44%、27%~44%、28%~44%、2%~43%、3%~43%、4%~43%、5%~43%、6%~43%、7%~43%、8%~43%、9%~43%、10%~43%、11%~43%、12%~43%、13%~43%、14%~43%、15%~43%、16%~43%、17%~43%、18%~43%、19%~43%、20%~43%、21%~43%、22%~43%、23%~43%、24%~43%、25%~43%、26%~43%、27%~43%、28%~43%、2%~42%、3%~42%、4%~42%、5%~42%、6%~42%、7%~42%、8%~42%、9%~42%、10%~42%、11%~42%、12%~42%、13%~42%、14%~42%、15%~42%、16%~42%、17%~42%、18%~42%、19%~42%、20%~42%、21%~42%、22%~42%、23%~42%、24%~42%、25%~42%、26%~42%、27%~42%、28%~42%、2%~41%、3%~41%、4%~41%、5%~41%、6%~41%、7%~41%、8%~41%、9%~41%、10%~41%、11%~41%、12%~41%、13%~41%、14%~41%、15%~41%、16%~41%、17%~41%、18%~41%、19%~41%、20%~41%、21%~41%、22%~41%、23%~41%、24%~41%、25%~41%、26%~41%、27%~41%、28%~41%、2%~40%、3%~40%、4%~40%、5%~40%、6%~40%、7%~40%、8%~40%、9%~40%、10%~40%、11%~40%、12%~40%、13%~40%、14%~40%、15%~40%、16%~40%、17%~40%、18%~40%、19%~40%、20%~40%、21%~40%、22%~40%、23%~40%、24%~40%、25%~40%、26%~40%、27%~40%、28%~40%、2%~39%、3%~39%、4%~39%、5%~39%、6%~39%、7%~39%、8%~39%、9%~39%、10%~39%、11%~39%、12%~39%、13%~39%、14%~39%、15%~39%、16%~39%、17%~39%、18%~39%、19%~39%、20%~39%、21%~39%、22%~39%、23%~39%、24%~39%、25%~39%、26%~39%、27%~39%、28%~39%、2%~38%、3%~38%、4%~38%、5%~38%、6%~38%、7%~38%、8%~38%、9%~38%、10%~38%、11%~38%、12%~38%、13%~38%、14%~38%、15%~38%、16%~38%、17%~38%、18%~38%、19%~38%、20%~38%、21%~38%、22%~38%、23%~38%、24%~38%、25%~38%、26%~38%、27%~38%、28%~38%、2%~37%、3%~37%、4%~37%、5%~37%、6%~37%、7%~37%、8%~37%、9%~37%、10%~37%、11%~37%、12%~37%、13%~37%、14%~37%、15%~37%、16%~37%、17%~37%、18%~37%、19%~37%、20%~37%、21%~37%、22%~37%、23%~37%、24%~37%、25%~37%、26%~37%、27%~37%、28%~37%、2%~36%、3%~36%、4%~36%、5%~36%、6%~36%、7%~36%、8%~36%、9%~36%、10%~36%、11%~36%、12%~36%、13%~36%、14%~36%、15%~36%、16%~36%、17%~36%、18%~36%、19%~36%、20%~36%、21%~36%、22%~36%、23%~36%、24%~36%、25%~36%、26%~36%、27%~36%、28%~36%、2%~35%、3%~35%、4%~35%、5%~35%、6%~35%、7%~35%、8%~35%、9%~35%、10%~35%、11%~35%、12%~35%、13%~35%、14%~35%、15%~35%、16%~35%、17%~35%、18%~35%、19%~35%、20%~35%、21%~35%、22%~35%、23%~35%、24%~35%、25%~35%、26%~35%、27%~35%、28%~35%、2%~34%、3%~34%、4%~34%、5%~34%、6%~34%、7%~34%、8%~34%、9%~34%、10%~34%、11%~34%、12%~34%、13%~34%、14%~34%、15%~34%、16%~34%、17%~34%、18%~34%、19%~34%、20%~34%、21%~34%、22%~34%、23%~34%、24%~34%、25%~34%、26%~34%、27%~34%、28%~34%、2%~33%、3%~33%、4%~33%、5%~33%、6%~33%、7%~33%、8%~33%、9%~33%、10%~33%、11%~33%、12%~33%、13%~33%、14%~33%、15%~33%、16%~33%、17%~33%、18%~33%、19%~33%、20%~33%、21%~33%、22%~33%、23%~33%、24%~33%、25%~33%、26%~33%、27%~33%、28%~33%、2%~32%、3%~32%、4%~32%、5%~32%、6%~32%、7%~32%、8%~32%、9%~32%、10%~32%、11%~32%、12%~32%、13%~32%、14%~32%、15%~32%、16%~32%、17%~32%、18%~32%、19%~32%、20%~32%、21%~32%、22%~32%、23%~32%、24%~32%、25%~32%、26%~32%、27%~32%、28%~32%、2%~31%、3%~31%、4%~31%、5%~31%、6%~31%、7%~31%、8%~31%、9%~31%、10%~31%、11%~31%、12%~31%、13%~31%、14%~31%、15%~31%、16%~31%、17%~31%、18%~31%、19%~31%、20%~31%、21%~31%、22%~31%、23%~31%、24%~31%、25%~31%、26%~31%、27%~31%、28%~31%、2%~30%、3%~30%、4%~30%、5%~30%、6%~30%、7%~30%、8%~30%、9%~30%、10%~30%、11%~30%、12%~30%、13%~30%、14%~30%、15%~30%、16%~30%、17%~30%、18%~30%、19%~30%、20%~30%、21%~30%、22%~30%、23%~30%、24%~30%、25%~30%、26%~30%、27%~30%、28%~30%、29%~30%、1%~29%、2%~29%、3%~、 29%、4%~29%、5%~29%、6%~29%、7%~29%、8%~29%、9%~29%、10%~29%、11%~29%、12%~29%、13%~29%、14%~29%、15%~29%、16%~29%、17%~29%、18%~29%、19%~29%、20%~29%、21%~29%、22%~29%、23%~29%、24%~29%、25%~29%、26%~29%、27%~29%、28%~29%、1%~28%、2%~28%、3%~28%、4%~28%、5%~28%、6%~28%、7%~28%、8%~28%、9%~28%、10%~28%、11%~28%、12%~28%、13%~28%、14%~28%、15%~28%、16%~28%、17%~28%、18%~28%、19%~28%、20%~28%、21%~28%、22%~28%、23%~28%、24%~28%、25%~28%、26%~28%、27%~28%、1%~27%、2%~27%、3%~27%、4%~27%、5%~27%、6%~27%、7%~27%、8%~27%、9%~27%、10%~27%、11%~27%、12%~27%、13%~27%、14%~27%、15%~27%、16%~27%、17%~27%、18%~27%、19%~27%、20%~27%、21%~27%、22%~27%、23%~27%、24%~27%、25%~27%、26%~27%、1%~26%、2%~26%、3%~26%、4%~26%、5%~26%、6%~26%、7%~26%、8%~26%、9%~26%、10%~26%、11%~26%、12%~26%、13%~26%、14%~26%、15%~26%、16%~26%、17%~26%、18%~26%、19%~26%、20%~26%、21%~26%、22%~26%、23%~26%、24%~26%、25%~26%、1%~25%、2%~25%、3%~25%、4%~25%、5%~25%、6%~25%、7%~25%、8%~25%、9%~25%、10%~25%、11%~25%、12%~25%、13%~25%、14%~25%、15%~25%、16%~25%、17%~25%、18%~25%、19%~25%、20%~25%、21%~25%、22%~25%、23%~25%、24%~25%、1%~24%、2%~24%、3%~24%、4%~24%、5%~24%、6%~24%、7%~24%、8%~24%、9%~24%、10%~24%、11%~24%、12%~24%、13%~24%、14%~24%、15%~24%、16%~24%、17%~24%、18%~24%、19%~24%、20%~24%、21%~24%、22%~24%、23%~24%、1%~23%、2%~23%、3%~23%、4%~23%、5%~23%、6%~23%、7%~23%、8%~23%、9%~23%、10%~23%、11%~23%、12%~23%、13%~23%、14%~23%、15%~23%、16%~23%、17%~23%、18%~23%、19%~23%、20%~23%、21%~23%、22%~23%、1%~22%、2%~22%、3%~22%、4%~22%、5%~22%、6%~22%、7%~22%、8%~22%、9%~22%、10%~22%、11%~22%、12%~22%、13%~22%、14%~22%、15%~22%、16%~22%、17%~22%、18%~22%、19%~22%、20%~22%、21%~22%、1%~21%、2%~21%、3%~21%、4%~21%、5%~21%、6%~21%、7%~21%、8%~21%、9%~21%、10%~21%、11%~21%、12%~21%、13%~21%、14%~21%、15%~21%、16%~21%、17%~21%、18%~21%、19%~21%、20%~21%、1%~20%、2%~20%、3%~20%、4%~20%、5%~20%、6%~20%、7%~20%、8%~20%、9%~20%、10%~20%、11%~20%、12%~20%、13%~20%、14%~20%、15%~20%、16%~20%、17%~20%、18%~20%、19%~20%、1%~19%、2%~19%、3%~19%、4%~19%、5%~19%、6%~19%、7%~19%、8%~19%、9%~19%、10%~19%、11%~19%、12%~19%、13%~19%、14%~19%、15%~19%、16%~19%、17%~19%、18%~19%、1%~18%、2%~18%、3%~18%、4%~18%、5%~18%、6%~18%、7%~18%、8%~18%、9%~18%、10%~18%、11%~18%、12%~18%、13%~18%, 14%~18%, 15%~18%, 16%~18%, 17%~18%, 1%~17%, 2%~17%, 3%~17%, 4%~17%, 5%~17%, 6%~17%, 7%~17%, 8%~17%, 9%~17%, 10%~17%, 11%~17%, 12%~1 7%, 13%~17%, 14%~17%, 15%~17%, 16%~17%, 1%~16%, 2%~16%, 3%~16%, 4%~16%, 5%~16%, 6%~16%, 7%~16%, 8%~16%, 9%~16%, 10%~16%, 11%~16%, 12%~16%, 13% ~16%, 14%~16%, 15%~16%, 1%~15%, 2%~15%, 3%~15%, 4%~15%, 5%~15%, 6%~15%, 7%~15%, 8%~15%, 9%~15%, 10%~15%, 11%~15%, 12%~15%, 13%~15%, 14%~15%, 1%~14%, 2%~14%, 3%~14%, 4%~14%, 5%~14%, 6%~14%, 7%~14%, 8%~14%, 9%~14%, 10%~14%, 11%~14%, 12%~14%, 13%~14%, 1%~13%, 2%~13%, 3%~13%, 4%~13%, 5% ~13%, 6%~13%, 7%~13%, 8%~13%, 9%~13%, 10%~13%, 11%~13%, 12%~13%, 1%~12%, 2%~12%, 3%~12%, 4%~12%, 5%~12%, 6%~12%, 7%~12%, 8%~12%, 9%~12%, 10%~ 12%, 11%~12%, 1%~11%, 2%~11%, 3%~11%, 4%~11%, 5%~11%, 6%~11%, 7%~11%, 8%~11%, 9%~11%, 10%~11%, 1%~10%, 2%~10%, 3%~10%, 4%~10%, 5%~10%, 6%~10%, 7%-10%, 8%-10%, 9%-10%, 1%-9%, 2%-9%, 3%-9%, 4%-9%, 5%-9%, 6%-9%, 7%-9%, 8%-9%, 1%-8%, 2%-8%, 3%-8%, 4%-8%, 5%-8%, 6%-8%, 7%-8%, 1%-7%, 2%-7%, 3%-7%, 4%-7%, 5%-7%, 6%-7%, 1%-6%, 2%-6%, 3%-6%, 4%-6%, 5%-6%, 1%-5%, 2%-5%, 3%-5%, 4%-5%, 1%-4%, 2%-4%, 3%-4%, 1%-3%, 2%-3%, or 1%-2% higher fermentation rate.

[0065]

[0069] In some embodiments, the yeast strains and derivatives thereof described herein have one or more defining characteristics described herein. In some embodiments, the yeast strains and derivatives thereof described herein have one or more defining characteristics described herein at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids. In some embodiments, the yeast strains and derivatives thereof described herein have one or more defining characteristics described herein at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at normal pH without organic acids. For example, in some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly higher ethanol yield, a statistically significantly lower glycerol yield, a statistically significantly higher fermentation rate, a statistically significantly higher temperature tolerance, and a statistically significantly higher organic acid tolerance than typical yeast strains used in fermentation (e.g., Saccharomyces strain Y1609). In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly higher fermentation rate, produce lower levels of glycerol, and have a statistically significantly higher ethanol yield than typical yeast strains used in fermentation (e.g., Saccharomyces strain Y1609). In some embodiments, the yeast strains and derivatives thereof described herein have a statistically significantly higher fermentation rate, and produce statistically significantly lower levels of glycerol than typical yeast strains used in fermentation (e.g., Saccharomyces strain Y1609).

[0066]

[0070] The yeast strains and derivatives thereof described herein may be in any viable form, including crushed, dried (including active dry and instant), compressed, creamed, etc. In a particular embodiment, the Saccharomyces cerevisiae yeast strain or derivative thereof is a dry yeast, such as an active dry yeast. In another particular embodiment, the Saccharomyces cerevisiae yeast strain or derivative thereof is a compressed yeast. In another embodiment, the Saccharomyces cerevisiae yeast strain or derivative thereof is a cream yeast.

[0067] A yeast strain

[0071] One embodiment described herein is a Saccharomyces yeast strain designated Y1912 (deposited under NRRL patent deposit designation number Y-68003); Y1913 (deposited under NRRL patent deposit designation number Y-68004); Y1914 (deposited under NRRL patent deposit designation number Y-68005); Y1919 (deposited under NRRL patent deposit designation number Y-68006); Y1923 (deposited under NRRL patent deposit designation number Y-68007); Y1927 (deposited under NRRL patent deposit designation number Y-68008); Y1929 (deposited under NRRL patent deposit designation number Y-68009). These yeast strains are referred to herein as "yeast strains," "Saccharomyces yeast strains," or designations thereof (i.e., "Y1912" or "H041"; "Y1913" or "H136"; "Y1914" or "H138"; "Y1919" or "H172"; "Y1923" or "H237"; "Y1927" or "MM2E2A3"; "Y1929" or "MM2E3A1"). The yeast strains (i.e., Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929) were produced from one or more different Saccharomyces yeast strains by one or more methods shown in FIG. 1. In one aspect, the yeast strains described herein include one or more defining characteristics including higher ethanol production, lower glycerol production, higher temperature tolerance, higher organic acid tolerance, and higher fermentation rate than other yeast strains and typical yeast strains used in fermentation, particularly compared to the yeast Y1609 strain used in Fali products. Representative samples of the yeast strains have been deposited at the Agricultural Research Service Patent Culture Collection (NRRL), Northern Regional Research Center, 1815 University Street, Peoria, Illinois, USA under the above accession numbers.

[0068] b. Yeast strain derivative

[0072] Another embodiment described herein is a derivative of a Saccharomyces yeast strain selected from the Saccharomyces yeasts designated Y1912 (deposited under NRRL patent deposit designation number Y-68003); Y1913 (deposited under NRRL patent deposit designation number Y-68004); Y1914 (deposited under NRRL patent deposit designation number Y-68005); Y1919 (deposited under NRRL patent deposit designation number Y-68006); Y1923 (deposited under NRRL patent deposit designation number Y-68007); Y1927 (deposited under NRRL patent deposit designation number Y-68008); Y1929 (deposited under NRRL patent deposit designation number Y-68009). In one aspect, this derivative comprises one or more defining characteristics, including higher ethanol production, lower glycerol production, higher temperature tolerance, higher organic acid tolerance, and higher fermentation rate than other yeast strains and typical yeast strains used in fermentation, particularly compared to the yeast Y1609 strain used in Fali products. In another aspect, this derivative is a parent strain and can be used to produce other derivatives. For example, Y1912 is a derivative of two strains, but is also a parent strain (e.g., Y1912 was crossed with a second strain to produce Y1913).

[0069] C. Mutant yeast and derivatives

[0073] Another embodiment described herein is a mutant or derivative of a Saccharomyces yeast strain selected from the Saccharomyces yeast strains designated Y1912 (deposited under NRRL Patent Deposit Designation No. Y-68003), Y1913 (deposited under NRRL Patent Deposit Designation No. Y-68004); Y1914 (deposited under NRRL Patent Deposit Designation No. Y-68005); Y1919 (deposited under NRRL Patent Deposit Designation No. Y-68006); Y1923 (deposited under NRRL Patent Deposit Designation No. Y-68007); Y1927 (deposited under NRRL Patent Deposit Designation No. Y-68008); Y1929 (deposited under NRRL Patent Deposit Designation No. Y-68009). In one embodiment, the Saccharomyces yeast strains designated Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929 can be derived from one or more different Saccharomyces yeast strains by the method shown in FIG. 1, so that one or more of Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929 can be mutant yeast strains. In one aspect, the mutant yeast strains and mutant derivatives include one or more defining characteristics including higher ethanol production, lower glycerol production, higher temperature tolerance, higher organic acid tolerance, and higher fermentation rate than other yeast strains and typical yeast strains used in fermentation, particularly compared to the yeast Y1609 strain used in Fali products. In another aspect, the mutant yeast strains and mutant derivatives can be used as parent strains to produce other derivatives. An example of mutagenesis is shown in FIG. 1D. In one embodiment, the mutant yeast strains and mutant derivatives described herein are derived from the method depicted in FIG. 1D.

[0070]

[0074] Mutant yeast strains and mutant derivatives can be made by contacting any of the yeast strains described herein with a mutagen. The mutagen can be any mutagen known in the art. For example, the mutagen can be ethyl methanesulfonate (EMS), ultraviolet light (UV), X-rays, methyl methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine mustard, 2-methoxy-6-chloro-9[3-(ethyl-2-chloroethyl)aminopropylamino]acridine·2 (ICR-170), nitrogen mustard, and the like.

[0071] D. Evolved yeasts and derivatives

[0075] Another embodiment described herein is an evolved yeast strain or derivative of a Saccharomyces yeast strain selected from the Saccharomyces yeast strains designated Y1912 (deposited under NRRL Patent Deposit Designation No. Y-68003); Y1913 (deposited under NRRL Patent Deposit Designation No. Y-68004); Y1914 (deposited under NRRL Patent Deposit Designation No. Y-68005); Y1919 (deposited under NRRL Patent Deposit Designation No. Y-68006); Y1923 (deposited under NRRL Patent Deposit Designation No. Y-68007); Y1927 (deposited under NRRL Patent Deposit Designation No. Y-68008); Y1929 (deposited under NRRL Patent Deposit Designation No. Y-68009). In one embodiment, the Saccharomyces yeast strains designated Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929 can be derived from one or more different Saccharomyces yeast strains by the method shown in FIG. 1C, so that one or more of Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929 can be evolved yeast strains. In one aspect, the evolved yeast strains and evolved derivatives include one or more defining characteristics including higher ethanol production, lower glycerol production, higher temperature tolerance, higher organic acid tolerance, and higher fermentation rate than other yeast strains and typical yeast strains used in fermentation, particularly compared to the yeast Y1609 strain used in Fali products. In another aspect, the evolved yeast strains and evolved derivatives can be used as parent strains to produce other derivatives. An example of evolution is shown in FIG. 1C. In one embodiment, the evolved yeast strains and evolved derivatives described herein are derived from the method depicted in FIG. 1C.

[0072]

[0076] The evolved yeast strains and evolved derivatives can be made by applying selective pressure to any of the yeast strains described herein. The selective pressure can be negative (reducing the occurrence of a trait) or positive (increasing the rate of a trait). The selective pressure can be constant or intermittent. The selective pressure can be applied by altering the presence of sources (e.g., starch and sugars) and / or by altering environmental conditions (e.g., temperature, presence of organic acids, pH, and length of fermentation).

[0073] e. Recombinant yeast and derivatives

[0077] Further embodiments described herein are recombinant yeast strains and recombinant derivatives. The recombinant yeast strains and derivatives may be derived from the Saccharomyces cerevisiae yeast strains described herein or their derivatives. The recombinant yeast strains may include modifications to suppress expression of a gene, enhance expression of a gene, introduce a gene, delete a gene, or modify the sequence of a gene. An aspect described herein is a method of making a recombinant yeast strain or its derivative. The method may include using recombinant DNA techniques to introduce a nucleic acid into a Saccharomyces yeast described herein. Methods of introducing nucleic acids into Saccharomyces yeast cells, particularly strains of Saccharomyces, are known in the art and are disclosed, for example, in Ausubel et al. (1997), Current Protocols in Molecular Biology, 2:13.7.1-13.7.7, and Yang and Blenner, (2020), Curr Opin Biotechnol., 66:255-266, both of which are incorporated herein by reference. The method may include the step of altering a nucleic acid sequence of a Saccharomyces yeast or derivative described herein using gene editing or similar techniques.

[0074] 3. Composition

[0078] Further provided herein is a composition comprising the yeast strain or derivative thereof. In one embodiment, the composition can comprise at least one of the yeast strains described herein, the derivatives described herein, or combinations thereof, and natural and / or non-natural ingredients. For example, the composition can comprise one or more ingredients selected from surfactants, emulsifiers, thickeners, leavening agents, antioxidants, starches, metabolites, and other processing aids. In one embodiment, the composition can comprise dry yeast of any yeast strain and / or derivatives thereof, starch, and an emulsifier. In another embodiment, the composition can comprise cream yeast of any yeast strain and / or derivatives thereof, glycerol, and xanthan gum. In one embodiment, a concentrated culture of any yeast strain described herein is provided, the concentrated culture being 90%-99% pure. In another embodiment, a pure culture of any yeast strain described herein is provided, the pure culture being 100% pure and therefore no additional yeast is present.

[0075]

[0079] The composition may include the Saccharomyces yeast described herein and any suitable surfactant, hi one embodiment, the surfactant is an anionic surfactant, a cationic surfactant, and / or a non-ionic surfactant.

[0076]

[0080] The composition may include the Saccharomyces yeast described herein and any suitable emulsifier. In one embodiment, the emulsifier is a fatty acid ester of sorbitan. In one embodiment, the emulsifier is selected from the group of sorbitan monostearate (SMS), citric acid esters of monodiglycerides, polyglycerol esters, and fatty acid esters of propylene glycol.

[0077]

[0081] The compositions may include the Saccharomyces yeast described herein and Olindronic SMS, Olindronic SK, or Olindronic SPL, which contain compositions related to European Patent No. 1,724,336. These products can be purchased as active dry yeast from Bussetti of Austria.

[0078]

[0082] The composition may include the Saccharomyces yeast described herein and any suitable thickening agent. In one embodiment, the thickening agent is gum acacia, particularly for cream yeast, compressed yeast and dry yeast.

[0079]

[0083] The composition may include the Saccharomyces yeast described herein and any suitable leavening agent, hi one embodiment, the leavening agent is methylcellulose or carboxymethylcellulose.

[0080]

[0084] The compositions may include the Saccharomyces yeast described herein and any suitable antioxidant. In one embodiment, the antioxidant is butylated hydroxyanisole (BHA) and / or butylated hydroxytoluene (BHT), or ascorbic acid (vitamin C), particularly for active dry yeast.

[0081]

[0085] The composition may include the Saccharomyces yeast described herein and any suitable starch, hi one embodiment, the starch is potato starch, corn starch, or pea starch.

[0082]

[0086] The composition may include a Saccharomyces yeast as described herein and any suitable yeast protectant, hi one embodiment, the protectant is glycerol.

[0083] a. Composition characteristics

[0087] In one aspect, the composition comprises one or more defining characteristics including higher ethanol production, lower glycerol production, higher temperature tolerance, higher organic acid tolerance, and higher fermentation rate than other yeast products and typical yeast products used in fermentation, particularly compared to yeast product Fali. Additionally, the compositions described herein can be readily distinguished from other yeast products used in the ethanol industry that do not have the ethanol production capabilities and defining characteristics of the compositions described herein.

[0084]

[0088] In some embodiments, the compositions described herein have a higher ethanol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In some embodiments, the compositions described herein have a higher ethanol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C. In a particular embodiment, the compositions described herein have a higher ethanol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at a temperature of 32° C. In another particular embodiment, the compositions described herein have a higher ethanol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at a temperature of 36° C.

[0085]

[0089] In some embodiments, the compositions described herein produce at least about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0%, or 21.0% more ethanol after 50 hours of fermentation compared to a typical yeast product used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In some embodiments, the compositions described herein produce up to about 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0%, or 21.0% more ethanol after 50 hours of fermentation compared to a typical yeast product used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). ... Alcohol®), after 50 hours of fermentation, the percentage of ethanol in the range of about 0.1% to 21% (i.e., from about 0.1% to about 21%), 0.2% to 21%, 0.3% to 21%, 0.4% to 21%, 0.5% to 21%, 0.6% to 21%, 0.7% to 21%, 0.8% to 21%, 0.9% to 21%, 1% ~21%, 2%~21%, 3%~21%, 4%~21%, 5%~21%, 6%~21%, 7%~21%, 8%~21%, 9%~21%, 10%~21%, 0.2%~20%, 0.3%~20%, 0.4%~20%, 0.5%~20%, 0.6%~20%, 0.7%~20%, 0.8%~20%, 0.9%~20%、1%~20%、2%~20%、3%~20%、4%~20%、5%~20%、6%~20%、7%~20%、8%~20%、9%~20%、10%~20%、0.2%~19%、0.3%~19%、0.4%~19%、0.5%~19%、0.6%~19%、0.7%~19%、0.8%~19%、0.9%~19%、1%~19%、2%~19%、3%~19%、4%~19%、5%~19%、6%~19%、7%~19%、8%~19%、9%~19%、10%~19%、0.2%~18%、0.3%~18%、0.4%~18%、0.5%~18%、0.6%~18%、0.7%~18%、0.8%~18%、0.9%~18%、1%~18%、2%~18%、3%~18%、4%~18%、5%~18%、6%~18%、7%~18%、8%~18%、9%~18%、10%~18%、0.2%~17%、0.3%~17%、0.4%~17%、0.5%~17%、0.6%~17%、0.7%~17%、0.8%~17%、0.9%~17%、1%~17%、2%~17%、3%~17%、4%~17%、5%~17%、6%~17%、7%~17%、8%~17%、9%~17%、10%~17%、0.2%~16%、0.3%~16%、0.4%~16%、0.5%~16%、0.6%~16%、0.7%~16%、0.8%~16%、0.9%~16%、1%~16%、2%~16%、3%~16%、4%~16%、5%~16%、6%~16%、7%~16%、8%~16%、9%~16%、10%~16%、0.2%~15%、0.3%~15%、0.4%~15%、0.5%~15%、0.6%~15%、0.7%~15%、0.8%~15%、0.9%~15%、1%~15%、2%~15%、3%~15%、4%~15%、5%~15%、6%~15%、7%~15%、8%~15%、9%~15%、10%~15%、0.1%~14%、0.2%~14%、0.3%~14%、0.4%~14%、0.5%~14%、0.6%~14%、0.7%~14%、0.8%~14%、0.9%~14%、1%~14%、2%~14%、3%~14%、4%~14%、5%~14%、6%~14%、7%~14%、8%~14%、9%~14%、10%~14%、0.1%~13%、0.2%~13%、0.3%~13%、0.4%~13%、0.5%~13%、0.6%~13%、0.7%~13%, 0.8%~13%, 0.9%~13%, 1%~13%, 2%~13%, 3%~13%, 4%~13%, 5%~13%, 6%~13%, 7%~13%, 8%~13%, 9%~13%, 10%~13%, 0.1%~12%, 0.2%~12%, 0.3%~12%, 0.4%~12%, 0.5%~12%, 0.6%~12%, 0.7%~12%, 0.8%~12%, 0.9%~12%, 1%~12%, 2%~12%, 3%~12%, 4%~12%, 5%~12%, 6%~12%, 7%~12%, 8%~12%, 9%~12%, 10%~12%, 0.1%~11%, 0.2%~11%, 0.3%~11%, 0.4%~11% , 0.5%~11%, 0.6%~11%, 0.7%~11%, 0.8%~11%, 0.9%~11%, 1%~11%, 2%~11%, 3%~11%, 4%~11%, 5%~11%, 6%~11%, 7%~11%, 8%~11%, 9%~11%, 10%~11%, 0.1%~0.2%, 0.1%~0.3%, 0.1%~ Produces 0.4%, 0.1%-0.5%, 0.1%-0.6%, 0.1%-0.7%, 0.1%-0.8%, 0.1%-0.9%, 0.1%-1%, 0.1%-2%, 0.1%-3%, 0.1%-4%, 0.1%-5%, 0.1%-6%, 0.1%-7%, 0.1%-8%, or 0.1%-9% more ethanol.

[0086]

[0090] In some embodiments, the compositions described herein have a lower glycerol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In some embodiments, the compositions described herein have a lower glycerol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at temperatures between 20°C and 40°C, preferably between 32°C and 36°C. In a particular embodiment, the compositions described herein have a lower glycerol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at a temperature of 32°C. In another particular embodiment, the compositions described herein have a lower glycerol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at a temperature of 36°C.

[0087]

[0091] In some embodiments, the compositions described herein produce at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, or 22% less glycerol after 50 hours of fermentation compared to typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In some embodiments, the compositions described herein produce up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30% less glycerol after 50 hours of fermentation compared to typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In some embodiments, the yeast strains and derivatives thereof described herein produce up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30% less glycerol after 50 hours of fermentation compared to typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). Alcohol®), after 50 hours of fermentation, about 1%-30% (i.e., from about 1% to about 30%), 2%-30%, 3%-30%, 4%-30%, 5%-30%, 6%-30%, 7%-30%, 8%-30%, 9%-30%, 10%-30%, 11%-30%, 12%-30%, 13%-30%, 14%-30%, 15%-30%, 16%-30%, 17%-30%, 18%-30%, 19%-30%, 20%-30%, 21%-30%, 22%-30% , 23%~30%, 24%~30%, 25%~30%, 26%~30%, 27%~30%, 28%~30%, 29%~30%, 1%~29%, 2%~29%, 3%~29%, 4%~29%, 5%~29%, 6%~29%, 7%~29%, 8%~29%, 9%~29%, 10%~29%, 11%~29%, 12%~29%, 13%~29%, 14%~29%, 15%~29%, 16%~29%, 17%~29%, 18%~29%, 19%~29%, 20%~29%, 21%~29%,22%~29%、23%~29%、24%~29%、25%~29%、26%~29%、27%~29%、28%~29%、1%~28%、2%~28%、3%~28%、4%~28%、5%~28%、6%~28%、7%~28%、8%~28%、9%~28%、10%~28%、11%~28%、12%~28%、13%~28%、14%~28%、15%~28%、16%~28%、17%~28%、18%~28%、19%~28%、20%~28%、21%~28%、22%~28%、23%~28%、24%~28%、25%~28%、26%~28%、27%~28%、1%~27%、2%~27%、3%~27%、4%~27%、5%~27%、6%~27%、7%~27%、8%~27%、9%~27%、10%~27%、11%~27%、12%~27%、13%~27%、14%~27%、15%~27%、16%~27%、17%~27%、18%~27%、19%~27%、20%~27%、21%~27%、22%~27%、23%~27%、24%~27%、25%~27%、26%~27%、1%~26%、2%~26%、3%~26%、4%~26%、5%~26%、6%~26%、7%~26%、8%~26%、9%~26%、10%~26%、11%~26%、12%~26%、13%~26%、14%~26%、15%~26%、16%~26%、17%~26%、18%~26%、19%~26%、20%~26%、21%~26%、22%~26%、23%~26%、24%~26%、25%~26%、1%~25%、2%~25%、3%~25%、4%~25%、5%~25%、6%~25%、7%~25%、8%~25%、9%~25%、10%~25%、11%~25%、12%~25%、13%~25%、14%~25%、15%~25%、16%~25%、17%~25%、18%~25%、19%~25%、20%~25%、21%~25%、22%~25%、23%~25%、24%~25%、1%~24%、2%~24%、3%~24%、4%~24%、5%~24%、6%~24%、7%~24%、8%~24%、9%~24%、10%~24%、11%~24%、12%~24%、13%~24%、14%~24%、15%~24%、16%~24%、17%~24%、18%~24%、19%~24%、20%~24%、21%~24%、22%~24%、23%~24%、1%~23%、2%~23%、3%~23%、4%~23%、5%~23%、6%~23%、7%~23%、8%~23%、9%~23%、10%~23%、11%~23%、12%~23%、13%~23%、14%~23%、15%~23%、16%~23%、17%~23%、18%~23%、19%~23%、20%~23%、21%~23%、22%~23%、1%~22%、2%~22%、3%~22%、4%~22%、5%~22%、6%~22%、7%~22%、8%~22%、9%~22%、10%~22%、11%~22%、12%~22%、13%~22%、14%~22%、15%~22%、16%~22%、17%~22%、18%~22%、19%~22%、20%~22%、21%~22%、1%~21%、2%~21%、3%~21%、4%~21%、5%~21%、6%~21%、7%~21%、8%~21%、9%~21%、10%~21%、11%~21%、12%~21%、13%~21%、14%~21%、15%~21%、16%~21%、17%~21%、18%~21%、19%~21%、20%~21%、1%~20%、2%~20%、3%~20%、4%~20%、5%~20%、6%~20%、7%~20%、8%~20%、9%~20%、10%~20%、11%~20%、12%~20%、13%~20%、14%~20%、15%~20%、16%~20%、17%~20%、18%~20%、19%~20%、1%~19%、2%~19%、3%~19%、4%~19%、5%~19%、6%~19%、7%~19%、8%~19%、9%~19%、10%~19%、11%~19%、12%~19%、13%~19%、14%~19%、15%~19%、16%~19%、17%~19%、18%~19%、1%~18%、2%~18%、3%~18%、4%~18%、5%~18%、6%~18%、7%~18%、8%~18%、9%~18%、10%~18%、11%~18%、12%~18%、13%~18%、14%~18%、15%~18%、16%~18%、17%~18%、1%~17%、2%~17%、3%~17%、4%~17%、5%~17%、6%~17%、7%~17%、8%~17%、9%~17%、10%~17%、11%~17%、12%~17%、13%~17%、14%~17%、15%~17%, 16%~17%, 1%~16%, 2%~16%, 3%~16%, 4%~16%, 5%~16%, 6%~16%, 7%~16%, 8%~16%, 9%~16%, 10%~16%, 11%~16%, 12%~16%, 13%~16%, 14%~16%, 15%~16%, 1%~15%, 2%~15%, 3%~15%, 4%~15%, 5%~15%, 6%~15%, 7%~15%, 8%~15%, 9%~15%, 10%~15%, 11%~15%, 12%~15%, 13%~15%, 14%~15%, 1%~14%, 2%~14%, 3%~14%, 4%~14%, 5%~14%, 6%~14%, 7%~14%, 8%~14%, 9%~14%, 10%~14%, 11%~14%, 12%~14%, 13%~14%, 1%~13%, 2%~13%, 3%~13%, 4%~13%, 5%~13%, 6%~13%, 7%~13%, 8%~13%, 9%~13%, 10%~13%, 11%~13%, 12%~13%, 1%~12%, 2%~12%, 3%~ 12%, 4%~12%, 5%~12%, 6%~12%, 7%~12%, 8%~12%, 9%~12%, 10%~12%, 11%~12%, 1%~11%, 2%~11%, 3%~11%, 4%~11%, 5%~11%, 6%~11%, 7%~11%, 8%~11%, 9%~11%, 10%~11%, 1%~10%, 2%~10%, 3%~10%, 4%~10%, 5%~10%, 6%~10%, 7%~10%, 8%~10%, 9%~10%, 1%~9%, 2%~9%, 3%~9%, Generate 4%-9%, 5%-9%, 6%-9%, 7%-9%, 8%-9%, 1%-8%, 2%-8%, 3%-8%, 4%-8%, 5%-8%, 6%-8%, 7%-8%, 1%-7%, 2%-7%, 3%-7%, 4%-7%, 5%-7%, 6%-7%, 1%-6%, 2%-6%, 3%-6%, 4%-6%, 5%-6%, 1%-5%, 2%-5%, 3%-5%, 4%-5%, 1%-4%, 2%-4%, 3%-4%, 1%-3%, 2%-3%, or 1%-2% less glycerol.

[0088]

[0092] In some embodiments, the compositions described herein have a higher fermentation rate than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In some embodiments, the compositions described herein have a higher fermentation rate than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at temperatures between 20°C and 40°C, preferably between 32°C and 36°C. In a particular embodiment, the compositions described herein have a higher fermentation rate than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at a temperature of 32°C. In another particular embodiment, the compositions described herein have a higher fermentation rate than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at a temperature of 36°C.

[0089]

[0093] In some embodiments, the compositions described herein have a fermentation rate after 24 hours of fermentation that is at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% higher than a typical yeast product used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In some embodiments, the compositions described herein have a fermentation rate that is up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% higher after 24 hours of fermentation than a typical yeast product used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In some embodiments, the compositions described herein, after 24 hours of fermentation, exhibit a yield of about 1%-50% (i.e., from about 1% to about 50%), 2%-50%, 3%-50%, 4%-50%, 5%-50%, 6%-50%, 7%-50%, 8%-50%, 9%-50%, 10%-50%, 11%-50%, 12%-50%, 13%-50%, 14%-50%, 15%-50%, 16%-50%, 17%-50%, 18%-50%, 19%-50%, 20%-50%, 21%-50%, 22%-50%, 23%-50%, 24%-50%, 25%-50%, 26%-50%, 27%-50%, 28%-50%, 29%-50%, 30%-50%, 31%-50%, 32%-50%, 33%-50%, 34%-50%, 35%-50%, 36%-50%, 37%-50%, 38%-50%, 39%-50%, 40%-50%, 41%-50%, 42%-50%, 43%-50%, 44%-50%, 45%-50%, 46%-50%, 47%-50%, 48%-50%, 49%-50%, 50%-50%, 50%-50%, 51%-50%, 52%-50%, 53%-50%, 54%-50%, 55%-50%, 56%-50%, 57%-50%, 58%-50%, 59%- %~50%, 22%~50%, 23%~50%, 24%~50%, 25%~50%, 26%~50%, 27%~50%, 28%~50%, 2%~49%, 3%~49%, 4%~49%, 5%~49%, 6%~49%, 7%~49%, 8%~49%, 9%~49%, 10%~49%, 11%~49%, 12%~49%, 13%~49%, 14%~49%, 15%~49%, 16%~49%, 17%~49%, 18%~49%,19%~49%、20%~49%、21%~49%、22%~49%、23%~49%、24%~49%、25%~49%、26%~49%、27%~49%、28%~49%、2%~48%、3%~48%、4%~48%、5%~48%、6%~48%、7%~48%、8%~48%、9%~48%、10%~48%、11%~48%、12%~48%、13%~48%、14%~48%、15%~48%、16%~48%、17%~48%、18%~48%、19%~48%、20%~48%、21%~48%、22%~48%、23%~48%、24%~48%、25%~48%、26%~48%、27%~48%、28%~48%、2%~47%、3%~47%、4%~47%、5%~47%、6%~47%、7%~47%、8%~47%、9%~47%、10%~47%、11%~47%、12%~47%、13%~47%、14%~47%、15%~47%、16%~47%、17%~47%、18%~47%、19%~47%、20%~47%、21%~47%、22%~47%、23%~47%、24%~47%、25%~47%、26%~47%、27%~47%、28%~47%、2%~46%、3%~46%、4%~46%、5%~46%、6%~46%、7%~46%、8%~46%、9%~46%、10%~46%、11%~46%、12%~46%、13%~46%、14%~46%、15%~46%、16%~46%、17%~46%、18%~46%、19%~46%、20%~46%、21%~46%、22%~46%、23%~46%、24%~46%、25%~46%、26%~46%、27%~46%、28%~46%、2%~45%、3%~45%、4%~45%、5%~45%、6%~45%、7%~45%、8%~45%、9%~45%、10%~45%、11%~45%、12%~45%、13%~45%、14%~45%、15%~45%、16%~45%、17%~45%、18%~45%、19%~45%、20%~45%、21%~45%、22%~45%、23%~45%、24%~45%、25%~45%、26%~45%、27%~45%、28%~45%、2%~44%、3%~44%、4%~44%、5%~44%、6%~44%、7%~44%、8%~44%、9%~44%、10%~44%、11%~44%、12%~44%、13%~44%、14%~44%、15%~44%、16%~44%、17%~44%、18%~44%、19%~44%、20%~44%、21%~44%、22%~44%、23%~44%、24%~44%、25%~44%、26%~44%、27%~44%、28%~44%、2%~43%、3%~43%、4%~43%、5%~43%、6%~43%、7%~43%、8%~43%、9%~43%、10%~43%、11%~43%、12%~43%、13%~43%、14%~43%、15%~43%、16%~43%、17%~43%、18%~43%、19%~43%、20%~43%、21%~43%、22%~43%、23%~43%、24%~43%、25%~43%、26%~43%、27%~43%、28%~43%、2%~42%、3%~42%、4%~42%、5%~42%、6%~42%、7%~42%、8%~42%、9%~42%、10%~42%、11%~42%、12%~42%、13%~42%、14%~42%、15%~42%、16%~42%、17%~42%、18%~42%、19%~42%、20%~42%、21%~42%、22%~42%、23%~42%、24%~42%、25%~42%、26%~42%、27%~42%、28%~42%、2%~41%、3%~41%、4%~41%、5%~41%、6%~41%、7%~41%、8%~41%、9%~41%、10%~41%、11%~41%、12%~41%、13%~41%、14%~41%、15%~41%、16%~41%、17%~41%、18%~41%、19%~41%、20%~41%、21%~41%、22%~41%、23%~41%、24%~41%、25%~41%、26%~41%、27%~41%、28%~41%、2%~40%、3%~40%、4%~40%、5%~40%、6%~40%、7%~40%、8%~40%、9%~40%、10%~40%、11%~40%、12%~40%、13%~40%、14%~40%、15%~40%、16%~40%、17%~40%、18%~40%、19%~40%、20%~40%、21%~40%、22%~40%、23%~40%、24%~40%、25%~40%、26%~40%、27%~40%、28%~40%、2%~39%、3%~39%、4%~39%、5%~39%、6%~39%、7%~39%、8%~39%、9%~39%、10%~39%、11%~39%、12%~39%、13%~39%、14%~39%、15%~39%、16%~39%、17%~39%、18%~39%、19%~39%、20%~39%、21%~39%、22%~39%、23%~39%、24%~39%、25%~39%、26%~39%、27%~39%、28%~39%、2%~38%、3%~38%、4%~38%、5%~38%、6%~38%、7%~38%、8%~38%、9%~38%、10%~38%、11%~38%、12%~38%、13%~38%、14%~38%、15%~38%、16%~38%、17%~38%、18%~38%、19%~38%、20%~38%、21%~38%、22%~38%、23%~38%、24%~38%、25%~38%、26%~38%、27%~38%、28%~38%、2%~37%、3%~37%、4%~37%、5%~37%、6%~37%、7%~37%、8%~37%、9%~37%、10%~37%、11%~37%、12%~37%、13%~37%、14%~37%、15%~37%、16%~37%、17%~37%、18%~37%、19%~37%、20%~37%、21%~37%、22%~37%、23%~37%、24%~37%、25%~37%、26%~37%、27%~37%、28%~37%、2%~36%、3%~36%、4%~36%、5%~36%、6%~36%、7%~36%、8%~36%、9%~36%、10%~36%、11%~36%、12%~36%、13%~36%、14%~36%、15%~36%、16%~36%、17%~36%、18%~36%、19%~36%、20%~36%、21%~36%、22%~36%、23%~36%、24%~36%、25%~36%、26%~36%、27%~36%、28%~36%、2%~35%、3%~35%、4%~35%、5%~35%、6%~35%、7%~35%、8%~35%、9%~35%、10%~35%、11%~35%、12%~35%、13%~35%、14%~35%、15%~35%、16%~35%、17%~35%、18%~35%、19%~35%、20%~35%、21%~35%、22%~35%、23%~35%、24%~35%、25%~35%、26%~35%、27%~35%、28%~35%、2%~34%、3%~34%、4%~34%、5%~34%、6%~34%、7%~34%、8%~34%、9%~34%、10%~34%、11%~34%、12%~34%、13%~34%、14%~34%、15%~34%、16%~34%、17%~34%、18%~34%、19%~34%、20%~34%、21%~34%、22%~34%、23%~34%、24%~34%、25%~34%、26%~34%、27%~34%、28%~34%、2%~33%、3%~33%、4%~33%、5%~33%、6%~33%、7%~33%、8%~33%、9%~33%、10%~33%、11%~33%、12%~33%、13%~33%、14%~33%、15%~33%、16%~33%、17%~33%、18%~33%、19%~33%、20%~33%、21%~33%、22%~33%、23%~33%、24%~33%、25%~33%、26%~33%、27%~33%、28%~33%、2%~32%、3%~32%、4%~32%、5%~32%、6%~32%、7%~32%、8%~32%、9%~32%、10%~32%、11%~32%、12%~32%、13%~32%、14%~32%、15%~32%、16%~32%、17%~32%、18%~32%、19%~32%、20%~32%、21%~32%、22%~32%、23%~32%、24%~32%、25%~32%、26%~32%、27%~32%、28%~32%、2%~31%、3%~31%、4%~31%、5%~31%、6%~31%、7%~31%、8%~31%、9%~31%、10%~31%、11%~31%、12%~31%、13%~31%、14%~31%、15%~31%、16%~31%、17%~31%、18%~31%、19%~31%、20%~31%、21%~31%、22%~31%、23%~31%、24%~31%、25%~31%、26%~31%、27%~31%、28%~31%、2%~30%、3%~30%、4%~30%、5%~30%、6%~30%、7%~30%、8%~30%、9%~30%、10%~30%、11%~30%、12%~30%、13%~30%、14%~30%、15%~30%、16%~30%、17%~30%、18%~30%、19%~30%、20%~30%、21%~30%、22%~30%、23%~30%、24%~30%、25%~30%、26%~30%、27%~30%、28%~30%、29%~30%、1%~29%、2%~29%、3%~29%、4%~29%、5%~29%、 、6%~29%、7%~29%、8%~29%、9%~29%、10%~29%、11%~29%、12%~29%、13%~29%、14%~29%、15%~29%、16%~29%、17%~29%、18%~29%、19%~29%、20%~29%、21%~29%、22%~29%、23%~29%、24%~29%、25%~29%、26%~29%、27%~29%、28%~29%、1%~28%、2%~28%、3%~28%、4%~28%、5%~28%、6%~28%、7%~28%、8%~28%、9%~28%、10%~28%、11%~28%、12%~28%、13%~28%、14%~28%、15%~28%、16%~28%、17%~28%、18%~28%、19%~28%、20%~28%、21%~28%、22%~28%、23%~28%、24%~28%、25%~28%、26%~28%、27%~28%、1%~27%、2%~27%、3%~27%、4%~27%、5%~27%、6%~27%、7%~27%、8%~27%、9%~27%、10%~27%、11%~27%、12%~27%、13%~27%、14%~27%、15%~27%、16%~27%、17%~27%、18%~27%、19%~27%、20%~27%、21%~27%、22%~27%、23%~27%、24%~27%、25%~27%、26%~27%、1%~26%、2%~26%、3%~26%、4%~26%、5%~26%、6%~26%、7%~26%、8%~26%、9%~26%、10%~26%、11%~26%、12%~26%、13%~26%、14%~26%、15%~26%、16%~26%、17%~26%、18%~26%、19%~26%、20%~26%、21%~26%、22%~26%、23%~26%、24%~26%、25%~26%、1%~25%、2%~25%、3%~25%、4%~25%、5%~25%、6%~25%、7%~25%、8%~25%、9%~25%、10%~25%、11%~25%、12%~25%、13%~25%、14%~25%、15%~25%、16%~25%、17%~25%、18%~25%、19%~25%、20%~25%、21%~25%、22%~25%、23%~25%、24%~25%、1%~24%、2%~24%、3%~24%、4%~24%、5%~24%、6%~24%、7%~24%、8%~24%、9%~24%、10%~24%、11%~24%、12%~24%、13%~24%、14%~24%、15%~24%、16%~24%、17%~24%、18%~24%、19%~24%、20%~24%、21%~24%、22%~24%、23%~24%、1%~23%、2%~23%、3%~23%、4%~23%、5%~23%、6%~23%、7%~23%、8%~23%、9%~23%、10%~23%、11%~23%、12%~23%、13%~23%、14%~23%、15%~23%、16%~23%、17%~23%、18%~23%、19%~23%、20%~23%、21%~23%、22%~23%、1%~22%、2%~22%、3%~22%、4%~22%、5%~22%、6%~22%、7%~22%、8%~22%、9%~22%、10%~22%、11%~22%、12%~22%、13%~22%、14%~22%、15%~22%、16%~22%、17%~22%、18%~22%、19%~22%、20%~22%、21%~22%、1%~21%、2%~21%、3%~21%、4%~21%、5%~21%、6%~21%、7%~21%、8%~21%、9%~21%、10%~21%、11%~21%、12%~21%、13%~21%、14%~21%、15%~21%、16%~21%、17%~21%、18%~21%、19%~21%、20%~21%、1%~20%、2%~20%、3%~20%、4%~20%、5%~20%、6%~20%、7%~20%、8%~20%、9%~20%、10%~20%、11%~20%、12%~20%、13%~20%、14%~20%、15%~20%、16%~20%、17%~20%、18%~20%、19%~20%、1%~19%、2%~19%、3%~19%、4%~19%、5%~19%、6%~19%、7%~19%、8%~19%、9%~19%、10%~19%、11%~19%、12%~19%、13%~19%、14%~19%、15%~19%、16%~19%、17%~19%、18%~19%、1%~18%、2%~18%、3%~18%、4%~18%、5%~18%、6%~18%、7%~18%、8%~18%、9%~18%、10%~18%、11%~18%、12%~18%、13%~18%、14%~18%、15%~18%, 16%~18%, 17%~18%, 1%~17%, 2%~17%, 3%~17%, 4%~17%, 5%~17%, 6%~17%, 7%~17%, 8%~17%, 9%~17%, 10%~17%, 11%~17%, 12%~17%, 13%~17%, 14%~17%, 15%~17%, 16%~17%, 1%~16%, 2%~16%, 3%~16%, 4%~16%, 5%~16%, 6%~16%, 7%~16%, 8%~16%, 9%~16%, 10%~16%, 11%~16%, 12%~16%, 13%~16%, 14%~16% , 15%~16%, 1%~15%, 2%~15%, 3%~15%, 4%~15%, 5%~15%, 6%~15%, 7%~15%, 8%~15%, 9%~15%, 10%~15%, 11%~15%, 12%~15%, 13%~15%, 14%~15%, 1%~14%, 2%~ 14%, 3%~14%, 4%~14%, 5%~14%, 6%~14%, 7%~14%, 8%~14%, 9%~14%, 10%~14%, 11%~14%, 12%~14%, 13%~14%, 1%~13%, 2%~13%, 3%~13%, 4%~13%, 5%~13%, 6%~ 13%, 7%~13%, 8%~13%, 9%~13%, 10%~13%, 11%~13%, 12%~13%, 1%~12%, 2%~12%, 3%~12%, 4%~12%, 5%~12%, 6%~12%, 7%~12%, 8%~12%, 9%~12%, 10%~12%, 11 %~12%, 1%~11%, 2%~11%, 3%~11%, 4%~11%, 5%~11%, 6%~11%, 7%~11%, 8%~11%, 9%~11%, 10%~11%, 1%~10%, 2%~10%, 3%~10%, 4%~10%, 5%~10%, 6%~10%, 7%~1 0%, 8%-10%, 9%-10%, 1%-9%, 2%-9%, 3%-9%, 4%-9%, 5%-9%, 6%-9%, 7%-9%, 8%-9%, 1%-8%, 2%-8%, 3%-8%, 4%-8%, 5%-8%, 6%-8%, 7%-8%, 1%-7%, 2%-7%, 3%-7%, 4%-7%, 5%-7%, 6%-7%, 1%-6%, 2%-6%, 3%-6%, 4%-6%, 5%-6%, 1%-5%, 2%-5%, 3%-5%, 4%-5%, 1%-4%, 2%-4%, 3%-4%, 1%-3%, 2%-3%, or 1%-2% higher fermentation rate.

[0090]

[0094] In some embodiments, the compositions described herein have higher temperature tolerance than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). Temperature tolerance may be indicated by increased ethanol production, decreased glycerol production, and / or increased fermentation rate. In certain embodiments, the compositions described herein may withstand temperatures of about 20°C, about 21°C, about 22°C, about 23°C, about 24°C, about 25°C, about 26°C, about 27°C, about 28°C, about 29°C, about 30°C, about 31°C, about 32°C, about 33°C, about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, and / or about 40°C.

[0091]

[0095] In some embodiments, the compositions described herein have a higher organic acid tolerance at low pH than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In some embodiments, the compositions described herein have a higher organic acid tolerance at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In a particular embodiment, the compositions described herein have a higher organic acid tolerance at a temperature of 32° C. than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In another particular embodiment, the compositions described herein have a higher organic acid tolerance at a temperature of 36° C. than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®).

[0092]

[0096] In some embodiments, the compositions described herein have a higher ethanol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids. In a particular embodiment, the compositions described herein have a higher ethanol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at temperatures between 32° C. and low pH with organic acids. In another particular embodiment, the compositions described herein have a higher ethanol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at temperatures between 36° C. and low pH with organic acids.

[0093]

[0097] In some embodiments, the compositions described herein can be grown at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., at low pH with organic acids, in the presence of typical yeast products used for fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super®, etc.). In some embodiments, the ethanol produced by the process produces at least about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0%, or 21.0% more ethanol after 50 hours of fermentation compared to ethanol produced by the process described above. In some embodiments, the compositions described herein produce up to about 1.0%, 2.0%, 3.0%, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10.0%, 11.0%, 12.0%, 13.0%, 14.0%, 15.0%, 16.0%, 17.0%, 18.0%, 19.0%, 20.0%, or 21.0% more ethanol after 50 hours of fermentation compared to typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids. In some embodiments, the compositions described herein can be grown at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., at low pH with organic acids, in the presence of typical yeast products used for fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super®, etc.). Alcohol®), after 50 hours of fermentation, the percentage of maltose in the range of about 0.1% to 21% (i.e., from about 0.1% to about 21%), 0.2% to 21%, 0.3% to 21%, 0.4% to 21%, 0.5% to 21%, 0.6% to 21%, 0.7% to 21%, 0.8% to 21%, 0.9% to 21%, 1% to 21%, 2% to 21%, 3% to 21%, 4% to 21%, 5% to 21%, 6% to 21%, 7% to 21%, 8% to 21%, 9% to 21%, 10% to 21%, 0.2% to 20%, 0.3% to 20%, 0.4%~20%、0.5%~20%、0.6%~20%、0.7%~20%、0.8%~20%、0.9%~20%、1%~20%、2%~20%、3%~20%、4%~20%、5%~20%、6%~20%、7%~20%、8%~20%、9%~20%、10%~20%、0.2%~19%、0.3%~19%、0.4%~19%、0.5%~19%、0.6%~19%、0.7%~19%、0.8%~19%、0.9%~19%、1%~19%、2%~19%、3%~19%、4%~19%、5%~19%、6%~19%、7%~19%、8%~19%、9%~19%、10%~19%、0.2%~18%、0.3%~18%、0.4%~18%、0.5%~18%、0.6%~18%、0.7%~18%、0.8%~18%、0.9%~18%、1%~18%、2%~18%、3%~18%、4%~18%、5%~18%、6%~18%、7%~18%、8%~18%、9%~18%、10%~18%、0.2%~17%、0.3%~17%、0.4%~17%、0.5%~17%、0.6%~17%、0.7%~17%、0.8%~17%、0.9%~17%、1%~17%、2%~17%、3%~17%、4%~17%、5%~17%、6%~17%、7%~17%、8%~17%、9%~17%、10%~17%、0.2%~16%、0.3%~16%、0.4%~16%、0.5%~16%、0.6%~16%、0.7%~16%、0.8%~16%、0.9%~16%、1%~16%、2%~16%、3%~16%、4%~16%、5%~16%、6%~16%、7%~16%、8%~16%、9%~16%、10%~16%、0.2%~15%、0.3%~15%、0.4%~15%、0.5%~15%、0.6%~15%、0.7%~15%、0.8%~15%、0.9%~15%、1%~15%、2%~15%、3%~15%、4%~15%、5%~15%、6%~15%、7%~15%、8%~15%、9%~15%、10%~15%、0.1%~14%、0.2%~14%、0.3%~14%、0.4%~14%、0.5%~14%、0.6%~14%、0.7%~14%、0.8%~14%、0.9%~14%、1%~14%、2%~14%、3%~14%、4%~14%、5%~14%、6%~14%、7%~14%、8%~14%、9%~14%、10%~14%、0.1%~13%、0.2%~13%, 0.3%~13%, 0.4%~13%, 0.5%~13%, 0.6%~13%, 0.7%~13%, 0.8%~13%, 0.9%~13%, 1%~13%, 2%~13%, 3%~13%, 4%~13%, 5%~13%, 6%~13%, 7%~13%, 8%~13%, 9%~13%, 10%~13%, 0.1%~12%, 0.2 %~12%, 0.3%~12%, 0.4%~12%, 0.5%~12%, 0.6%~12%, 0.7%~12%, 0.8%~12%, 0.9%~12%, 1%~12%, 2%~12%, 3%~12%, 4%~12%, 5%~12%, 6%~12%, 7%~12%, 8%~12%, 9%~12%, 10%~12%, 0.1%~11%, 0.2% ~11%, 0.3%~11%, 0.4%~11%, 0.5%~11%, 0.6%~11%, 0.7%~11%, 0.8%~11%, 0.9%~11%, 1%~11%, 2%~11%, 3%~11%, 4%~11%, 5%~11%, 6%~11%, 7%~11%, 8%~11%, 9%~11%, 10%~11%, 0.1%~0.2%, 0.1% Produces ~0.3%, 0.1%-0.4%, 0.1%-0.5%, 0.1%-0.6%, 0.1%-0.7%, 0.1%-0.8%, 0.1%-0.9%, 0.1%-1%, 0.1%-2%, 0.1%-3%, 0.1%-4%, 0.1%-5%, 0.1%-6%, 0.1%-7%, 0.1%-8%, or 0.1%-9% more ethanol.

[0094]

[0098] In some embodiments, the compositions described herein have a lower glycerol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids. In a particular embodiment, the compositions described herein have a lower glycerol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at temperatures between 32° C., and at low pH with organic acids. In another particular embodiment, the compositions described herein have a lower glycerol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at temperatures between 36° C., and at low pH with organic acids.

[0095]

[0099] In some embodiments, the compositions described herein produce at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, or 22% less glycerol after 50 hours of fermentation compared to typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids. In some embodiments, the compositions described herein produce up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30% less glycerol after 50 hours of fermentation compared to typical yeast products used for fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids. ... Alcohol®), after 50 hours of fermentation, about 1%-30% (i.e., from about 1% to about 30%), 2%-30%, 3%-30%, 4%-30%, 5%-30%, 6%-30%, 7%-30%, 8%-30%, 9%-30%, 10%-30%, 11%-30%, 12%-30%, 13%-30%, 14%-30%, 15%-30%, 16%-30% , 17%~30%, 18%~30%, 19%~30%, 20%~30%, 21%~30%, 22%~30%, 23%~30%, 24%~30%, 25%~30%, 26%~30%, 27%~30%, 28%~30%, 29%~30%, 1%~29%, 2%~29%, 3%~29%, 4%~29%, 5%~29%, 6%~29%, 7%~29%, 8%~29%, 9%~29%,10%~29%、11%~29%、12%~29%、13%~29%、14%~29%、15%~29%、16%~29%、17%~29%、18%~29%、19%~29%、20%~29%、21%~29%、22%~29%、23%~29%、24%~29%、25%~29%、26%~29%、27%~29%、28%~29%、1%~28%、2%~28%、3%~28%、4%~28%、5%~28%、6%~28%、7%~28%、8%~28%、9%~28%、10%~28%、11%~28%、12%~28%、13%~28%、14%~28%、15%~28%、16%~28%、17%~28%、18%~28%、19%~28%、20%~28%、21%~28%、22%~28%、23%~28%、24%~28%、25%~28%、26%~28%、27%~28%、1%~27%、2%~27%、3%~27%、4%~27%、5%~27%、6%~27%、7%~27%、8%~27%、9%~27%、10%~27%、11%~27%、12%~27%、13%~27%、14%~27%、15%~27%、16%~27%、17%~27%、18%~27%、19%~27%、20%~27%、21%~27%、22%~27%、23%~27%、24%~27%、25%~27%、26%~27%、1%~26%、2%~26%、3%~26%、4%~26%、5%~26%、6%~26%、7%~26%、8%~26%、9%~26%、10%~26%、11%~26%、12%~26%、13%~26%、14%~26%、15%~26%、16%~26%、17%~26%、18%~26%、19%~26%、20%~26%、21%~26%、22%~26%、23%~26%、24%~26%、25%~26%、1%~25%、2%~25%、3%~25%、4%~25%、5%~25%、6%~25%、7%~25%、8%~25%、9%~25%、10%~25%、11%~25%、12%~25%、13%~25%、14%~25%、15%~25%、16%~25%、17%~25%、18%~25%、19%~25%、20%~25%、21%~25%、22%~25%、23%~25%、24%~25%、1%~24%、2%~24%、3%~24%、4%~24%、5%~24%、6%~24%、7%~24%、8%~24%、9%~24%、10%~24%、11%~24%、12%~24%、13%~24%、14%~24%、15%~24%、16%~24%、17%~24%、18%~24%、19%~24%、20%~24%、21%~24%、22%~24%、23%~24%、1%~23%、2%~23%、3%~23%、4%~23%、5%~23%、6%~23%、7%~23%、8%~23%、9%~23%、10%~23%、11%~23%、12%~23%、13%~23%、14%~23%、15%~23%、16%~23%、17%~23%、18%~23%、19%~23%、20%~23%、21%~23%、22%~23%、1%~22%、2%~22%、3%~22%、4%~22%、5%~22%、6%~22%、7%~22%、8%~22%、9%~22%、10%~22%、11%~22%、12%~22%、13%~22%、14%~22%、15%~22%、16%~22%、17%~22%、18%~22%、19%~22%、20%~22%、21%~22%、1%~21%、2%~21%、3%~21%、4%~21%、5%~21%、6%~21%、7%~21%、8%~21%、9%~21%、10%~21%、11%~21%、12%~21%、13%~21%、14%~21%、15%~21%、16%~21%、17%~21%、18%~21%、19%~21%、20%~21%、1%~20%、2%~20%、3%~20%、4%~20%、5%~20%、6%~20%、7%~20%、8%~20%、9%~20%、10%~20%、11%~20%、12%~20%、13%~20%、14%~20%、15%~20%、16%~20%、17%~20%、18%~20%、19%~20%、1%~19%、2%~19%、3%~19%、4%~19%、5%~19%、6%~19%、7%~19%、8%~19%、9%~19%、10%~19%、11%~19%、12%~19%、13%~19%、14%~19%、15%~19%、16%~19%、17%~19%、18%~19%、1%~18%、2%~18%、3%~18%、4%~18%、5%~18%、6%~18%、7%~18%、8%~18%、9%~18%、10%~18%、11%~18%、12%~18%、13%~18%、14%~18%、15%~18%、16%~18%、17%~18%、1%~17%、2%~17%, 3%~17%, 4%~17%, 5%~17%, 6%~17%, 7%~17%, 8%~17%, 9%~17%, 10%~17%, 11%~17%, 12%~17%, 13%~17%, 14%~17%, 15%~17%, 16%~17%, 1%~16%, 2%~16%, 3%~16%, 4%~16%, 5%~16%, 6%~16%, 7%~16%, 8%~16%, 9%~16%, 10%~16%, 11%~16%, 12%~16%, 13%~16%, 14%~16%, 15%~16%, 1%~15%, 2%~15%, 3 %~15%, 4%~15%, 5%~15%, 6%~15%, 7%~15%, 8%~15%, 9%~15%, 10%~15%, 11%~15%, 12%~15%, 13%~15%, 14%~15%, 1%~14%, 2%~14%, 3%~14%, 4%~14%, 5%~14%, 6%~14%, 7%~14%, 8%~14%, 9%~14%, 10%~14%, 11%~14%, 12%~14%, 13%~14%, 1%~13%, 2%~13%, 3%~13%, 4%~13%, 5%~13%, 6%~13%, 7%~13%, 8%~13%, 9%~13%, 10%~13%, 11%~13%, 12%~13%, 1%~12%, 2%~12%, 3%~12%, 4%~12%, 5%~12%, 6%~12%, 7%~12%, 8%~12%, 9%~12%, 10%~12%, 11%~12%, 1%~11%, 2%~11%, 3%~11%, 4%~11%, 5%~11%, 6%~11%, 7%~11%, 8%~11%, 9%~11%, 10%~11%, 1%~10%, 2%~10%, 3%~10%, 4%~10%, 5%~10%, 6%~10%, 7%~10%, 8%~10%, 9 Produces %-10%, 1%-9%, 2%-9%, 3%-9%, 4%-9%, 5%-9%, 6%-9%, 7%-9%, 8%-9%, 1%-8%, 2%-8%, 3%-8%, 4%-8%, 5%-8%, 6%-8%, 7%-8%, 1%-7%, 2%-7%, 3%-7%, 4%-7%, 5%-7%, 6%-7%, 1%-6%, 2%-6%, 3%-6%, 4%-6%, 5%-6%, 1%-5%, 2%-5%, 3%-5%, 4%-5%, 1%-4%, 2%-4%, 3%-4%, 1%-3%, 2%-3%, or 1%-2% less glycerol.

[0096] [000100] In some embodiments, the compositions described herein have a higher fermentation rate at temperatures between 20°C and 40°C, preferably between 32°C and 36°C, and at low pH with organic acids than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In a particular embodiment, the compositions described herein have a higher fermentation rate at temperatures between 32°C, and at low pH with organic acids than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In another particular embodiment, the compositions described herein have a higher fermentation rate at temperatures between 36°C, and at low pH with organic acids than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®).

[0097] [000101] In some embodiments, the compositions described herein have a fermentation rate at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% higher than a typical yeast product used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®) after 24 hours of fermentation at temperatures between 20°C and 40°C, preferably between 32°C and 36°C, and at low pH with organic acids. ... In some embodiments, the fermentation rate is up to about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% higher than that of ethanol. In some embodiments, the compositions described herein are prepared at temperatures between 20° C. and 40° C., preferably between 32° C. and 36° C., and at low pH with organic acids, after 24 hours of fermentation, using typical yeast products used for fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super®, etc.). Alcohol (registered trademark)) by about 1% to 50% (i.e., from about 1% to about 50%), 2% to 50%, 3% to 50%, 4% to 50%, 5% to 50%, 6% to 50%, 7% to 50%, 8% to 50%, 9% to 50%, 10% to 50%, 11% to 50%, 12% to 50%, 13% to 50%, 14% to 50%, 15% to 50%, 16% to 50%, 17% to 50%, 18% to 50%, 19% to 50%, 20% to 50%, 21% to 50%, 22% to 50%, 23% to 50%, 24% to 50%, 25% to 50%, 26% to 50%, 27% to 50%, 28% to 50%, 2% to 49%,3%~49%、4%~49%、5%~49%、6%~49%、7%~49%、8%~49%、9%~49%、10%~49%、11%~49%、12%~49%、13%~49%、14%~49%、15%~49%、16%~49%、17%~49%、18%~49%、19%~49%、20%~49%、21%~49%、22%~49%、23%~49%、24%~49%、25%~49%、26%~49%、27%~49%、28%~49%、2%~48%、3%~48%、4%~48%、5%~48%、6%~48%、7%~48%、8%~48%、9%~48%、10%~48%、11%~48%、12%~48%、13%~48%、14%~48%、15%~48%、16%~48%、17%~48%、18%~48%、19%~48%、20%~48%、21%~48%、22%~48%、23%~48%、24%~48%、25%~48%、26%~48%、27%~48%、28%~48%、2%~47%、3%~47%、4%~47%、5%~47%、6%~47%、7%~47%、8%~47%、9%~47%、10%~47%、11%~47%、12%~47%、13%~47%、14%~47%、15%~47%、16%~47%、17%~47%、18%~47%、19%~47%、20%~47%、21%~47%、22%~47%、23%~47%、24%~47%、25%~47%、26%~47%、27%~47%、28%~47%、2%~46%、3%~46%、4%~46%、5%~46%、6%~46%、7%~46%、8%~46%、9%~46%、10%~46%、11%~46%、12%~46%、13%~46%、14%~46%、15%~46%、16%~46%、17%~46%、18%~46%、19%~46%、20%~46%、21%~46%、22%~46%、23%~46%、24%~46%、25%~46%、26%~46%、27%~46%、28%~46%、2%~45%、3%~45%、4%~45%、5%~45%、6%~45%、7%~45%、8%~45%、9%~45%、10%~45%、11%~45%、12%~45%、13%~45%、14%~45%、15%~45%、16%~45%、17%~45%、18%~45%、19%~45%、20%~45%、21%~45%、22%~45%、23%~45%、24%~45%、25%~45%、26%~45%、27%~45%、28%~45%、2%~44%、3%~44%、4%~44%、5%~44%、6%~44%、7%~44%、8%~44%、9%~44%、10%~44%、11%~44%、12%~44%、13%~44%、14%~44%、15%~44%、16%~44%、17%~44%、18%~44%、19%~44%、20%~44%、21%~44%、22%~44%、23%~44%、24%~44%、25%~44%、26%~44%、27%~44%、28%~44%、2%~43%、3%~43%、4%~43%、5%~43%、6%~43%、7%~43%、8%~43%、9%~43%、10%~43%、11%~43%、12%~43%、13%~43%、14%~43%、15%~43%、16%~43%、17%~43%、18%~43%、19%~43%、20%~43%、21%~43%、22%~43%、23%~43%、24%~43%、25%~43%、26%~43%、27%~43%、28%~43%、2%~42%、3%~42%、4%~42%、5%~42%、6%~42%、7%~42%、8%~42%、9%~42%、10%~42%、11%~42%、12%~42%、13%~42%、14%~42%、15%~42%、16%~42%、17%~42%、18%~42%、19%~42%、20%~42%、21%~42%、22%~42%、23%~42%、24%~42%、25%~42%、26%~42%、27%~42%、28%~42%、2%~41%、3%~41%、4%~41%、5%~41%、6%~41%、7%~41%、8%~41%、9%~41%、10%~41%、11%~41%、12%~41%、13%~41%、14%~41%、15%~41%、16%~41%、17%~41%、18%~41%、19%~41%、20%~41%、21%~41%、22%~41%、23%~41%、24%~41%、25%~41%、26%~41%、27%~41%、28%~41%、2%~40%、3%~40%、4%~40%、5%~40%、6%~40%、7%~40%、8%~40%、9%~40%、10%~40%、11%~40%、12%~40%、13%~40%、14%~40%、15%~40%、16%~40%、17%~40%、18%~40%、19%~40%、20%~40%、21%~40%、22%~40%、23%~40%、24%~40%、25%~40%、26%~40%、27%~40%、28%~40%、2%~39%、3%~39%、4%~39%、5%~39%、6%~39%、7%~39%、8%~39%、9%~39%、10%~39%、11%~39%、12%~39%、13%~39%、14%~39%、15%~39%、16%~39%、17%~39%、18%~39%、19%~39%、20%~39%、21%~39%、22%~39%、23%~39%、24%~39%、25%~39%、26%~39%、27%~39%、28%~39%、2%~38%、3%~38%、4%~38%、5%~38%、6%~38%、7%~38%、8%~38%、9%~38%、10%~38%、11%~38%、12%~38%、13%~38%、14%~38%、15%~38%、16%~38%、17%~38%、18%~38%、19%~38%、20%~38%、21%~38%、22%~38%、23%~38%、24%~38%、25%~38%、26%~38%、27%~38%、28%~38%、2%~37%、3%~37%、4%~37%、5%~37%、6%~37%、7%~37%、8%~37%、9%~37%、10%~37%、11%~37%、12%~37%、13%~37%、14%~37%、15%~37%、16%~37%、17%~37%、18%~37%、19%~37%、20%~37%、21%~37%、22%~37%、23%~37%、24%~37%、25%~37%、26%~37%、27%~37%、28%~37%、2%~36%、3%~36%、4%~36%、5%~36%、6%~36%、7%~36%、8%~36%、9%~36%、10%~36%、11%~36%、12%~36%、13%~36%、14%~36%、15%~36%、16%~36%、17%~36%、18%~36%、19%~36%、20%~36%、21%~36%、22%~36%、23%~36%、24%~36%、25%~36%、26%~36%、27%~36%、28%~36%、2%~35%、3%~35%、4%~35%、5%~35%、6%~35%、7%~35%、8%~35%、9%~35%、10%~35%、11%~35%、12%~35%、13%~35%、14%~35%、15%~35%、16%~35%、17%~35%、18%~35%、19%~35%、20%~35%、21%~35%、22%~35%、23%~35%、24%~35%、25%~35%、26%~35%、27%~35%、28%~35%、2%~34%、3%~34%、4%~34%、5%~34%、6%~34%、7%~34%、8%~34%、9%~34%、10%~34%、11%~34%、12%~34%、13%~34%、14%~34%、15%~34%、16%~34%、17%~34%、18%~34%、19%~34%、20%~34%、21%~34%、22%~34%、23%~34%、24%~34%、25%~34%、26%~34%、27%~34%、28%~34%、2%~33%、3%~33%、4%~33%、5%~33%、6%~33%、7%~33%、8%~33%、9%~33%、10%~33%、11%~33%、12%~33%、13%~33%、14%~33%、15%~33%、16%~33%、17%~33%、18%~33%、19%~33%、20%~33%、21%~33%、22%~33%、23%~33%、24%~33%、25%~33%、26%~33%、27%~33%、28%~33%、2%~32%、3%~32%、4%~32%、5%~32%、6%~32%、7%~32%、8%~32%、9%~32%、10%~32%、11%~32%、12%~32%、13%~32%、14%~32%、15%~32%、16%~32%、17%~32%、18%~32%、19%~32%、20%~32%、21%~32%、22%~32%、23%~32%、24%~32%、25%~32%、26%~32%、27%~32%、28%~32%、2%~31%、3%~31%、4%~31%、5%~31%、6%~31%、7%~31%、8%~31%、9%~31%、10%~31%、11%~31%、12%~31%、13%~31%、14%~31%、15%~31%、16%~31%、17%~31%、18%~31%、19%~31%、20%~31%、21%~31%、22%~31%、23%~31%、24%~31%、25%~31%、26%~31%、27%~31%、28%~31%、2%~30%、3%~30%、4%~3、 0%、5%~30%、6%~30%、7%~30%、8%~30%、9%~30%、10%~30%、11%~30%、12%~30%、13%~30%、14%~30%、15%~30%、16%~30%、17%~30%、18%~30%、19%~30%、20%~30%、21%~30%、22%~30%、23%~30%、24%~30%、25%~30%、26%~30%、27%~30%、28%~30%、29%~30%、1%~29%、2%~29%、3%~29%、4%~29%、5%~29%、6%~29%、7%~29%、8%~29%、9%~29%、10%~29%、11%~29%、12%~29%、13%~29%、14%~29%、15%~29%、16%~29%、17%~29%、18%~29%、19%~29%、20%~29%、21%~29%、22%~29%、23%~29%、24%~29%、25%~29%、26%~29%、27%~29%、28%~29%、1%~28%、2%~28%、3%~28%、4%~28%、5%~28%、6%~28%、7%~28%、8%~28%、9%~28%、10%~28%、11%~28%、12%~28%、13%~28%、14%~28%、15%~28%、16%~28%、17%~28%、18%~28%、19%~28%、20%~28%、21%~28%、22%~28%、23%~28%、24%~28%、25%~28%、26%~28%、27%~28%、1%~27%、2%~27%、3%~27%、4%~27%、5%~27%、6%~27%、7%~27%、8%~27%、9%~27%、10%~27%、11%~27%、12%~27%、13%~27%、14%~27%、15%~27%、16%~27%、17%~27%、18%~27%、19%~27%、20%~27%、21%~27%、22%~27%、23%~27%、24%~27%、25%~27%、26%~27%、1%~26%、2%~26%、3%~26%、4%~26%、5%~26%、6%~26%、7%~26%、8%~26%、9%~26%、10%~26%、11%~26%、12%~26%、13%~26%、14%~26%、15%~26%、16%~26%、17%~26%、18%~26%、19%~26%、20%~26%、21%~26%、22%~26%、23%~26%、24%~26%、25%~26%、1%~25%、2%~25%、3%~25%、4%~25%、5%~25%、6%~25%、7%~25%、8%~25%、9%~25%、10%~25%、11%~25%、12%~25%、13%~25%、14%~25%、15%~25%、16%~25%、17%~25%、18%~25%、19%~25%、20%~25%、21%~25%、22%~25%、23%~25%、24%~25%、1%~24%、2%~24%、3%~24%、4%~24%、5%~24%、6%~24%、7%~24%、8%~24%、9%~24%、10%~24%、11%~24%、12%~24%、13%~24%、14%~24%、15%~24%、16%~24%、17%~24%、18%~24%、19%~24%、20%~24%、21%~24%、22%~24%、23%~24%、1%~23%、2%~23%、3%~23%、4%~23%、5%~23%、6%~23%、7%~23%、8%~23%、9%~23%、10%~23%、11%~23%、12%~23%、13%~23%、14%~23%、15%~23%、16%~23%、17%~23%、18%~23%、19%~23%、20%~23%、21%~23%、22%~23%、1%~22%、2%~22%、3%~22%、4%~22%、5%~22%、6%~22%、7%~22%、8%~22%、9%~22%、10%~22%、11%~22%、12%~22%、13%~22%、14%~22%、15%~22%、16%~22%、17%~22%、18%~22%、19%~22%、20%~22%、21%~22%、1%~21%、2%~21%、3%~21%、4%~21%、5%~21%、6%~21%、7%~21%、8%~21%、9%~21%、10%~21%、11%~21%、12%~21%、13%~21%、14%~21%、15%~21%、16%~21%、17%~21%、18%~21%、19%~21%、20%~21%、1%~20%、2%~20%、3%~20%、4%~20%、5%~20%、6%~20%、7%~20%、8%~20%、9%~20%、10%~20%、11%~20%、12%~20%、13%~20%、14%~20%、15%~20%、16%~20%、17%~20%、18%~20%、19%~20%、1%~19%、2%~19%、3%~19%、4%~19%、5%~19%、6%~19%、7%~19%、8%~19%、9%~19%、10%~19%、11%~19%、12%~19%、13%~19%、14%~19%、15%~19%、16%~19%、17%~19%、18%~19%、1%~18%、2%~18%、3%~18%、4%~18%、5%~18%、6%~18%、7%~18%、8%~18%、9%~18%、10%~18%、11%~18%、12%~18%、13%~18%、14%~18%、15%~18%、16%~18%、17%~18%、1%~17%、2%~17%、3%~17%、4%~17%、5%~17%、6%~17%、7%~17%、8%~17%、9%~17%、10%~17%、11%~17%、12%~17%、13%~17%、14%~17%、15%~17%、16%~17%、1%~16%、2%~16%、3%~16%、4%~16%、5%~16%、6%~16%、7%~16%、8%~16%、9%~16%、10%~16%、11%~16%、12%~16%、13%~16%、14%~16%、15%~16%、1%~15%、2%~15%、3%~15%、4%~15%、5%~15%、6%~15%、7%~15%、8%~15%、9%~15%、10%~15%、11%~15%、12%~15%、13%~15%、14%~15%、1%~14%、2%~14%、3%~14%、4%~14%、5%~14%、6%~14%、7%~14%、8%~14%、9%~14%、10%~14%、11%~14%、12%~14%、13%~14%、1%~13%、2%~13%、3%~13%、4%~13%、5%~13%、6%~13%、7%~13%、8%~13%、9%~13%、10%~13%、11%~13%、12%~13%、1%~12%、2%~12%、3%~12%、4%~12%、5%~12%、6%~12%、7%~12%、8%~12%、9%~12%、10%~12%、11%~12%、1%~11%、2%~11%、3%~11%、4%~11%、5%~11%、6%~11%、7%~11%、8%~11%、9%~11%、10%~11%、1%~10%、2%~10%、3%~10%、4%~10%、5%~10%、6%~10%、7%~10%、8%~10%、9%~10%、1%~9%、2%~9%、3%~9%、4%-9%, 5%-9%, 6%-9%, 7%-9%, 8%-9%, 1%-8%, 2%-8%, 3%-8%, 4%-8%, 5%-8%, 6%-8%, 7%-8%, 1%-7%, 2%-7%, 3%-7%, 4%-7%, 5%-7%, 6%-7%, 1%-6%, 2%-6%, 3%-6%, 4%-6%, 5%-6%, 1%-5%, 2%-5%, 3%-5%, 4%-5%, 1%-4%, 2%-4%, 3%-4%, 1%-3%, 2%-3%, or 1%-2% higher fermentation rate.

[0098] [000102] In some embodiments, the compositions described herein have one or more defining characteristics described herein. In some embodiments, the compositions described herein have one or more defining characteristics described herein at temperatures between 20°C and 40°C, preferably between 32°C and 36°C, and at low pH with organic acids. In some embodiments, the compositions described herein have one or more defining characteristics described herein at temperatures between 20°C and 40°C, preferably between 32°C and 36°C, and at normal pH without organic acids. For example, in some embodiments, the compositions described herein have a higher ethanol yield, lower glycerol yield, higher fermentation rate, higher temperature tolerance, and higher organic acid tolerance than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In some embodiments, the compositions described herein have a higher fermentation rate, produce lower levels of glycerol, and have higher ethanol yield than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®). In some embodiments, the compositions described herein have a higher fermentation rate and produce lower levels of glycerol than typical yeast products used in fermentation (e.g., Fali, Ethanol Red®, Thermosacc®, Angel Super Alcohol®).

[0099] 4. Ethanol production [000103] Described herein is a method for producing ethanol from a substrate by contacting the substrate with a fermenting organism or a composition comprising a fermenting organism. The fermenting organism is selected from the yeast strains described herein and derivatives thereof. Fermenting organisms having substantially the same characteristics as the yeast strains described herein, such as Saccharomyces cerevisiae Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or yeast strains having defining characteristics or derivatives of yeast strains described herein, can be used in the methods described herein. Also described herein is a fermentation product comprising any of the yeasts described herein. Further described herein is a fermentation product obtained by the methods described herein. Fermentation products include, but are not limited to, fuel ethanol, industrial ethanol, potable ethanol, bioethanol, fermented foods, such as alcoholic beverages, fermented milk and yogurt, wine, beer, cider, tempeh, miso, kimchi, sauerkraut, and fermented sausages.

[0100] [000104] Fermentation is carried out in a fermentation medium. The fermentation medium includes a fermentation substrate, i.e., a carbohydrate source that is metabolized by the fermenting organism, such as biomass. The fermentation medium may include nutrients for the fermenting organism. Nutrients are widely used in the field of fermentation and include nitrogen sources, vitamins, minerals, or combinations thereof.

[0101] [000105] In one embodiment, the strain or derivative or composition described herein is incubated with a substrate containing fermentable sugars from biomass, such as plant biomass from forests, and / or agricultural or food processing products and / or co-products that constitute a significant amount of carbon source for producing molecules of interest. The strain or derivative is incubated with the substrate under conditions that allow the fermentable sugars to be fermented. The fermentable sugars can be glucose, galactose, maltose, fructose, sucrose, mannose, or combinations thereof. Typically, the fermentable sugars are glucose and sucrose. The source of fermentable sugars in the substrate can be any source that contains fermentable sugars. The fermentable sugars in the substrate may be derived, for example, from any one or more of the following sources: hydrolyzed starch, hydrolyzed cellulose, molasses (from sugar cane or sugar beet), sugar cane juice, agave, sugar beet juice, grape juice, fruit juice, glucose, hydrolyzed maltodextrin, raw sugar juice, galactose, sucrose, other forms of fermentable sugars, or combinations thereof. The starch may be obtained from any crop rich in starch. Examples of crops rich in starch include, but are not limited to, corn, wheat, barley, cassava, sorghum, sweet potato, millet, rice, or other starch-rich crops. In preparing the substrate, the crop is typically milled and mixed with water and hydrolytic enzymes under conditions that result in hydrolysis of starch and liberation of fermentable sugars such as glucose. Exemplary enzymes for hydrolyzing starch include α-amylase, amyloglucosidase, pullulanase, β-amylase, glucoamylase, or mixtures thereof.

[0102] [000106] In general, fermenting organisms such as yeast, including Saccharomyces cerevisiae yeast, require an appropriate nitrogen source for growth and fermentation. Many nitrogen sources can be used and are well known in the art. The nitrogen source may be organic, such as urea or corn mash, or inorganic, such as ammonia, ammonium hydroxide, or ammonium salts.

[0103] [000107] In certain embodiments, the biomass may include or be derived from sugarcane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, cassava, or combinations thereof. In certain embodiments, the substrate is provided in the form of corn mash or synthetic corn medium (SCM). Methods for preparing corn mash are known in the art and are described, for example, in Thomas et al., (2001) Journal of Applied Microbiology, 90, 819-828. Methods for preparing substrates similar in function to SCM are known in the art and are described, for example, in U.S. Patent No. 10,106,823, which is incorporated herein by reference in its entirety. Methods for preparing starch-based substrates are also described, for example, in PCT Publication No. 2006 / 113683 or US Patent Publication No. 2007 / 0014905.

[0104] [000108] The sugar content of the fermentation medium can be adjusted so that it is as high in sugar as possible while at the same time converting the sugars to ethanol as quickly and completely as possible. It is preferred that the yeast convert all the sugars in the medium to ethanol so that the overall yield of conversion of consumed sugars to ethanol is as high as possible, and as a result, co-products such as glycerol are minimally produced during fermentation.

[0105] [000109] Fermentation is carried out at a temperature that allows for the fermentation of fermentable sugars. Generally, the higher the temperature at which fermentation can be carried out, the more economical the industrial process will be. Typically, the temperature at which fermentation is carried out is 25-42°C (i.e., from 25°C to 42°C). Suitable temperature ranges include 25-41°C, 26-40°C, 27-40°C, 28-40°C, 29-40°C, 30-40°C, 25-39°C, 26-39°C, 27-39°C, 28-39°C, 29-39°C, 30-39°C, 31-39°C, 32-39°C, 33-39°C, 25-38°C, 26-38°C, 27-38°C, 28-38°C, 29-38°C, 30-38°C, 31-38°C, 32-38°C, 33-38°C, 25-27°C, 26 These include ~37°C, 27-37°C, 28-37°C, 29-37°C, 30-37°C, 31-37°C, 32-37°C, 33-37°C, 25-36°C, 26-36°C, 27-36°C, 28-36°C, 29-36°C, 30-36°C, 31-36°C, 32-36°C, 33-36°C, 25-35°C, 26-35°C, 27-35°C, 28-35°C, 29-35°C, 30-35°C, 31-35°C, 32-35°C, or 33-35°C.

[0106] [000110] Methods of fermentation and distillation are known in the art and are described, for example, in WO2006 / 113683 or US2007 / 0014905. In particular, the ethanol production described herein can be carried out using simultaneous saccharification and fermentation ("SSF"), batch fermentation, or continuous fermentation.

[0107] a. Simultaneous Saccharification and Fermentation ("SSF") [000111] SSF is widely used in industrial scale processes for producing fermentation products, particularly in processes for producing ethanol. When performing SSF, the saccharification and fermentation steps are carried out simultaneously. There is no maintenance step for saccharification, which means that the fermenting organism, such as yeast, and the enzymes can be added together. However, it is also conceivable that the fermenting organism and the enzymes are added separately (i.e., separated hydrolysis fermentation (SHF)). SSF can be carried out at a temperature between 25°C and 40°C, such as between 28°C and 35°C, for example between 30°C and 34°C, preferably around about 32°C. In one embodiment, the fermentation continues for 6 to 120 hours, in particular between 24 to 96 hours, preferably around 50 hours. In one embodiment, the pH is about 3.0 to 6.0, preferably about 4.0 to 5.0.

[0108] [000112] After fermentation, e.g., SSF, ethanol can be separated from the spent fermentation medium or beer. The beer can be rectified or distilled to recover / extract the desired fermentation products (i.e., ethanol and higher alcohols). Alternatively, the desired fermentation products (i.e., ethanol) can be extracted from the fermentation medium by microfiltration or membrane filtration techniques known in the art. The fermentation products (i.e., ethanol) can also be recovered by stripping or other methods known in the art.

[0109] [000113] In some embodiments, ethanol is not recovered / extracted from the fermentation medium or beer, for example for the production of alcoholic beverages.

[0110] B. Batch fermentation [000114] Batch fermentation is when fermentation is performed in separate batches. Batch fermentation is a process in which a fermentation medium is fed to a fermenter at the start, the fermenter is inoculated with the desired microorganism (i.e., yeast, yeast product), and the fermentation process is carried out until a predetermined condition is reached, typically exhaustion of the substrate in the fermentation medium, and cessation of ethanol production caused by exhaustion. Once the process is complete, the product is removed from the fermenter and the fermenter is sterilized before the next fermentation is performed. The contents can then be made into a final product (e.g., wine) or rectified / distilled (e.g., fuel ethanol and whiskey).

[0111] [000115] Fed-batch methods may also be used. A fed-batch method is a fermentation in which a portion of the fermentation medium is provided from the beginning of the fermentation process where an inoculum is added, and at a certain point after the start of the fermentation, an additional substrate, a feed, is fed to the fermenter at a rate that may be predetermined or determined by the conditions in the fermenter, until a maximum amount is reached. The feed may or may not have the same composition as the initial fermentation medium. The contents may then be made into a final product (e.g. wine) or rectified / distilled (e.g. fuel ethanol and whiskey).

[0112] C. Continuous fermentation [000116] Continuous fermentation allows fermentation to occur over an extended period of time without separate batch fermentations. A continuous fermentation method is one in which the volume in the fermenter is constant, as new growth medium is continuously fed to the fermenter and fermented material is simultaneously removed from the fermenter at the same rate. The contents can then be made into a final product (e.g., wine) or rectified / distilled (e.g., fuel ethanol and whiskey).

[0113] 5. Ethanol [000117] The ethanol produced by the yeast and its derivatives described herein can be fuel ethanol, industrial ethanol, and / or potable ethanol. Fuel ethanol, industrial ethanol, potable alcohol can be produced from starch-containing biomass, including starch found in cereals (e.g., corn, wheat, rice, sorghum / milo, barley, etc.), and from starch in tubers and root crops (e.g., potato, cassava, etc.); from vegetative parts of plants that contain the sugars sucrose, glucose, and fructose (e.g., sugar cane, sweet sorghum, sugar beet, agave, etc.); and from fruits and berries of plants that contain sucrose, glucose, and fructose (e.g., grapes, oranges, peaches, cherries, etc.).

[0114] [000118] Fuel and industrial ethanol can be produced from plant biomass containing cellulose and hemicellulose, such as cereal crop residues (e.g., wheat and rice straw, corn stover, corn cobs, etc.), from corn fiber, from so-called energy crops such as switchgrass and poplar, from woody material waste including residues from sawmills (e.g., sawdust and wood chips), from residues from pulp and paper manufacturing, and from waste paper and cardboard.

[0115] A. Ethanol for fuel [000119] Fuel grade ethanol is produced for use in internal combustion engines and can be produced as anhydrous or hydrous fuel grade ethanol. Anhydrous fuel grade ethanol can be mixed with gasoline to form an ethanol / gasoline mixture or with diesel to form an ethanol / diesel mixture. Hydrous fuel grade ethanol can be used directly as a fuel for internal combustion engines.

[0116] b. Industrial ethanol [000120] Industrial ethanol is produced for use in a variety of applications, including as a solvent in pharmaceuticals, cosmetics, detergents, household cleaners and disinfectants, and paints and inks; and as a chemical intermediate in the production of ethyl acetate, ethyl acrylate, polyethylene, acetic acid, and other industrially important organic molecules.

[0117] C. Potable ethanol [000121] Potable ethanol is that which is produced for human consumption and includes the ethanol found in wine, beer, cider, sake, mead, kombucha, as well as distilled spirits including whiskey, bourbon, cachaça, Chinese baijiu, baijiu, etc.

[0118] 6. Yeast Strain Preparation a. Direct crossbreeding [000122] Provided herein is a method for producing derivatives of the Saccharomyces yeast strains described herein. The method may include providing a first yeast strain selected from Saccharomyces Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929 strains, and a second yeast strain that is any yeast strain, such as a yeast strain of the phylogenetic clade of Saccharomyces sensu stricto, e.g., a Saccharomyces cerevisiae strain. The second strain may also be any of the yeast strains described herein. The method may further include inducing sporulation of the first yeast strain and the second yeast strain. The method may also include screening and selecting spores from the first yeast strain and spores from the second yeast strain. Further, the method may include crossing selected spores of the first yeast strain with selected spores of the second yeast strain, and screening or selecting derivative strains. The method may include screening or selecting spores that exhibit one or more defining characteristics of the Saccharomyces strains described herein. The method may further include screening or selecting hybrids that exhibit one or more defining characteristics of the Saccharomyces strains described herein. An example of direct crossing is provided in FIG. 1. Thus, the parents (i.e., the donors of the "a" haploid and "alpha" haploid) that generate the hybrids are known. In one embodiment, the yeast strains described herein and their derivatives are produced from the method shown in FIG. 1. In one embodiment, the Saccharomyces yeast strains designated Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929 are derived from one or more different Saccharomyces yeast strains by the method shown in Figure 1, e.g., by the method shown in Figure 1A, such that one or more of Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929 are the product of direct mating. Methods of direct mating are known in the art and are described in U.S. Patent Nos. 10,308,963 and 10,106,823, which are incorporated herein by reference.

[0119] B mass mating [000123] Provided herein is a method for producing derivatives of the Saccharomyces yeast strains described herein. The method may include providing a first yeast strain selected from Saccharomyces Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929 strains, and one or more additional yeast strains that are any yeast strain, such as a yeast strain of the phylogenetic clade of Saccharomyces sensu stricto, e.g., a Saccharomyces cerevisiae strain. The one or more additional yeast strains may also be any yeast strain described herein. The method may further include inducing sporulation of the first yeast strain and the one or more additional yeast strains. The method may also include mixing all spores so that the spores can cross and screening or selecting derivative strains. The method may include screening or selecting hybrids that exhibit one or more defining characteristics of the Saccharomyces strains described herein. An example of a mass mating is provided in FIG. 1B. Thus, the parents (i.e., the "a" haploid and "alpha" haploid donors) that produce the hybrids are unknown. In one embodiment, the yeast strains described herein and derivatives thereof are derived from the method shown in FIG. 1. In one embodiment, the Saccharomyces yeast strains designated Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, and Y1929 are derived from one or more different Saccharomyces yeast strains by the method shown in FIG. 1, e.g., by the method shown in FIG. 1B, such that one or more of Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929 are the product of a mass mating. EXAMPLES

[0120] [000124] The foregoing may be better understood with reference to the following examples, which are presented for purposes of illustration and are not intended to limit the scope of the invention. The present disclosure has multiple aspects and embodiments, which are illustrated by the accompanying non-limiting examples.

[0121] Example 1 material and method [000125] Synthetic corn medium (SCM) was used in the following examples. SCM simulates corn mash commonly used in industrial bioethanol production. SCM is designed to reflect the chemical composition of Yellow Dent Corn #2 in terms of sugar and nutrient availability. SCM reflects a high gravity model corn line with 32% solids and approximately 72% starch, which should result in final ethanol of 13-14% w / v. The synthetic nature of the medium allows for close reproduction of test conditions for all experiments.

[0122] [000126] This medium is used to monitor the performance of yeast in a simultaneous saccharification and fermentation (SSF) process, in which, over the same period (simultaneously), starch in the medium is broken down into glucose by using the enzyme glucoamylase (saccharification) and the glucose is used by the yeast to produce ethanol (fermentation).

[0123] [000127] Preparation of SCM. 1 L of SCM complete medium was made by adding 500 g water to a 2 L beaker. The amounts of ingredients in Table 1 were added to the beaker using less than 100 mL total water. The solution was stirred at 300 rpm and gradually heated to 75°C. Once the solution was at 75°C, the heat was removed and allowed to cool to ambient temperature while mixing. While mixing, 100.0 mL of 10x SCM Vitamin Mix (Table 2) and 10.0 mL of 100x SCM Mineral Mix (Table 3) were added to the solution. Then, 25.0 mL of SCM Amino Acid Slurry (Table 4), 1.2 g of urea (550 ppm nitrogen equivalent), 0.400 mL of Virginiamycin Stock Solution (1.0 ppm), and 0.1 g of Penicillin G Powder (100 ppm) were added to the solution while stirring. The solution was transferred to a 1 L volumetric flask and made up to 1 L with water. The pH of the solutions was adjusted to 5.0-5.2 with 28% sulfuric acid or 28% ammonium / potassium hydroxide. The solutions were mixed by inversion and further stirred at 300-450 rpm. Representative samples of the solutions were withdrawn for specific gravity measurement and HPLC analysis.

[0124] [Table 1]

[0125] [Table 2]

[0126] [Table 3]

[0127] [Table 4]

[0128] [000128] 100.00 ± 0.10 mL of SCM was added to each test sample vessel for each yeast strain. At least three replicates were performed per test sample. All test sample vessels with SCM were incubated at 32°C and 150 rpm for 1-2 hours to allow for temperature equilibration. The specific gravity of each sample vessel was measured using a densitometer. Samples were removed from the sample vessels for HPLC analysis. The sample vessels were swirled to ensure all cells were suspended, and 1.9 mL of sample vessel medium was placed into a 2 mL microcentrifuge tube. Samples were centrifuged at 3,000 rpm for 10 minutes. The supernatant was decanted into a syringe with a pre-attached 0.2 μm filter, and the filtrate was collected in a glass HPLC vial. The HPLC vials containing the samples can be stored at +4°C for up to one month prior to HPLC analysis or can be analyzed immediately.

[0129] [000129] Isolation of Saccharomyces cerevisiae strains. Yeast strains were isolated from agar plates, and methods for isolation are known in the art.

[0130] [000130] Yeast culture. 2.5 mL of yeast culture (i.e., yeast in general yeast growth medium; e.g., glucose-yeast-peptone medium (GYP)) was cultured at 100°C for a final cell concentration of 1.0x10 6 The sample vessels containing SCM were inoculated to reach 100 cells / mL.

[0131] [000131] Colonies on yeast slants or plates. A 10 μL loopful of cells was inoculated into 50 mL of sterile 2% GYP. Cultures were incubated overnight at 32°C or 36°C, 150 rpm, and the cell concentration was increased to 1x10 8 The 2.5 mL culture was diluted to a final cell concentration of 2.5x10 6The sample vessels containing SCM were inoculated to reach 100 cells / mL. Immediately after all yeast samples were added, a 10-fold diluted glucoamylase was added to the sample vessels. For Spirizyme Fuel HS, 140 μL was added, whereas for Spirizyme Ultra, 224 μL was added. If an alternative glucoamylase is used, the enzyme dosage must be adjusted accordingly to 100 mL based on 32% dry corn solids. Typically, dosage is expressed in % w / w units, so the SCM solution density must be taken into account.

[0132] [000132] The sample container was closed with an airlock cap. The pressurized airlock was filled with 9 mL of sterile water. The best way to do this is to place the tip of a pipette in the airlock opening and dispense as close to the bubble as possible. The sample container was incubated at 32°C or 36°C, 150 rpm and 80% humidity for 50 hours. The weight of the sample container was measured before incubation and at the following time points after the start of incubation: 18, 24, 44, and 50 hours. After the 50 hour incubation, the sample container was removed from the incubator and sampled using HPLC analysis as described above. The specific gravity of the sample container was measured by density meter. The pH of the sample remaining in the sample container was then measured. After the pH measurement, the yeast cell concentration and viability were measured using methylene blue in a hemocytometer.

[0133] [000133] Statistical analysis. Analysis of variance (ANOVA) was performed on the final mass loss, ethanol, glycerol and glucose values ​​and 95% confidence intervals (95% CI) for each analyte were calculated. A program such as SAS JMP is preferred.

[0134] Example 2 Strain description [000134] Representative samples of the Saccharomyces yeast strains described herein, as well as those strains deposited under NRRL Patent Deposit Designation No. Y-68003 (Y1912), NRRL Patent Deposit Designation No. Y-68004 (Y1913), NRRL Patent Deposit Designation No. Y-68005 (Y1914), NRRL Patent Deposit Designation No. Y-68006 (Y1919), NRRL Patent Deposit Designation No. Y-68007 (Y1923), NRRL Patent Deposit Designation No. Y-68008 (Y1927), and NRRL Patent Deposit Designation No. Y-68009 (Y1929), were determined to have the following characteristics based on four replicate tests performed at 32°C or 36°C, at normal pH without organic acid, or at low pH with organic acid. Y1609 is shown for comparison and was analyzed under similar conditions.

[0135] [000135] Notable advantageous characteristics of the yeast strains described herein include increased metabolic ethanol, decreased metabolic glycerol, and increased initial fermentation rate compared to controls. The advantageous fermentation characteristics of the yeast strains described herein are provided in the examples below.

[0136] Example 3 Ethanol production, glycerol production, and fermentation rate [000136] The yeast strains described herein were evaluated for ethanol production, glycerol production, and fermentation rate in synthetic corn starch medium (SCM) at 32°C and pH 5.2 (Table 5 and Figure 2). A control (Y1609) is shown for comparison.

[0137] [Table 5]

[0138] [000137] The parent yeast strains described herein were evaluated for ethanol production, glycerol production, and fermentation rate at 32°C and pH 4.0 in SCM exogenously supplemented with 1% w / v lactic acid and 0.05% w / v acetic acid (Table 6 and Figure 3). A control (Y1609) is shown for comparison.

[0139] [Table 6]

[0140] [000138] The parent yeast strains described herein were evaluated for ethanol production, glycerol production, and fermentation rate at 36°C and pH 5.2 in SCM without organic acids (Table 7 and Figure 4). A control (Y1609) is shown for comparison.

[0141] [Table 7]

[0142] [000139] The parent yeast strains described herein were evaluated for ethanol production, glycerol production, and fermentation rate at 36°C and pH 4.0 in SCM exogenously supplemented with 1% w / v lactic acid and 0.05% w / v acetic acid (Table 8 and Figure 5). A control (Y1609) is shown for comparison.

[0143] [Table 8]

[0144] [000140] The foregoing description of the specific embodiments sufficiently reveals the general nature of the present invention, so that others, by applying the knowledge of those skilled in the art, can easily modify and / or adapt such specific embodiments to various applications without departing from the general concept of the disclosure and without undue experimentation. Such adaptations and modifications are therefore intended to be within the meaning and range of equivalents of the disclosed embodiments, based on the teaching and guidance presented herein. It is to be understood that the phraseology or terminology used herein is for the purpose of description and not for the purpose of limitation, and thus should be interpreted by those skilled in the art in light of the teaching and guidance.

[0145] [000141] The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.

[0146] [000142] All publications, patents, patent applications, and / or other documents cited in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, and / or other document was individually indicated to be incorporated by reference for all purposes.

[0147] [000143] For completeness, various aspects of the invention are described in the following numbered paragraphs.

[0148] [000144] Paragraph 1. (a) Saccharomyces strain Y1912, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68003; (b) Saccharomyces strain Y1913, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68004; (c) Saccharomyces strain Y1914, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68005; (d) Saccharomyces strain Y1915, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68006. (e) Saccharomyces strain Y1923, which is a representative representative of the strains deposited under NRRL Patent Deposit Designation No. Y-68007; (f) Saccharomyces strain Y1927, which is a representative representative of the strains deposited under NRRL Patent Deposit Designation No. Y-68008; and (g) Saccharomyces strain Y1929, which is a representative representative of the strains deposited under NRRL Patent Deposit Designation No. Y-68009.

[0149] [000145] Section 2. (a) Saccharomyces strain Y1912, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68003; (b) Saccharomyces strain Y1913, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68004; (c) Saccharomyces strain Y1914, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68005; (d) a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68006. (e) Saccharomyces strain Y1923, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68007; (f) Saccharomyces strain Y1927, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68008; and (g) Saccharomyces strain Y1929, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68009.

[0150] [000146] Paragraph 3. The yeast strain of paragraph 1 or the derivative of paragraph 2, wherein the yeast strain or derivative comprises one or more defining characteristics selected from: (a) a higher ethanol yield than Saccharomyces Y1609 strain under the same fermentation conditions; (b) increased temperature tolerance compared to Saccharomyces Y1609 strain at temperatures between 32°C and 36°C; (c) a lower glycerol yield than Saccharomyces Y1609 strain under the same fermentation conditions; (d) increased organic acid tolerance compared to Saccharomyces Y1609 strain at a pH of about 4.0 to about 5.2 in the presence of an organic acid; and (e) an increased fermentation rate compared to Saccharomyces Y1609 strain under the same fermentation conditions.

[0151] [000147] Item 4. A yeast strain or derivative according to any one of the preceding items, wherein the yeast strain or derivative produces at least about 0.5% more ethanol after 50 hours of fermentation compared to Saccharomyces cerevisiae Y1609 strain.

[0152] [000148] Item 5. A yeast strain or derivative according to any one of the preceding items, wherein the yeast strain or derivative produces at least about 2% less glycerol after 50 hours of fermentation compared to Saccharomyces cerevisiae Y1609 strain.

[0153] [000149] Item 6. A yeast strain or derivative according to any one of the preceding items, wherein the yeast strain or derivative has a fermentation rate at least about 2% higher than Saccharomyces cerevisiae Y1609 strain after 24 hours of fermentation.

[0154] [000150] Item 7. A yeast strain or derivative described in any one of items 3 to 6, wherein the temperature is 32°C.

[0155] [000151] Item 8. The yeast strain or derivative of item 3, wherein the yeast strain or derivative produces at least about 5% more ethanol after 50 hours of fermentation compared to Saccharomyces cerevisiae Y1609 strain.

[0156] [000152] Item 9. The yeast strain or derivative of item 3, wherein the yeast strain or derivative produces at least about 3% less glycerol after 50 hours of fermentation compared to Saccharomyces cerevisiae Y1609 strain.

[0157] [000153] Item 10. The yeast strain or derivative of item 3, wherein the yeast strain or derivative has a fermentation rate at least about 10% higher than Saccharomyces cerevisiae Y1609 strain after 24 hours of fermentation.

[0158] [000154] Item 11. A yeast strain or derivative described in any one of items 3 or 8 to 11, wherein the temperature is 36°C.

[0159] [000155] Item 12. The yeast strain or derivative of any one of items 3 to 11, wherein the organic acid comprises lactic acid, acetic acid, succinic acid, citric acid, malic acid, fumaric acid, or a combination thereof.

[0160] [000156] Paragraph 13. A method for producing a derivative of a Saccharomyces yeast strain described in paragraph 2, comprising the steps of: (a) providing (i) a first yeast strain selected from Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, and derivatives thereof, and (ii) a second yeast strain belonging to the phylogenetic clade of Saccharomyces sensu stricto; (b) inducing sporulation of the first yeast strain and the second yeast strain; (c) screening and selecting spores from the first yeast strain and spores from the second yeast strain; (d) crossing the selected spores of the first yeast strain with the selected spores of the second yeast strain; and (e) screening or selecting the derivative strain.

[0161] [000157] Clause 14. The method of clause 13, wherein step (c) comprises screening or selecting spores that exhibit one or more defining characteristics of Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or derivatives thereof.

[0162] [000158] Clause 15. The method of clause 13, wherein step (e) comprises screening or selecting hybrids exhibiting one or more defining characteristics of Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or derivatives thereof.

[0163] [000159] Paragraph 16. A method for producing a derivative of a Saccharomyces yeast strain described in paragraph 2, comprising the steps of: (a) providing (i) a first yeast strain selected from Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, and derivatives thereof, and (ii) one or more additional yeast strains belonging to the phylogenetic clade of Saccharomyces sensu stricto; (b) inducing sporulation of the first yeast strain and the one or more additional yeast strains to produce spores; (c) mixing all of the spores from step (b) to allow for spore crossing; and (d) screening or selecting a derivative strain.

[0164] [000160] Clause 17. The method of clause 16, wherein step (d) comprises screening or selecting hybrids that exhibit one or more defining characteristics of Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929.

[0165] [000161] Item 18. A mutant yeast of the yeast strain described in item 1 or a derivative described in item 2.

[0166] [000162] Item 19. A method for producing the mutant yeast of item 18, wherein the mutant yeast is mutated by contacting the yeast strain with a mutagen.

[0167] [000163] Item 20. The method of item 19, wherein the mutagen is ethyl methanesulfonate (EMS), ultraviolet light (UV), X-rays, methyl methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine mustard, 2-methoxy-6-chloro-9[3-(ethyl-2-chloroethyl)aminopropylamino]acridine-2 (ICR-170), or nitrogen mustard.

[0168] [000164] Item 21. A method for producing the mutant yeast of item 18, wherein the mutant yeast is mutated by contacting the derivative with a mutagen.

[0169] [000165] Item 22. The method of item 21, wherein the mutagen is ethyl methanesulfonate (EMS), ultraviolet light (UV), X-rays, methyl methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine mustard, 2-methoxy-6-chloro-9[3-(ethyl-2-chloroethyl)aminopropylamino]acridine-2 (ICR-170), or nitrogen mustard.

[0170] [000166] Item 23. An evolved yeast of the yeast strain described in item 1 or a derivative described in item 2.

[0171] [000167] Paragraph 24. A method for producing an evolved yeast according to paragraph 23, wherein the evolution is induced by applying selective pressure to a yeast strain.

[0172] [000168] Item 25. A method for producing an evolved yeast according to item 23, wherein the evolution is induced by applying selective pressure to the derivative.

[0173] [000169] Item 26. A genetically modified yeast of the yeast strain described in item 1 or a derivative described in item 2.

[0174] [000170] Paragraph 27. The genetically modified yeast of paragraph 26, wherein the nucleic acid sequence of the genetically modified yeast is altered using gene editing.

[0175] [000171] Item 28. A recombinant yeast strain of the yeast strain described in item 1 or a derivative of the yeast strain described in item 2.

[0176] [000172] 29. The recombinant yeast of claim 28, comprising a modification to suppress expression of a gene, enhance expression of a gene, introduce a gene, or delete a gene.

[0177] [000173] Item 30. A method for producing ethanol from a substrate by contacting the substrate with a fermenting organism, the fermenting organism being: (a) Saccharomyces strain Y1912, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68003, or a derivative thereof; (b) Saccharomyces strain Y1913, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68004, or a derivative thereof; (c) Saccharomyces strain Y1914, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68005, or a derivative thereof; (d) Saccharomyces strain Y1915, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68006, or a derivative thereof; (e) Saccharomyces strain Y1923, or a derivative thereof, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68007; (f) Saccharomyces strain Y1927, or a derivative thereof, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68008; and (g) Saccharomyces strain Y1929, or a derivative thereof, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68009.

[0178] [000174] Clause 31. The method of clause 30, wherein the substrate comprises or is derived from sugar cane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, cassava, or combinations thereof.

[0179] [000175] Paragraph 32. The method of paragraph 30, wherein the yeast strain comprises one or more defining characteristics selected from: (a) a higher ethanol yield than Saccharomyces Y1609 strain under the same fermentation conditions; (b) increased temperature tolerance compared to Saccharomyces Y1609 strain at temperatures between 32°C and 36°C; (c) a lower glycerol yield than Saccharomyces Y1609 strain under the same fermentation conditions; (d) increased organic acid tolerance compared to Saccharomyces Y1609 strain at a pH of about 4.0 to about 5.2 in the presence of an organic acid; and (e) an increased fermentation rate compared to Saccharomyces Y1609 strain under the same fermentation conditions.

[0180] [000176] Item 33. The method of item 30, wherein the ethanol is used as fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof.

[0181] [000177] Item 34. The method of item 30, wherein ethanol is produced using starch.

[0182] [000178] Item 35. The method of item 34, wherein ethanol is produced using simultaneous saccharification and fermentation (SSF) or continuous fermentation.

[0183] [000179] Item 36. The method of item 30, wherein ethanol is produced using sugar.

[0184] [000180] Item 37. The method of item 36, wherein ethanol is produced using batch fermentation or continuous fermentation.

[0185] [000181] Item 38. The method of item 30, wherein ethanol is produced using lignocellulosic sugars.

[0186] [000182] Item 39. The method of item 38, wherein ethanol is produced using simultaneous saccharification and fermentation (SSF) or separate hydrolysis and fermentation (SHF).

[0187] [000183] Item 40. A composition comprising the yeast strain described in item 1 or the derivative described in item 2 and one or more ingredients selected from surfactants, emulsifiers, thickeners, swelling agents, protectants, and antioxidants.

[0188] [000184] Paragraph 41. The composition of paragraph 40, comprising one or more defining characteristics selected from: (a) a higher ethanol yield than Fali under the same fermentation conditions; (b) increased temperature tolerance compared to Fali at temperatures between 32°C and 36°C; (c) a lower glycerol yield than Fali under the same fermentation conditions; (d) increased organic acid tolerance compared to Fali at a pH between about 4.0 and about 5.2 in the presence of an organic acid; and (e) an increased fermentation rate compared to Fali under the same fermentation conditions.

[0189] [000185] Item 42. A method for producing ethanol from biomass by contacting the biomass with the composition of item 40.

[0190] [000186] Item 43. The method of item 42, wherein the ethanol is used as fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof.

[0191] [000187] Item 44. The method of item 42, wherein ethanol is produced using starch.

[0192] [000188] 45. The method of claim 44, wherein ethanol is produced using simultaneous saccharification and fermentation (SSF) or continuous fermentation.

[0193] [000189] Item 46. The method of item 42, wherein ethanol is produced using sugar.

[0194] [000190] Item 47. The method of item 46, wherein ethanol is produced using batch fermentation or continuous fermentation.

[0195] [000191] Item 48. The method of item 42, wherein ethanol is produced using lignocellulosic sugars.

[0196] [000192] 49. The method of claim 48, wherein ethanol is produced using simultaneous saccharification and fermentation (SSF) or separate hydrolysis and fermentation (SHF).

[0197] [000193] Item 50. A method for producing a fermented product from a substrate by contacting the substrate with a fermenting organism, the fermenting organism being: (a) Saccharomyces strain Y1912, a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68003, or a derivative thereof; (b) Saccharomyces strain Y1913, a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68004, or a derivative thereof; (c) Saccharomyces strain Y1914, a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68005, or a derivative thereof; (d) Saccharomyces strain Y1915, a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68006, or a derivative thereof; (e) Saccharomyces strain Y1923, or a derivative thereof, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68007; (f) Saccharomyces strain Y1927, or a derivative thereof, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68008; and (g) Saccharomyces strain Y1929, or a derivative thereof, which is a representative specimen of the strain deposited under NRRL Patent Deposit Designation No. Y-68009.

[0198] [000194] Clause 51. The method of clause 50, wherein the substrate comprises or is derived from sugar cane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, cassava, or combinations thereof.

[0199] [000195] Item 52. The method of item 50, wherein the fermentation product is ethanol.

[0200] [000196] Item 53. The method of item 52, wherein the ethanol is used as fuel ethanol, industrial ethanol, potable ethanol, or a combination thereof.

[0201] [000197] Item 54. The method of item 50, wherein the fermentation product is produced using batch fermentation, continuous fermentation, simultaneous saccharification and fermentation (SSF), or separate hydrolysis and fermentation (SHF).

[0202] 8. Biological deposit [000198] Representative samples of the Saccharomyces yeast strains described herein were deposited at the Agricultural Research Service Patent Culture Collection Northern Regional Research Center (NRRL), 1815 University Street, Peoria, Illinois, USA on December 10, 2020, and were assigned NRRL Patent Deposit Designation No. Y-68003 (Y1912), NRRL Patent Deposit Designation No. Y-68004 (Y1913), NRRL Patent Deposit Designation No. Y-68005 (Y1914), NRRL Patent Deposit Designation No. Y-68006 (Y1919), NRRL Patent Deposit Designation No. Y-68007 (Y1923), NRRL Patent Deposit Designation No. Y-68008 (Y1927), and NRRL Patent Deposit Designation No. Y-68009 (Y1929) on December 23, 2020. The deposit will be kept at the NRRL depository under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure for at least 30 years and for at least 5 years after the depository accepts the most recent application to submit a sample of the deposit. Applicant has met all requirements of 37 C.FR §§ 1.801-1.809, including providing a representation of the viability of the sample. Additional deposits will be made at the NRRL as necessary to ensure availability, subject to the conditions set forth herein. Applicant will not impose any restrictions on the availability of the deposited material from the NRRL after the issuance of a patent on this application. Applicant is not authorized to wave any restrictions imposed by law on the transfer of biological material or its transportation in worldwide commerce. Applicant does not waive any rights granted under a patent issued on this application in any country.

Claims

1. (a) Saccharomyces strain Y1912, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68003; (b) Saccharomyces strain Y1913, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68004; (c) Saccharomyces strain Y1914, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68005; (d) Saccharomyces strain Y1919, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68006; (e) Saccharomyces strain Y1923, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68007; (f) Saccharomyces strain Y1927, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68008; and (g) Saccharomyces strain Y1929, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68009. A Saccharomyces yeast strain selected from the group consisting of:

2. (a) Saccharomyces strain Y1912, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68003; (b) Saccharomyces strain Y1913, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68004; (c) Saccharomyces strain Y1914, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68005; (d) Saccharomyces strain Y1919, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68006; (e) Saccharomyces strain Y1923, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68007; (f) Saccharomyces strain Y1927, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68008; and (g) Saccharomyces strain Y1929, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68009. A derivative of a Saccharomyces yeast strain selected from:

3. The yeast strain or derivative is (a) higher ethanol yield than Saccharomyces strain Y1609 under the same fermentation conditions; (b) increased temperature tolerance compared to Saccharomyces strain Y1609 at temperatures between 32°C and 36°C; (c) lower glycerol yield than Saccharomyces strain Y1609 under the same fermentation conditions; (d) increased organic acid tolerance compared to Saccharomyces strain Y1609 at a pH of about 4.0 to about 5.2 in the presence of organic acid; and (e) Increased fermentation rate compared to Saccharomyces strain Y1609 under the same fermentation conditions.

3. The yeast strain of claim 1 or a derivative of claim 2, comprising one or more defining characteristics selected from:

4. The yeast strain or derivative produces at least about 0.5% more ethanol after 50 hours of fermentation compared to Saccharomyces cerevisiae Y1609 strain; and / or The yeast strain or derivative produces at least about 2% less glycerol after 50 hours of fermentation compared to Saccharomyces cerevisiae Y1609 strain; and / or 4. The yeast strain or derivative of claim 3, wherein the derivative has a fermentation rate at least about 2% higher than the Saccharomyces cerevisiae Y1609 strain after 24 hours of fermentation.

5. 4. The yeast strain or derivative of claim 3, wherein the organic acid comprises lactic acid, acetic acid, succinic acid, citric acid, malic acid, fumaric acid, or a combination thereof.

6. A method for producing the derivative according to claim 2, comprising the steps of: (1) (a) (i) a first yeast strain selected from Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, and derivatives thereof; (ii) a second yeast strain belonging to the Saccharomyces sensu stricto phylogenetic clade; and providing a (b) inducing sporulation of the first yeast strain and the second yeast strain; (c) screening and selecting spores from the first yeast strain and spores from the second yeast strain; (d) crossing the selected spores of the first yeast strain with the selected spores of the second yeast strain; (e) screening or selecting derivative strains; Contains; or (2) (a) (i) a first yeast strain selected from Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, and derivatives thereof; (ii) one or more additional yeast strains belonging to the phylogenetic clade of Saccharomyces sensu stricto; and providing a (b) inducing sporulation of the first yeast strain and the one or more additional yeast strains to produce spores; (c) mixing all the spores from step (b) to allow cross-pollination of the spores; (d) screening or selecting derivative strains; A method comprising:

7. step (1)(c) comprising screening or selecting spores exhibiting one or more defining characteristics of Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or derivatives thereof; and 7. The method of claim 6, wherein step (e) comprises screening or selecting hybrids that exhibit one or more defining characteristics of Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, Y1929, or derivatives thereof.

8. 7. The method of claim 6, wherein step (2)(d) comprises screening or selecting hybrids that exhibit one or more defining characteristics of Saccharomyces strains Y1912, Y1913, Y1914, Y1919, Y1923, Y1927, or Y1929.

9. A mutant, evolved, genetically modified or recombinant yeast of the yeast strain according to claim 1 or a derivative according to claim 2.

10. (i) A method for producing the mutant yeast of claim 9, wherein the mutant yeast is mutated by contacting the yeast strain or derivative with a mutagen, optionally the mutagen being ethyl methanesulfonate (EMS), ultraviolet light (UV), X-rays, methyl methanesulfonate (MMS), nitrous acid, nitrosoguanidine (NNG), acridine mustard, 2-methoxy-6-chloro-9[3-(ethyl-2-chloroethyl)aminopropylamino]acridine.2 (ICR-170), or nitrogen mustard; (ii) a method for producing an evolved yeast according to claim 9, wherein the evolution is induced by applying selective pressure to a yeast strain or derivative; (iii) A method for producing the genetically modified yeast of claim 9, wherein the nucleic acid sequence of the genetically modified yeast is altered using gene editing; or (iv) A method for producing a recombinant yeast according to claim 9, comprising the step of introducing a nucleic acid into a yeast strain or derivative by recombinant DNA techniques.

11. 1. A method for producing ethanol from a substrate by contacting the substrate with a fermentation organism, the fermentation organism comprising: (a) Saccharomyces strain Y1912, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68003, or a derivative thereof; (b) Saccharomyces strain Y1913, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68004, or a derivative thereof; (c) Saccharomyces strain Y1914, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68005, or a derivative thereof; (d) Saccharomyces strain Y1919, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68006, or a derivative thereof; (e) Saccharomyces strain Y1923, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68007, or a derivative thereof; (f) Saccharomyces strain Y1927, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68008, or a derivative thereof; and (g) Saccharomyces strain Y1929, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68009, or a derivative thereof. A method selected from the group consisting of

12. A composition comprising the yeast strain according to claim 1 or the derivative according to claim 2 and one or more ingredients selected from surfactants, emulsifiers, thickeners, swelling agents, protectants and antioxidants, the composition comprising: (a) higher ethanol yield than Fali under the same fermentation conditions; (b) increased temperature tolerance compared to Fali at temperatures between 32°C and 36°C; (c) lower glycerol yield than Fali under the same fermentation conditions; (d) increased organic acid tolerance compared to Fali at a pH of about 4.0 to about 5.2 in the presence of an organic acid; and (e) Increased fermentation rate compared to Fali under the same fermentation conditions A composition comprising one or more defining features selected from:

13. 13. A method for producing a fermentation product, optionally ethanol, from a substrate by contacting the substrate with a fermenting organism or the composition of claim 12, the method comprising the steps of: (a) Saccharomyces strain Y1912, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68003, or a derivative thereof; (b) Saccharomyces strain Y1913, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68004, or a derivative thereof; (c) Saccharomyces strain Y1914, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68005, or a derivative thereof; (d) Saccharomyces strain Y1919, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68006, or a derivative thereof; (e) Saccharomyces strain Y1923, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68007, or a derivative thereof; (f) Saccharomyces strain Y1927, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68008, or a derivative thereof; and (g) Saccharomyces strain Y1929, which is a representative sample of the strain deposited under NRRL Patent Deposit Designation No. Y-68009, or a derivative thereof. The method of claim 1, wherein the

14. 14. The method of claim 13, wherein the substrate comprises or is derived from sugar cane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, cassava, or combinations thereof.

15. The ethanol is used as fuel ethanol, industrial ethanol, drinking ethanol, or a combination thereof; and / or 14. The method of claim 13, wherein the fermented product is produced using batch fermentation, continuous fermentation, simultaneous saccharification and fermentation (SSF), or separate hydrolysis and fermentation (SHF).

16. The method of claim 11, wherein the substrate comprises or is derived from sugar cane, sugar beet, sweet sorghum, agave, corn, wheat, rice, barley, rye, sorghum, triticale, potato, sweet potato, cassava, or combinations thereof.

17. The ethanol is used as fuel ethanol, industrial ethanol, drinking ethanol, or a combination thereof; and / or 12. The method of claim 11, wherein the fermented product is produced using batch fermentation, continuous fermentation, simultaneous saccharification and fermentation (SSF), or separate hydrolysis and fermentation (SHF).

Images