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
  • C12N1/08—Reducing the nucleic acid content
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
  • A23J3/00—Working-up of proteins for foodstuffs
  • A23J3/22—Working-up of proteins for foodstuffs by texturising
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
  • A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
  • A23J1/008—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from microorganisms
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
  • A23J3/00—Working-up of proteins for foodstuffs
  • A23J3/22—Working-up of proteins for foodstuffs by texturising
  • A23J3/225—Texturised simulated foods with high protein content
  • A23J3/227—Meat-like textured foods
• • 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
  • A23L13/00—Meat products; Meat meal; Preparation or treatment thereof
  • A23L13/40—Meat products; Meat meal; Preparation or treatment thereof containing additives
  • A23L13/42—Additives other than enzymes or microorganisms in meat products or meat meals
  • A23L13/424—Addition of non-meat animal protein material, e.g. blood, egg, dairy products, fish; Proteins from microorganisms, yeasts or fungi
• • 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
  • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
  • A23L29/065—Microorganisms
• • 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
• • 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
  • C12N1/145—Fungal isolates
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
  • C12R2001/00—Microorganisms ; Processes using microorganisms
  • C12R2001/645—Fungi ; Processes using fungi
  • C12R2001/77—Fusarium

Definitions

• the invention relates to edible fungus and particularly, although not exclusively, relates to a process for reducing the ribonucleic acid (RNA) content of an edible fungus and edible fungi with reduced RNA content per se.
• RNA ribonucleic acid
• Mycoprotein or fungal protein, is used in many food products as a meat substitute.
• Mycoprotein is often derived from the fungus Fusarium venenatum (formerly classified as Fusarium graminearum) as described in W095/23843.
• F.venenatum is grown by continuous or batch aerobic fermentation in a sterilised fermenter in a water and glucose solution with a continuous feed of nutrients.
• the rapidly growing fungal cells produce nucleic acids, such as RNA. If consumed in food products there is a concern that RNA may be broken down into purines and metabolised to uric acid which may lead to the development of gout and kidney stones.
• RNAase ribonuclease
• W095/23843 discloses growing Fusarium Graminaerum at 28 in the presence of a growth medium in a continuous fermenter, with continuous aeration with sterile air containing ammonia gas. Then, to remove RNA, the culture is passed into a continuous stirred tank reactor and steam is injected into the culture to heat shock the culture and increase its temperature to an especially preferred temperature of above 72°C. As a consequence, RNA passes from the fungus to the growth medium and treated fungus, with reduced RNA, can be isolated downstream.
• a method of reducing the level of RNA in a biomass comprising filamentous fungus comprising the following steps: (i) heating said biomass, downstream of a fermenter in which filamentous fungus is produced, to a first temperature;
• step (ii) separating said biomass comprising filamentous fungus from other components, for example from other components in a mixture produced in step (i) or downstream thereof which suitably contains said filamentous fungus having a reduced level of RNA compared to the level in the biomass heated in step (i).
• the first heating of said biomass comprising filamentous fungus downstream of said fermenter suitably comprises heating said biomass to said first temperature as described.
• the method preferably does not involve any heating (e.g. active heating) of said biomass downstream of said fermenter prior to heating of it to said first temperature.
• Heating said biomass in step (i) preferably uses a first heating device.
• the first heating of said biomass comprising filamentous fungus downstream of said fermenter suitably comprises heating said biomass to said first temperature using said first heating device.
• no other heating device is positioned between an outlet of said fermenter (via which said biomass of filamentous fungus passes downstream) and said first heating device.
• the first heating of said biomass in step (i) preferably does not involve direct contact of said biomass with any heated fluid; the first heating preferably does not involve direct contact of said biomass with steam.
• said biomass may be heated to increase its temperature by at least 20°C, for example by at least 25°C.
• said biomass may be heated to increase its temperature by at least 30°C or at least 33 °C.
• the temperature may be increased by less than 50 °C, for example by less than 45 °C.
• said biomass may take at least 2 minutes, for example at least 2.5 minutes to reach said first temperature, after contact with said first heating device.
• Said biomass may reach said first temperature in less than 20 minutes, preferably in less than 12 minutes, after contact with said first heating device.
• Said first temperature may be at least 40 °C, preferably at least 50 °C, more preferably at least 55 °C, especially at least 60 °C.
• Said first temperature may be less than 80 °C, preferably less than 75 °C, more preferably less than 71 °C.
• Step (i) may be undertaken in a receptacle (A), for example a pre-heat vessel, which suitably has an inlet positioned downstream of said fermenter, there being a conduit (A) positioned between said fermenter and said inlet for transfer of biomass from said fermenter to said receptacle (A).
• Heating in step (i) suitably involves contact of the biomass with a heat source which has a maximum temperature which is less than 100 S C, less than 95°C or less than 91 °C. Heating may involve contact of biomass with a heat source which has a temperature of at least 60 °C, preferably at least 76 °C.
• Heating in step (i) suitably involves subjecting biomass to a temperature which is no greater than 99°C, preferably no greater than 95 °C, especially no greater than 91 °C. Heating may involve subjecting biomass to a temperature of at least 55 °C, preferably at least 75 °C.
• step (i) comprises heating said biomass using a solid body which suitably contacts said biomass.
• Said solid body is preferably arranged to conduct heat to said biomass.
• a heat exchanger is suitably associated with said receptacle (A) for transferring heat to said biomass.
• Step (i) of said method preferably does not involve contact of said biomass with a fluid, for example steam, for heating the biomass to said first temperature.
• step (i) said biomass is preferably heated, at least in part, by a material, for example fluid, which is generated in said process, for example downstream of step (i) and/or downstream of receptacle (A).
• a material for example fluid
• heat is conducted from said fluid to said biomass; and suitably the fluid does not directly contact and/or is not mixed with said biomass.
• said biomass may be heated by a material which is produced by and/or subsequent to heating of the biomass to said first temperature in step (i).
• said material, for example fluid, which is generated in said process is introduced into a heat exchanger which is suitably associated with receptacle (A) as described.
• no additive e.g.
• the method suitably includes a step (i) * between step (i) and step (ii), wherein step (i) * comprises subsequently heating the biomass to a second temperature greater than the first temperature to facilitate release of RNA from cells of said filamentous fungus.
• said biomass may be heated to increase its temperature by at least 2 °C, preferably at least 4°C, more preferably at least 6°C. It may be heated to increase its temperature by less than 20 °C, preferably less than ⁇ 5 °C, more preferably less than 12°C.
• Said second temperature may be at least 60°C, preferably at least 62°C, more preferably at least 64°C. It may be less than 68°C, for example less than 67°C.
• Heating of the biomass in step (i) * may involve contact (e.g. direct contact) of a heated fluid, for example steam with the biomass.
• a heated fluid for example steam with the biomass.
• heating in step (i) * suitably takes place downstream of said receptacle (A). It may take place in a conduit (B) which is downstream of and communicates with receptacle (A).
• Conduit (B) may be arranged to deliver biomass to a RNA removal receptacle.
• Heating of biomass in step (i) * preferably utilises steam at greater than 100 for example at greater than 120 or greater than 140°C at a pressure of greater than 3barg or greater than 5barg.
• said biomass may be held at said second temperature (e.g. within the range 60 to 67 ⁇ €, for example 64 to 66°C), for at least 10, at least 20 or, preferably at least 30 minutes. It may be maintained at said temperature for less than 2 hours for example less than 1 hour.
• said second temperature e.g. within the range 60 to 67 ⁇ €, for example 64 to 66°C
• RNA RNA from cells of the filamentous fungus.
• a receptacle e.g. a or said RNA removal receptacle
• the only heating of the biomass involves heating to said first temperature.
• said biomass is not contacted with any heated fluid; preferably it is not contacted with steam.
• said biomass may be heated (e.g. by steam) downstream of receptacle (X), for example for hygienic reasons, although this is not preferred.
• the only heating of said biomass prior to step (ii) is heating in step (i) as described, to said first temperature.
• the first temperature is at least 60 °C, for example in the range 60-70 ; and/or in step (i), the biomass may be heated to increase its temperature by at least 30 °C and suitably 30-50 °C.
• said biomass may be contacted with a heated fluid, for example steam, for example for hygienic reasons.
• the heated fluid may be used to increase the temperature of the biomass.
• said biomass may be treated to remove fluid (e.g. primarily water), thereby to produce a biomass which includes a lower level of fluid (e.g. water).
• said biomass may include less than 40wt%, preferably less than 30wt%, for example less than 25wt% water.
• Step (ii) may comprise centrifuging said biomass.
• Step (ii) may comprise isolating dewatered biomass.
• the fluid (e.g. comprising water) removed in step (ii) may be relatively hot.
• it may have a temperature of greater than 50°C, greater than 60°C or greater than 70°C.
• the temperature of the fluid (e.g. comprising water) may be less than 90°C.
• the fluid (e.g. comprising water) removed is used to heat the biomass in step (i). It is suitably arranged to heat the biomass to said first temperature.
• no heat source other than fluid (e.g. comprising water) removed in step (ii) is used to heat said biomass to said first temperature.
• fluid removed in step (ii) may be heated prior to the fluid being used to heat the biomass in step (i).
• said heating of said fluid removed in step (ii) may involve contact (e.g. direct contact) of a heated fluid, for example steam, with said fluid removed in step (ii).
• the fluid removed in step (ii) may be heated to a temperature of at least 70 °C, for example at least 80 °C. It may be heated to a temperature of less than 95 °C.
• fluid (e.g. comprising water) removed in step (ii) may be a component of said first heating device.
• fluid (e.g. comprising water) removed in step (ii) is preferably directed into said heat exchanger for transferring heat to said biomass. Downstream of said heat exchanger, fluid (e.g. comprising water) removed in step (ii) may be discarded as waste.
• Said biomass may, after separation in step (ii), include less than 82 wt%, for example less than 80 wt% water; and suitably includes at least 18 wt%, preferably at least 20 wt% of mycoprotein (on a dry matter basis).
• said biomass e.g. dewatered biomass
• said biomass may include less than 2 wt% of
• Said filamentous fungus preferably comprises fungal mycelia and suitably at least 80 wt%, preferably at least 90 wt%, more preferably at least 95 wt% and, especially, at least 99 wt% of the fungal particles in said formulation comprise fungal mycelia.
• Some filamentous fungi may include both fungal mycelia and fruiting bodies.
• Said fungal particles preferably comprise a filamentous fungus of a type which does not produce fruiting bodies.
• Said filamentous fungus preferably comprises fungus selected from fungi imperfecti.
• filamentous fungus comprise, and preferably consist essentially of, cells of Fusarium species, especially of Fusarium venenatum A3/5 (formerly classified as Fusarium graminearum) (IMI 145425; ATCC PTA-2684 deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, VA.).
• Fusarium species especially of Fusarium venenatum A3/5 (formerly classified as Fusarium graminearum) (IMI 145425; ATCC PTA-2684 deposited with the American Type Culture Collection, 10801 University Boulevard, Manassas, VA.).
• Said filamentous fungus in said biomass heated in step (i) may comprise filaments having lengths of less than 1000 ⁇ , preferably less than 800 ⁇ . Said filaments may have a length greater than 100 ⁇ , preferably greater than 200 ⁇ . Preferably, fewer than 5wt%, preferably substantially no, filaments in said biomass have lengths of greater than 5000 ⁇ ; and preferably fewer than 5wt%, preferably substantially no filaments have lengths of greater than 2500 ⁇ . Preferably, values for the number average of the lengths of said filamentous fungus in said formulation are also as stated above.
• Said filamentous fungus in said biomass heated in step (i) may comprise filaments having diameters of less than 20 ⁇ , preferably less than 10 ⁇ , more preferably 5 ⁇ or less. Said filaments may have diameters greater than 1 ⁇ , preferably greater than 2 ⁇ . Preferably, values for the number average of said diameters of said fungal particles in said formulation are also as stated above.
• Said filamentous fungus in said biomass heated in step (i) may comprise filaments having an aspect ratio (length/diameter) of less than 1000, preferably less than 750, more preferably less than 500, especially of 250 or less. The aspect ratio may be greater than 10, preferably greater than 40, more preferably greater than 70.
• values for the average aspect ratio of said filamentous fungus i.e. the average of the lengths of the particles divided by the average of the diameters of the said filamentous fungus in said formulation) are also as stated above.
• Said filamentous fungus may be grown in said fermenter. It may be grown in a continuous or batch aerobic fermentation. Fermentation may utilise water and glucose and other nutrients.
• step (i) of the method heating of said biomass is suitably undertaken with said filamentous fungus in the presence of its growth medium. Immediately prior to said heating, the filamentous fungus is preferably in a viable state.
• the pH of the biomass is not adjusted by addition of any pH adjusting material, for example acidic or alkaline material.
• the pH of the biomass is not adjusted by addition of any pH adjusting material, for example acidic or alkaline material
• the pH of the biomass is not adjusted by addition of any pH adjusting material, for example acidic or alkaline material.
• the pH of the biomass is not adjusted by addition of any pH adjusting material, for example acidic or alkaline material.
• steam/water per se is not regarded as a pH adjusting material.
• Said method is preferably carried out using an apparatus comprising:
• step (b) a said first heating device for heating said biomass in step (i);
• a said receptacle (A) for example a pre-heat vessel, there being a said conduit (A) between said fermenter and said receptacle (A) for transferring fluid from said fermenter to receptacle (A);
• step (g) a conduit for circulating centrate removed from biomass (e.g. by centrifugation) to said first heating device, for example a heat-exchanger, for heating said biomass in step (i).
• said first heating device for example a heat-exchanger
• the biomass separated and/or isolated in the method of the first aspect is different compared to biomass produced in a comparative process.
• the size of filamentous fungus in said biomass is greater than that produced in a comparative process.
• use of the method leads to an improved yield of biomass compared to the yield produced in a comparative process.
• the invention extends, in a second aspect, to a product obtained by the process of the first aspect.
• a biomass produced in said method of said first aspect there is suitably provided a biomass produced in said method of said first aspect.
• Said biomass suitably has any feature of the biomass of the first aspect.
• Said biomass may include filamentous fungus with filament lengths which are, on average, longer than may be produced in processes different from the comparative process of the first aspect and/or as discussed herein.
• a biomass per se comprising a filamentous fungus, wherein said biomass is isolated from a mixture comprising said biomass and a liquid, wherein said liquid comprises particles of filamentous fungus, wherein particles in said liquid have one or more of the following characteristics (measured by laser diffraction as herein described): - a median size of less than 1 .3 ⁇ , for example less than 1 .1 ⁇ ;
• Particles in said liquid preferably may include at least two, preferably at least four, more preferably each of the characteristics described.
• the median size may be at least 0.5 ⁇ .
• the mean size may be at least 0.5 ⁇ . ⁇ mode size may be at least 0.5 ⁇ .
• the D (v, 0.1 ) size may be at least 0.4 ⁇ .
• the D (v, 0.5) size may be at least 0.4 ⁇ .
• the D (v, 0.9) size may be at least 0.5 ⁇ .
• the median, mean and/or mode particle sizes of filaments of said filamentous fungus in said biomass is greater than the median, mean and/or mode particle sizes in said liquid.
• the total weight of RNA in said liquid is greater than the total weight of RNA in said biomass.
• the total weight of filamentous fungus in said biomass divided by the total weight of filamentous fungus in said liquid is suitably greater than 2.3.
• Said biomass may include at least 20wt% water; it may include 30wt% water or less.
• said biomass includes at least 5kg, for example at least 50kg of filamentous fungus.
• Said biomass, said mixture and said liquid may independently have any features described in any of the preceding aspects.
• a foodstuff for human consumption comprising:
• Said foodstuff is suitably a meat-substitute. It may be in the form of, for example, mince, burger, sausage or meat-like pieces or strips. Said other ingredients may comprise any meat-free food ingredients. Said ingredients may include one or more meat-free fillers, flavours, oils, fats, proteins or vegetables.
• Said method may comprise packaging the foodstuff, for example in a substantially airtight and/or fluid tight package.
• At least 5kg, for example at least 100kg of said foodstuff may be made.
• the method of the fourth aspect is believed to be novel.
• the invention extends, in a fifth aspect, to a foodstuff obtained by the method of the fourth aspect.
• a foodstuff comprising a biomass as described in the second or third aspects and other ingredients, suitably being as described in the fourth aspect.
• Said foodstuff may include at least 1 wt%, for example at least 5wt% of other ingredients.
• Said foodstuff may include at least 10wt% water.
• Figure 1 is a schematic diagram showing a currently used process for producing mycoprotein paste with reduced RNA levels by direct steam injection;
• Figure 2 is a schematic diagram showing the steps involved in producing mycoprotein paste with reduced RNA levels using an alternative process
• Figure 3 is a graph comparing yields of mycoprotein obtained using the processes of Figures 1 and 2 over a period of days;
• Figure 4 is a particle size analysis graph for waste centrate produced in the process of Figure 1 ;
• Figure 5 is a particle size analysis graph for waste centrate produced in the process of Figure 2; and Figure 6 is a schematic diagram showing steps involved in producing mycoprotein paste with reduced RNA levels using a further alternative process.
• Mycoprotein paste - refers to a visco-elastic material comprising a mass of edible filamentous fungus derived from Fusarium venenatum A3/5 (formerly classified as Fusarium graminearum Schwabe) (IMI 145425; ATCC PTA-2684 deposited with the American type Culture Collection, 12301 Parklawn Drive, Rockville Md. 20852). It typically comprises about 23-25wt % solids (the balance being water) made up of non-viable RNA reduced fungal hyphae of approximately 400-750 ⁇ length, 3-5 ⁇ in diameter and a branching frequency of 2-3 tips per hyphal length.
• particle size analysis is undertaken by laser diffraction, for example using a Horiba LA950WET particle size analyser.
• an existing current, commercially-used process 100 for producing a mycoprotein paste involves growing a fungal culture in a pressure cycle fermenter 1 10 at 27°C in the presence of a growth medium.
• the culture broth passes from the fermenter 1 10 through a conduit 1 1 1 into an RNA reduction vessel 120.
• Steam (at 7barg and 160 S C) is injected into the culture broth via a steam injection port 1 12 in the conduit 1 1 1 .
• Steam injection raises the temperature of the culture broth to 60-70 °C.
• Steam injection is performed to reduce the RNA content of the final mycoprotein paste 140.
• the RNA reduction vessel 120 is a continuously stirred tank reactor.
• the culture broth is held in the RNA reduction vessel 120 at the RNA reduction temperature for at least 30 minutes.
• the culture broth then passes from the RNA reduction vessel 120 to centrifuges 130 via a conduit 121 .
• Steam is injected into the culture broth via a steam injection port 122 in the conduit 121 . This injection of steam increases the temperature of the culture broth to 80-90 °C for hygienic purposes.
• the centrifuges 130 are run at 5000g for a period of time.
• the centrifuges 130 separate the mycoprotein paste 140 and waste liquid centrate.
• the mycoprotein paste leaves the centrifuges 130 via conduit 131 .
• the waste liquid centrate contains RNA and digestion products of RNA that have passed out of the fungal cells into the surrounding aqueous media.
• the waste liquid centrate which at this stage has a temperature of 80-90°C, passes through conduit 132 to a cooler 1 50 in which it is cooled to 30°C. It then travels through conduit 151 to an effluent treatment plant (ETP) 1 60 for disposal.
• ETP effluent treatment plant
• the final mycoprotein paste 140 has a nucleic acid content of less than 2% on a dry weight basis.
• An alternative process for reducing the level of RNA is shown in Figure 2. Referring to the figure, a fungal culture is grown in a pressure cycle fermenter 210 at 27°C in the presence of a growth medium.
• the culture broth passes through a conduit 21 1 from the fermenter 210 to a pre-heat vessel 220.
• the culture is pre-heated to 55-60 (and maintained at the temperature for about 3 to 8 minutes) by waste liquid centrate which is produced further downstream in the process 200, as described below.
• the waste liquid centrate is passed through a heat exchanger (not shown) associated with vessel 220; the waste liquid centrate does not contact the culture directly.
• the pre-heated culture broth then passes along a conduit 221 to an RNA reduction vessel 230.
• steam at 7barg and 160 °
• Steam injection raises the temperature of the culture broth to 64-66 °C.
• the culture broth is held at this temperature in the vessel 230 for at least 30 minutes.
• the culture broth then passes from the RNA reduction vessel 230 to centrifuges 240 via a conduit 231 .
• steam (at 7barg and 160°) is injected into the culture broth via a steam injection port 232 in the conduit 231 . This injection of steam increases the temperature of the culture broth to 80-90 °C for hygienic purposes.
• the centrifuges 240 are run at approximately 5000g and are arranged to separate the mycoprotein paste 250 and waste liquid centrate. Downstream of the centrifuges 240, the mycoprotein paste 250 passes through conduit 241 and is isolated. The waste liquid centrate, which has a temperature of 80-90 °C, passes through conduit 242 to a heat exchanger associated with the pre-heat vessel 220. Thus, the centrate is used to heat the culture broth in the pre-heat vessel 220. After heating the pre-heat vessel via the heat exchanger, the waste liquid centrate, which at this stage has a temperature of 40-50 , passes through conduit 243 to a cooler 260.
• the centrate, containing waste RNA and waste RNA digestion products, is cooled to 30°C and travels through conduit 261 to an effluent treatment plant (ETP) 270 for disposal.
• ETP effluent treatment plant
• Example 1 Assessment of yields
• the yields of mycoprotein paste 140 and 250 achieved using the Figure 1 and Figure 2 processes respectively were assessed.
• the respective pastes 140 and 250 were dried after isolation by heating to cause evaporation of water.
• the % solid mycoprotein recovered was calculated by comparing the weight of mycoprotein recovered at 140 and 250 to the weight exiting the fermenters 1 10 and 21 0 respectively. Results are presented graphically in Figure 3, wherein the % solids recovered is plotted on the y axis and the time of sample collection (in days) is plotted on the x axis.
• Example 1 (a) (comparative) illustrates the yield obtained using the process described with reference to Figure 1 ; and example 1 (b) illustrates the yield obtained using the process described with reference to Figure 2.
• Figure 3 illustrates the increased yield achieved through use of the Figure 2 process
• Example 1 (b) compared to the Figure 1 process (Example 1 (a)).
• the increase in yield is about a 5% increase which is commercially very significant.
• the RNA content is less than 1 .7% RNA, on a dry weight basis.
• use of the Figure 2 process is able to reduce the level of RNA in the mycoprotein paste to an acceptable level and is found to produce a significant increase in yield.
• centrates produced by centrifugation in Figures 1 and 2 were isolated downstream of centrifuges 130 and 240 and particle size analysis of mycoprotein fragments in the centrate was undertaken.
• steam is not directly injected into the fungal culture broth, at any stage, but only into recirculated waste centrate.
• a fungal culture is grown in a pressure cycle fermenter 310 at 27°C in the presence of a growth medium.
• the culture broth passes through a conduit 31 1 from the fermenter 310 to a pre-heat vessel 320.
• the culture is pre-heated to 60-70°C by waste liquid centrate which passes through conduit 345.
• the waste liquid centrate is produced further downstream in the process 300, and heats the culture via a heat exchanger (not shown), as described in the Figure 2 embodiment.
• the pre-heated culture broth passes along a conduit 321 to RNA reduction vessel 330.
• the culture broth is held at 60-70 in the RNA reduction vessel 330 for at least 30 minutes.
• the culture broth then passes from the RNA reduction vessel 330 to centrifuges 340 via a conduit 331 .
• the temperature of the culture broth in conduit 331 remains at about 60-70°C.
• the centrifuges 340 are run at approximately 5000g. Heat is applied directly to the mycoprotein paste 350 in the centrifuges 340 to maintain hygienic conditions.
• the centrifuges 340 separate the mycoprotein paste 350 and waste liquid centrate.
• the mycoprotein paste 350 passes through conduit 342.
• the waste liquid centrate passes into conduit 343 from centrifuges 340. Steam is injected into the centrate in conduit 343 via steam injection port 344.
• the heated centrate passes through conduit 345 to the pre-heat vessel 320.
• the centrate in conduit 345 has a temperature of 80-90°C.
• the waste liquid centrate is used to indirectly heat the culture broth in the pre-heat vessel 320. Thereafter, the waste liquid centrate passes through conduit 346 to a cooler 360.
• the waste liquid centrate in conduit 361 has a temperature of 40-50 and then travels to an effluent treatment plant (ETP) 370 for disposal.
• ETP effluent treatment plant

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• 2016
  • 2016-06-27 GB GB1611143.7A patent/GB2557886A/en not_active Withdrawn
• 2017
  • 2017-06-16 WO PCT/GB2017/051761 patent/WO2018002581A1/en active Application Filing
  • 2017-06-26 TW TW106121266A patent/TW201809261A/zh unknown

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