US20150250839A1 - Nutritional supplement containing iron - Google Patents

Nutritional supplement containing iron Download PDF

Info

Publication number
US20150250839A1
US20150250839A1 US14/425,493 US201314425493A US2015250839A1 US 20150250839 A1 US20150250839 A1 US 20150250839A1 US 201314425493 A US201314425493 A US 201314425493A US 2015250839 A1 US2015250839 A1 US 2015250839A1
Authority
US
United States
Prior art keywords
iron
nutritional supplement
product
ferrous
filamentous fungi
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US14/425,493
Other languages
English (en)
Inventor
J. Bruce WICKING
Yilin Bian
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Cura Global Health BVI Ltd
Original Assignee
DAKOTA STAR CAPITAL LLC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by DAKOTA STAR CAPITAL LLC filed Critical DAKOTA STAR CAPITAL LLC
Priority to US14/425,493 priority Critical patent/US20150250839A1/en
Assigned to DAKOTA STAR CAPITAL, LLC reassignment DAKOTA STAR CAPITAL, LLC ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: BIAN, YILIN, WICKING, J. BRUCE
Publication of US20150250839A1 publication Critical patent/US20150250839A1/en
Assigned to CURA GLOBAL HEALTH (BVI) LIMITED reassignment CURA GLOBAL HEALTH (BVI) LIMITED ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: DAKOTA STAR CAPITAL, LLC
Abandoned legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/062—Ascomycota
    • A23L1/0017—
    • A23L1/0029—
    • A23L1/0035—
    • A23L1/28—
    • A23L1/304—
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/52—Adding ingredients
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L31/00—Edible extracts or preparations of fungi; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • 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
    • A23L33/16—Inorganic salts, minerals or trace elements
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/20—Agglomerating; Granulating; Tabletting
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/30—Encapsulation of particles, e.g. foodstuff additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/40—Shaping or working of foodstuffs characterised by the products free-flowing powder or instant powder, i.e. powder which is reconstituted rapidly when liquid is added
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A61K33/26—Iron; Compounds thereof
    • 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
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L7/00—Cereal-derived products; Malt products; Preparation or treatment 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

Definitions

  • the technology relates to nutritional supplements containing iron and processes for making the supplements using filamentous fungi.
  • anemia is a public health problem that affects populations in both rich and poor countries,’ in particular pregnant women, non-pregnant women of reproductive age, and preschool-age children.
  • WHO World Health Organization
  • anemia is a public health problem that affects populations in both rich and poor countries,’ in particular pregnant women, non-pregnant women of reproductive age, and preschool-age children.
  • the most significant contributor to the onset of anemia is iron deficiency anemia (IDA). It has been estimated that over 1 billion people (approximately 15% of the global population) suffer from IDA. Additionally 1 billion people suffer from iron deficiency (ID) or iron depletion.
  • ID iron deficiency
  • Iron is essential to most life because it is an integral part of many proteins and enzymes that maintain good health. In humans, iron is an essential component of the red blood cells involved in oxygen transport, cell growth and other functions. Generally, ID limits oxygen delivery to the cell, resulting in fatigue, poor work performance and decreased disease immunity. But in children aged 0-5 years, ID impairs physical and cognitive development and thus may persist into adulthood. In pregnancy, IDA can result in 20-30% of maternal deaths, premature births and infant loss.
  • Type 1 Major types of iron that are considered important in human nutritional applications
  • Type Major Source Heme Iron Hemoglobin Animal livers, red meat, poultry, and seafood
  • Non-Heme Plant Based Lentils, grains and some Iron Food vegetables Inorganic Ferrous Ferrous sulfate, ferrous fumarate, Salt/Iron Salts ferrous citrate, ferrous gluconate, Supplements ferric ammoniums citrate, ferrous citrate and ethylenediaminetetraacetic acid (EDTA) iron
  • Soluble compounds of iron salt are desirable as iron supplements but cannot be used in many fortified foods due to sensory issues.
  • Other forms such as ferrous fumarate can be microencapsulated and added to semi-liquid food but costs are significantly higher.
  • Iron fortification of food is the preferred method to provide iron to regions of the world.
  • successful forms include ferrous sulfate or NAFeEDTA fortified fish sauce and soy sauce with ascorbic acid.
  • the impact of fortification on the prevalence of ID at the population level has not proven successful.
  • One of the reasons is due to iron enhancers vs inhibitors in the food delivery vehicles, for example cereal flours.
  • the iron fortification and supplements need different products to fit into many food systems. Therefore, more non-heme iron products in organic form are needed to meet demand across diverse dietary situations.
  • filamentous fungi can be cultivated on an agricultural by-product or a food processing by-product and harvested to obtain a nutritional product high in iron.
  • Nutritional supplements containing iron and methods for forming nutritional supplements containing iron are disclosed.
  • a nutritional supplement comprising:
  • fungal biomass having at least about 100 mg/kg iron.
  • the nutritional supplement contains from about 200 to 40000 mg/kg iron.
  • the nutritional supplement containing iron may include diluents or other co-ingredients such as selenium and zinc.
  • the supplement may contain selenium from 10 to 400 mg/kg; and zinc from 20 to 20,000 mg/kg.
  • the nutritional supplement containing iron may be formulated as a powder, solution, drink, capsule, tablet, caplet.
  • the powdered form of product can also be added to food and used as a food fortification ingredient.
  • the fortification of food includes, but is not limited to condiments, salt, baby formula, and flours of wheat, corn and beans.
  • the nutritional supplement contains natural organic iron derived from the by-products.
  • the nutritional supplement may be formulated to further contain phytase and other enzymes naturally produced by the filamentous fungi.
  • the nutritional supplement may contain additional inorganic iron salts or other high iron compounds added during growth of the filamentous fungi.
  • additional inorganic iron salts or other high iron compounds include ferrous sulfate, ferrous fumarate, ferrous citrate, ferrous gluconate, ferric ammoniums citrate, ferrous citrate and EDTA iron.
  • a process for forming a nutritional supplement containing iron comprising:
  • the filamentous fungi is selected from Aspergillus oryzae or Aspergillus niger.
  • the agricultural by-product can be from waste derived from corn, wheat, sugar beet, cane sugar, soybean, stillage and solid waste from alcohol production. Examples of such products are sugar cane and beet pulps, soybean hull, soybean process whey, wheat hull, spent grain and stillage.
  • the agricultural by-product is condensed corn soluble (Syrup), corn, wheat and soybean process by-products. More preferably, the agricultural by-product is Syrup.
  • the food processing by-product is selected from corn steeping liquor, corn stillage, soybean whey, sugar cane and beet molasses, soybean hull and wheat bran and wheat hull.
  • Additional growth media can be provided to the agricultural by-product or a food processing by-product to assist growth and accumulation of iron by the filamentous fungi during culture.
  • Additional inorganic iron salts and other high iron compounds such as ferric sulfate, ferrous sulfate, ferric ammonium citrate, ferrous citrate, but not limited to these salts maybe added during growth of the filamentous fungi to further increase iron content of the nutritional supplement.
  • the nutritional supplement may further include other minerals such at selenium and zinc or other minerals such as magnesium, calcium and chromium.
  • selenium and zinc compounds may be added to the culture. Suitable compounds include sodium selenite, zinc sulfate, calcium sulphate, chromium chloride, and magnesium sulphate.
  • the nutritional supplement may further include selenium and zinc.
  • the nutritional supplement contains selenium from 10 to 400 mg/kg; and zinc from 20 to 20,000 mg/kg.
  • Additional culture media or nutrients may be provided to assist in growth of the filamentous fungi.
  • yeast extract ammonium salts, urea, and potassium phosphorus.
  • the filamentous fungi may be cultured in any suitable environment such as fermentation vessels used in both solid and liquid fermentations.
  • Culture of the filamentous fungi may be carried out at room temperature or elevated temperatures such as 25 to 55° C.
  • the filamentous fungi can be harvested by any suitable means. Examples include filtration, such as filter press, belt press; centrifugation, such as decanter, drying, such as rotary drier, steam drier.
  • the drying temperature is typically lower than about 90° C. to avoid any unwanted heat damage of the product.
  • the harvested filamentous fungi may be further processed to form the nutritional supplement containing iron. Further processing may include separating, crushing, grinding, fractionation, extraction, washing with cold and hot water to remove excess salts, or mild acid with pH of 2 or alkaline wash with pH of 9-10 to remove other soluble compounds.
  • the nutritional supplement contains at least about 200 mg/kg iron.
  • the nutritional supplement contains from about 200 to 40000 mg/kg iron.
  • the iron content can be higher than 40000 mg/kg, but the yield of fungi biomass may be reduced and may not be economical in practice.
  • the nutritional supplement containing iron may include diluents or other co-ingredients such as selenium and zinc.
  • the supplement may contain selenium from 10 to 400 mg/kg; and zinc from 20 to 20,000 mg/kg.
  • the nutritional supplement may contain additional inorganic iron salts or other high iron compounds added during growth of the filamentous fungi.
  • additional inorganic iron salts or other high iron compounds include ferrous sulfate, ferrous fumarate, ferrous citrate, ferrous gluconate, ferric ammoniums citrate, ferrous citrate and EDTA iron.
  • the nutritional supplement may be formulated for human or animal use.
  • the technology relates to a nutritional supplement containing iron produced by the process described.
  • the technology relates to a nutritional supplement containing iron derived or obtained from filamentous fungi cultured in an agricultural by-product or a food processing by-product.
  • FIG. 1 is a schematic of a preferred process for the present technology.
  • FIG. 2 is a schematic of another preferred process for the present technology.
  • FIG. 3 shows results of the effect of addition of inorganic iron salts on fungal biomass iron content.
  • FIG. 4 shows results of Caco-2 Cell ferritin formation as % of ferrous sulfate.
  • Yeast has been reported to accumulate many microelements including iron on Yeast Extract Peptone Dextrose (YEPD) plates containing FeSO 4 . Certain yeast strains can uptake iron to 25 mg per gram of cell and as high as 95% of this iron was considered as being an organic form in the cell structure. Yeast biomass enriched with iron is used as an animal feed supplement and is considered less toxic as compared to iron salt. In other non-yeast mycelia forming fungi, the Fe3+, Fe2+, Mg2+ and Zn2+ are able to stimulate glucan formation. However, the use of filamentous fungi to produce an iron enriched product has not been practiced.
  • the fundamental difference between the current technology and the iron enrichment of yeast is the recognition by the present inventors that the filamentous fungi, such as Aspergillus oryzae , have the ability to accumulate organic iron from natural sources such as agricultural and food process wastes and by-products without iron fortification when such wastes and by-products are used as a growth media.
  • the filamentous fungi that have Generally Recognized as Safe (GRAS) status, such as Aspergillus oryzae and Aspergillus niger that can be used in production of iron rich and enriched products.
  • GRAS Generally Recognized as Safe
  • a further advantage is that the filamentous fungi product has no off flavor as compared to yeast products.
  • Table 2 lists some of the iron rich agricultural by products in North America that could be used for the production of iron rich fungal products. Other by products such wheat bran and hull, rice hull, sorghum hull and potato skin also have potential.
  • the present technology relates to the use of selected fungal species as the means to concentrate iron into an organic form suitable for animal or human consumption.
  • testing is preferably carried out for microbial spoilage, mycotoxins and heavy metal contamination.
  • dilution may be necessary as the optimal total solids for liquid fermentation of fungal species, such as Aspergillus oryzae or Aspergillus niger is about 3-10%.
  • Dry raw materials may be ground, soaked or cooked to release natural iron and reduce anti-fermentation factors.
  • enzymes such as cellulase, hemicellulase and phytase, may be used during soaking and cooking. Multiple raw materials may be used at the same time depending upon availability and pricing.
  • FIG. 1 General steps of a preferred process using an agricultural by-product or a food processing by-product are set out in FIG. 1 .
  • Strains of Aspergillus oryzae used were the same strains that are approved and employed commercially for soy sauce and miso manufacture, including Aspergillus oryzae 2355 and 40151 from Chinese Center of Industrial Culture Collection (CICC); Aspergillus oryzae 22787 from American Type Culture Collection (ATCC) and Aspergillus niger var. 2206 and 10557 for citric acid production from CICC and Aspergillus niger 66876 for phytase production from ATCC.
  • CICC Chinese Center of Industrial Culture Collection
  • ATCC American Type Culture Collection
  • ATCC American Type Culture Collection
  • Aspergillus niger var. 2206 and 10557 for citric acid production from CICC
  • Aspergillus niger 66876 for phytase production from ATCC.
  • Stains of Aspergillus oryzae and Aspergillus niger were cultured and maintained in media composed of ground whole corn, wheat bran, soybean hulls, molasses of beet, cane and fruits juice process by-product, and any other food process by-product consist of starch, sugar and protein.
  • Such raw materials can be pretreated by enzymes, including amylases, gluco-amylases, phytase and protease.
  • Inorganic iron salts were added to the fungal culture media, such inorganic iron salts include ferrous sulfate, ferrous fumarate, ferrous citrate, ferrous gluconate, ferric ammoniums citrate, ferrous citrate and ethylenediaminetetraacetic acid (EDTA) iron at concentration of 300-2000 mg of iron element per liter of media.
  • EDTA ethylenediaminetetraacetic acid
  • Fungal spores were prepared by inoculating a solid media, such as cooked rice, soybean, and sorghum and the combination of them with moisture of 40-70%. In 2-3 weeks, the spores germinated and were ready to be collected. The fungal spores were collected into sterilized distill water.
  • the pre-cultures fermenters were prepared with 1-10% volume of the final production fermenters. The media for the pre-cultures can be the same as the production media as described above. Incubation for 18-28 hours of pre-culture fermentation time is suitable to generate healthy pre-cultures after the spores were introduced into the pre-culture media. The pre-culture is added to, the production fermenter and fungus is allowed to grow to produce the desired fungal mass containing iron.
  • Large scale fermentation can be carried out in any suitable fermentation vessel or apparatus.
  • the fermentation is preferably carried out under aerobic conditions for 48-72 hours.
  • Sterilized or filtered air can be pumped into the fermenter at 0.5 to 1.0 vvm during the fermentation period to improve growth and yield.
  • the culture is preferably agitated or stirred during fermentation. The combination of air, agitation and design of the fermentation vessel is well understood for commercial microbial culture.
  • Fermentation can be carried out for 48-72 hours or until cell autolysis begins at a temperature of 28-30° C. It will be appreciated that incubation times and temperature may vary depending on the fungus type and strain used.
  • nutrients may be needed to supplement the growth media for an aerated fungal fermentation.
  • These nutrients may include organic and inorganic nitrogen sources, phosphors source and micro minerals.
  • Fungi including filamentous fungi, have the ability to further uptake relatively bio-unavailable and strongly cytotoxic iron. It should be noted that, direct supplementation of soluble inorganic iron salt in human diets can result in a cytotoxic reaction. Therefore, using fungi to uptake the inorganic iron salts and transform to an organic form may reduce the side effects of the direct consumption of iron salts.
  • Inorganic salts can be added during the fermentation.
  • the common choice of iron salts includes ferric sulfate, ferrous sulfate, and ferric citrate.
  • the inorganic iron is converted to an organic form by the fungi.
  • a given iron salt can be incrementally fed during the fermentation.
  • the dosing of iron salts depends on the type of salt used but the dosing level needs to not compromise the growth of fungi.
  • the fungal mycelium can be thoroughly washed to remove excess iron salts. A mild acid, pH 2-3, wash can be effective in this regard. Detailed steps of the process are described in FIG. 2 .
  • fungal biomass containing iron can be harvested by a dewatering machine such as a centrifuge, belt press etc. Washing with water and/or mild acid such as hydrochloric acid 0.01 M can be used to remove inorganic iron residues.
  • the iron enriched fungal product can then be dried at 60-80° C. using forced air, fluid bed dryer, etc. The final moisture of the product is preferably less than about 10%.
  • the compounds may be formulated for oral delivery.
  • suitable acceptable excipients or carriers include tablets, capsules, caplets, powders, granules, ampoules, vials, ready-to-use solutions or suspensions, drinks, and lyophilized materials.
  • the solid formulations such as the tablets or capsules may contain any number of suitable acceptable excipients or carriers.
  • the nutritional supplement comprises fungal biomass having at least about 100 mg/kg iron.
  • the nutritional supplement typically contains from about 100 to 40000 mg/kg iron.
  • the supplement can have at least about 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10000, 11000, 12000, 13000, 14000, 15000, 16000, 17000, 18000, 19000, 20000, 21000, 22000, 23000, 24000, 25000, 26000, 27000, 28000; 29000, 30000, 31000, 32000, 33000, 34000, 35000, 36000, 37000, 38000, 39000, or 40000 mg/
  • the nutritional supplement containing iron may also include diluents or other co-ingredients such as selenium and zinc.
  • the supplement may contain selenium from 10 to 400 mg/kg; and zinc from 20 to 20,000 mg/kg.
  • the supplement may contain 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390 or 400 mg/kg selenium.
  • the supplement may also contain 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10000, 11000, 12000, 13000, 14000, 15000, 16000, 17000, 18000, 19000 or 20000 mg/kg zinc.
  • the nutritional supplement containing iron may be formulated as a powder, solution, drink, capsule, tablet, caplet.
  • the powdered form of product can also be added to food and used as a food fortification ingredient.
  • the fortification of food includes, but is not limited to condiments, salt, baby formula, and flours of wheat, corn and beans.
  • the nutritional supplement contains natural organic iron derived from the by-products.
  • the nutritional supplement may be formulated to further contain phytase and other enzymes naturally produced by the filamentous fungi.
  • the nutritional supplement may contain additional inorganic iron salts or other high iron compounds added during growth of the filamentous fungi.
  • additional inorganic iron salts or other high iron compounds include ferrous sulfate, ferrous fumarate, ferrous citrate, ferrous gluconate, ferric ammoniums citrate, ferrous citrate and EDTA iron.
  • the nutritional supplement can be formulated for human or animal use.
  • Condensed corn distillers soluble (Syrup), a major by-product, is generated from the dry grind of corn to produce ethanol.
  • the fermentation process converts corn starch to ethanol but leaves a major portion of non-starch nutrients, including iron from the corn, behind as stillage.
  • the stillage is then concentrated to Syrup and is used primarily for animal feed.
  • Syrup has now been found to contain viable nutrients for the production of fungal biomass, such as Aspergillus oryzae and Aspergillus niger.
  • iron content in corn is determined by geographic location and weather conditions. Consequently the iron residue in Syrup varies from corn-ethanol plant to plant. As a result, iron in fungal biomass produced from DS of different ethanol plants exhibited a range from 180 to 320 mg/kg.
  • iron rich product can be fed to livestock, and is especially suited to young swine (Kornievicz d, et. Al. 2007 Effect of dietary yeast enriched with Cu, Fe and Mn on digestibility of main nutrients and absorption of minerals by growing pigs. Am. J. Agril. Biol. Sci. 2(4), 267-275.
  • Ferric sulfate Fe 2 (SO 4 ) 3 .2H 2 O
  • Fe3+ Fe3+ of 25.69%
  • the dose of iron was calculated as a concentration of iron elements rather than the concentration of salt; to enable comparison with the iron content in the fungal biomass. All fungal samples were thoroughly washed with deionizer water prior to testing by the ICP-OES method.
  • the results indicated that the iron content in the fungal biomass increased dramatically as the Fe3+ increased from 1-200 mg/L. Also interesting is that ferric sulfate was not toxic to Aspergillus oryzae at the given doses, but increased fungal yield numerically.
  • FIG. 3 illustrates that Aspergillus oryzae could use both Fe3+ and Fe2+ inorganic salts and uptake both of them until the doses of iron reached over 1000 mg (Fe)/L. Both iron salts resulted in iron content in fungal biomass close to or over 20,000 mg/kg.
  • ferrous sulfate is more water soluble and often used as an iron supplement, it did not appear to be different from ferric sulphate results.
  • the mechanism of iron uptake by Aspergillus oryzae is capable of increasing iron content from 257 mg/kg to over 20,000 mg/kg; a 40 fold increase. When comparing these iron levels with the foods contained in Table 6, the fungal product is able to provide a very heavily enriched iron content product.
  • an iron supplement has high iron content, but more importantly that such iron can be absorbed by humans.
  • the iron enriched fungal product (Ao-Iron) was produced with the process illustrated in FIG. 2 using ingredients of corn, wheat bran and soybean hulls. The iron content in the final product ranged from 2-4% by the different combinations of the raw ingredients and fermentation optimization.
  • the Ao-iron was ground to powder and subjected to an in vitro digestion/Caco-2 Cell Culture Model (Cell Test) detailed by Glahn (Glahn, R. P. et. al. 1998. Caco-2 cell ferritin formation predicts nonradiolabeled food iron availability in an In Vitro digestion/Caco-2 cell culture model).
  • Caco-2 cells were obtained from the ATCC at passage 17, and used in experiments at passage 25-33. Cells were seeded at a density of 1.9 ⁇ 10 5 in collagen-treated 6-well plates. The cells were grown in Eagle's medium with 10% fetal calf serum and maintained at 37° C. in an environmental with 5% CO 2 . The medium was changed every 2 days. In vitro digestion of the iron enriched fungal was following the steps also detailed by Glahn. The Cell Test measures the ferritin, an iron containing protein, formation in the cell incubated with the digested iron product; therefore has been used as a predictor of iron availability in humans.
  • the basic steps of the Cell Test are enzymatic digestion (pepsin and pancreatin-bile) of the sample, culture on Caco-2 cell layer (from human large intestine) and measurement of ferritin (an iron containing protein) formation within the cell.
  • enzymatic digestion pepsin and pancreatin-bile
  • Caco-2 cell layer from human large intestine
  • ferritin an iron containing protein
  • the sample cannot be diluted like inorganic salts during the enzymatic digestion.
  • a modified sample preparation procedure has been developed to be used in the Cell Test for the high iron content of organic materials. It was noted that the weight of such Ao-iron sample be kept low in a given digestion volume, in this case, it was 22.5 ml. Table 7 shows that an over loading of the sample during digestion led to low iron solubility. Such low solubility was due to incomplete digestion. However, when sample weight is ultra low, it is likely that an error may occur during weighing owing to the sensitivity of the weighing balance. It was determined that 0.005-0.01 grams of sample was optimal for the digestion volume of 20-25 ml. A larger sample size can be used but the enzyme dose during the digestion must also be adjusted.
  • ferritin response of Ao-iron was compared to ferrous sulfate, the most widely used inorganic iron supplement.
  • ferrous sulfate being very absorbable by humans, it can also cause severe side effects in humans, especially in children. Because it is very reactive, ferrous sulfate added to food can cause food oxidation and as a result food develops off flavour and a short shelf life.
  • ferrous sulfate supplements have been suggested to be taken with food.
  • iron in Ao-iron is in an organic form that can be taken without food.
  • ferritin response of ferrous sulfate was comparable to ferrous sulfate with corn ( FIG. 5 ).
  • the effect of corn meal on the ferritin response was more dramatic to ferrous sulfate than to Ao-iron.
  • Ao-iron is tasteless and can be added to food directly, but ferrous sulfate is mostly taken in a pill or capsule form.
  • the ferritin response in Caco-2 Cell was 52% of the ferrous sulfate, but it was better than iron in beef and corn with 25% and less than 20% respectively.
  • Aspergillus oryzae ATCC 22787 was selected to grow in a media made of corn flour, soybean hulls and wheat bran.
  • the corn flour and wheat bran were treated with enzymes of amylases, gluco-amylases, phytase.
  • Initial mix of the total flour and bran in the media ranged from 10-25%.
  • Amylase was added at neutral pH around 6 at temperature of 105-107° C. via autoclave or jet cooking, and then held at 95° C. for 2 hours.
  • the next step is to apply gluco-amylase for saccharification.
  • phytase may or may not needed to treat the mixture of corn and wheat bran at the temperature and pH required by the specification of the enzyme manufacturer.
  • soy bean hulls are used, the preferred practice would be to apply protease to the soybean hull in water at the pH required by the enzyme manufacturer.
  • the protease with optimal pH at acid range is preferred.
  • a mixture of these ingredients can be formed and used for both fungal pre-culture and product media.
  • a filter made of double layers of nylon cloth was used to harvest the fungal biomass in the laboratory and product was dried in 60° C. in a oven for over night.
  • the composition of final iron enriched product fungal is shown in Table 8.
  • the Aspergillus oryzae strain used is a non mycotoxin producing microorganism.
  • the toxin tests by LC/MS/MS method by a commercial laboratory showed no detectable mycotoxins present (Table 9).
  • inorganic minerals such as selenium and zinc have been used to produce selenium or zinc enriched yeast, but the use of filamentous fungi has not been reported. Both forms of enrichment provide better functional properties than the inorganic equivalents.
  • a similar process ( FIG. 2 ) was used for growing Aspergillus oryzae fortified with either sodium selenite or zinc sulfate. The dose of either inorganic salt was calculated as a concentration of the elements rather than the concentration of salt to enable comparison with the mineral content in the fungal biomass. Increases in the selected mineral contents in the fungal product are listed in Table 10 and demonstrate that Aspergillus oryzae can be used to produce organic selenium or zinc enriched products. However, as the dose of Se increased to 15 mg/L, the fungal yield significantly reduced. Zinc, however, increased fungal yield and over 67% of the added zinc element was converted to organic zinc.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Mycology (AREA)
  • Nutrition Science (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Epidemiology (AREA)
  • Zoology (AREA)
  • Botany (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Organic Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Inorganic Chemistry (AREA)
  • Virology (AREA)
  • Biomedical Technology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medical Informatics (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
US14/425,493 2012-09-11 2013-09-10 Nutritional supplement containing iron Abandoned US20150250839A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US14/425,493 US20150250839A1 (en) 2012-09-11 2013-09-10 Nutritional supplement containing iron

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201261699623P 2012-09-11 2012-09-11
PCT/AU2013/001028 WO2014040122A1 (en) 2012-09-11 2013-09-10 Nutritional supplement containing iron
US14/425,493 US20150250839A1 (en) 2012-09-11 2013-09-10 Nutritional supplement containing iron

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/AU2013/001028 A-371-Of-International WO2014040122A1 (en) 2012-09-11 2013-09-10 Nutritional supplement containing iron

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US16/557,293 Division US11478519B2 (en) 2012-09-11 2019-08-30 Nutritional supplement containing iron

Publications (1)

Publication Number Publication Date
US20150250839A1 true US20150250839A1 (en) 2015-09-10

Family

ID=50277406

Family Applications (2)

Application Number Title Priority Date Filing Date
US14/425,493 Abandoned US20150250839A1 (en) 2012-09-11 2013-09-10 Nutritional supplement containing iron
US16/557,293 Active 2033-09-16 US11478519B2 (en) 2012-09-11 2019-08-30 Nutritional supplement containing iron

Family Applications After (1)

Application Number Title Priority Date Filing Date
US16/557,293 Active 2033-09-16 US11478519B2 (en) 2012-09-11 2019-08-30 Nutritional supplement containing iron

Country Status (6)

Country Link
US (2) US20150250839A1 (zh)
EP (1) EP2895179A4 (zh)
JP (1) JP6573551B2 (zh)
CN (1) CN104717968A (zh)
AU (1) AU2013315341B2 (zh)
WO (1) WO2014040122A1 (zh)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20180035704A1 (en) * 2015-02-26 2018-02-08 Cura Global Health (Bvi) Limited Mineral enriched natural supplements
CN109561722A (zh) * 2016-06-01 2019-04-02 库拉全球健康(英属维尔京群岛)有限公司 形成富含铁的营养产品的方法
CN112220046A (zh) * 2020-10-28 2021-01-15 青岛鹏洋生物工程有限公司 一种含硒化卡拉胶的复配营养强化剂及其制备方法

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20170360075A1 (en) * 2014-12-11 2017-12-21 The Penn State Research Foundation Medical food for the treatment of malaria and/or iron deficiency
JP6890960B2 (ja) * 2016-12-08 2021-06-18 太平洋セメント株式会社 脱窒反応促進材組成物
WO2020031141A1 (en) * 2018-08-10 2020-02-13 Cura Global Health (Bvi) Limited Method for treating iron deficiency

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060134227A1 (en) * 2004-12-22 2006-06-22 Bortz Jonathan D Compositions including iron
US20070190209A1 (en) * 2006-02-10 2007-08-16 Mannatech, Inc. All natural multivitamin and multimineral dietary supplement formulations for enhanced absorption and biological utilization
US20120208893A1 (en) * 2009-09-14 2012-08-16 Nestec S.A. Nutritional compositions for modulating inflammation including exogenous vitamin k2

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS53148587A (en) * 1977-05-31 1978-12-25 Sapporo Breweries Production serene containing microorganism fungus body
JPS5768783A (en) * 1981-07-10 1982-04-27 Sapporo Breweries Ltd Preparation of microbial cell
JP4673071B2 (ja) * 2005-01-21 2011-04-20 ヒガシマル醤油株式会社 鉄吸着性高分子物質及び鉄含有高分子物質ならびにこれらの製造方法
US8481295B2 (en) * 2007-06-20 2013-07-09 Johannes van Leeuwen Fungi cultivation on alcohol fermentation stillage for useful products and energy savings
KR20090066410A (ko) * 2007-12-20 2009-06-24 (주) 비엔씨바이오팜 바실러스 서브틸리스 ｂｃ1212 균주를 포함하는 사료첨가용 조성물
RU2422043C1 (ru) * 2010-03-04 2011-06-27 Общество с ограниченной ответственностью "Альбит" Селенсодержащий препарат для животноводства
CN102356829B (zh) * 2011-10-31 2013-02-06 湖南创新生物科技有限公司 生长肥育猪后期微生物发酵无抗生素饲料

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060134227A1 (en) * 2004-12-22 2006-06-22 Bortz Jonathan D Compositions including iron
US20070190209A1 (en) * 2006-02-10 2007-08-16 Mannatech, Inc. All natural multivitamin and multimineral dietary supplement formulations for enhanced absorption and biological utilization
US20120208893A1 (en) * 2009-09-14 2012-08-16 Nestec S.A. Nutritional compositions for modulating inflammation including exogenous vitamin k2

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
Paknikar et al, Bioremediation of Metalliferous Wastes and Products using InactivatedMicrobial Biomass, 2003, Indian Journal of Biotechnology, 2: 426-443 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20180035704A1 (en) * 2015-02-26 2018-02-08 Cura Global Health (Bvi) Limited Mineral enriched natural supplements
US10863762B2 (en) * 2015-02-26 2020-12-15 Cura Global Health (Bvi) Limited Mineral enriched natural supplements
CN109561722A (zh) * 2016-06-01 2019-04-02 库拉全球健康(英属维尔京群岛)有限公司 形成富含铁的营养产品的方法
EP3462915A4 (en) * 2016-06-01 2019-04-10 Cura Global Health (BVI) Limited PROCESS FOR FORMATION OF IRON-ENHANCED NUTRITION PRODUCTS
CN112220046A (zh) * 2020-10-28 2021-01-15 青岛鹏洋生物工程有限公司 一种含硒化卡拉胶的复配营养强化剂及其制备方法

Also Published As

Publication number Publication date
AU2013315341B2 (en) 2016-02-11
AU2013315341A1 (en) 2015-03-05
US20200030393A1 (en) 2020-01-30
JP2015532827A (ja) 2015-11-16
US11478519B2 (en) 2022-10-25
EP2895179A1 (en) 2015-07-22
EP2895179A4 (en) 2016-04-06
JP6573551B2 (ja) 2019-09-11
CN104717968A (zh) 2015-06-17
WO2014040122A1 (en) 2014-03-20

Similar Documents

Publication Publication Date Title
US11478519B2 (en) Nutritional supplement containing iron
Steinkraus Nutritional significance of fermented foods
Svanberg et al. Fermentation and nutrient availability
CA2977287C (en) Mineral enriched natural supplements
Thierry et al. Effect of pure culture fermentation on biochemical composition of Moringa oleifera Lam leaves powders
WO2020247363A1 (en) Nutritional compositions from brewers' spent grain and methods for making the same
Amadou Millet Based Functional Foods: Bio‐Chemical and Bio‐Functional Properties
Jankorazov et al. Chemical Composition Of Soy Plant And Improving Technology Of Oil Processing In Industry
Akinsola et al. Nutritional evaluation of maize-millet based complementary foods fortified with soybean.
CA3026029C (en) Process for forming iron enriched nutritional products
Winarno Fermented vegetable protein and related foods of Southeast Asia with special reference to Indonesia
Chude et al. Effect of bioprocess on nutritional quality and chemical properties of Bambara groundnut (Vigna Subterranean (L) Verdc.) Flour
Sriherwanto Studies on the solid state fermentation of cassava bagasse for animal feed
Raju et al. Pearl Millet–a Minor Millet with Many Benefits
Hoover The effect of fermentation and soybean and peanut supplementation on corn protein
Ottaway et al. 8 Fortification Involving

Legal Events

Date Code Title Description
AS Assignment

Owner name: DAKOTA STAR CAPITAL, LLC, IOWA

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:WICKING, J. BRUCE;BIAN, YILIN;REEL/FRAME:035825/0497

Effective date: 20150420

AS Assignment

Owner name: CURA GLOBAL HEALTH (BVI) LIMITED, VIRGIN ISLANDS,

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:DAKOTA STAR CAPITAL, LLC;REEL/FRAME:043685/0192

Effective date: 20151231

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: FINAL REJECTION MAILED

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION