Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
  • C12N1/14—Fungi; Culture media therefor
  • C12N1/16—Yeasts; Culture media therefor
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K10/00—Animal feeding-stuffs
  • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
  • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
  • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K20/00—Accessory food factors for animal feeding-stuffs
  • A23K20/10—Organic substances
  • A23K20/174—Vitamins
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K20/00—Accessory food factors for animal feeding-stuffs
  • A23K20/20—Inorganic substances, e.g. oligoelements
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K20/00—Accessory food factors for animal feeding-stuffs
  • A23K20/20—Inorganic substances, e.g. oligoelements
  • A23K20/30—Oligoelements
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K50/00—Feeding-stuffs specially adapted for particular animals
  • A23K50/40—Feeding-stuffs specially adapted for particular animals for carnivorous animals, e.g. cats or dogs
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
  • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
  • A23L33/14—Yeasts or derivatives thereof
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
  • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
  • Y02E50/00—Technologies for the production of fuel of non-fossil origin
  • Y02E50/10—Biofuels, e.g. bio-diesel

Definitions

• the present invention relates to a method of preparing nutritionally fortified yeast, a nutritionally fortified yeast composition and a food composition.
• Yeast is often used in the food manufacturing industry, and particularly in the pet food manufacturing industry as a source of protein, nutraceutical supplement, texture modifier, and aroma enhancer. Furthermore, yeast is often used as a palatability enhancer due to the high inherent content of free glutamate and nucleotides.
• yeast species that is used in the food industry.
• Saccharomyces cerevisiae also known as baker's yeast or brewer's yeast
• Torulopsis utilis Torula utilis
• Candida utilis also known as Torula yeast
• yeast In addition to the abundant source of protein, glutamate and nucleotides, yeast also contains micronutrients such as B vitamins, potassium, and selenium, which can be used to supplement the diets of pets. It is preferable to utilize yeast as a source of micronutrients for pet foods rather than incorporating micronutrients individually into pet foods, to simplify the manufacturing process and to simplify long ingredient lists on pet food labels, which may deter pet owners.
• micronutrients such as B vitamins, potassium, and selenium
• the present invention provides a method of preparing nutritionally fortified yeast comprising: a) incubating yeast with at least one fermentable substrate under conditions suitable for achieving fermentation;
• the isolated yeast may be combined with at least one vitamin and/or mineral.
• the isolated yeast is combined with at least one vitamin in the absence of any minerals.
• the isolated yeast may be combined with at least one mineral, in the absence of any vitamins.
• isolated yeast is combined with one or more minerals that are chelated.
• the chelant comprises a peptide or partially hydrolysed peptide.
• the one or more minerals are selected from a proteinate of calcium, potassium, sodium, magnesium, iron, copper, manganese, zinc, iodine, cobalt and selenium.
• the vitamins comprise vitamin A, vitamin C, vitamin D, vitamin E, vitamin Bl (thiamin), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B6 (pyridoxine), vitamin B9 (folic acid), vitamin B12 (cobalamin), and choline.
• the isolated yeast is combined with at least one micronutrient selected from vitamins, minerals, amino acids and antioxidants at a temperature of 3°C to 45°C for a period of 30 minutes to 48 hours.
• the minerals comprise calcium, potassium, sodium, magnesium, iron, copper, manganese, zinc, iodine, cobalt and selenium.
• the minerals are chelated as defined above.
• the amino acids comprise threonine, isoleucine, lysine, methionine, cystine, phenylalanine, tyrosine, valine, arginine, histidine, alanine, and aspartic acid.
• the antioxidants comprise taurine, lipoic acid, glutathione, N-acetyl cysteine, vitamin E, vitamin C and beta-carotene.
• the isolated yeast is combined with at least one vitamin and/or mineral. Further optionally, the isolated yeast is combined with a heat- labile vitamin such as vitamin B 1 (thiamin) and/or vitamin C. Still further optionally, the isolated yeast is combined with at least one mineral selected from iron, zinc, copper, and manganese.
• a heat- labile vitamin such as vitamin B 1 (thiamin) and/or vitamin C.
• the isolated yeast is combined with at least one mineral selected from iron, zinc, copper, and manganese.
• the isolated yeast comprises a yeast cream.
• the yeast cream comprises 15 wt.% to 25 wt.% dry matter.
• the isolated yeast is combined with at least one vitamin in a total amount of 0.1 wt.% to 2 wt.% vitamins by total weight of the solid content of the isolated yeast. More preferably, the isolated yeast is combined with vitamins in a total amount of 1 wt.% to 1.5 wt.% vitamins by total weight of the solid content of the isolated yeast.
• the isolated yeast is combined with minerals in a total amount of 0.01 wt.% to 2 wt.% minerals by total weight of the solid content of the isolated yeast. More preferably, the isolated yeast is combined with minerals in a total amount of 0.1 wt.% to 1 wt.% minerals by total weight of the solid content of the isolated yeast.
• the minerals are chelated as defined above.
• the yeast is isolated by centrifugation.
• the isolated yeast is allowed to autolyse prior to combining with the at least one micronutrient. More preferably, the autolysed yeast comprises 10 to 25 wt.% dry matter.
• the nutritionally fortified yeast is pasteurized by heating to a temperature of 60°C to 135°C.
• the nutritionally fortified yeast is dried to a moisture content of 1 wt.% to 10 wt.%.
• the yeast is dried by spray-drying or freeze-drying.
• the fermentable substrate comprises ethanol.
• the yeast is Torulopsis utilis, Torula utilis or Candida utilis.
• the present invention provides a nutritionally fortified yeast composition obtained by the method as defined herein.
• the present invention provides a food composition comprising nutritionally fortified yeast obtained by the method as defined herein at least one food ingredient.
• the food composition comprises the nutritionally fortified yeast in an amount of up to 2 wt.%.
• the present invention provides a use of a nutritionally fortified yeast composition obtained by the method as defined herein as a palatability enhancer.
• ranges are used as shorthand for describing each and every value that is within the range. Any value within the range can be selected as the terminus of the range.
• the term "food” may refer not only to a food product which typically provides most, if not all, the nutrient value for an animal, but may also refer to such items as a snack, treat, and supplement.
• the present invention provides a method of preparing nutritionally fortified yeast comprising:
• yeast A variety of species of yeast may be used in the methods of the present invention. These include without limitation, Saccharomcyes, Kluyveromyces, Candida and Torulaspora.
• the yeast is Saccharomcyes cerevisiae.
• the yeast is Torulopsis utilis, Torula utilis or Candida utilis (Torula yeast).
• Torula yeast is a good candidate for the methods of the present invention due to its inherently low selenium level (which can cause toxicity in the consumer).
• yeast is fermented according to standard methods that would be known to those skilled in the art.
• Fermentable substrates include dextrose, glucose, maltose, fructose and sucrose.
• the fermentable substrate is ethanol.
• Ethanol is particularly suitable for Torula yeast fermentation.
• the yeast is isolated.
• isolation it is meant that the yeast is separated from a major proportion of the fermentation broth.
• the yeast is preferably isolated by centrifugation.
• the centrifugate may be washed and re-centrifuged to yield isolated yeast.
• the isolated yeast is a yeast cream having a solid content of about 15 wt.% to about 25 wt.%.
• the solid content of the yeast cream is from about 15 wt.%, 16 wt.%, 17 wt.% or 18 wt.% to about 22 wt.%.
• yeast isolated from the fermentation broth for example, in the form of a yeast cream
• yeast extract may be prepared from the isolated yeast.
• Conventional methods of preparing yeast extract would be known to the person skilled in the art of food manufacturing. Typically methods involve removing cells walls and isolating the cellular contents.
• the isolated yeast is allowed to autolyse.
• Yeast autolysis is a complex and slow phenomenon which involves the yeast's endogenous hydrolytic enzymes such as proteases, nucleases, lipases and glycanases degrading cellular components. Unlike the preparation of yeast extract, cell wall components are not removed during autolysis.
• the solid content (dry matter) of the autolysed yeast is from 10 wt.% or 15 wt.% to 25 wt.%. Often the solid content (dry matter) of the autolysed yeast is from about 15 wt.%>, 16 wt.%>, 17 wt.%> or 18 wt.%> to about 22 wt.%>.
• the autolysed yeast extract may subsequently be combined with one or more micronutrients for fortification of the yeast as described below. In one embodiment, a mixture of (unautolysed) yeast cream and autolysed yeast is fortified with micronutrients. Micronutrients
• the isolated yeast is typically combined with one or more micronutrients selected from vitamins, minerals, amino acids and antioxidants.
• the isolated yeast is combined with at least one vitamin and/or at least one mineral.
• Vitamins include without limitation, vitamin A, vitamin Bl (thiamine), vitamin B2 (riboflavin), vitamin B3 (niacin), vitamin B5 (pantothenic acid), vitamin B6 (pyridoxine), vitamin B9 (folic acid), vitamin B12, (cobalamin), biotin, choline vitamin C, vitamin D and vitamin E.
• the isolated yeast is combined with a heat-labile vitamin such as vitamin Bl (thiamine) and/or vitamin C.
• a heat-labile vitamin such as vitamin Bl (thiamine) and/or vitamin C.
• One or more vitamins may be combined with the isolated yeast in a total amount of 0.1 wt.% to 5 wt.%, or 0.1 wt.% to 4 wt.%, or 0.1 wt.% to 3 wt.%, or 0.1 wt.% to 2 wt.% vitamins by total weight of the solid content(dry matter) of the isolated yeast.
• the total amount of vitamins is 0.5 wt.% to 3 wt.%, or 0.5 wt.% to 2 wt.%, or 0.5 wt.%) to 1 wt.%), or 1 wt.%) to 3 wt.%, or 1 wt.% to 2 wt.% by total weight of the solid content (dry matter) of the isolated yeast.
• Minerals include without limitation, calcium, potassium, sodium, magnesium, iron, copper, manganese, zinc, iodine, cobalt and selenium.
• the isolated yeast is combined with at least one mineral selected from iron, zinc, copper and manganese.
• One or more minerals may be combined with the isolated yeast in a total amount of 0.1 wt.% to 3 wt.%, 0.1 wt.% to 2 wt.%, or 0.1 wt.% to 1 wt.%, or 0.1 wt.% to 0.5 wt.% minerals by total weight of the solid content (dry matter) of the isolated yeast.
• the total amount of minerals is 0.5 wt.% to 3 wt.%, or 0.5 wt.% to 2 wt.%, or 0.5 wt.% to 1 wt.% by total weight of the solid content (dry matter) of the isolated yeast.
• the minerals are chelated.
• the isolated yeast may be combined with one or more chelated minerals.
• the minerals may be chelated by one or more amino acids, a partially hydrolysed peptide, or a peptide.
• Chelated minerals that may be used in the methods of the present invention include without limitation, a proteinate of calcium, phosphorus, potassium, sodium, chloride, magnesium iron, copper, manganese, zinc, iodine, and selenium. Chelated minerals are commercially available from various sources such as Alltech ® (Nicholasville KY).
• at least two minerals may be chelated by the same chelant as a complex structure.
• One example of such a multimineral-chelant complex is Bioplex ® zinc/copper/manganese.
• the present inventors have found that when conventional (unchelated) minerals are used in the methods of the present invention, the minerals have a tendency to coagulate and precipitate with yeast. This significantly reduces the level of fortification. Such coagulation and precipitation is not observed with chelated minerals, and the level of fortification is also increased with chelated minerals.
• Amino acids include without limitation asparagine, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, alanine, arginine, aspartic acid cysteine, glutamic acid, glutamine, glycine, tyrosine, proline, valine and serine.
• the amino acid is selected from threonine, isoleucine, lysine, methionine, cystine, phenylalanine, tyrosine, valine, arginine, histidine, alanine, and aspartic acid.
• One or more amino acids may be combined with the isolated yeast in a total amount of up to 1 wt.% by total weight of the solid content (dry matter) of the isolated yeast. In some embodiments, one or more amino acids are combined with the isolated yeast in a total amount of 0.1 wt.% to 1 wt.%, or 0.1 wt.% to 0.5 wt.% or 0.1 wt.% to 0.3 wt.% by total weight of the solid content (dry matter) of the isolated yeast.
• one or more amino acids are combined with the isolated yeast in a total amount of 0.3 wt.% to 1 wt.%, 0.5 to 1 wt.% or 0.5 wt.% to 0.8 wt.% by total weight of the solid content (dry matter) of the isolated yeast.
• Antioxidants include without limitation, taurine, lipoic acid, glutathione, N-acetyl cysteine, vitamin E, vitamin C and beta-carotene.
• the antioxidant is taurine.
• One or more antioxidants may be combined with the isolated yeast in a total amount of up to 0.2 wt.% by total weight of the solid content (dry matter) of the isolated yeast. In one embodiment, one or more antioxidants are combined with the isolated yeast in a total amount of 0.01 wt.% to 0.2 wt.%, or 0.01 wt.% to 0.1 wt.% or 0.01 wt.% to 0.05 wt.% by total weight of the solid content (dry matter) of the isolated yeast.
• one or more antioxidants are combined with the isolated yeast in a total amount of 0.05 wt.% to 0.2 wt.%, or 0.05 wt.% to 0.1 wt.% or 0.05 wt.% to 0.08 wt.% by total weight of the solid content (dry matter) of the isolated yeast.
• the micronutrients are combined with the isolated yeast at a temperature of 3°C to 45°C for a period of 0.5 hours to 48 hours. In some embodiments, the micronutrients are combined with the isolated yeast at a temperature of 5°C to 25°C or 5°C to 10°C.
• the micronutrients are combined with the isolated yeast at a temperature of 25°C to 45°C or 10°C to 20°C.
• the micronutrients are combined with the isolated yeast for a period of from 0.5 hours, 1 hour, 5 hours, 10 hours or 20 hours up to 30 hours.
• the micronutrients are combined with the isolated yeast for a period of 10 hours to 20 hours.
• the isolated yeast typically has a solid content ranging from about 15 wt.% to about 22 wt.%. Fermentation media constitutes the remainder of the isolated yeast.
• micronutrients are combined directly with the isolated yeast.
• micronutrients may be combined directly with the isolated yeast in solid form, without being made into a solution first.
• step c) as described herein the yeast and micronutrients may be mixed continuously to ensure optimal fortification.
• micronutrients may become adhered to or encapsulated by yeast cellular components, resulting in the fortification of the yeast.
• the nutritionally fortified yeast may be pasteurized and/or dried. Methods of pasteurization and drying would be known to those skilled in the art of food manufacturing. Typically, the nutritionally fortified yeast is heated to a temperature of 60°C to 135°C, or from 80°C to 100°C, or from 90°C to 95°C. The time period over which the nutritionally fortified yeast is heated may be from 5 seconds to 200 seconds, or from 5 seconds to 120 seconds, or from 30 seconds to 120 minutes.
• the nutritionally fortified yeast may be dried by conventional techniques such as spray-drying or freeze-drying. Typically, the nutritionally fortified yeast is dried to a moisture content of 0.5 wt.% to 10 wt.%, or from 1 wt.% to 5 wt.% or from 1 wt.% to 2 wt.% by total weight of the yeast.
• yeast fortified according to the methods of the present invention unexpectedly retain high levels of micronutrients.
• retention refers to an association or combination of micronutrients with the yeast or yeast cellular components. Therefore, nutritionally fortified yeast obtained by the methods of the present invention is useful for incorporation into food compositions, and in particular, pet food compositions.
• the present invention provides a food composition comprising nutritionally fortified yeast obtained according to the methods of the present invention.
• a food composition comprising nutritionally fortified yeast obtained according to the methods of the present invention.
• an amount as low as 4 wt.%, 3 wt.% or 2 wt.% of the nutritionally fortified yeast may be incorporated into food compositions to achieve adequate micronutrient supplementation of the food.
• the food composition comprises a nutritionally fortified yeast obtained by the methods of the present invention in amount of up to 4 wt.%), up to 3 wt.%), or up to 2 wt.% by total weight of the composition.
• the food composition comprises a nutritionally fortified yeast obtained by the methods of the present invention in amount of 1 wt.% to 3 wt.% or 1 wt.% to 2 wt.% by total weight of the composition.
• the food composition of the present invention typically comprises nutritionally fortified yeast and at least one food ingredient.
• the at least one food ingredient may be selected from protein (for example, meat, meat-by products, dairy products, eggs, wheat protein, soy protein and potato concentrate), fat (for example, animal fat, fish oil, vegetable oil, meat and meat by-products), and carbohydrate (for example, grains such as wheat, corn, barley and rice).
• Other food ingredients include, without limitation, fiber (for example cellulose, beet pulp, peanut hulls and soy fiber), vitamins, minerals and preservatives.
• the food ingredient may be any food ingredient defined herein.
• the food compositions of the present invention comprise nutritionally fortified yeast obtained by the method as defined herein and at least one food ingredient selected from protein, fat and carbohydrate.
• Food compositions supplemented with nutritionally fortified yeast according to the present invention may comprise 0.01 wt.% to 0.2 wt.% vitamin C, 0.01 wt.% to 0.2 wt.% vitamin E, 0.005 wt.% to 0.05 wt.% thiamine, 0.005 wt.% to 0.2 wt.% zinc and 0.002 wt.% to 0.1 wt.% iron.
• Table 1 indicates a typical micronutrient profile of a food composition comprising nutritionally fortified yeast obtained by the method of the present invention, wherein the micronutrients are derived from the nutritionally fortified yeast.
• Table 1 Micronutrient profile of food composition comprising up to 2 wt.% nutritionally fortified yeast.
• VITAMIN A IU/Kg 1,000 500,000
• VIT B12 ppm 0.01 1000
• the food composition of the present invention may be suitable for consumption by any animal.
• the food composition is for consumption by non-human animals, which include, without limitation, avians, bovines, canines, equines, felines, murines, ovines, and porcines.
• the food composition is for consumption by a pet. Most preferably, the food composition is for consumption by a companion animal, including a canine or a feline.
• Food compositions of the present invention can be prepared in a dry or wet form using conventional processes.
• the food composition as defined herein is free of further supplemented or exogenously incorporated micronutrients (for example, vitamins and minerals).
• supplied or exogenously incorporated micronutrients refers to micronutrients that are not contained with the at least one food ingredient of the food composition or the nutritionally modified yeast.
• the nutritionally fortified yeast obtained by the methods of the present invention may be used to enhance the palatability of a food composition.
• the present invention further provides a palatability enhancer comprising nutritionally fortified yeast obtained by the method defined herein, a use of nutritionally fortified yeast obtained by the method defined herein for enhancing the palatability of a food composition, and a method of enhancing the palatability of a food composition comprising incorporation nutritionally fortified yeast obtained by the method defined herein into the food composition.
• Torula yeast was fermented in the presence of ethanol. Post-fermentation, the yeast was centrifuged to 15 % to 22% solids, cooled, and placed in a process tank. A portion of the yeast was subjected to autolysis in this tank and the remainder was retained in the tank as (unautolysed) yeast cream. When the autolysis process was completed, the yeast cream and autolysed yeast were mixed to form an isolated yeast mixture. Subsequently, the isolated yeast mixture was combined and mixed with four different amounts of a vitamin blend comprising amongst other vitamins, 4.35 wt.% thiamine, as indicated in Table 2.
• the mixing took place in a high speed, high sheer mixer over a period of at least 30 minutes and at a temperature in the range of 20°C-30°C. Subsequently, the fortified yeast was pasteurized. [00067] The amount of thiamine retained in the yeast after fortification was subsequently determined by using acid hydrolysis to promote the release of the thiamine from the yeast matrix, followed by HPLC determination for separation and quantitative determination of the thiamine. The determined amount of thiamine was subsequently compared to expected thiamine retention levels (assuming no losses). Thiamine was selected as the prototype vitamin as it is susceptible to thermal degradation, and thus it is likely to be more susceptible to a reduced recovery. The results are illustrated in Table 2. As detailed below, all calculations are conducted on a dry matter basis to account for moisture variability.
• Example 1 The method set out in Example 1 was repeated with various minerals as set out in Table 3.
• the minerals were combined with yeast either alone, or with the vitamin pre-mix.
• the amount of minerals retained during the fortification was determined by converting the target elements into ashes by combustion of organic substances in the yeast matrix, followed by quantitatively determining the target elements using an Inductively Coupled Plasma instrument.
• Example 1 The method set out in Example 1 was repeated using thiamine and vitamin C (as individual vitamins rather than a vitamin blend), and various chelated minerals.
• Chelated minerals (Bioplex ® zinc/manganese/copper and Bioplex ® iron) were purchased from Alltech ® (NicholasviUe KY). The fortification process was carried out using the minerals alone, vitamins alone, and both vitamins and minerals. The retention of vitamins and minerals was quantified as described in Examples 1 and 2, respectively.

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