Process for producing products with amino acid and taste retention

Abstract

A process for the preparation of umami taste components in a fermentation product includes producing glutamate, N-acetyl glutamine and respective salts thereof. The process produces at least glutamate.

Description

PROCESS FOR PRODUCING PRODUCTSWITH AMINO ACID AND TASTE RETENTIONField of the Invention

[0001] The field relates to processes for producing products in which an end product retaining amino acids such as glutamate and certain taste flavors is achieved.Background

[0002] U.S. Patent No. 3,087,763 discloses a process for generating glutamic acid.

[0003] WO2021 -045968 discloses a “process for preparing a purified human milk oligosaccharide ("HMO") from an HMO-containing solution (e.g., a fermentation broth) by a process comprising removal of an antifoam agent, and a product of such a process.” However, such purification processes remove antifoam either before or after removal of cell biomass and are intended for production of a purified product.

[0004] EP86112599A discloses a method for removing antifoaming agents during processing of microbial fermentations.

[0005] There remains a need for a process that leads to retention of certain amino acids and taste flavors in a final product.Summary of the Invention

[0006] In one example, a process for the preparation of umami taste components in a fermentation product comprises at least glutamate, the process comprising: preparing a seed medium comprising: a fruit juice or a fruit juice concentrate; a first set of inorganic salts of iron, magnesium and phosphates; inoculating the seed medium using an inoculum comprising a microorganism of a species from the genus of Corynebacterium to produce an inoculated seed medium; preparing a main fermentation medium comprising: a carbohydrate being sucrose, glucose, hydrolyzed starch or maltodextrin at a concentration from 0% to 25% of the main fermentation medium; a second fruit juice or a second fruit concentrate; and a second set of inorganic salts of iron, magnesium and phosphates; and a fruit pomace; mixing the inoculated seed medium with the main fermentation medium;performing fermentation in order to yield a fermentation product, wherein an antifoaming agent for controlling foaming is added (1 ) in a step prior to performing fermentation; or (2) in a step performing fermentation, or (3) in both of the foregoing steps; removing cellular biomass and the antifoaming agent from the fermentation product by filtration; removing brown color and residual amounts of the antifoaming agent in the fermentation product by using activated carbon as a percentage of a dry solid weight content of the fermentation product on a weight by weight basis; and drying the fermentation product. In one example, Corynebacterium glutamicum ATCC 15990 is the microorganism. In another example, Corynebacterium glutamicum ATCC 13032 is the microorganism. In one example, the food carrier is maltodextrin. In another example, the food carrier is gum acacia.

[0007] In another example, a process for the preparation of umami taste components in a fermentation product comprising glutamate, aspartate, N-acetyl glutamine, GMP (guanosine monophosphate), IMP (inosine monophosphate), or respective salts thereof, comprises preparing a seed medium comprising: a first fruit juice or a first fruit juice concentrate having a concentration from 0.4 to 30% of the seed medium; a first set of inorganic salts of iron, magnesium and phosphates having a concentration from 0% to 15%; inoculating the seed medium using an inoculum comprising a microorganism of a species from the genus of Corynebacterium to produce an inoculated seed medium; preparing a main fermentation medium comprising: a carbohydrate being sucrose, glucose, hydrolyzed starch or hydrolyzed maltodextrin at a concentration from 0% to 25% of the main fermentation medium; a second fruit juice or a second fruit concentrate having a concentration from 0.4 % to 30% of the seed medium and a biotin concentration in a range of 1 to 1000 pg / kg of the second fruit juice or the second juice concentrate; a second set of inorganic salts of iron, magnesium and phosphates having a concentration from 0.2% to 15%; a fruit pomace having a concentration up to 4%; mixing the inoculated seed medium with the main fermentation medium; performing fermentation, to yield a fermentation product, wherein an antifoaming agent for controlling foaming is added (1 ) in a step prior to performing fermentation; or (2) in a step performing fermentation, or (3) bothof the foregoing steps; performing either (1 ) a step of removing cellular biomass and the antifoaming agent from the fermentation product by filtration or (2) removing the cellular biomass in the fermentation product by centrifugation and removing the antifoaming agent in the fermentation product by filtration; removing brown color and residual amounts of the antifoaming agent in the fermentation product by using activated carbon in an amount ranging from 1 -100% of a dry solid weight content of the fermentation product; and drying the fermentation product.

[0008] In a further example, activated carbon may be used in an amount ranging from 1 -30% of a dry solid weight content of the fermentation product. The process produces at least glutamate. The process can also produce aspartate, N-acetyl glutamine, GMP (guanosine monophosphate), IMP (inosine monophosphate), or respective salts thereof. The process can also produce combinations of glutamate, aspartate, N-acetyl glutamine, GMP (guanosine monophosphate), IMP (inosine monophosphate), and respective salts thereof.

[0009] In another example, a process for the preparation of umami taste components in a fermentation product comprising glutamate, aspartate and N-acetyl glutamine, GMP and IMP and respective salts thereof, comprises preparing a seed medium comprising: a fruit juice or a fruit juice concentrate; a first set of inorganic salts of iron, magnesium and phosphates; inoculating the seed medium using an inoculum comprising a microorganism of a species from the genus of Corynebacterium to produce an inoculated seed medium; preparing a main fermentation medium comprising: a carbohydrate being sucrose, glucose, hydrolyzed starch or maltodextrin at a concentration from 0% to 25% of the main fermentation medium; a second fruit juice or a second fruit concentrate; and a second set of inorganic salts of iron, magnesium and phosphates; and a fruit pomace; mixing the inoculated seed medium with the main fermentation medium; performing fermentation, to yield a fermentation product, wherein an antifoaming agent for controlling foaming is added (1 ) in a step prior to performing fermentation; or (2) in a step performing fermentation, or (3) both of the foregoing steps; removing cellular biomass and the antifoaming agent from the fermentation product by filtration; removing brown color and residual amounts of the antifoaming agent in thefermentation product by using activated carbon as a percentage of a dry solid weight content of the fermentation product; and drying the fermentation product.Brief Description of the Figures

[0010] The above objects and other advantages of the invention will become more readily apparent upon reading the following description and drawings, in which:

[0011] FIG. 1 shows an example of the claimed process, where a first filtration step is utilized after a fermentation step.

[0012] FIG. 2 shows an example of the claimed process, where a centrifugation step is utilized after a fermentation step.

[0013] Figure 3 shows turbidity of different antifoams at the concentration of 2.5 g / L in umami flavored fermentation liquid as a function of temperature.

[0014] Figure 4 is a graph of flux versus time for exemplary ceramic and polymer membranes.Detailed Description of the Invention

[0015] The examples provided in the detailed description are merely examples and should not be used to limit the scope of the claims in any claim construction or interpretation.Abbreviations and terms

[0016] In one non-limiting example, “semi-purified” includes a product from a fermentation where the biomass, antifoam, color and off-taste products have been removed but otherwise contains all raw materials and metabolites from the fermentation. The claimed process will yield fermentation products including semipurified products.

[0017] Terms. The term “AC” means “activated carbon.” The term “AF” means “antifoam.” The term “DS” means “dry solids.” For purposes of claim interpretation, the term “glutamate” as understood in the specification and in the priority application is intended to include glutamate anions in combination with any type of cations or free glutamic acid or both. The term “glutamate” thus includes salts thereof, such as ammonium glutamate. Free “glutamic acid” as mentioned in the priority application is included within the scope of “glutamate” as previously defined herein. The term “MF” indicates “microfiltration.” The term “product” is intended to include singular or plural.The term “umami components” and the term “umami taste components” are used interchangeable and have the same meaning.

[0018] The following describes an example of a general process as claimed.Example 1 -General Process for producing the claimed products Fermentation steps

[0019] Raw materials. Carrot juice was obtained from Austria Juice in concentration of 70 Brix and used. In addition, carrot juices from Florida Food Products, Van Rijsingen Ingredients and Dbhler in ranges from 34 to 70 °Bx were tested (i.e. approximately 34-70% (w / w) dry matter). Wikipedia states that “’degrees Brix’ is a measure of the dissolved solids in a liquid and is commonly used to measure dissolved sugar content of an aqueous solution. One degree Brix is 1 gram of sucrose in 100 grams of solution and represents the strength of the solution as percentage by mass.”

[0020] Carrot pomace was obtained from Ernteband Fruchtsaft GmbH. In addition, pomace from Dbhler was tested. These are exemplary examples of raw materials but other materials known in the art may be utilized.

[0021] Carrot Juice characteristics. It is important that the fruit juice in the main fermentation medium, described later in the specification, have the biotin characteristics as the following examples will show. In one example, the carrot juices had biotin levels from 20 to 250 pg / kg. In another example, the carrot juice had biotin level from 106 to 250 pg / kg. In yet another example, a carrot juice can have a biotin concentration ranging from 1 to 1000 pg / kg.

[0022] Other juices known in the art and sources of biotin may be used for the process as disclosed in the specification. In one example, a juice that has similar features as the mentioned carrot juices may be used in the described exemplary process.Production organism / strain

[0023] The production organism, Corynebacterium glutamicum ATCC 15990 was used (American Type Culture Collection Deposit Number 15990 as previously mentioned. The product sheet provided in the ATCC website on November 21 , 2024describes this strain with a strain designation as “NRRL B-2243” and as originally deposited as “Corynebacterium lilium.” See Product Sheet retrieved at https: / / www.atcc.org / products / 15990, November 21 , 2024. In another experiment, Corynebacterium glutamicum ATCC 13032 was used. This strain has the American Type Culture Collection Deposit Number 13032 as previously mentioned. The product sheet provided in the ATCC website on November 21 , 2024 describes this strain with a strain designation of “534 [NCIB 10025]” and as originally deposited as Micrococcus glutamicus, See Product sheet retrieved at https: / / www.atcc.org / products / 13032, November 21 , 2024. However, a person of ordinary skill in the art may use any other microorganism from the genus of Corynebacterium so long as it produces at least glutamate and can additionally produce one or more of the following: aspartate, N-acetyl glutamine, GMP, IMP or respective salts thereof. For example, a salt of N-acetyl glutamine can be an ammonium salt.Seed fermentation

[0024] Seed medium preparation. As shown in the following table, the seed medium was prepared where end concentration is defined by a given weight of component / kg of final seed medium preparation.Table 1

[0025] For the seed medium, the total salt concentration can range from as low as 0% to a high of 15% of the seed medium. After preparing the seed medium containingthe components of Table 1 , the seed medium was adjusted to pH 7.0+1-0.1 using 25% ammonium hydroxide (NH4OH).

[0026] The seed medium was autoclaved at 121 °C for 20 minutes.

[0027] After autoclaving, four kilograms of the seed medium was poured into a Wave bag from Wave Biotech in a sterile bench and inoculated with 1 .8 grams of vial containing the bacterial strain Corynebactehum glutamicum ATCC 15990 and then incubated at 30°C for 18 hours. Alternatively, shake flasks or fermenters could be used instead of the Wave bag.Main fermentation medium

[0028] The main fermentation medium, 220 kg, has the following components as shown in the following Table 2. The preparation of the main fermentation medium will be described as follows. The concentrations of the following components are placed in a fermenter prior to inoculation with the seed medium.

[0029] The main fermentation medium was prepared as follows, where the end concentration is defined by a given component in weight / kg of final seed medium preparation. For example, for 1 kg of the final seed medium, 130 g of sucrose is added in a weight-by-weight ratio. Unless otherwise mentioned, all parts and percentages are by weight.Table 2

[0030] In addition to a phosphate salts, suitable salts include, for example, KH2PO4, KH2PO4, Na2HPO4, NaH2PO4, or (NH4)2HPC>4. Phosphoric acid may also be used.

[0031] Alternatives to sucrose. In addition to sucrose, other alternative carbohydrates include glucose, hydrolyzed starch or hydrolyzed maltodextrin. In one alternative example, sucrose, glucose, hydrolyzed starch or hydrolyzed maltodextrin or combinations thereof can be added. One of more of the mentioned carbohydrates can be added in a range from 0 to 25% of the main fermentation medium. Since the process can be operated as a fed-batch system and the carrot juices contain carbohydrates, one or more of the mentioned carbohydrates can be added at a later point in the process even if no carbohydrate is added to the main fermentation medium initially.

[0032] For the main fermentation medium, an antifoaming agent is used. Exemplary antifoaming agents used for this process include Foam Blast® 882 from Dystar, Struktol J673 (Schill and Seilacher) and Pluronic PE6100 from BASF. Other alternatives known in the art may be used by a person of ordinary skill in the art.

[0033] As known in the art, Foam Blast 882® is a polypropylene glycol based nonionic antifoam. It’s a clear liquid with a cloud point of 19-21 °C. The antifoam is added on as-needed basis to prevent foaming during fermentation and broth transfer to the harvest tank. As shown in FIG. 3, turbidity is the highest at a cloud point of a given antifoam.

[0034] To avoid Maillard reactions, the sucrose was sterilized separately from the remaining components of the final seed medium, the carrot juice was sterilized separately from the remaining components of the final seed medium and the other components from Table 2 were sterilized in the fermenter.

[0035] The fermenter was sterilized at 125°C for 60 minutes with steam and then cooled to 30°C.

[0036] After cooling of the fermenter, sucrose and carrot juice were added. The following describes the preparation of carrot juice and sucrose prior to adding both of them into the fermenter.

[0037] Preparation of carrot juice. Carrot juice was diluted in water at room temperature, and filter-sterilized (0.22 pm cartridge filter) and added into the fermenter at a concentration of 34 g / kg.

[0038] Preparation of sucrose. The sucrose solution, 60% (w / w) was sterilized at 124-131 °C for 24-39 minutes. After sterilization, sucrose was added to the fermenter at a concentration of 130 g / kg.Main fermentation process conditions

[0039] The fermentation was started with 220 kg of the main fermentation medium prior to inoculation with the seed medium and a measurement of 224 kg of the resulting medium was noted after inoculation with the seed medium. An antifoaming agent can be added at a time when the seed medium is mixed with the main fermentation medium or alternatively can be added when fermentation is performed or in yet another example, an antifoaming agent be added for both, a time when the seed medium is mixed with the main fermentation medium and can be added when fermentation is performed. Since a step of incorporation of an antifoaming agent can be variable, Figures 1 and 2, for purposes of simplicity, omit the step of including an antifoaming agent. In view of the previous teachings, an antifoaming agent can be added at a step prior to step 102, concurrently at step 102, or both of the foregoing in Figure 1. Similarly, an antifoaming agent can be added at a step prior to step 202, concurrently at step 202, or both of the foregoing in Figure 2.

[0040] The headspace pressure of the fermenter was controlled at 7 PSI.

[0041] At the start of the fermentation process, the airflow started at 100 standard liters per minute (SLPM) and was increased to 150 SLMP after 9 hours, and thereafter kept at 150 SLPM. The pH was controlled at pH 7.0 using 19% NHs-water.

[0042] The pO2 electrode was calibrated to 100% at a headspace pressure of 7 PSI before the fermentation started. During the fermentation the pO2 level was controlled at least 30% by controlling the agitation.

[0043] The temperature of the process was 30°C.

[0044] After 17 hours, the feed of 60% sucrose was started and kept at 2.45 kg / h throughout the run.Harvest

[0045] At the end of fermentation, i.e., 45 hours, the feed was stopped, and the airflow was reduced to 100 SLPM.

[0046] Subsequently, the fermenter was sampled approximately every hour for the next five hours to follow the consumption of sucrose and glucose. After the first one hour, the airflow was further reduced to 50 SLPM (standard liters per minute).Recovery of the fermentation product

[0047] The recovery of the fermentation product to produce an umami taste base including glutamate was conducted immediately after the fermentation was stopped. The umami components of the product included glutamate, aspartate and N-acetyl glutamine. The umami products also included GMP and IMP.Particle removal in the umami product by straining

[0048] The harvested fermentation broth containing the cellular biomass was first strained through an exemplary automatic mechanically cleaned strainer (model MCF 824 by Eaton) equipped with a 230-micron straining basket to remove the coarse pomace solids. Coarse pomace solids were removed to protect the downstream microfiltration membranes. For purposes of simplicity, Figure 1 omits the step of straining and straining occurs after fermentation step 102 but before a filtration step 104.

[0049] In an alternative step to straining, centrifugation can be used for removing particles such as coarse pomace solids. The step of centrifugation is shown as centrifugation step 204 in Figure 2.Cellular biomass and antifoam removal by filtration including microfiltration

[0050] After the straining step, a preferred step is to remove the cellular biomass and antifoam by a form of filtration such as microfiltration. Filtration is a preferred method as it can be used to remove both biomass and antifoam in one step when operating above the cloud point of the Anti-Foam. Centrifugation predominantly removes biomass.

[0051] Three different Micro Filtration elements were used to remove the biomass: (1) 0.1 -micron rated PES-based membrane from Koch Membrane systems. Model K603 3838 / 80, 3.5m2 area; (2) 0.1 -micron rated PVDF-based spiralwound elementfrom Solecta. Model PV1 3838 / 65. 4.2 m2 area; (3) 0.2-micron rated ceramic tubular membrane. Membralox model 1940. 0.72 m2 area.

[0052] Simultaneous biomass removal and antifoam removal. Microfiltration is a preferred method of filtration as it is used to remove both biomass and antifoam in one step when operating above the cloud point of the antifoam utilized. In Figure 1 , where microfiltration, a form of filtration is used, both biomass and antifoam are shown as removed in step 114 after a step of filtration 104.

[0053] Alternatively, or in combination with microfiltration, ultrafiltration may be used. Alternatively, or in combination with microfiltration, nanofiltration may be used. Alternatively, or in combination with microfiltration, rotary / vacuum drum filter or filter press with filter aid may be used. In yet another alternative, ultrafiltration and nanofiltration may be used.

[0054] Alternative to microfiltration. Alternatively, centrifugation can be used instead of microfiltration but centrifugation predominantly removes biomass. Subsequently microfiltration and activated carbon are used to remove the remaining fermentation solids and antifoam. Figure 2 shows another example of the general process where centrifugation step 204, after fermentation 202, is used to remove biomass. When centrifugation is used in step 204, biomass is removed as shown in step 214.

[0055] Antifoam cloud point. Cloud point of a particular antifoam in a particular process liquid is determined by the temperature where the antifoam has the lowest solubility in the liquid. Cloud point can be determined by measuring turbidity from such a process liquid as a function of temperature, for example. Cloud points of four (4) antifoaming agents in umami flavored fermentation liquid were determined.First, the original antifoam was removed from the process liquid by means of ultrafiltration and activated carbon treatment. Then each of the studied antifoams was added into the process liquid to match antifoam concentration of 2.5 g / L in the liquid. Turbidities as a function of temperature were measured from the process solutions with HACH 21 OOP Turbidimeter. Table 3 shows turbidity of different antifoams at the concentration of 2.5 g / L in umami flavored fermentation liquid as a function of temperature.Table 3

[0056] The same data is illustrated in Figure 3. The cloud point of each antifoam can be determined from the Figure 3 as the temperature with the highest turbidity. At this temperature, the antifoam has the lowest solubility in the measured sample matrix. Based on the data, Foam Blast® 882, Pluronic PE6100, Struktol J673 and Struktol J674 had cloud points approximately at 28°C, 32°C, 8°C and 25°C, respectively. By working at the temperature corresponding to the cloud point of the antifoam, the most efficient antifoam removal can be achieved in the means of conventional filtration technologies, such as microfiltration. Spiral wound and tubular MF elements were used.

[0057] Ultrafiltration and nanofiltration as alternatives to microfiltration. Alternatively, instead of microfiltration, simultaneous biomass and antifoam removal can be achieved with ultrafiltration or nanofiltration or both. Depending on the selected nanofiltration membrane and the antifoam in question, the antifoam removal can also be achieved by running the process below the cloud point of the antifoam. In yetanother alternative, microfiltration can be utilized followed by a step of ultrafiltration or a step of nanofiltration or both steps of ultrafiltration and nanofiltration.

[0058] Alternatively, depth filtration (filter presses and rotary vacuum drum filters) can be used as an alternative to membranes to remove biomass. For depth filtration filter aids (like diatomaceous earth, perlite) and flocculants may be used. The filter aid may be applied to as a precoat on the filtration media or be added as a body feed (also called admix), or both.

[0059] In one example, a filtration or centrifugation step is used to remove biomass. In an example of filtration, microfiltration, or ultrafiltration is used.

[0060] Optional diafiltration. The fermentation broth was microfiltered at 40-50°C using a 0.1 -micron PES membrane, mentioned above, first concentrating the cellular biomass five-fold.

[0061] Optionally, a diafiltration step using water was utilized to further increase the yield of glutamate and other umami-tasting components.

[0062] Alternatively, antifoam can be removed in a separate filtration step after biomass removal with for example, centrifugation, filter press or vacuum drum filter.

[0063] Concentrating and pH adjustment. After biomass removal, the described exemplary process stream typically has a dry solids content between 8-15% depending on the diafiltration factor. This dilute process stream can optionally be concentrated to higher dry solids content by e.g., evaporation or reverse osmosis. If reverse osmosis is used, it is an optional dewatering step of the MF permeate to increase dry solids of the evaporator feed to approximately 15% in case a low dry solid amount (8%, for example) is yielded. The concentrating can be done either before or after activated carbon treatment. Thus, in Figure 1 , a step of optional concentrating can be done before step 106 of utilizing activated carbon. Alternatively, a step of optional concentrating can be done after step 108, when the activated carbon has been removed. Similarly, in Figure 2, a step of optional concentrating can be done before step 206 of utilizing activated carbon. Alternatively, a step of optional concentrating can be done after step 208, when the activated carbon has been removed. In the described exemplary process, the process stream has been concentrated to dry solids content between 45-60%.

[0064] To minimize color and off-taste component generation in the process stream during the evaporation process, the boiling point of water can be lowered by running the evaporation process under vacuum. Exemplary evaporator types include wiped film evaporator, falling film evaporator, plate evaporator but are not limited to such evaporators.

[0065] Evaporation in the described exemplary process has been conducted under vacuum of 40-320 mbar.

[0066] pH criticality. Certain components, e.g. glutamate, in the described exemplary process stream can have a tendency to precipitate or crystallize out from the solution upon concentrating and possible change in the pH during the process. To minimize precipitation and yield loss, the pH is kept at the range where the umami components, e.g., glutamate, have the highest solubility. In the described exemplary process, the pH has been adjusted to between 6.4-7.2 after the concentration step. Even if concentrating is not utilized, pH adjustment, as readily known by a person of ordinary skill can be utilized to minimize precipitation and yield loss of certain components such as glutamate, for example. Figures 1 and 2, for simplicity, also omits the optional step of concentrating and a step of pH adjustment as it can vary depending on a specific process. In some cases, pH adjustment may not be needed.Color and antifoam removal

[0067] Use of activated carbon. Color, antifoam and other off-taste components can be removed from the described exemplary process stream with the use of activated carbon. Possible activated carbon types include, but are not restricted to, powdered activated carbon, granular activated carbon and immobilized activated carbon. The amount of activated carbon is based on the amount of dry solids in the fermentation product. For example, a 100 kg product with 5% dry solids will have 5 kg solids in the product. If the activated carbon dosage is e.g., 6% of the dry solids, we should add 100 kg x .05 x .06, which is 0.3 kg activated carbon. In the described exemplary process, activated carbon amounts ranging between 3% to 24% of the dry solids have been used.

[0068] Using activated carbon as a percentage of dry solid weight of product.The term “using carbon as percentage of a dry solid weight of a product” for purposesof claim interpretation can be interpreted exemplarily as follows. The following two examples are not limiting for purposes of claim interpretation. In a first example, activated carbon can be added as a percentage of a dry solid weight of a product on a weight by weight basis. In a second example, activated carbon can be used in the form of granular activated carbon and as mentioned later, a process stream was run through a column with granular activated carbon, where granular activated carbon was immobilized, for example, and was bound to a column. In this example, bound granular activated carbon is not added and is hence stationary. The amount of immobilized activated carbon is still a percentage of a dry solid weight of the product on a weight by weight basis. In the mentioned later example, 24% granular activated carbon (Chemviron CPG LF 12x40 from Calgon Carbon) was used. However, various ranges can be used similarly to the following examples for adding activated carbon. The following describes further examples of the addition of activated carbon. For example, activated carbon can be added in an amount ranging from 3% to 24% of the dry solid weight of a product. However, in another example, activated carbon can be added in an amount ranging from 1 % to 100% of the dry solid weight of a product. In still another example, activated carbon can be added in an amount being 400% of the dry solid weight of a product. In general, the selection of activated carbon as a percentage of a dry solid weight of a product is dependent on the knowledge of a person of ordinary skill in the art depending on considerations such as cost and other factors.

[0069] In one example, after the foregoing steps of microfiltration and an optional step of diafiltration, the resulting permeate was mixed with 6% powdered activated carbon (Nuchar ® SA-20 from Ingevity) based on dry solids in the permeate and kept at 50°C with continuous stirring for one hour.

[0070] Optionally, a step of concentration of the MF permeate can be performed before the activated carbon treatment. Concentration is a thickening step of the MF permeate prior to activated carbon treatment and drying such as spray drying. In one example of thickening, the resulting dilute microfiltration permeate was concentrated in the means of evaporation to dry solids of 52%. The concentrated process stream was pH adjusted to 7.2. The resulting process stream was mixed with 6 % powderedactivated carbon (Nuchar ® SA-20 from Ingevity) based on dry solids in the permeate and kept at 50°C for one hour.

[0071] Removal of powdered activated carbon in general. Powdered activated carbon was then removed from the process stream in the means of depth filtration by using a plate and frame filter. Other means of removal known to a person of ordinary skill in the art may be used.

[0072] In another example of concentrating, the resulting dilute microfiltration permeate was concentrated in the means of evaporation to dry solids of 58%. The concentrated process stream was pH adjusted to 6.9. The resulting process stream was run through a column with total of 24% granular activated carbon (Chemviron CPG LF 12x40 from Calgon Carbon) based on dry solids in the process stream.

[0073] Granular activated carbon treatment was conducted at temperature of 40°C with flowrate of 0.3 bed volumes per hour. Granular carbon does not need a separate removal step as it is bound to a column.

[0074] In another example of using activated carbon, powdered activated carbon (PAC) was used to remove color and antifoam from the MF permeate. A powdered activated carbon_Nuchar ® SA-20 from Ingevity was added to the MF permeate hold tank at a dosage of 9% on a dry solids basis of the MF permeate. The mixture of PAC and MF permeate was kept at 50°C for 60 min under mild agitation. The PAC was removed from the MF permeate by a 40x40 filter press using FW12 filter aid (admix of 1.5% on MF permeate weight basis) and K300 filter pads by Seitz. Powdered activated carbon was then removed from the process stream in the means of depth filtration by using plate and frame filter.

[0075] The following describes in detail the removal of powdered activated carbon mentioned above.Filtration to remove powdered activated carbon

[0076] Removal of powdered activated carbon. 6% activated carbon (SA-20) was used for one hour and then a filtration step was used to remove the carbon.

[0077] In one example of removing the activated carbon, polish filtration on a plate and frame filter with a precoat of diatomaceous earth was applied. In an alternative to filtration to remove the activated carbon, centrifugation can be utilized.

[0078] SA-20 is a powder activated carbon (PAC) manufactured by Ingevity 5255Virginia Ave. North Charleston, SC 29406. SA-20 is made from a chemically activated wood and has a high adsorptive capacity for many organics because of its high surface area.

[0079] In another example of removing the activated carbon, a plate-and-frame filter with filter sheets (Seitz T 2600 from Pall) was utilized to remove the powdered activated carbon from the process stream. After treating the process stream with powdered activated carbon, it was circulated through the plate and frame filter for half an hour to collect the powdered carbon on the filter sheets. After circulation, the carbon free process stream was collected through the filter.

[0080] Sterile filtration. Optionally, the product was then sterile filtered using 0.45 / 0.2 micron dual-layer pleated cartridges at 40-50°C temperature through a cartridge filter and filled into jerry cans. In one example, these pleated cartridges can be obtained from MDI.Carrier addition and Pasteurization step

[0081] After using filtration to remove the activated carbon, steps of (1 ) mixing with a food carrier and thickener such as maltodextrin or gum acacia and (2) pasteurization may be performed. For purposes of simplicity, Figures 1 and 2 omit the step of pasteurization. Pasteurization can be done concurrently with carrier addition step 110 or after carrier addition step 110 as shown in Figure 1. Pasteurization can be done concurrently with carrier addition step 210 or after carrier addition step 210 as shown in Figure 2. On a weight-to-weight basis, a carrier may be exemplarily added in an amount ranging from 45% to 67% based on the dry weight of a composition (e.g., dry powder) which includes umami components and which is recovered after a removal step of activated carbon. For example, if a carrier is 45% on a weight-by-weight basis, this means that the final dry powder has 45% carrier and 55% umami components. Other weight percentages as known to a person of ordinary skill are contemplated.Final step of drying or evaporation

[0082] A final step includes a step of drying, which can be accomplished by spray drying, as shown schematically in step 112 of Figure 1 and in step 212 of Figure 2. Other methods of drying known in the art, e.g., freeze drying can be utilized. In yet another example, vacuum drying can be utilized.

[0083] The results of the antifoam measurements in the following Table 4 demonstrated that removal of more than 99% of the antifoam was achieved. In Table 4, it is seen that there is a drop in dry matter concentration (DM%) and glutamate after the microfiltration. Without being bound by theory, the drop is believed to have been caused by the additional water added during an optional step of diafiltration. Additional water added during the diafiltration process further dilutes the process.

[0084] As seen from the results of Table 4, both microfiltration and the activated carbon treatment remove antifoam. The brown color and “brown taste” are removed by the activated carbon treatment.

[0085] In an alternative to a drying step, evaporation may be utilized.

[0086] The following table describes the results of dry matter, color, antifoam, glucose, and glutamate from fermentation through recovery.Table 4

[0087] “Strained broth” is defined as the harvested broth at the end of fermentation after it was passed through the inline strainer between the fermenter and the harvest tank.

[0088] The following is a description of the ICUMSA values stated above.

[0089] United States Patent No. 11 ,447,805, discloses the following:

[0090] An “ICUMSA” (International Commission for Uniform Methods of Sugar Analysis) value, or “standard ICUMSA” value, is an international unit for expressing the purity of a sugar sample in solution, and is directly related to the color of the dissolved sugar. The greater the ICUMSA value of a sugar sample, the darker the sugar sample is. Methods of determining ICUMSA values for sugar samples are well known in the art and are disclosed by the International Commission for Uniform Methods of Sugar Analysis in ICUMSA Methods of Sugar Analysis: Official and Tentative Methods Recommended by the International Commission for Uniform Methods of Sugar Analysis (ICUMSA) (Ed. H. C. S. de Whalley, Elsevier Pub. Co., 1964), for example, which is incorporated herein by reference. ICUMSA can be measured, for example, by ICUMSA Method GS1 / 3-7 as described by R. J. McCowage, R. M. Urquhart and M. L. Burge (Determination of the Solution Colour of Raw Sugars, Brown Sugars and Colored Syrups at pH 7.0 — Official, Verlag Dr Albert Bartens, 2011 revision), which is incorporated herein by reference.

[0091] The ‘805 patent also discloses that “ICUMSA values can be expressed in “reference base units” (RBU), for example.”

[0092] Thus, for the subject disclosure, a lowered ICUMSA value is indicative of lower “brown” color.

[0093] The ICUMSA color was reduced from approximately 10,000 (exact number: 10,358 in Table 4) to 1800 (1785 as seen in Sterile Filtered A22) by the activated carbon treatment.

[0094] The residual amount of glucose was low, but measurable. The amount of retrieved glucose was about 25 mg / L of glucose.

[0095] The following discusses various examples of the claimed process. The following Examples 2-9, 11 and related Tables 5-17 and 20 utilized Corynebacterium glutamicum ATCC 15990 as the production organism in theprocess. Example 10 and related tables 18-19 utilized Corynebacterium glutamicum ATCC 13032 as the production organism in the process.Other ExamplesExample 2- Combined antifoam and cell removal from umami flavor fermentation broth by means of ultrafiltration and nanofiltration

[0096] Umami flavor fermentation broth, containing the cellular biomass and Foam Blast® 882 as antifoam, was filtered with LabStak (Alfa Laval) flatsheet module.

[0097] Two membrane types were tested: Ultrafiltration membrane K-131 (Koch Membrane Systems) and nanofiltration membrane UA60 (TriSep). Both membranes were tested at process temperature of 9 °C. UA60 was also tested at process temperature of 50 °C. Antifoam was measured from all samples with Hach LCK 433 .

[0098] The properties of the umami flavor process stream after ultra- and nanofiltrations are shown in the table below. Below the cloud point temperature of Foam Blast® 882, the ultrafiltration membrane K-131 (Koch Filtration Systems) had very low if any retention for the antifoam. Instead, the nanofiltration membrane UA60 (TriSep) showed a high retention for the antifoam at both below and above the cloud point. UA60 also retained much more color at 9 °C compared to other tested alternatives.

[0099] At a lower temperature below the cloud point of the antifoaming agent, nanofiltration is needed. Alternatively, or in combination with nanofiltration, activated carbon below the cloud point of the antifoaming agent can be used.Table 5Example 3- Combined antifoam and cell removal from umami flavor fermentation broth by means of microfiltration

[0100] Umami flavor fermentation broth, containing the cellular biomass and Foam Blast® 882 as antifoam, was filtered with ceramic microfiltration module, Membralox T1 -70 (PALL). The membrane pore size given by the manufacturer was 0.2 pm. The microfiltration process was conducted at temperature of 50 °C. As a reference, a sample of whole broth was centrifuged to compare the antifoam removal efficiency to microfiltration .

[0101] Antifoam was measured from all samples with Hach LCK 433 assay kit. Properties of umami flavor fermentation broth before the processing, supernatant from centrifuged fermentation broth and microfiltration permeate are shown in the below table.Table 6

[0102] Microfiltration above the cloud point of Foam Blast® 882 antifoam decreased the amount of antifoam in the cell removed process stream to half as compared to using centrifugation. It is expected that part of the antifoam is adsorbed to cellular biomass and is also partly reduced in centrifugation above the cloud point of the antifoam.Example 4-Use of activated carbon: Taste improvement of microfiltered umami flavor ferment with different activated carbon grades

[0103] Microfiltration permeate from umami flavor ferment was used as a feed.Four different activated carbon grades were tested for taste improvement of the process stream. One powdered activated carbon: Nuchar SA-20 (Ingevity) and three granular activated carbon: Chemviron CPG LF 12x40 (Calgon Carbon), ColorsorbH620 (Jacobi) and Colorsorb H150-LF (Jacobi). The properties of the umami flavor microfiltration permeate are shown in the table below.Table 7

[0104] Use of activated carbon. Each carbon treatment was conducted in a sample bottle with continuous stirring at temperature of 50°C. The tested carbon dosages and contact times are shown in the table below.Table 8

[0105] Depth filtration. After each activated carbon treatment, the carbon was removed from the process solution by means of filtration that is Buchner filtration in this case. The aroma and taste of each sample was evaluated and scored. Thescoring was made on scale from 5 to 9, where 5 was considered as unacceptable, 6 and 7 were considered as marginal, and 8 and 9 were considered as acceptable flavor for an umami taste base product. Antifoam was measured with Hach LCK 333 assay. The properties of the corresponding microfiltered and carbon treated umami flavor ferments are shown in the table below.Table 9

[0106] As evidenced by different levels of scoring ranging from a low of 5 to a high of 9, different activated carbon grades showed a high difference in efficiency in removal of color and off-taste components from the umami flavor microfiltration permeate.

[0107] Nuchar SA-20 (Ingevity) with 6 % dose (of the feed total dry solids weight) with 1 -hour contact time and Chemviron CPG LF 12x40 with 24 % dose (of the feed total dry solids weight) with 3-hour contact time showed the best performance in terms of color and off-taste removal. The antifoam agent used was Foam Blast® 882. Other alternatives to Nuchar Sa-20 and Foam Blast® 882 can be used by a person of ordinary skill in the art.Example 5-Use of activated carbon: Antifoam and color removal from umami flavor ferment by treatment with powdered activated carbon

[0108] Microfiltration permeate from umami flavor ferment was used as a feed. Nuchar SA-20 (Ingevity) powdered activated carbon was used for antifoam and color reduction. The feed material was concentrated by means of evaporation at 100 mbar vacuum to dry solids content of 52%. The pH of the resulted concentrated process stream was adjusted from 6.1 to 7.2 with 30 wt.% potassium hydroxide solution.

[0109] The concentrated and pH adjusted process stream was divided into two fractions and treated at two different temperatures: 30°C and 50°C. Both fractions were heated up to their corresponding process temperatures in a jacketed reactorvessel with stirring. 6% (of the feed total dry solids weight) of powdered activated carbon was added into the reactor. The materials were incubated at their corresponding temperatures for about 1 hour. The materials were then circulated through an exemplary depth filter (Seitz plate and frame filter with Seitz T 2600 filter sheets) for about 0.5 hours. Then the materials were filtered once through the depth filter and collected.

[0110] Properties of umami flavor microfiltration permeate before and after concentration are shown in the table below. Antifoam Foam Blast® 882 concentration of the feed solution was measured with Hach LCK 443 assay and antifoam concentration of carbon treated materials were measured with Hach LCK 333 assay.Table 10

[0111] The properties of concentrated umami flavor microfiltration permeates after Nuchar SA-20 powdered activated carbon treatment are shown in the table below.Table 11- Activated Carbon treatment of microfiltration permeates

[0112] Color and antifoam removal was at the same level at both 30°C and 50°C process temperatures. 69-71 % of the original color was removed. Antifoam Foam Blast® 882 was removed below the detection limit of the assay. Some color may havebeen generated during the processing at 50°C, which is then shown as higher color after the treatment. Glutamate concentration was not affected by the carbon treatment. Other alternatives to Nuchar Sa-20 and Foam Blast® 882 can be used by a person of ordinary skill in the art.Example 6- Use of Activated Carbon: Antifoam and color removal from umami flavor ferment by treatment with granular activated carbon

[0113] Microfiltration permeate from umami flavor ferment was used as a feed. Chemviron CPG LF 12x40 (Calgon Carbon) granular activated carbon was used for antifoam Foam Blast® 882 and color reduction. The feed material was concentrated by means of evaporation at 20 mbar vacuum to dry solids content of 58 %. The pH of the resulted concentrated process stream was adjusted from 6.0 to 6.9 with 45 wt.% potassium hydroxide solution. Other alternatives to Chemviron CPG LF 12x40 (Calgon Carbon and Foam Blast® 882 can be used by a person of ordinary skill in the art.

[0114] 24% (of the feed total dry solids weight) of granular activated carbon was packed into jacketed glass column. The carbon was backwashed with ion-exchanged water for removal of fine carbon dust from the column. The column was then heated to process temperature of 40°C. The concentrated and pH adjusted process stream was also heated up to 40°C before the start of the column operation. The material was then run through the column with a flowrate of 0.3 bed volumes per hour. Properties of umami flavor microfiltration permeate are shown below. Antifoam concentration of the feed solution was measured with Hach LCK 443 assay and antifoam concentration of carbon treated materials were measured with Hach LCK 333 assay.Table 12

[0115] The properties of concentrated umami flavor microfiltration permeates after Chemviron CPG LF 12x40 granular activated carbon treatment are shown below.Table 13-Activated carbon treatment of microfiltration permeate

[0116] 70% of the color was removed in the granular activated carbon treatment. Antifoam was removed below the detection limit of the assay. Glutamate concentration was not affected by the carbon treatment.Example 7-Use of Microfiltration and Activated carbon: Biomass and antifoam removal from Corynebacterium glutamicum fermentation broth using Microfiltration Membranes, Powdered Activated Carbon and Depth Filtration. (500 L scale piloting)

[0117] After the fermentation was completed, the broth was pumped through a self-cleaning strainer (Eaton MCF 824, filter area 0.39 m2, 230 micron wedge-wire screen) strained to remove pomace particles. 4 different types of Micro Filtration (MF) membranes were evaluated to simultaneously remove cell mass and antifoam Foam Blast® 882, polypropylene glycol based nonionic antifoam). Table 14 lists the properties of the MF membranes tested.Table 14 Membrane Types (properties and run IDs of 500 L piloting)

[0118] Other alternatives to the membranes above and Foam Blast® 882 can be used by a person of ordinary skill in the art.

[0119] The operating temperature of the MF step was kept between 40-50°C, which is above the cloud point of the antifoam agent at a temperature of 20°C. Above its cloud point, the homogeneous aqueous solution of the nonionic antifoam separates into a dilute phase containing a low antifoam concentration and colloidal phase with a high antifoam concentration. The colloidal droplets will be rejected by the MF membrane. Some of the antifoam colloids could also attach to the biomass and be rejected by the MF membrane this way. Running at 50°C has the added benefit of reducing the risk of microbial contamination during the microfiltration. As shown in Figure 4, ceramic membranes have greater flux over run time and polymeric membranes have larger membrane surface area and hence have greater flow over similar period of run time.

[0120] The MF steps involved two steps: (1 ) a five-fold concentration step, and (2) a two times diafiltration step to recover (dilution washing) extra glutamate from the biomass. The MF permeate of these two steps is collected in a tank as the composite MF permeate.Table 15: Fluxes, Biomass and Antifoam removal from broth using different MF membranes** The antifoam levels were measure on a HACH DR1900 assay using the TNT875 / 876 assay kits for nonionic surfactants; “AF” is an abbreviation for antifoam. “Eff” is an abbreviation for “efficiency.” “EOF” is an abbreviation for “end of fermentation.”

[0121] As shown in Table 15, microfiltration above the cloud point of the antifoam Foam Blast® 882 used is effective in removing biomass and antifoam, while allowing glutamate to pass through. Other alternatives to the MF membranes and Foam Blast® 882 can be used by a person of ordinary skill in the art.Example 8-Use of Activated Carbon: Antifoam removal from MF permeate using Powdered Activated Carbon (PAC) followed by Depth Filtration

[0122] Composite MF permeate (21 RA182) was treated with Powdered Activated Carbon (SA-20) and sterile filtered. The MF permeate was mixed with 9% activated carbon on a dry-solids basis and kept at 50°C for 60 minutes. After that the AC (activated carbon) was removed by depth filtration using a Seitz 40cmx40cm filter press equipped with 2 end-plates and 3 frames, holding 8 Pall K300 filter pads. The filter pads were precoated with 2,400 grams of diatomaceous earth (FW-12) by pumping a DE slurry over the filter press. A body feedof 1 ,800 grams of FW-12 (1 .5% of MF permeate mass) was added to the mixture of AC and MF permeate mixture prior to polishing.

[0123] The results of the antifoam measurements (Foam Blast® 882) shown in Table 16 below indicate that removal of about 99% of the antifoam present at the end of fermentation was achieved.Table 16Example 9- Biomass and antifoam removal from fermentation broth using a combination of Microfiltration, Ultrafiltration, Activated Carbon and Depth Filtration

[0124] Fermentation broth from a 14L-pilot fermenter was strained using a 230- micron screen to remove coarse fermentation solids. The biomass of the strained broth was removed by microfiltration (MF). The MF permeate was then filtered through an ultra-filtration (UF) membrane to remove additional antifoam. The operating temperatures during the MF and UF filtration steps were 45°C.

[0125] The MF filtration and subsequent UF filtration were done on a Pellicon-2 benchtop membrane system (EMD Millipore). The MF membrane used was a WPP cassette (0.1 pm pore size, hydrophilic PVDF). The UF membrane used was a Biomax-5 cassette (5kD, modified polyethersulfone).

[0126] 1.5 wt% of Powdered Activated Carbon (SA-20) was added on a weight basis to the final UF permeate. The mixture of AC powder and UF filtrate was agitated at 50C for 60 min.

[0127] For the final UF permeate, the dry solids of the final UF permeate was 8.6 wt%. Thus, for example, for 100 g of UF permeate (liquid), the dry solids weight percentage was 8.6% or 8.6 g. If 1.5% of activated carbon is added to 100 g of UF Permeate (liquid), the amount of activated carbon added to the liquid UF permeate is 1.5 g.

[0128] In relation to the amount of activated carbon added to the amount of dry solids in the fermentation product, the activated carbon dosage was 17.4 wt% on a dry solids basis of the final UF permeate because the same amount of 1.5 g of activated carbon distributed over 8.6 grams of dry solids equals 17.4%.

[0129] The AC powder was removed with a pressure filtration holder having a filter diameter of 142 mm and a filtration area of 113 cm2 (Advantec 302300 304 SS Pressure Filtration Holder w / Reservoir, 142 mm) using a filter pad having an average particle retention range of 6-12 microns and a flow velocity of 850 liters per square meter per hour at a maximum differential pressure of 3 bar / 43.5 psi

[0130] K300 filter pad. The following filter aid was used: (1 ) The filter pad was precoated with 1.5% diatomaceous earth (FW-12), and 1.5wt% of diatomaceous earth (FW-12) was added as a body feed to the AC-treated MF permeate.

[0131] The glutamate concentration, total protein and residual antifoam concentration of the polished UF permeate are listed in Table 17 below.Table 17: Glutamate concentration, total protein and antifoam concentration of the polished UF permeate** Antifoam concentration of incoming broth was 1 ,425 mg / L.Example 10- Lab-scale fermentation with Corynebacterium glutamicum ATCC 13032

[0132] The following describes a performed lab-scale fermentation example using Corynebacterium glutamicum ATCC 13032.

[0133] A lab-scale fermentation was run with Corynebacterium glutamicum ATCC 13032 as production organism. The fermentation was carried out in 3 L Sartorius fermenters with 1 .5 L of the following medium.Seed medium

[0134] A seed medium, 100 mL, has the following components as shown in the following Table 18.Table 18

[0135] The seed medium was adjusted to pH 9 using 25% NH4OH and added to a 500 mL baffled shake flask and autoclaved (121 °C / 20 min).

[0136] The shake flask was inoculated with 0.1 mL of glycerol stock of Corynebacterium glutamicum ATCC 13032 and incubated at 30°C and 250 RPM for 16 hours.Main fermentation medium

[0137] The main fermentation medium, 1.5 kg, has the following components as shown in the following Table 19. The preparation of the main fermentation medium will be described as follows. The concentrations of the following components are placed in a fermenter prior to inoculation with the seed medium.

[0138] The main fermentation medium was prepared as follows, where the end concentration is defined by a given component in weight / kg of final seed medium preparation. For example, for 1 kg of the final seed medium, 130 g of sucrose is added in a weight-by-weight ratio. Unless otherwise mentioned, all parts and percentages are by weight.

[0139] The standard main fermentation medium eventually developed, 1.5 kg, included the following, as shown in Table 19.Table 19

[0140] All concentrations mentioned in the above table are the concentrations in the fermenter prior to inoculation.

[0141] The medium was prepared by adding all components except pomace, and sucrose to distilled water and mixing for 10 minutes.

[0142] The dissolved media components were added to the fermenters (Sartorius Biostat B, 3 L nominal volume) followed by pomace which was best handled dry.

[0143] The fermenter was autoclaved at 121 °C for 90 minutes (autoclave setting) which ensured that the medium was at 121 °C for 20 minutes.

[0144] After cooling to the fermentation temperature (30°C), the sucrose was added as a solution containing 650 g / kg sucrose which had previously been UHT sterilized (142°C / 30 sec).

[0145] Before inoculation, a sample was drawn for external pH control and possible re-calibration of the fermenter pH meter.

[0146] The pH was adjusted to 7.0 using 25% NHs-water. The aeration was set to 1 .5 SLPM (1 WM) and the agitation was set to 500 RPM. The pO2 electrode was calibrated to 100%.

[0147] The fermenter was inoculated with 60 mL of seed culture resulting from the seed medium and the fermentation was started.

[0148] The fermentation was controlled at pH 7.0 using 25% NHs-water. The pO2 was controlled at 30% by the agitation. The aeration was 1.5 SLPM throughout the fermentation.

[0149] Sucrose (650 g / kg) was added as a feed, starting at 15 hours of fermentation, before the sucrose from the batch medium was exhausted.

[0150] The feed rate of the 650 g / kg sucrose solution was kept constant at 9 g / kg / h based on the 0+ weight from 18 hours to the end of fermentation.

[0151] When a temperature ramp was used, the fermentation ran at 30°C from the start to 14 hours, whereupon the temperature was increased linearly to 38°C at 25 hours, and then kept constant at 38°C until the end of fermentation.

[0152] There was some tendency for foaming in the first 10 hours of fermentation, which was controlled by manually pumping in small doses of sterile antifoam. The antifoam used in this fermentation was Pluronic PE6100 from BASF.

[0153] Glutamate yield. The fermentation gave 33 g / L of glutamate after 47 hours of fermentation.Example 11 -Composition of products produced with Corynebacterium glutamicum ATCC 15990

[0154] For this example, umami components such as glutamate and N-acetyl- glutamine were produced as described by the general process of Example 1 which included the steps of fermentation and recovery as previously described.

[0155] The following table reports the recovery of umami components glutamate and N-acetyl-glutamine in a carrier-free composition recovered after the step of removal of activated carbon.

[0156] For purposes of simplification, the recovery results of the mentioned umami components in the table are before the addition of a carrier, which would affect the final amounts recovered. As carrier addition can be variable (e.g., 45 to 67% of a composition recovered after the step of removal of activated carbon), calculations of the umami components of the recovered composition would vary, when taking the umami components as a weight percentage of the combined amounts of the recovered composition and the carrier.

[0157] The following table reports the results of three runs of a carrier-free composition on a dry matter basis prior to spray-drying:Table 20

[0158] For each of the runs, a composition is given. Thus, for Run 1 , the composition glutamate of 55.6% would be 55.6 g of glutamate / 100 g of final product. Respectively, for run 1 , a 11 % of N-acetyl-glutamine would be 11 g of N-acetyl- glutamine per 100 g. The foregoing analyses also apply to runs 2 and 3.

[0159] When a carrier is subsequently used, a carrier, on a weight-to-weight basis, may be exemplarily added in an amount ranging from 45% to 67% based on the dry weight of a composition which includes umami components and which is recovered after a removal step of activated carbon. For the three runs mentioned, a carrier percentage used was 50%.

[0160] For this exemplary process, the vast majority of the cations for the glutamate was ammonium. This results indicates that the major product recovered was substantially and primarily ammonium glutamate.

[0161] The range of recited numerical values disclosed in the specification includes values, e.g., + / — 5-10% of the recited value, that a person of ordinary skill in the art would consider equivalent to the recited value, e.g., having the same function or result.

[0162] The claims are not limited by the preferred embodiments and examples but will cover many modifications and equivalents consistent with the written description as a whole.

Claims

What is claimed is:1 . A process for the preparation of umami taste components in a fermentation product comprising at least glutamate, the process comprising: preparing a seed medium comprising: a fruit juice or a fruit juice concentrate; a first set of inorganic salts of iron, magnesium and phosphates inoculating the seed medium using an inoculum comprising a microorganism of a species from the genus of Corynebacterium to produce an inoculated seed medium; preparing a main fermentation medium comprising: a carbohydrate being sucrose, glucose, hydrolyzed starch or maltodextrin at a concentration from 0% to 25% of the main fermentation medium; a second fruit juice or a second fruit concentrate; and a second set of inorganic salts of iron, magnesium and phosphates; and a fruit pomace; mixing the inoculated seed medium with the main fermentation medium; performing fermentation in order to yield a fermentation product, wherein an antifoaming agent for controlling foaming is added (1 ) in a step prior to performing fermentation; or (2) in a step performing fermentation, or (3) in both of the foregoing steps; removing cellular biomass and the antifoaming agent from the fermentation product by filtration; removing brown color and residual amounts of the antifoaming agent in the fermentation product by using-activated carbon as a percentage of a dry solid weight content of the fermentation product on a weight by weight basis; and drying the fermentation product.

2. The process of Claim 1 , wherein the fermentation product further comprises one or more of the following: N-acetyl glutamine or respective salts thereof.

3. The process of Claim 1 , wherein the seed medium comprises: the first fruit juice or a first fruit juice concentrate having a concentration from 0.4 to 30% of the seed medium and the first set of inorganic salts of iron, magnesium and phosphates having a concentration from 0% to 15%; and the second fruit juice or a second fruit concentrate having a concentration from 0.4 % to 30% of the seed medium and a biotin concentration in a range of 1 to 1000 pg / kg of the second fruit juice or the second juice concentrate; the second set of inorganic salts of iron, magnesium and phosphates having a concentration from 0.2% to 15%; the fruit pomace having a concentration up to 4%; wherein the percentage is 1 -100%, and the activated carbon is used in an amount ranging from the 1 -100% of the dry solid weight content of the fermentation product on the weight by weight basis.

4. The process of Claim 1 , wherein the step of inoculating the seed medium includes inoculating the seed medium with the inoculum at a 0.008% to 10% of the seed medium.

5. The process of Claim 1 , wherein the first and second fruit juice or the first and second fruit juice concentrate and the fruit juice pomace are carrot.

6. The process of Claim 1 , further comprising a step of removing particles by straining after the fermentation product is produced from the step of mixing the inoculated seed medium with the main fermentation medium.

7. The process of Claim 1 , further comprising a step of removing particles by centrifugation after the fermentation product is produced from the step of mixing the inoculated seed medium with the main fermentation medium.

8. The process of Claim 1 , wherein the step of removing cellular biomass and the antifoaming agent from the fermentation product by filtration utilizes microfiltration.

9. The process of Claim 1 , further comprising a step of diafiltration after the step of removing cellular biomass and the antifoaming agent from the fermentation product by filtration utilizing microfiltration is performed.

10. The process of Claim 1 , further comprising a step of removing the activated carbon by filtration after the step of removing brown color and residual amounts of the antifoaming agent in the fermentation product.11 .The process of Claim 1 , further comprising a step of removing the activated carbon by centrifugation after the step of removing brown color and residual amounts of the antifoaming agent in the fermentation product.

12. The process of Claim 1 , further comprising a step of mixing with a food carrier and a step of pasteurization.

13. The process of Claim 12, wherein the food carrier is maltodextrin.

14. The process of Claim 1 , wherein the step of removing cellular biomass and the antifoaming agent from the fermentation product by filtration uses nanofiltration.

15. The process of Claim 1 , wherein the step of removing cellular biomass and the antifoaming agent from the fermentation product by filtration uses microfiltration and the step of preparing a main fermentation medium includes providing the antifoaming agent, and the microfiltration is performed above the cloud point of the antifoaming agent.

16. The process of Claim 1 , wherein the step of preparing a main fermentation medium includes using the antifoam agent and the step of removing cellular biomass and the antifoaming agent from the fermentation product by filtration uses microfiltration.

17. The process of Claim 16, wherein the microfiltration is above a cloud point of the antifoaming agent and decreased the amount of foam.

18. The process of Claim 1 , wherein the step of removing cellular biomass and the antifoaming agent from the fermentation product by filtration uses ultrafiltration.

19. The process of Claim 1 , wherein the step of preparing a main fermentation includes providing sucrose as the carbohydrate.

20. The process of Claim 1 , wherein the step of inoculating the seed medium uses Corynebacterium glutamicum ATCC 15990 as the microorganism.

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