Direct vat set functional bacterial agent as well as preparation method and application thereof
By preparing direct-injection functional bacterial agents and using kimchi fermentation broth for multi-step treatment, the problems of low comprehensive utilization rate and poor flavor quality in the kimchi industry are solved, and efficient utilization and flavor improvement are achieved.
Patent Information
- Application Number
- CN202510108859.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-23
- Publication Date
- 2025-04-08
AI Technical Summary
The existing kimchi fermentation technology is difficult to achieve effective comprehensive utilization of waste liquid in large-scale production, resulting in waste of resources, and the fermented flavor quality is different from traditional kimchi.
By preparing a direct-injected functional bacterial agent, using the kimchi fermentation broth for pretreatment, centrifugation, membrane separation and ion exchange, a kimchi fermentation concentrate containing a high number of viable bacteria was prepared, and mixed with the precipitate to form a direct-injected functional bacterial agent.
It improves the flavor quality and nutritional value of kimchi, enhances the comprehensive utilization rate of kimchi fermentation waste liquid, reduces resource waste, and maintains microbial activity.
Smart Images
Figure CN119776149A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of vegetable deep processing, and in particular to a direct-injection functional bacterial agent and a preparation method and application thereof. Background Art
[0002] During the kimchi fermentation process, the number and composition of microorganisms continue to change, and finally reach a relatively stable stage of microorganism number, abundance, composition, and characteristic physical and chemical indicators. Steady-state fermentation is the core of processing and producing high-quality kimchi. After the kimchi is mature and taken out, the remaining kimchi fermentation liquid contains a large number of stable microbial communities, including lactic acid bacteria, yeast, acetic acid bacteria and other microorganisms. In addition, the kimchi fermentation liquid is rich in organic components such as lactic acid, organic acids, amino acids, reducing sugars, etc. formed by the decomposition of carbohydrates and proteins by microorganisms, as well as various flavor substances such as esters, alcohols, aldehydes and ketones, as well as inorganic substances and minerals such as calcium and phosphorus.
[0003] In the traditional kimchi fermentation process, the kimchi fermentation liquid is recycled and used. This fermentation liquid is called kimchi mother water, but kimchi mother water is only suitable for small family workshops. With the large-scale development of the kimchi industry, the fermentation liquid of kimchi is generally discharged directly as wastewater after simple treatment in industry. At present, there are also related studies on the reuse of brine after treatment, and its application in soy sauce or seasoning liquid production, but the comprehensive utilization rate is low. In order to realize the artificial control of kimchi fermentation, people began to achieve the rapid replication of dominant bacteria in the steady-state fermentation system by preparing direct-injection bacterial agents. At present, most of them mainly screen out dominant strains from kimchi fermentation liquid, and then prepare direct-injection bacterial agents by compounding single, two or three strains. After inoculating and fermenting kimchi, the fermentation environment can be controlled, but its flavor quality is still somewhat different from traditional kimchi. Summary of the invention
[0004] In view of the above problems, the present invention provides a direct-injection functional bacterial agent and a preparation method and application thereof, which can effectively improve the flavor quality of kimchi, and can also improve the comprehensive utilization rate of kimchi fermentation waste liquid in kimchi enterprises and reduce resource waste.
[0005] In order to achieve the purpose of the present invention, the following scheme is adopted: A method for preparing a direct-injection functional bacterial agent comprises the following steps: S100: pretreatment: using kimchi fermented liquid after kimchi fermentation, firstly coarsely filtering the kimchi fermented liquid through a coarse grid with a grid pitch of 10 mm to 20 mm, and then finely filtering the mixed liquid that has passed the coarse filtration through a fine grid with a grid pitch of 0.5 mm to 1 mm; S200: centrifugation: centrifuging the kimchi fermented liquid that has been finely filtered in S100, with the speed of the centrifuge being 8000 rpm to 10000 rpm, to obtain a precipitate and a supernatant, respectively; S300: membrane separation: the supernatant obtained in S200 is subjected to membrane separation to obtain a cutoff liquid, and the molecular weight cutoff of the separation membrane is 100 Daltons to 10,000 Daltons; S400: Preparation: The intercepted liquid obtained in S300 is subjected to ion exchange for adsorption of lactic acid, and the lactic acid content is adjusted to 2 g / L-2.5 g / L at the end of the ion exchange; and then 0.4%-0.8% of glucose, 0.2%-0.6% of peptone, and 0.1%-0.2% of dipotassium hydrogen phosphate are added in an amount equivalent to the mass of the intercepted liquid obtained in S300 to prepare a kimchi fermentation concentrate; S500: Mixing: The precipitate obtained in S200 and the kimchi fermentation concentrate obtained in S400 are mixed to prepare a direct-injection functional bacterial agent, wherein the number of live bacteria in the direct-injection functional bacterial agent is 1.0×10 8 CFU / mL~1.0×10 9 CFU / mL.
[0006] Furthermore, in S100, the coarse grid pitch is 15 mm, and the fine grid pitch is 0.7 mm.
[0007] Furthermore, in S200, the rotation speed of the centrifuge is 9000 rpm.
[0008] Furthermore, when membrane separation is performed in S300, the temperature is controlled to be below 30°C.
[0009] Furthermore, the molecular weight cutoff of the separation membrane used in S300 is 200 Daltons to 5000 Daltons.
[0010] Furthermore, in S400, ion exchange is performed using a weakly basic anion exchange resin, and at the end of the ion exchange, the lactic acid content is set to 2.2 g / L.
[0011] Furthermore, in S400, 0.6% of glucose, 0.3% of peptone, and 0.15% of dipotassium hydrogen phosphate are added in a mass equivalent to that of the intercepted liquid obtained in S300.
[0012] A direct-injection functional bacterial agent is prepared by adopting the preparation method of the direct-injection functional bacterial agent.
[0013] An application of a direct-injection functional bacterial agent, wherein the direct-injection functional bacterial agent is used in kimchi fermentation.
[0014] The beneficial effects of this technical solution are: 1. The direct-injection functional bacterial agent prepared by the method of the present application can be directly used in kimchi fermentation, which can not only accelerate the fermentation of kimchi, but also contain nutritional factors and flavor substances in the kimchi liquid in the bacterial agent, thereby improving the nutritional value and flavor of the kimchi product.
[0015] 2. Using ion exchange adsorption to reduce lactic acid in the retentate can effectively reduce the acidity of the retentate, reduce the stress of acid on microorganisms, and is beneficial to the maintenance of microbial activity.
[0016] 3. The kimchi fermentation liquid is prepared into a direct-injection functional bacterial agent to improve the comprehensive utilization rate of kimchi wastewater and reduce resource waste. BRIEF DESCRIPTION OF THE DRAWINGS
[0017] Figure 1 The main flow chart of the method for preparing the direct-injection functional bacterial agent according to the embodiment of the present application is shown. DETAILED DESCRIPTION
[0018] In order to make the purpose, technical solutions and advantages of the present application clearer, the implementation methods of the present application are described in detail below, but the embodiments described in this application are only part of the embodiments of the present application, rather than all the embodiments.
[0019] In such Figure 1 Based on the main flow chart shown, the direct injection functional bacterial agent is prepared according to the following specific steps: S100: Pretreatment: The fermented kimchi liquid after Sichuan kimchi fermentation is firstly coarsely filtered through a coarse grid with a grid pitch of 10 mm to 20 mm, and then the mixed liquid that has passed the coarse filtration is finely filtered through a fine grid with a grid pitch of 0.5 mm to 1 mm. Preferably, the grid pitch of the coarse grid is 15 mm, and the grid pitch of the fine grid is 0.7 mm. The coarse filtration is used to remove larger impurities such as vegetable leaves and vegetable residues, and the fine filtration is used to remove smaller impurities such as particles and suspended matter. The Sichuan kimchi described in this step is usually a kimchi product with red radish, green bamboo shoots, carrots, etc. as main raw materials; S200: centrifugation: centrifuging the kimchi fermented liquid that has been finely filtered in S100, specifically, using a disc centrifuge, the rotation speed of the centrifuge is 8000 rpm to 10000 rpm, preferably, the rotation speed is 9000 rpm, to obtain a precipitate and a supernatant respectively; S300: membrane separation: performing membrane separation on the supernatant obtained in S200 to obtain a cutoff liquid. Specifically, the temperature is controlled below 30° C. during membrane separation, and a nanofiltration membrane is used as the separation membrane. The molecular weight cutoff of the separation membrane is 100 Dalton to 10,000 Dalton, preferably 200 Dalton to 5,000 Dalton, and more preferably 300 Dalton to 3,000 Dalton. S400: preparation: using weakly alkaline anion exchange resin to perform ion exchange on the intercepted liquid obtained in S300 for adsorbing lactic acid, and making the lactic acid content range from 2 g / L to 2.5 g / L at the end of the ion exchange, preferably, making the lactic acid content range from 2 g / L to 2.5 g / L; then adding 0.4% to 0.8% of glucose, 0.2% to 0.6% of peptone, and 0.1% to 0.2% of dipotassium hydrogen phosphate equivalent to the intercepted liquid obtained in S300 to prepare kimchi fermentation concentrate, preferably, adding 0.6% of glucose, 0.3% of peptone, and 0.15% of dipotassium hydrogen phosphate equivalent to the intercepted liquid obtained in S300; S500: Mixing: The precipitate obtained in S200 and the kimchi fermentation concentrate obtained in S400 are mixed to prepare a direct-injection functional bacterial agent, wherein the number of live bacteria in the direct-injection functional bacterial agent is 1.0×10 8 CFU / mL~1.0×10 9 CFU / mL.
[0020] The prepared direct-injection functional bacterial agent is used in kimchi fermentation, that is, inoculation is carried out in a fermentation container.
[0021] In order to verify the effect of the direct-injection functional bacterial agent prepared by the above method in the actual kimchi fermentation, the kimchi preparation test was carried out according to the following same process: 300g of red radish, 100g of green bamboo shoots, 100g of carrots, 1000g of water, 10g of white sugar, 20g of ginger slices, 5g of red chili pepper, 0.3g of pepper, and 42g of salt were taken; the surface impurities of the above raw materials were removed, and then washed with pure water and drained; the red radish, green bamboo shoots and carrots were cut into 3cm×1.5cm×1.5cm rectangular blocks, the ginger was cut into slices of uniform thickness, the red chili pepper was cut evenly, and other raw materials were not cut; the pottery jar was scalded with boiling water 3 times in advance, all the raw materials were added to the jar, and then the prepared direct-injection functional bacterial agent or a commercial kimchi fermentation agent available on the market was inoculated, and the inoculation amount was 5.0×10 5 CFU / mL, mix well, add water to the edge of the jar and seal it, and ferment at a constant temperature of 20℃ for 7 days.
[0022] Six preparations were performed, of which the ones inoculated with direct-injection functional bacterial agents were recorded as Experimental Examples 1 to 5, and the one inoculated with commercial kimchi fermentation agents was recorded as a comparative example.
[0023] In Experimental Examples 1 to 3, the same batch of kimchi fermentation liquid was used in S100, and the test results of various indicators are shown in Table 1: Table 1 Test results of various indicators of kimchi fermentation liquid project unit Measurement results pH / 3.6 Total acid (calculated as lactic acid) g / 100mL 0.62 Salinity (as NaCl) g / 100mL 3.55 Nitrite mg / L 0.51 glucose mg / L 138.11 lactic acid g / L 4.0 Citric Acid g / L - Succinic acid g / L 0.9 Acetic acid g / L 1.1 Lactic acid bacteria count CFU / mL <![CDATA[1.2×10 8 ]]> Yeast count CFU / mL <![CDATA[6.8×10 6 ]]> Coliform bacteria CFU / mL - Mold CFU / mL - Pathogens CFU / mL - Test Example 1: S100: The kimchi fermented liquid is firstly coarsely filtered through a coarse grid with a grid pitch of 15 mm, and then the mixed liquid that has passed the coarse filtration is finely filtered through a fine grid with a grid pitch of 0.7 mm; S200: centrifuging the kimchi fermented liquid that has been finely filtered in S100 using a disc centrifuge at a speed of 9000 rpm to obtain a precipitate and a supernatant respectively; S300: using a nanofiltration membrane to perform membrane separation on the supernatant obtained in S200 to obtain a cutoff liquid, the temperature during membrane separation is controlled below 30°C, and the molecular weight cutoff of the separation membrane is 300 Daltons to 3000 Daltons; S400: using a weakly alkaline anion exchange resin to perform ion exchange on the intercepted liquid obtained in S300, so that the lactic acid content is 2.2 g / L at the end of the ion exchange, and then adding 0.6% of glucose, 0.3% of peptone, and 0.15% of dipotassium hydrogen phosphate in an amount equivalent to the mass of the intercepted liquid obtained in S300 to prepare a kimchi fermentation concentrate; S500: The precipitate obtained in S200 and the kimchi fermentation concentrate obtained in S400 are mixed to prepare a direct-injection functional bacterial agent, wherein the number of live bacteria in the direct-injection functional bacterial agent is 1.0×10 9 CFU / mL.
[0024] The direct-injection functional bacterial agent is prepared according to the above steps, and then used to prepare kimchi.
[0025] Test Example 2: S100: The kimchi fermented liquid is firstly coarsely filtered through a coarse grid with a grid pitch of 10 mm, and then the mixed liquid that has passed the coarse filtration is finely filtered through a fine grid with a grid pitch of 0.5 mm; S200: centrifuging the kimchi fermented liquid that has been finely filtered in S100 using a disc centrifuge at a speed of 8000 rpm to obtain a precipitate and a supernatant respectively; S300: using a nanofiltration membrane to perform membrane separation on the supernatant obtained in S200 to obtain a cutoff liquid, the temperature during membrane separation is controlled below 30°C, and the molecular weight cutoff of the separation membrane is 100 Daltons to 10,000 Daltons; S400: using a weakly alkaline anion exchange resin to perform ion exchange on the intercepted liquid obtained in S300, so that the lactic acid content is 2 g / L at the end of the ion exchange, and then adding 0.4% of glucose, 0.2% of peptone, and 0.1% of dipotassium hydrogen phosphate in an amount equivalent to the mass of the intercepted liquid obtained in S300 to prepare a kimchi fermentation concentrate; S500: The precipitate obtained in S200 and the kimchi fermentation concentrate obtained in S400 are mixed to prepare a direct-injection functional bacterial agent, wherein the number of live bacteria in the direct-injection functional bacterial agent is 1.0×10 8CFU / mL.
[0026] The direct-injection functional bacterial agent is prepared according to the above steps, and then used to prepare kimchi.
[0027] Test Example 3: S100: The kimchi fermented liquid is firstly coarsely filtered through a coarse grid with a grid pitch of 20 mm, and then the mixed liquid that has passed the coarse filtration is finely filtered through a fine grid with a grid pitch of 1 mm; S200: centrifuging the kimchi fermented liquid that has been finely filtered in S100 using a disc centrifuge at a speed of 10,000 rpm to obtain a precipitate and a supernatant respectively; S300: using a nanofiltration membrane to perform membrane separation on the supernatant obtained in S200 to obtain a cutoff liquid, the temperature during membrane separation is controlled below 30°C, and the molecular weight cutoff of the separation membrane is 200 Daltons to 5000 Daltons; S400: using a weakly alkaline anion exchange resin to perform ion exchange on the intercepted liquid obtained in S300, so that the lactic acid content is 2.5 g / L at the end of the ion exchange, and then adding 0.8% of glucose, 0.6% of peptone, and 0.2% of dipotassium hydrogen phosphate in an amount equivalent to the mass of the intercepted liquid obtained in S300 to prepare a kimchi fermentation concentrate; S500: The precipitate obtained in S200 and the kimchi fermentation concentrate obtained in S400 are mixed to prepare a direct-injection functional bacterial agent, wherein the number of live bacteria in the direct-injection functional bacterial agent is 6.0×10 8 CFU / mL.
[0028] The direct-injection functional bacterial agent is prepared according to the above steps, and then used to prepare kimchi.
[0029] Test example 4: The second batch of kimchi fermented liquid was used, and then the direct injection functional bacterial agent was prepared according to the same steps as in Experimental Example 1, and then used to prepare kimchi. The test results of various indicators of the kimchi fermented liquid used in S100 are shown in Table 2: Table 2 Test results of various indicators of kimchi fermentation liquid project unit Measurement results pH / 3.5 Total acid (calculated as lactic acid) g / 100mL 0.69 Salinity (as NaCl) g / 100mL 3.45 Nitrite mg / L 0.41 glucose mg / L 108.85 lactic acid g / L 4.3 Citric Acid g / L - Succinic acid g / L 0.8 Acetic acid g / L 1.3 Lactic acid bacteria count CFU / mL <![CDATA[5.5×10 8 ]]> Yeast count CFU / mL <![CDATA[1.0×10 7 ]]> Coliform bacteria CFU / mL - Mold CFU / mL - Pathogens CFU / mL - Test Example 5: The third batch of kimchi fermented liquid was used, and then the direct injection functional bacterial agent was prepared according to the same steps as in Experimental Example 1, and then used to prepare kimchi. The test results of various indicators of the kimchi fermented liquid used in S100 are shown in Table 3: Table 3 Test results of various indicators of kimchi fermentation liquid project unit Measurement results pH / 3.9 Total acid (calculated as lactic acid) g / 100mL 0.49 Salinity (as NaCl) g / 100mL 3.48 Nitrite mg / L 0.63 glucose mg / L 157.19 lactic acid g / L 3.6 Citric Acid g / L - Succinic acid g / L 0.25 Acetic acid g / L 0.68 Lactic acid bacteria count CFU / mL <![CDATA[3.4×10 7 ]]> Yeast count CFU / mL <![CDATA[9.4×10 5 ]]> Coliform bacteria CFU / mL - Mold CFU / mL - Pathogens CFU / mL - Comparative Example: Commercial kimchi starter culture was directly used for kimchi preparation.
[0030] After the kimchi of Experimental Examples 1 to 5 and the comparative example was prepared, 20 people conducted sensory evaluation on the kimchi, and evaluated them from five aspects, namely color, taste, acidity, aroma, and shape. The scores were integers of 1 to 5 points, and each score corresponded to: negligible, small, medium, strong, and very strong. The sensory evaluation results were averaged and are shown in Table 4: Table 4 Sensory evaluation score table project Color taste acidity aroma form Test Example 1 4.2 4.2 3.5 4.6 4.4 Test Example 2 3.9 4.3 4.0 4.1 4.3 Test Example 3 4.0 4.0 3.2 4.3 4.4 Test Example 4 3.9 4.1 3.8 3.6 4.2 Test Example 5 3.8 3.9 3.0 3.8 4.3 Comparative Example 3.6 3.9 4.5 3.0 4.4 From the sensory evaluation, it can be seen that the kimchi inoculated with direct-injection functional bacteria agents in each group has significantly higher color and aroma scores than the kimchi inoculated with commercial kimchi starter agent, and the taste is slightly better. The sourness score is significantly lower, that is, the sourness is softer and not too strong, and the morphology scores are similar.
[0031] The above are only some of the embodiments listed in this application and are not intended to limit this application.
Claims
1. A method for preparing a direct-injection functional bacterial agent, characterized in that: The following steps are involved: S100： Pretreatment: using kimchi fermented liquid after kimchi fermentation, firstly filtering the kimchi fermented liquid through a coarse grid with a grid pitch of 10 mm to 20 mm, and then finely filtering the mixed liquid that has passed the coarse filtration through a fine grid with a grid pitch of 0.5 mm to 1 mm; S200: centrifugation: centrifuging the kimchi fermented liquid that has been finely filtered in S100, with the speed of the centrifuge being 8000 rpm to 10000 rpm, to obtain a precipitate and a supernatant, respectively; S300: membrane separation: the supernatant obtained in S200 is subjected to membrane separation to obtain a cutoff liquid, and the molecular weight cutoff of the separation membrane is 100 Daltons to 10,000 Daltons; S400: Preparation: The intercepted liquid obtained in S300 is subjected to ion exchange for adsorption of lactic acid, and the lactic acid content is adjusted to 2 g / L-2.5 g / L at the end of the ion exchange; and then 0.4%-0.8% of glucose, 0.2%-0.6% of peptone, and 0.1%-0.2% of dipotassium hydrogen phosphate are added in an amount equivalent to the mass of the intercepted liquid obtained in S300 to prepare a kimchi fermentation concentrate; S500: Mixing: The precipitate obtained in S200 and the kimchi fermentation concentrate obtained in S400 are mixed to prepare a direct-injection functional bacterial agent, wherein the number of live bacteria in the direct-injection functional bacterial agent is 1.0×10 8 CFU / mL~1.0×10 9 CFU / mL.
2. The method for preparing the direct-administered functional bacterial agent according to claim 1, characterized in that: In S100, the coarse grid pitch is 15 mm and the fine grid pitch is 0.7 mm.
3. The method for preparing the direct-administered functional bacterial agent according to claim 1, characterized in that: In S200, the rotation speed of the centrifuge is 9000 rpm.
4. The method for preparing the direct-administered functional bacterial agent according to claim 1, characterized in that: When membrane separation is performed in S300, the temperature is controlled below 30°C.
5. The method for preparing the direct-administered functional bacterial agent according to claim 1, characterized in that: The molecular weight cutoff of the separation membrane used in S300 is 200 Daltons to 5000 Daltons.
6. The method for preparing the direct-administered functional bacterial agent according to claim 1, characterized in that: In S400, ion exchange is performed using a weakly basic anion exchange resin, and at the end of the ion exchange, the lactic acid content is set to 2.2 g / L.
7. The method for preparing the direct-administered functional bacterial agent according to claim 1, characterized in that: In S400, 0.6% of glucose, 0.3% of peptone, and 0.15% of dipotassium hydrogen phosphate were added in a mass equivalent to that of the intercepted liquid obtained in S300.
8. A direct-injection functional bacterial agent, characterized in that: The method is adopted to prepare the direct-administered functional bacterial agent as claimed in any one of claims 1 to 7.
9. An application of a direct injection functional bacterial agent, characterized in that: The direct-injection functional bacterial agent described in claim 8 is used in kimchi fermentation.