EP3570690A1 - Composition and method of production thereof - Google Patents
Composition and method of production thereofInfo
- Publication number
- EP3570690A1 EP3570690A1 EP18702173.8A EP18702173A EP3570690A1 EP 3570690 A1 EP3570690 A1 EP 3570690A1 EP 18702173 A EP18702173 A EP 18702173A EP 3570690 A1 EP3570690 A1 EP 3570690A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- food composition
- microbial biomass
- composition according
- protein
- dry cell
- 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.)
- Pending
Links
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Classifications
-
- 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
- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
- A23L33/12—Fatty acids or derivatives thereof
-
- 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
-
- 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
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
-
- 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
- A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
-
- 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
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
- C12P7/6434—Docosahexenoic acids [DHA]
Definitions
- the invention relates to improved food compositions comprising microbial biomass.
- the present invention also relates to a method for growing a microbial biomass.
- Omega-3 fatty acids are of significant commercial interest due to their recognition as important dietary compounds that can prevent arteriosclerosis and coronary heart disease, alleviate inflammatory conditions and retard the growth of tumour cells. They are also very important for brain health and cognitive development and it has been suggested they may reduce delinquency and mediate autism in specific cases. These fatty acids are not made by the fish de novo but are derived from their diet.
- EAAs essential amino acids
- Microbial based feeds offer advantages compared to terrestrial animal and plant based feeds due to continuous harvesting, low water and energy requirements, cost- effectiveness and protein rich constituency.
- Microorganisms can provide a natural, sustainable supply of feeds comprising proteins, carbohydrates, oils and vitamins. Some microorganisms comprise a rich source of natural omega-3 fatty acids and provide a better omega-3/omega-6 ratio than soy and corn based feeds and derived vegetable oils.
- Thraustochytrids are microorganisms of the order Thraustochytriales. Thraustochytrids include the genera Schizochytrium and Thraustochytrium which have been recognised as a potential source of omega-3 fatty acids as discussed in US 5130242.
- Omega-3 fatty acid producing microorganisms can be grown and the oils produced used in food formulations.
- exposure of fatty acids to air, for example during processing, extraction or product storage, can result in a loss of activity, decomposition and oxidation.
- Decomposition of fatty acids can produce high levels of free fatty acids (FFA) which are highly undesirable, and generally speaking, no more than 3% FFA content is permitted in foodstuffs and feedstocks.
- FFA free fatty acids
- contact with light and certain metals can also accelerate decomposition and oxidation of fatty acids.
- any decomposition products of the fatty acids can result in an undesirable taste and these products have the potential to be carcinogenic.
- Antioxidants are often added to omega-3 oils to slow down oxidation of the oil.
- antioxidants are expensive and can substantially increase the cost of the oil. If higher levels of FFAs are present in the oil, then the oil will need to be refined which is not only costly but also results in up to 10% of the oil being lost.
- oils may be protected by encapsulation prior to incorporation into food products as detailed in WO2007150047. However, encapsulation procedures can be complex and expensive.
- the present invention is based, in part, on studies undertaken by the inventors in which they show that surprisingly, it is possible to develop microbial biomass having particular contents of protein, oil and/or DHA which therefore make such biomass suitable for use in food compositions, particularly in feed formulations for fish, for example.
- a food composition comprising a microbial biomass, wherein the microbial biomass comprises:
- the microbial biomass of the present invention comprises 15-95 dry cell wt% protein.
- the microbial biomass comprises around 15-65 dry cell wt%, 25- 65 dry cell wt%, 30-65 dry cell wt%, 34-65 dry cell wt%, 45-65 dry cell wt% or 55-65 dry cell wt% protein.
- the microbial biomass comprises around 30-95 dry cell wt%, 50-95 dry cell wt%, or 65-95 dry cell wt% protein. Most preferably the microbial biomass comprises around 75-95dry cell wt% protein.
- the microbial biomass may comprise essential amino acids and non-essential amino acids.
- the microbial biomass of the present invention may comprise 0.5-75 dry cell wt% oil. Therefore, in such embodiments, the microbial biomass may comprise:
- the microbial biomass of the present invention may comprise 7-75 dry cell wt% oil. Therefore, in such embodiments, the microbial biomass may comprise:
- the microbial biomass comprises around 0.5-65 dry cell wt%, 0.5-55 dry cell wt% or 0.5-45 dry cell wt% oil. Most preferably, the microbial biomass comprises around 0.5-25 dry cell wt% oil.
- An oil content of less than around 25 dry cell wt% can reduce the likelihood of cells bursting and releasing oil during feed formulation processes, for example an extrusion process. The release of oil could cause problems with shear during extrusion.
- a low ratio of oil to protein in the microbial biomass provides a food composition which can meet all of a consumer's protein requirements. This is especially advantageous in the provision of, for instance, fish feed for farmed fish. Generally the diets of wild fish contain far higher amounts of protein than those of farmed fish. This is because of the economic pressure on fish feed producers to produce more cost effective and balanced diets thus resulting in feed with an increased proportion of oil to protein to provide energy and as well as essential lipids.
- the microbial biomass of the present invention can provide a cost-effective and complete source of protein in consumer's diets.
- the oil comprises around 95-97% fatty acids. Therefore, the present invention also provides a food composition comprising a microbial biomass, wherein the microbial biomass comprises:
- the present invention also provides a food composition comprising a microbial biomass, wherein the microbial biomass comprises:
- the microbial biomass comprises 25-55% DHA as a percentage of total fatty acids. More preferably, the microbial biomass comprises around 30-55% DHA as a percentage of total fatty acids, more preferably around 35-55% or 40-55% DHA as a percentage of total fatty acids.
- farmed fish require preformed DHA and the essential amino acids in their diets for each phase of production and critically during early developmental stages.
- the present inventors have found that it is possible to provide a microbial biomass with between 25-55% DHA as a percentage of total fatty acids, which makes the microbial biomass particularly suitable for use as a fish feed composition.
- the microbial biomass may comprise or additionally comprise around 6-1 1 % docosapentaenoic acid (DPA) as a percentage of total fatty acids. More preferably, the microbial biomass comprises around 8-10% DPA as a percentage of total fatty acids.
- the microbial biomass may comprise or additionally comprise around 15-55% 16:0 fatty acids as a percentage of total fatty acids. More preferably, the microbial biomass comprises around 25-50% 16:0 fatty acids, more preferably around 28-40% 16:0 fatty acids as a percentage of total fatty acids.
- the microbial biomass may comprise cultures of Thraustochytrids, Thraustochytrium, Aurantiochytrium or Schizochytrium.
- the microbial biomass comprises a culture of Schizochytrium mangrovei.
- the present inventors have surprisingly found that it is possible to produce food compositions in accordance with the first aspect of the invention in particularly high yields when the microbial biomass comprises a culture of Schizochytrium mangrovei. This results in a highly economic product which can be used throughout the life cycle of fish, for example.
- the microbial biomass is in the form of microbial whole cell biomass.
- the consumption of intact, whole microbial cells and their post-consumption digestion protects oils within microbial cells from oxidation.
- oxidised oils can be damaging to consumers.
- cell wall residues can have positive effects on gut morphology and overall function in many fish species.
- the microbial biomass may further comprise vitamins and minerals.
- the microbial biomass may comprise vitamin E a-tocopherol, vitamin D, vitamin A and vitamin K.
- the microbial biomass may further comprise iron, manganese, zinc, copper and selenium.
- the food composition may further comprise starch, vitamins, minerals and binding agents, such as soybean meal.
- the food composition may additionally comprise one or more essential amino acids selected from: methionine, arginine, threonine, tryptophan, histidine, isoleucine, lysine, leucine, valine and phenylalanine.
- the quantities of the one or more essential amino acids provided in the composition will be in accordance with usual animal or fish food compositions and will largely be dependent upon the desired application.
- the food composition may form part of the diet of fish, dogs, cats, chickens, other domestic animals and/or of humans.
- the food composition forms the whole diet for fish, dogs, cats, chickens, other domestic animals and/or humans.
- the food composition is for consumption by fish and in particular high value species such as salmon, trout, sea bass and seabreams.
- the food composition may be adapted for fish to meet their essential fatty acids requirements and provide enhanced fatty acid (Omega-3) for the consumer.
- the food composition is for consumption by sea bream, sea bass, salmonids, flounder, turbot, sole and rainbow trout at various production stages but especially from grow-out to harvest.
- the food composition may be employed for consumption by crustaceans such as penaid shrimp and to provide enhanced HUFA (highly unsaturated fatty acids) in the flesh for species like tilapia as an added value niche market for this important fish species.
- the oil and protein concentrations of the microbial biomass may be selected based on the age of the consumer. For example, rapidly growing fish and shellfish (juvenile to pre- harvest) require a higher protein to fat (oil) ratio in their diets typically 45-50% protein: 18- 20% oil. When fish such as salmon approach the end of their growth phase, they require relatively more fat (oil) (>25%) than protein (35%) in their diet. Suitably they also need a particular omega-3 fatty acid profile in their flesh for subsequent consumption.
- the food composition is adapted for juvenile fish and shellfish and comprises: a) 7-25 dry cell wt% oil; and
- the composition may preferably comprise: a) 45-50 dry cell wt% protein;
- the food composition is adapted for adult fish and shellfish and comprises: a) 15-25 dry cell wt% oil; and
- the composition may preferably comprise: a) around 25 dry cell wt% oil;
- the food composition of the present invention may be in any suitable form, for example pellets, micro-pellets, granules, crumbles, powders, microcapsules or flakes.
- the composition may also be in a liquid (such as a solution or suspension) or in a dried form.
- the food composition may be in the form of a pellet.
- the food composition may be extruded.
- a method for growing a microbial biomass wherein the microbial biomass comprises:
- nitrogen and carbon are fed to the microbial biomass during fermentation.
- Nitrogen and carbon feed rates can of course be adjusted to change microbial cell growth rate and microbial biomass oil and protein content.
- the present inventors have shown that increasing the feed rate of carbon (for example in the form of glucose or glycerol) during fermentation results in high cell growth rate and high oil content and conversely, increasing the feed rate of nitrogen (for example in the form of yeast extract, corn steep liquor (CSL) and/or (NhU ⁇ SCU) during fermentation favours cell growth and results in high yields of microbial biomass and a high protein content.
- carbon for example in the form of glucose or glycerol
- nitrogen for example in the form of yeast extract, corn steep liquor (CSL) and/or (NhU ⁇ SCU
- the mass ratio of total carbon to nitrogen sources is less than around 5.
- the mass ratio of total carbon to nitrogen sources is between around 3 and 5, preferably around 4.
- the mass ratio of total carbon to nitrogen sources may be in the range of around 1 .8 to 4.2, around 1 .9 to 4.2, around 2.2 to 4.2, around 2.4 to 4.2, around 2.7 to 4.2, around 2.9 to 4.2, around 3.2 to 4.2, around 3.4 to 4.2, around 3.5 to 4.3 or around 3.8 to 4.2.
- the mass ratio of total carbon to nitrogen sources may be in the range of around 1 .8 to 4.2, around 1 .8 to 3.8, around 1 .8 to 3.5, around 1.8 to 3.4, around 1 .8 to 3.2, around 1.8 to 2.9, around 1 .8 to 2.7, around 1 .8 to 2.4, around 1 .8 to 2.4, around 1.8 to 2.2, or around 1 .8 to 1 .9. Most preferably, the mass ratio of total carbon to nitrogen sources is between around 4.1 and 4.2.
- the present inventors have surprisingly shown that by having a mass ratio of total carbon to nitrogen sources as described above, microbial biomass with high protein and high levels of DHA can be produced, making such biomass highly suited for use in feed compositions, particularly fish feed compositions.
- the method of the present invention may be grown in aerobic fermentation conditions.
- the oil to protein ratio is in the region of around 0.02 to 1 .24.
- the oil to protein ratio may be in the region of around 0.03 to 1 .24, around 0.04 to 1 .24, around 0.06 to 1 .24, around 0.08 to 1 .24, around 0.1 1 to 1 .24, around 0.14 to 1 .24 or around 0.61 to 1 .24.
- the oil to protein ratio may be in the region of around 0.02 to 0.61 , around 0.02 to 0.14, around 0.02 to 0.1 1 , around 0.02 to 0.08, around 0.02 to 0.06, around 0.02 to 0.04 or around 0.02 to 1 .03.
- the oil to protein ratio is in the region of around 0.02 to 0.61.
- the set-point of the dissolved oxygen concentration in the fermentation medium is controlled between around 20% and 30% of the saturation concentration.
- the microbial biomass grown by the method of the present invention is in accordance with the microbial biomass of the first aspect of the present invention.
- Figure shows the mass ratio of total carbon to nitrogen correlating with the oil to protein ratio of the product biomass produced in accordance with the invention.
- Microbial cells believed to be Schizochytrium mangrovei, were initially grown on a culture plate before being transferred to a shaker flask and fermentation tank. Twelve trials were undertaken, each using different fermentation conditions, as shown in table 1 .
- Trial 1 was a batch fermentation reaction, whereas trials 2-12 were fed-batch fermentations. The components of the additional feed used in trials 2-12 are shown in table 2.
- the mass ratio of total carbon to nitrogen sources used in the twelve trials is shown 4.
- the present inventors have surprisingly shown that high protein contents can be achieved in the resultant microbial biomass.
- the present inventors have surprisingly shown that as shown in Figure 1 a mass ratio of total carbon to nitrogen sources correlates with the oil to protein ratio of the product biomass achieved.
- the microbial biomass produced in the twelve trials was spray dried and the protein, oil, carbohydrate and moisture content determined.
- Carbohydrate content was determined by Dubois, M., Gilles, K.A., Hamilton, J.K., Rebers, P.A., Smith, F. (1956) Colorimetric method for determination of sugars and related substances Anal. Chem. 28, 350-356. The results (along with analytical methods used) are shown in table 5.
- the fatty acid composition (% total fatty acids) of total lipid from the spray dried biomass are shown in table 6.
- DHA, DPA and 16:0 are highlighted in the table.
- the limit of quantification for fatty acid analysis is 0.06%.
- the relatively high proportions of DHA in the biomass demonstrate the suitability of microbial biomass to be used in a food composition which can supply health improving omega-3 fatty acids to consumers.
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US5130242A (en) | 1988-09-07 | 1992-07-14 | Phycotech, Inc. | Process for the heterotrophic production of microbial products with high concentrations of omega-3 highly unsaturated fatty acids |
US5407957A (en) | 1990-02-13 | 1995-04-18 | Martek Corporation | Production of docosahexaenoic acid by dinoflagellates |
ES2668640T5 (en) | 2006-06-22 | 2022-05-12 | Dsm Ip Assets Bv | Compositions of encapsulated labile compounds and methods for preparing the same |
US7989195B2 (en) * | 2008-02-20 | 2011-08-02 | Washington State University Research Foundation | Heterotrophic algal high cell density production method and system |
FR3001736B1 (en) * | 2013-02-06 | 2016-03-04 | Roquette Freres | BIOMASS OF MICROALGUE SCHIZOCHYTRIUM MANGROVEI AND METHOD FOR PREPARING THE SAME |
US11122817B2 (en) | 2014-07-25 | 2021-09-21 | Smallfood Inc. | Protein rich food ingredient from biomass and methods of production |
FR3031985B1 (en) | 2015-01-26 | 2017-02-17 | Roquette Freres | PROCESS FOR OBTAINING PEPTIDE ISOLATE FROM BIOMASS OF MICROALGUES ENRICHED IN PROTEINS |
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