Purifying and recycling device for probiotic fermentation liquor

Abstract

The utility model relates to a purifying and recycling device for probiotic fermentation liquor, and belongs to the field of probiotic production. Comprising a fermentation tank used for fermentation of probiotics; the solid-liquid separation equipment is used for removing suspended impurities in the fermentation liquor; the ceramic membrane is connected to the filtrate side of the solid-liquid separation equipment and is used for concentrating probiotics in the filtrate; the nanofiltration membrane is connected to the filtrate side of the ceramic membrane and is used for filtering the filtrate of the ceramic membrane; the drying equipment is connected to the concentration side of the ceramic membrane and is used for drying the concentrated solution to obtain bacterial powder; and the filtrate side of the nanofiltration membrane is connected to the fermentation tank.

Description

TECHNICAL FIELD

[0001] The utility model relates to a kind of processing and recycling methods of probiotic fermentation broth, more particularly to a kind of membrane separation technology method of fermentation broth, belong to probiotic production field. BACKGROUND

[0002] Probiotics are well known for their high safety, no drug resistance and influence on host health. Probiotics have functions such as regulating intestinal flora, maintaining intestinal mucosal barrier, antibacterial and immune regulation, and can synthesize digestive enzymes. Together with the digestive enzymes synthesized by the animal body, they participate in the digestion of nutrients in the intestine, stimulate the secretion of digestive enzymes in the animal body and promote the absorption of nutrients in the intestine. The components such as peptidoglycan and lipoteichoic acid in the structure of probiotics can directly play an immune activation role as antigens, or stimulate the host immune system by secreting immune activators to enhance the activity of innate immune cells and natural killer cells. Probiotics have been widely used in food processing, human health and livestock breeding and other fields.

[0003] Chinese patent CN106261000A discloses a concentration and drying method and process of probiotic fermentation broth. The process adds a protective agent to the fermentation broth and then concentrates under reduced pressure. The process has low filtration precision, low concentration ratio and needs additional protective agent, which is not suitable for continuous operation and industrial production. SUMMARY

[0004] The technical problem to be solved by the utility model is to provide a production method that is easy to industrialize continuously, has low energy consumption and high product purity, and utilizes probiotic fermentation broth as a resource to produce probiotic powder preparation.

[0005] The technical solution of the utility model mainly utilizes membrane separation technology. First, the fermentation broth containing probiotics is filtered, decolorized and concentrated by membrane separation technology to obtain probiotic powder preparation. The specific technical solution is as follows:

[0006] (1) Probiotic fermentation: inoculate probiotic strains into corresponding culture medium to obtain probiotic fermentation broth;

[0007] (2) Impurity removal: remove large-particle impurities in the probiotic fermentation broth by centrifuge;

[0008] (3) Concentration: further filter the centrifugal supernatant obtained in step (2) by ceramic membrane filter, and obtain ceramic membrane concentrate after concentration;

[0009] (4) Drying: freeze-dry the ceramic membrane concentrate obtained in step (3) and grind it into powder to obtain probiotic powder finished product.

[0010] (5) removing fermentation metabolites: the ceramic membrane filtrate obtained in step (3) is filtered through a nanofiltration membrane to remove small molecule fermentation metabolites, obtaining a nanofiltration membrane filtrate;

[0011] (6) recycling: the nanofiltration dialysis filtrate obtained in step (5) is recycled and added to the new culture medium in a certain proportion during the next fermentation.

[0012] In step (1), the probiotic strain used is activated. The optional process of activating the probiotic is: after dissolving the probiotic, inoculate it in a liquid culture medium to culture and activate the expanded culture, and after recovering the bacterial concentrate in the culture medium through a ceramic membrane, the activated probiotic strain is obtained.

[0013] In step (1), the fermentation temperature is preferably below 30°C; the probiotic species for fermentation include but are not limited to yeast, lactobacillus, bifidobacterium, streptococcus, etc.

[0014] In step (3), the pore size of the ceramic membrane is 20-500 nm, and further preferably, the membrane pore size can be 40-200 nm, and most preferably, the membrane pore size can be 200 nm. The operating pressure of the ceramic membrane is 0.1-0.5 MPa, and the membrane surface flow rate is 1-6 m / s, and preferably, the operating pressure of the ceramic membrane can be 0.2-0.4 MPa, and the membrane surface flow rate can be 3-5 m / s, and further preferably, the operating pressure of the ceramic membrane can be 0.2 MPa, and the membrane surface flow rate can be 4 m / s. The concentration multiple is 10-30, and preferably, the concentration multiple can be 15-25, and further preferably, the concentration multiple can be 20. The operating temperature is preferably 20-40°C.

[0015] In step (5), the nanofiltration membrane is used to remove pigment, multivalent anion and cation, and small molecule metabolite impurities; the temperature is preferably 20-40°C, the operating pressure is preferably 0.8-2.5 MPa, and preferably, the operating pressure can be 1-2 MPa, and further preferably, the operating pressure can be 1.5 MPa; the concentration multiple is preferably 10-30, and preferably, the concentration multiple is preferably 15-25, and further preferably, the concentration multiple can be 20.

[0016] In step (6), the ratio of the nanofiltration filtrate to the fresh culture medium is 1-100.

[0017] A purification and recycling device for probiotic fermentation broth, comprising:

[0018] a fermentation tank for carrying out fermentation of probiotics;

[0019] A solid-liquid separation device is used to remove suspended impurities in the fermentation liquor.

[0020] A ceramic membrane is connected to the filtrate side of the solid-liquid separation device and used to concentrate probiotic bacteria in the filtrate.

[0021] A nanofiltration membrane is connected to the filtrate side of the ceramic membrane and used to filter the filtrate of the ceramic membrane.

[0022] A drying device is connected to the concentrated side of the ceramic membrane and used to dry the concentrated liquor to obtain probiotic powder.

[0023] The filtrate side of the nanofiltration membrane is connected to the fermentation tank.

[0024] The ceramic membrane is an ultrafiltration membrane or a microfiltration membrane.

[0025] The ceramic membrane has a pore size ranging from 20 to 500 nm.

[0026] The nanofiltration membrane has a molecular weight cut-off of 300 to 1000 Da.

[0027] The drying device is a freeze dryer.

[0028] The advantages of the present application are as follows:

[0029] The method for processing and recycling probiotic fermentation liquor according to the present application has the following advantages compared with the prior art.

[0030] 1. The present application produces probiotics with high added value, and the remaining fermentation liquor can be recycled, which not only reduces the pollution caused by waste liquid discharge to the environment, but also realizes resource utilization.

[0031] 2. The method of the present application uses ceramic membrane filtration, which changes the conventional fermentation liquor treatment method of using a filter press for pressure filtration. The filtration is clear and transparent, and the impurity removal effect is much higher than that of pressure filtration, flocculation and other methods. This method has the characteristics of simple operation and good selectivity, and can purify the fermentation liquor while recovering the bacterial strain. It overcomes the defects of low yield, large sewage discharge and high production labor intensity of the prior art, and significantly improves the quality of probiotics.

[0032] 3. The ceramic membrane has good chemical and mechanical properties, is resistant to high temperature, acid and alkali, and is easy to clean and maintain. Compared with other separation devices, the concentration multiple is improved and the filtration effect is optimized.

[0033] 4. The nanofiltration decolorization is higher in efficiency than the existing activated carbon adsorption decolorization or resin adsorption decolorization, can remove some small molecule metabolic impurities at the same time, can significantly improve the filtrate quality, and makes the product purity higher. The nanofiltration decolorization can be reused to a front-end fermentation process, saves production cost, avoids the inhibition of metabolic products on the fermentation process, and improves the product viable count in the overall circulation process.

[0034] 5. The process can be continuously operated, has long operation time, low operation cost, compact equipment, and is easy to realize automation and suitable for industrial large production. BRIEF DESCRIPTION OF DRAWINGS

[0035] Figure 1 is the flow chart of the patent;

[0036] Figure 2 is the device diagram of the patent.

[0037] 1, fermentation tank; 2, solid-liquid separation equipment; 3, ceramic membrane; 4, nanofiltration membrane; 5, drying equipment. DETAILED DESCRIPTION

[0038] The purification and reuse method of the probiotic fermentation liquor provided by the utility model, comprising the following steps:

[0039] (1) probiotic fermentation: inoculate probiotic strains into corresponding culture medium, and ferment to obtain probiotic fermentation liquor;

[0040] (2) impurity removal: first pass the probiotic fermentation liquor through a centrifugal machine to remove large-particle fermentation liquor impurities;

[0041] (3) concentration: further filter the centrifugal supernatant obtained in the step (2) through a ceramic membrane filter, and obtain ceramic membrane concentrated liquor after concentration;

[0042] (4) drying: freeze-dry the ceramic membrane concentrated liquor obtained in the step (3), grind into powder, and obtain probiotic powder finished product;

[0043] (5) fermentation metabolite removal: filter the ceramic membrane supernatant obtained in the step (3) through a nanofiltration membrane, remove small molecule fermentation metabolites in the ceramic membrane supernatant, and obtain nanofiltration membrane supernatant;

[0044] (6) reuse: reuse the nanofiltration dialysis supernatant obtained in the step (5), and add the nanofiltration dialysis supernatant to new culture solution in a certain proportion during next fermentation.

[0045] The average pore size of the ceramic membrane in the step (3) is 20-500 nm; the operation pressure of the ceramic membrane is 0.1-0.5 MPa; the membrane surface flow rate is 1-6 m / s; and the concentration multiple is 10-30 times;

[0046] The molecular weight cut-off of the nanofiltration membrane in the step (5) is 300-1000 Da; the operating pressure is 0.5-2.5 MPa; and the concentration multiple is 10-30 times.

[0047] The average pore size of the ceramic membrane in the step (3) is 40-200 nm, the operating pressure of the ceramic membrane is 0.2-0.4 MPa, the membrane surface flow rate is 3-5 m / s, and the concentration multiple is 15-25 times.

[0048] The operating pressure in the step (5) is 1-2 MPa, and the concentration multiple is 15-25 times.

[0049] The average pore size of the ceramic membrane in the step (3) is 200 nm, the operating pressure is 0.2 MPa, the membrane surface flow rate is 4 m / s, and the concentration multiple is 20 times.

[0050] The operating pressure in the step (5) is 1.5 MPa, and the concentration multiple is 20 times.

[0051] The probiotic bacteria used in the fermentation in the step (1) include, but are not limited to, yeast, lactobacillus, bifidobacterium, streptococcus, etc.

[0052] The types of metabolites removed in the step of removing metabolites include extracellular polysaccharides, bacteriocins and other substances affecting cell growth.

[0053] The ratio of the nanofiltration clear liquid to the fresh culture medium in the step of recycling is 1-100.

[0054] A device for purifying and recycling probiotic bacteria fermentation liquor, comprising:

[0055] A fermentation tank 1 for carrying out fermentation of probiotic bacteria;

[0056] A solid-liquid separation device 2 for removing suspended impurities in the fermentation liquor;

[0057] A ceramic membrane 3 connected to the filtrate side of the solid-liquid separation device 2 for concentrating probiotic bacteria in the filtrate;

[0058] A nanofiltration membrane 4 connected to the filtrate side of the ceramic membrane 3 for filtering the filtrate of the ceramic membrane 3;

[0059] A drying device 5 connected to the concentrated side of the ceramic membrane 3 for drying the concentrated liquid to obtain bacterial powder;

[0060] The filtrate side of the nanofiltration membrane 4 is connected to the fermentation tank 1.

[0061] The ceramic membrane 3 is an ultrafiltration membrane or a microfiltration membrane.

[0062] The ceramic membrane 3 has a pore size ranging from 20 to 500 nm.

[0063] The nanofiltration membrane 4 has a molecular weight cut-off of 300 to 1000 Da.

[0064] The drying device 5 is a freeze dryer.

[0065] The solid-liquid separation device 2 is a centrifuge.

[0066] The probiotic viable count detection method in this patent is: using a sterile pipette to suck 1 mL of culture solution that needs to be counted, adding it to a test tube containing 9 mL of sterile normal saline, and shaking well. In this way, the bacterial solution is diluted to 10 -1 The dilution liquid is then sequentially diluted at different concentration gradients. According to the dilution, three appropriate dilution gradients are selected. When counting, first melt the MRS culture medium in the anaerobic tube and cool it to about 45°C, then suck 1 mL of the diluted bacterial solution into it, shake gently, and pay attention not to generate bubbles. After the medium cools and solidifies, place it in a 37°C constant temperature incubator for 48 hours for counting. Example 1

[0067] The Bifidobacterium bifidum bacterial solution is inoculated into the culture medium and incubated for 24 hours to obtain seed culture solution. Then the activated seed culture solution is expanded and cultured using MRS culture medium, the fermentation temperature is 20°C, and the fermentation time is 24 hours. The fermentation broth is first removed by centrifuge, and then the supernatant is concentrated by ceramic membrane. The ceramic membrane has a pore size of 200 nm, the operating pressure is 0.3 MPa, the membrane surface flow rate is 4 m / s, and the concentration is 20 times. The ceramic membrane clear liquid is further removed by nanofiltration membrane, the nanofiltration membrane has a molecular weight cut-off of 800, and the operating pressure is 1.5 MPa. After concentration, the nanofiltration membrane concentrate appears brown in color, and the clear liquid appears light yellow in color. The ceramic membrane concentrate is freeze-dried to obtain probiotic finished product. The test shows that the viable count of the bacterial powder is 2.3 x 10 9 cfu / g, the survival rate reaches 2.6%, the particles are uniform, the color is light yellow, and the dispersibility is good. The clear liquid produced by the nanofiltration membrane is added to fresh culture medium for further fermentation. Example 2

[0068] Lactobacillus broth was inoculated in the culture medium and incubated for 24 h to obtain seed culture. Then the activated seed culture was expanded, MRS medium was used, the fermentation temperature was 20℃, and the fermentation time was 24 h. The fermentation temperature was 25℃. The fermentation broth was first removed by centrifuge, and then the supernatant was removed by ceramic membrane. The pore size of the ceramic membrane was 50 nm, the operating pressure was 0.2 MPa, the membrane surface flow rate was 4 m / s, and the concentration was 30 times. The ceramic membrane clear liquid was further removed by nanofiltration membrane, the nanofiltration membrane molecular weight cut-off was 600, the operating pressure was 1.5 MPa, and after concentration, the appearance of the nanofiltration membrane concentrate was brown, and the clear liquid was light yellow. The ceramic membrane concentrate was freeze-dried to obtain the probiotic finished product. The test showed that the viable count of the bacterial powder was 2.1×10 9 cfu / g, the survival rate reached 2.6%, the particles were uniform, the color was light yellow, and the dispersibility was good. The nanofiltration membrane clear liquid was added to fresh culture medium for next step fermentation.

[0069] Comparative Example 1

[0070] The difference from Example 1 is that the ceramic membrane clear liquid is not filtered by nanofiltration membrane, but is directly returned to the fermentation tank for reuse.

[0071] Lactobacillus bifidus broth was inoculated in the culture medium and incubated for 24 h to obtain seed culture. Then the activated seed culture was expanded, MRS medium was used, the fermentation temperature was 20℃, and the fermentation time was 24 h. The fermentation broth was first removed by centrifuge, and then the supernatant was removed by ceramic membrane. The pore size of the ceramic membrane was 200 nm, the operating pressure was 0.3 MPa, the membrane surface flow rate was 4 m / s, and the concentration was 20 times. The ceramic membrane concentrate was freeze-dried to obtain the probiotic finished product. The test showed that the viable count of the bacterial powder was 1.8×10 9 cfu / g, the survival rate reached 2.2%, the particles were uniform, the color was light yellow, and the dispersibility was good. The ceramic membrane clear liquid was added to fresh culture medium for next step fermentation.

Claims

1. A device for purification and reuse of a probiotic fermentation broth, characterized in that, The application relates to a fermentation system for producing probiotic powder, comprising: a fermentation tank (1) for carrying out fermentation of probiotics; a solid-liquid separation device (2) for removing suspended impurities in a fermentation liquor; a ceramic membrane (3) connected to the filtrate side of the solid-liquid separation device (2) and used for concentrating probiotic bodies in the filtrate; a nanofiltration membrane (4) connected to the filtrate side of the ceramic membrane (3) and used for filtering the filtrate of the ceramic membrane (3); and a drying device (5) connected to the concentrated side of the ceramic membrane (3) and used for drying the concentrated liquor to obtain probiotic powder. The filtrate side of the nanofiltration membrane (4) is connected to the fermentation tank (1). The ceramic membrane (3) is an ultrafiltration membrane or a microfiltration membrane. The ceramic membrane (3) has a pore size range of 20-500 nm. The nanofiltration membrane (4) has a molecular weight cut-off of 300-1000 Da. The drying device (5) is a freeze dryer. The solid-liquid separation device (2) is a centrifuge.

2. The apparatus for purification and reuse of probiotic fermentation broth according to claim 1, characterized in that, ​ 3. The apparatus for purification and reuse of probiotic fermentation broth according to claim 1, characterized in that, ​ 4. The apparatus for purification and reuse of probiotic fermentation broth according to claim 1, characterized in that, ​ 5. The apparatus for purification and reuse of probiotic fermentation broth according to claim 1, characterized in that, ​ 6. The apparatus for purification and reuse of probiotic fermentation broth according to claim 1, characterized in that, ​

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