Method for preparing seasoning material by mixed fermentation of heterogeneous microbes

Abstract

The present invention relates to a method for preparing a seasoning material using a process of mixing two or more species of microbes that produce products different from each other and fermenting the microbes at the same time. In the method for preparing a seasoning material, heterogeneous microbes that produce different products, that is, different amino acids, nucleic acids, and / or organic acids, are mixed and fermented, thereby enabling the preparation of the seasoning material that increases the taste and flavor of foods through a fermented solution containing the amino acids, nucleic acids, and / or organic acids and which improves overall sensory properties of foods. Such a seasoning material can find applications in the various food fields.

Description

[00192] Sensory evaluation was performed on the glutamic acid-arginine fermented powders obtained through individual 10 fermentation in Preparation Examples 4 and 5 and the glutamic acid-arginine fermented powder obtained through mixed fermentation in Preparation Example 6.

[00193] Sensory evaluation was performed in the same manner as in Example 2-3. The results are shown in Table 10 below. 15

[00194] [Table 10] Item Preparation Example 4 (individual fermentation) Preparation Example 5 (individual fermentation) Preparation Example 6 (mixed fermentation) Initial Umami taste ++ +++ ++++ Umami + ++ ++++ persistence Salty taste + ++ Sour taste + + Bitter taste ++ ++ Sweet taste

[00195] Referring to Table 10 above, due to the difference in components of the glutamic acid-arginine fermented powder as shown in Table 9 above, the sample prepared by the mixing process after individual fermentation showed a weak umami taste and an increased bitter taste compared to the sample prepared by mixed fermentation. In addition, since the increase in by-products such as organic acids and the increase in ions as shown in Table 9 above also affect the sensory properties of the sample, the mixed fermentation process is more effective in terms of taste or process simplification than mixing after individual fermentation.

[00196] Example 5. Mixed fermentation of glutamic acid and inosinic acid

[00197] 5-1. Seed culture

[00198] Corynebacterium glutamicum NFG6 (KCCM13164P) was used as a glutamic acid-producing microorganism, and Corynebacterium ammoniagenes NFI545 (KCCM13162P) that produces IMP was used as an inosinic acid-producing microorganism.

[00199] A glutamic acid seed culture broth of the glutamic acid-producing microorganism was prepared in the same manner as in Example 1-1.

[00200] For seed culture of the inosinic acid-producing microorganism, the inosinic acid-producing microorganism 5 was inoculated into a 2-L flask containing 0.3 L of a seed culture medium, followed by primary culture for 20 to 24 hours at 31°C and 150 rpm (OD610 = 15 to 20). Next, 1% of the primary culture was inoculated into a 5-L jar fermenter, and 2 to 2.5 L of a seed culture medium was added thereto, 10 followed by secondary culture at 31°C, pH 7.1, 600 rpm and an aeration rate of 1.0 vvm for 21 to 24 hours (OD610 = 20 to 40), thereby preparing an inosinic acid seed culture broth. The composition of the seed culture medium used is shown in Table 11 below. 15

[00201] [Table 11] Composition Seed culture medium for inosinic acidproducing microorganism 4 to 6% glucose, 2 to 4% yeast extract paste, 0.3% (NHJ2SO4, 0.2% KH2PO4, 0.2% K2HPO4, 200 to 300 ppm adenine, 200 to 300 ppm guanine, 0.15% MgSO4^7H2O, 10 ppm nicotinic acid, 100 ppm Ca-pantothenate, 15 ppm cysteine, 1 ppm thiamine HCl, 5 ppm ZnSO4^7H2O, 10 ppm MnSO4^5H2O, 0.1 ppm biotin, 15 ppm FeSO4^7H2O, and 0.01% antifoaming agent for food

[00202] 5-2. Main fermentation

[00203] In order to examine the ratio of glutamic acid to inosinic acid in the fermentation broth depending on the inoculums of the glutamic acid seed culture broth and the inosinic acid seed culture broth, the glutamic acid seed 5 culture broth and the inosinic acid seed culture broth were inoculated at various ratios and fermented.

[00204] In main fermentation, 14 to 18 L of a fermentation medium was added to a 50-L fermenter and inoculated with 1.2 to 1.8 L of the total seed culture broth at an 10 inoculation ratio of 0.05 to 99.95 (glutamic acid seed culture broth): 99.95 to 0.05 (inosinic acid seed culture broth), followed by mixed fermentation by fed-batch culture for 30 to 90 hours. The fermentation medium composition and fermentation conditions used here are shown in Table 12 15 below.

[00205] [Table 12] Fermentation medium composition 1 to 3% molasses, 5 to 7% raw sugar, 2 to 3% yeast extract paste, 0.6 to 1.2% H3PO4, 0.05 to 0.1% betaine, 100 to 200 ppm adenine, 50 to 150 ppm guanine, 0.2 to 0.5% MgSO4^7H2O, 50 to 100 ppm Ca-pantothenate, 5 to 15 ppm vitamin B3, 5 to 20 ppm thiamine HCl, 0.4 to 0.8% NaOH, 5 to 10 ppm FeSO4, 10 to 20 ppm MnSO4, 10 to 20 ppm ZnSO4, and 0.005% antifoaming agent for food Fermentation Temperature 31 to 32°C, pH 6.5 to 7.5, aeration conditions rate 0.8 to 1.2 vvm, internal pressure 0.6 to 1.0 kg / cm3,  agitation speed 320 to 350 rpm, and dissolved oxygen (DO) concentration 20 to 70%

[00206] This mixed fermentation was performed a total of three times, the average value was calculated, and the results are shown in Table 13 below.

[00207] [Table 13] Inoculation ratio between seed culture broths (GA: IMP) Fermentation time GA: IMP ratio in fermentation broth 99.95 : 0.05 33 hours 99.8 : 1 80 : 20 42 hours 51.3 : 1 70 : 30 48 hours 31.2 : 1 50 : 50 60 hours 16.8 : 1 20 : 80 83 hours 1.04 : 1 0.05 : 99.95 90 hours 0.02 : 1 5

[00208] Referring to Table 13 above, it was confirmed that, when the seed culture broths of the microorganisms were inoculated at a ratio of 0.05 to 99.95 (glutamic acidproducing microorganism): 99.95 to 0.05 (inosinic acidproducing microorganism), L-glutamic acid and IMP in the 10 fermentation broth were produced at a ratio of 0.02 to 99.8: 1.

[00209] Example 6. Comparison of glutamic acid-inosinic acid fermented powders between fermentation processes

[00210] 6-1. Production of glutamic acid-inosinic acid fermented powder

[00211] Conventionally, a flavor containing glutamic acid and inosinic acid was produced by individually fermenting a glutamic acid-producing microorganism and an inosinic acidproducing microorganism and then mixing the fermentation broths or dried products thereof at a suitable ratio. To compare the differences in taste between this conventional individual fermentation method and the method based on mixed fermentation of the glutamic acid-producing microorganism and the inosinic acid-producing microorganism, the components of glutamic acid-inosinic acid (GA-IMP) fermented powders obtained by the production methods were compared (see FIG. 3).

[00212] The glutamic acid seed culture broth and inosinic acid seed culture broth used here were prepared in the same manner as in Example 5-1.

[00213] @ Individual fermentation (Preparation Example 7)

[00214] In the method of mixing individual dried products among the conventional methods, each of the glutamic acid seed culture broth and the inosinic acid seed culture broth was transferred into a 50-L fermenter and then individually subjected to main fermentation. Next, cells were separated from each fermentation broth, and then decolorization and filtration processes were performed. The filtrates were concentrated and dried to obtain dried products. The obtained dried glutamic acid and isosinic acid products were mixed together so that the ratio of glutamic acid to inosinic acid was 1:1, thereby preparing fermented powder containing glutamic acid and inosinic acid.

[00215] @ Mixing of fermentation broths after individual fermentation (Preparation Example 8)

[00216] In the method of mixing individual fermentation broths among the conventional methods, each of the glutamic acid seed culture broth and the inosinic acid seed culture broth was transferred into a 50-L fermenter and then individually subjected to main fermentation. The fermentation broths obtained in the main fermentation were mixed together so that the ratio of glutamic acid to inosinic acid was 1: 1. Next, cells were separated from the fermentation broth mixture, and then decolorization and filtration processes were performed. The filtrate was concentrated and dried to obtain glutamic acid-inosinic acid fermented powder.

[00217] @ Mixed fermentation (Preparation Example 9)

[00218] In mixed fermentation, the glutamic acid seed culture broth and the inosinic acid seed culture broth were inoculated at a ratio of 20: 80 in the same manner as in Example 5-2 so that the ratio of glutamic acid to inosinic acid in the fermentation broth was about 1:1 in the same manner as in Preparation Examples 7 and 8, followed by mixed fermentation. Next, cells were separated from the fermentation broth, and decolorization and filtration processes were performed. The filtrate was concentrated and dried to obtain glutamic acid-inosinic acid fermented powder.

[00219] 6-2. Comparison of components between glutamic acid-inosinic acid fermented powders

[00220] Component analysis was performed on the glutamic acid-inosinic acid fermented powder obtained through individual fermentation in each of Preparation Examples 7 and 8 and the glutamic acid-inosinic acid fermented powder obtained through mixed fermentation in Preparation Example 9.

[00221] L-glutamic acid and IMP were measured by HPLC analysis (GA - 210 nm, UV detector, flow rate 0.9 ml / min; IMP - 254 nm, UV detector, flow rate 0.9 ml / min). Organic acid and ions were measured in the same manner as in Example 2-2. The results are shown in Table 14 below.

[00222] [Table 14] Item Preparation Example 7 Preparation Example 8 Preparation Example 9 GA 22% 31% 37% IMP 20% 31% 36% (GA+IMP) / TS* 42% 62% 73% Total nitrogen 8 . 6% 9.7% 10.5% Organic acids 4.0% 3.3% 3.1% Ions 10.4% 8.8% 4.2% Ammonium 4.0% 3.0% 1.7% *(GA+IMP) / TS: Proportion of product relative to total solid content

[00223] 6-3. Sensory comparison between individual fermentation and mixed fermentation

[00224] Sensory evaluation was performed on the glutamic acid-inosinic acid fermented powders obtained through 5 individual fermentation in Preparation Examples 7 and 8 and the glutamic acid-inosinic acid fermented powder obtained through mixed fermentation in Preparation Example 9.

[00225] Sensory evaluation was performed in the same manner as in Example 2-3. The results are shown in Table 15 below. 10

[00226] [Table 15] Item Preparation Example 7 (individual fermentation) Preparation Example 8 (individual fermentation) Preparation Example 9 (mixed fermentation) Initial Umami taste +++ ++++ ++++ Umami + +++ ++++ persistence Kokumi + ++ +++ Salty taste + ++ ++ Sour taste ++ ++ + Bitter taste ++ + Sweet taste + + ++

[00227] Referring to Table 15 above, due to the difference in components of the glutamic acid-inosinic acid fermented powder as in Table 14 above, the sample prepared by the mixing process after individual fermentation showed a weak 5 umami taste and umami persistence and an increased bitter taste compared to the sample prepared by mixed fermentation. In addition, since the increase in by-products such as organic acids and the increase in ions as shown in Table 14 above also affect the sensory properties of the sample, the 10 mixed fermentation process is more effective in terms of taste or process simplification than mixing after individual fermentation.

[00228] Example 7. Mixed fermentation of inosinic acid and lysine 15

[00229] 7-1. Seed culture

[00230] Corynebacterium ammoniagenes NFI545 (KCCM13162P) was used as an inosinic acid-producing microorganism, and Corynebacterium glutamicum NFL21 (KCCM13163P) was used as a lysine-producing microorganism.

[00231] An inosinic acid seed culture broth of the inosinic acid-producing microorganism and a lysine seed culture broth of the lysine-producing microorganism were prepared in the same manner as in Examples 5-1 and 1-1, respectively.

[00232] 7-2. Main fermentation

[00233] In order to examine the ratio of inosinic acid to lysine in the fermentation broth depending on the inoculums of the inosinic acid seed culture broth and the lysine seed culture broth, the inosinic acid seed culture broth and the lysine seed culture broth were inoculated at various ratios and fermented.

[00234] In main fermentation, 14 to 18 L of a fermentation medium was added to a 50-L fermenter and inoculated with 1.2 to 1.8 L of the total seed culture broth at an inoculation ratio of 0.05 to 99.95 (inosinic acid seed culture broth): 99.95 to 0.05 (lysine seed culture broth), followed by mixed fermentation by fed-batch culture for 45 to 90 hours. The fermentation medium composition and fermentation conditions used here are shown in Table 16 below.

[00235] [Table 16] Fermentation medium composition 1 to 3% molasses, 5 to 7% raw sugar, 2 to 3% yeast extract paste, 0.6 to 1.2% H3PO4, 0.05 to 0.1% betaine, 100 to 200 ppm adenine, 50 to 150 ppm guanine, 0.2 to 0.5% MgSO4^7H2O, 50 to 100 ppm Ca-pantothenate, 5 to 15 ppm vitamin B3,  5 to 20 ppm thiamine HCl, 0.4% (NH4)2SO4, 0.4 to 0.8% NaOH, 5 to 10 ppm FeSO4, 10 to 20 ppm MnSO4, 10 to 20 ppm ZnSO4, and 0.005% antifoaming agent for food Fermentation conditions Temperature 31 to 32°C, pH 6.5 to 7.5, aeration rate 0.8 to 1.2 vvm, internal pressure 0.6 to 1.0 kg / cm3, agitation speed 320 to 350 rpm, and dissolved oxygen (DO) concentration 20 to 70%

[00236] This mixed fermentation was performed a total of three times, the average value was calculated, and the results are shown in Table 17 below.

[00237] [Table 17] Inoculation ratio between seed culture Fermentation time IMP: LYS fermentat ratio in ion ratio broths (IMP: LYS) 99.95 : 0.05 90 hours 92.3 : 1 80 : 20 73 hours 1.01 : 1 50 : 50 61 hours 0.05 : 1 20 : 80 48 hours 0.02 : 1 0.05 : 99.95 45 hours 0.01 : 1 5

[00238] Referring to Table 17 above, it was confirmed that, when the seed culture broths of the microorganisms were inoculated at a ratio of 0.05 to 99.95 (inosinic acidproducing microorganism): 99.95 to 0.05 (lysine-producing microorganism), IMP and lysine in the fermentation broth were produced at a ratio of 0.01 to 92: 1.

[00239] Example 8. Comparison of inosinic acid-lysine fermented powders between fermentation processes

[00240] 8-1. Production of inosinic acid-lysine fermented powder

[00241] Conventionally, a flavor containing inosinic acid and lysine was produced by individually fermenting an inosinic acid-producing microorganism and a lysine-producing microorganism and then mixing the fermentation broths or dried products thereof at a suitable ratio. To compare the differences in taste between this conventional individual fermentation method and the method based on mixed fermentation of the inosinic acid-producing microorganism and the lysine-producing microorganism, the components of inosinic acid-lysine (IMP-LYS) fermented powders obtained by the production methods were compared (see FIG. 4).

[00242] The inosinic acid seed culture broth and lysine seed culture broth used here were prepared in the same manner as in Example 7-1.

[00243] @ Individual fermentation (Preparation Example 10)

[00244] In the method of mixing individual dried products among the conventional methods, each of the inosinic acid seed culture broth and the lysine seed culture broth was transferred into a 50-L fermenter and then individually subjected to main fermentation. Next, cells were separated from each fermentation broth, and then decolorization and filtration processes were performed. The filtrates were concentrated and dried to obtain dried products. The obtained dried inosinic acid and lysine products were mixed together so that the ratio of inosinic acid to lysine was 1:1, thereby preparing fermented powder containing inosinic acid and lysine.

[00245] @ Mixing of fermentation broths after individual fermentation (Preparation Example 11)

[00246] In the method of mixing individual fermentation broths among the conventional methods, each of the inosinic acid seed culture broth and the lysine seed culture broth was transferred into a 50-L fermenter and then individually subjected to main fermentation. The fermentation broths obtained in the main fermentation were mixed together so that the ratio of inosinic acid to lysine was 1: 1. Next, cells were separated from the fermentation culture broth, and then decolorization and filtration processes were performed. The filtrate was concentrated and dried to obtain inosinic acid-lysine fermented powder.

[00247] @ Mixed fermentation (Preparation Example 12)

[00248] In mixed fermentation, the inosinic acid seed culture broth and the lysine seed culture broth were inoculated at a ratio of 80: 20 in the same manner as in Example 7-2 so that the ratio of inosinic acid to lysine in the fermentation broth was about 1:1 in the same manner as in Preparation Examples 10 and 11, followed by mixed fermentation. Next, cells were separated from the fermentation broth, and decolorization and filtration processes were performed. The filtrate was concentrated and dried to obtain inosinic acid-lysine fermented powder.

[00249] 8-2. Comparison of components between lysine-inosinic acid fermented powders

[00250] Component analysis was performed on the inosinic acid-lysine fermented powder obtained through individual fermentation in each of Preparation Examples 10 and 11 and the inosinic acid-lysine fermented powder obtained through mixed fermentation in Preparation Example 12.

[00251] IMP and L-lysine were measured by HPLC analysis (IMP - 254 nm, UV detector, flow rate 0.9 ml / min; LYS - 214 nm, UV detector, flow rate 0.8 ml / min). Organic acid and ions were measured in the same manner as in Example 2-2. The results are shown in Table 18 below.

[00252] [Table 18] Item Preparation Example 10 Preparation Example 11 Preparation Example 12 LYS 26% 32% 38% IMP 24% 30% 33% (LYS+IMP) / TS* 45% 62% 71% Total nitrogen 9.7% 11.3% 12.5% Organic acids 5.0% 4.1% 3.7% Ions 9.2% 8.2% 5.6% Ammonium 3.9% 3.5% 1. 9% *(LYS+IMP) / TS: proportion of product relative to total solid

[00253] Referring to Table 18 above, mixed fermentation (Preparation Example 12) showed a higher LYS+IMP proportion than individual fermentation (Preparation Examples 10 and 11) and showed significant decreases in the contents of 5 organic acids, ions and ammonium.

[00254] 8-3. Sensory comparison between individual fermentation and mixed fermentation

[00255] Sensory evaluation was performed on the inosinic acid-lysine fermented powders obtained through individual 10 fermentation in Preparation Examples 10 and 11 and the inosinic acid-lysine fermented powder obtained through mixed fermentation in Preparation Example 12.

[00256] Sensory evaluation was performed in the same manner as in Example 2-3. The results are shown in Table 19 below. 15

[00257] [Table 19] Item Preparation Example 10 Preparation Example 11 Preparation Example 12 (individual fermentation) (individual fermentation) (mixed fermentation) Initial Umami taste + ++ ++ Umami persistence + +++ ++++ Kokumi + ++ +++ Salty taste + ++ ++ Sour taste ++ ++ + Bitter taste ++ ++ Sweet taste + + ++

[00258] Referring to Table 19 above, due to the difference in components of the inosinic acid-lysine fermented powder as shown in Table 18 above, the sample prepared by the mixing process after individual fermentation showed weak umami 5 persistence and an increased bitter taste compared to the sample prepared by mixed fermentation. In addition, since the increase in by-products such as organic acids and the increase in ions as shown in Table 18 above also affect the sensory properties of the sample, the mixed fermentation 10 process is more effective in terms of taste or process simplification than mixing after individual fermentation.

[00259] Example 9. Mixed fermentation of inosinic acid and arginine

[00260] 7-1. Seed culture

[00261] Corynebacterium ammoniagenes NFI545 (KCCM13162P) was used as an inosinic acid-producing microorganism, and Corynebacterium glutamicum NFA40 (KCCM13165P) was used as an arginine-producing microorganism.

[00262] An inosinic acid seed culture broth of the inosinic acid-producing microorganism and an arginine seed culture broth of the arginine-producing microorganism were prepared in the same manner as in Examples 5-1 and 3-1, respectively.

[00263] 9-2. Main fermentation

[00264] In order to examine the ratio of inosinic acid to arginine in the fermentation broth depending on the inoculums of the inosinic acid seed culture broth and the arginine seed culture broth, the inosinic acid seed culture broth and the arginine seed culture broth were inoculated at various ratios and fermented.

[00265] In main fermentation, 14 to 18 L of a fermentation medium was added to a 50-L fermenter and inoculated with 1.2 to 1.8 L of the total seed culture broth at an inoculation ratio of 65 to 99.95 (inosinic acid seed culture broth): 35 to 0.05 (arginine seed culture broth), followed by mixed fermentation by fed-batch culture for 45 to 80 hours. The fermentation medium composition and fermentation conditions used here are shown in Table 20 below.

[00266] [Table 20] Fermentation medium composition 1 to 3% molasses, 5 to 7% raw sugar, 2 to 3% yeast extract paste, 0.6 to 1.2% H3PO4, 0.05 to 0.1% betaine, 100 to 200 ppm adenine, 50 to 150 ppm guanine, 0.2 to 0.5% MgSO4^7H2O, 50 to 100 ppm Ca-pantothenate, 5 to 15 ppm vitamin B3, 5 to 20 ppm thiamine HCl, 0.4% (NH4)2SO4, 0.4 to 0.8% NaOH, 5 to 10 ppm FeSO4, 10 to 20 ppm MnSO4, 10 to 20 ppm ZnSO4, and 0.005% antifoaming agent for food Fermentation conditions Temperature 31 to 32°C, pH 6.5 to 7.5, aeration rate 0.8 to 1.2 vvm, internal pressure 0.6 to 1.0 kg / cm3, agitation speed 320 to 350 rpm, and dissolved oxygen (DO) concentration 20 to 70%

[00267] This mixed fermentation was performed a total of three times, the average value was calculated, and the results are shown in Table 21 below.

[00268] [Table 21] Inoculation ratio between seed culture Fermentation time IMP: ARG fermentat ratio in ion broth broths (IMP: ARG) 99.95 : 0.05 80 hours 95.1 : 1 80 : 20 46 hours 5.3 : 1 70 : 30 45 hours 2.45 : 1 65 : 35 45 hours 1.04 : 1

[00269] Referring to Table 21 above, it was confirmed that, when the seed culture broths of the microorganisms were inoculated at a ratio of 65 to 99.95 (inosinic acidproducing microorganism): 35 to 0.05 (arginine-producing microorganism), IMP and L-arginine in the fermentation broth were produced at a ratio of 1.04 to 95.1: 1.

[00270] Example 10. Comparison of inosinic acid-arginine fermented powders between fermentation processes

[00271] 10-1. Production of inosinic acid-arginine fermented powder

[00272] Conventionally, a flavor containing inosinic acid and arginine was produced by individually fermenting an inosinic acid-producing microorganism and an arginineproducing microorganism and then mixing the fermentation broths or dried products thereof at a suitable ratio. To compare the differences in taste between this conventional individual fermentation method and the method based on mixed fermentation of the inosinic acid-producing microorganism and the arginine-producing microorganism, the components of inosinic acid-arginine (IMP-ARG) fermented powders obtained by the production methods were compared (see FIG. 5).

[00273] The inosinic acid seed culture broth and arginine seed culture broth used here were prepared in the same manner as in Example 9-1.

[00274] @ Individual fermentation (Preparation Example 13)

[00275] In the method of mixing individual dried products among the conventional methods, each of the inosinic acid seed culture broth and the arginine seed culture broth was transferred into a 50-L fermenter and then individually subjected to main fermentation. Next, cells were separated from each fermentation broth, and then decolorization and filtration processes were performed. The filtrates were concentrated and dried to obtain dried products. The obtained dried inosinic acid and arginine products were mixed together so that the ratio of inosinic acid to arginine was 1:1, thereby preparing fermented powder containing inosinic acid and arginine.

[00276] @ Mixing of fermentation broths after individual fermentation (Preparation Example 14)

[00277] In the method of mixing individual fermentation broths among the conventional methods, each of the inosinic acid seed culture broth and the arginine seed culture broth was transferred into a 50-L fermenter and then individually subjected to main fermentation. The fermentation broths obtained in the main fermentation were mixed together so that the ratio of inosinic acid to arginine was 1: 1. Next, cells were separated from the fermentation broth mixture, and then decolorization and filtration processes were performed. The filtrate was concentrated and dried to obtain inosinic acid-arginine fermented powder.

[00278] @ Mixed fermentation (Preparation Example 15)

[00279] In mixed fermentation, the inosinic acid seed culture broth and the arginine seed culture broth were inoculated at a ratio of 65: 35 in the same manner as in Example 9-2 so that the ratio of inosinic acid to arginine in the fermentation broth was about 1:1 in the same manner as in Preparation Examples 13 and 14, followed by mixed fermentation. Next, cells were separated from the fermentation broth, and decolorization and filtration processes were performed. The filtrate was concentrated and dried to obtain inosinic acid-arginine fermented powder.

[00280] 10-2. Comparison of components between inosinic acid-arginine fermented powders

[00281] Component analysis was performed on the inosinic acid-arginine fermented powder obtained through individual fermentation in each of Preparation Examples 13 and 14 and the inosinic acid-arginine fermented powder obtained through mixed fermentation in Preparation Example 15.

[00282] L-arginine and IMP were measured by HPLC analysis (ARG - 195 nm, UV detector, flow rate 1 ml / min; IMP - 254 nm, UV detector, flow rate 0.9 ml / min). Organic acid and ions were measured in the same manner as in Example 2-2. The results are shown in Table 22 below.

[00283] [Table 22] Item Preparation Example 13 Preparation Example 14 Preparation Example 15 ARG 24% 31% 34% IMP 24% 31% 33% (ARG+IMP) / TS* 48% 62% 76% Total nitrogen 9.7% 11.3% 14.1% Organic acids 4.7% 3.8% 2.5% Ions 7.5% 6.2% 3.8% Ammonium 3.1% 2.5% 0.8% *(ARG+IMP) / TS: Proportion of product relative to total solid content

[00284] 10-3. Sensory comparison between individual fermentation and mixed fermentation

[00285] Sensory evaluation was performed on the inosinic acid-arginine fermented powders obtained through individual 5 fermentation in Preparation Examples 13 and 14 and the inosinic acid-arginine fermented powder obtained through mixed fermentation in Preparation Example 15.

[00286] Sensory evaluation was performed in the same manner as in Example 2-3. The results are shown in Table 23 below. 10

[00287] [Table 23] Item Preparation Example 13 (individual fermentation) Preparation Example 14 (individual fermentation) Preparation Example 15 (mixed fermentation) Initial Umami + ++ ++ taste Umami persistence + +++ ++++ Kokumi + ++ +++ Salty taste + ++ ++ Sour taste ++ ++ + Bitter taste ++ ++ Sweet taste

[00288] Referring to Table 23 above, due to the difference in components of the inosinic acid-arginine fermented powder as shown in Table 22 above, the sample prepared by the mixing process after individual fermentation showed weak 5 umami persistence and an increased bitter taste compared to the sample prepared by mixed fermentation. In addition, since the increase in by-products such as organic acids and the increase in ions as shown in Table 22 above also affect the sensory properties of the sample, the mixed 10 fermentation process is more effective in terms of taste or process simplification than mixing after individual fermentation.

[00289] So far, the present invention has been described with reference to the preferred embodiments. Those of ordinary 15 skill in the art to which the present invention pertains will appreciate that the present invention may be embodied in modified forms without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative point of view, not from a restrictive point of view. The scope of the present invention is defined by the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the scope of the present invention.

[00290] [Accession Numbers]

[00291] Depository authority: Korean Culture Center of Microorganisms (KCCM)

[00292] Accession number: KCCM13162P

[00293] Deposit date: April 21, 2022

[00294]

[00295] Depository authority: Korean Culture Center of Microorganisms (KCCM)

[00296] Accession number: KCCM13163P

[00297] Deposit date: April 21, 2022

[00298]

[00299] Depository authority: Korean Culture Center of Microorganisms (KCCM)

[00300] Accession number: KCCM13164P

[00301] Deposit date: April 21, 2022

[00302]

[00303] Depository authority: Korean Culture Center of Microorganisms (KCCM)

[00304] Accession number: KCCM13165P

[00305] Deposit date: April 21, 2022 5

Claims

1. A method for producing a flavor containing L-glutamic acid and L-arginine, themethod comprising a step of inoculating a fermentation medium with a glutamic acid-producing microorganism and an arginine-producing microorganism at a ratio of 60 to 70 : 30 to 40 and then producing a fermentation broth containing L-glutamic acid and L-arginine at a ratio of 1.96 to 2.41 : 1 by fermentation of the microorganisms,wherein the glutamic acid-producing microorganism and the arginine-producing microorganism are Corynebacterium glutamicum.

2. The method of claim 1, wherein the glutamic acid-producing microorganism andthe arginine-producing microorganism are in a seed culture broth state obtained by individual culture or co-culture.

3. A flavor containing L-glutamic acid and L-arginine produced by the method ofclaim 1.

4. The flavor of claim 3, wherein the flavor contains L-glutamic acid and L-arginine,in an amount of 3 to 90 wt% based on the total solid content.

5. A food composition containing the flavor containing L-glutamic acid and L-arginine according to claim 3.

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