Membrane integrated treatment process of probiotics and probiotic polypeptide

Abstract

The membrane integrated treatment process mainly comprises the following steps: fermenting a probiotic strain to obtain a probiotic fermentation solution, centrifuging to obtain a probiotic concentrated solution and a probiotic clear solution, and freeze-drying and packaging the probiotic concentrated solution to obtain a probiotic product, the probiotic clear liquid passes through a silicon carbide membrane treatment system, a ceramic membrane treatment system and a nanofiltration membrane treatment system, an obtained nanofiltration membrane concentrated solution is conveyed and packaged through a metering pump, a probiotic polypeptide product is obtained, and nanofiltration membrane permeate liquid is conveyed to a biochemical treatment system through a frequency conversion conveying pump to be treated. According to the method, the probiotic clear liquid is obtained through conventional fermentation and centrifugation processes, and is subjected to integrated treatment through a silicon carbide membrane system, a ceramic membrane system and a nanofiltration membrane system, and freeze-drying and packaging processes are combined, so that graded purification and productization of probiotics and probiotic polypeptide products are realized, and meanwhile, the biochemical treatment load of the probiotic clear liquid is effectively reduced; the method can be widely applied to treatment of various probiotics such as bacillus, bifidobacterium and lactic acid bacteria.

Description

Technical Field

[0001] The present invention belongs to the field of biomaterial preparation and relates to a membrane integrated processing process for probiotics and probiotic polypeptides. Background Art

[0002] Probiotics (such as lactic acid bacteria and bifidobacteria) produce a variety of bioactive peptides during metabolism, including antimicrobial peptides, immunomodulatory peptides, antioxidant peptides, and functional regulatory peptides. Antimicrobial peptides can inhibit the growth of pathogens and maintain a balanced intestinal flora; immunomodulatory peptides can regulate the host immune response, alleviate inflammation, or enhance immunity; antioxidant peptides play an important role in scavenging free radicals and reducing oxidative stress damage; and antihypertensive peptides help regulate blood pressure.

[0003] Currently, commonly used industrial fine purification processes for peptide products include: reverse-phase high-performance liquid chromatography (RP-HPLC), which uses a C18 or C8 column to separate peptides by gradient elution from the organic phase; gel filtration chromatography, which separates peptides by molecular weight; and affinity chromatography, which selectively adsorbs specific tags or functional groups. Patent CN202411310009 reports a method for preparing a cyclic dipeptide as a potential aroma component. After centrifugation of the fermentation broth, the harvested supernatant is extracted and separated by chromatography to obtain the target fragment. Patent CN202410678057 reports a method for preparing walnut kernel composite active peptides by adding Bifidobacterium longum to fresh walnut kernel slurry. Ethanol is added to the fermentation broth, which is then allowed to stand and centrifuged to obtain a supernatant. The supernatant is separated using an ultrafiltration membrane, and the permeate is collected and freeze-dried to obtain a walnut kernel mixed peptide. This method inactivates the bacteria, and the product obtained after ultrafiltration has a complex composition and poor purity. Reversed-phase high-performance liquid chromatography (RP-HPLC) has drawbacks such as reliance on organic solvents, low recovery rates for highly hydrophobic peptides, and high costs. Gel filtration chromatography suffers from low resolution, low throughput, and a significant dilution effect. Affinity chromatography has specificity limitations, high ligand costs, and potential contamination risks. Membrane separation technology, a physical separation process encompassing ultrafiltration, nanofiltration, and reverse osmosis, boasts a small footprint and high efficiency, and has been widely used in water treatment and material separation.

[0004] Therefore, seeking a new processing technology for probiotics and probiotic peptides is a technical problem that urgently needs to be solved in the field of probiotics production. Summary of the Invention

[0005] The purpose of the present invention is to provide a membrane-integrated processing process for probiotics and probiotic polypeptides.

[0006] The technical solution of the present invention is: A membrane integrated processing process for probiotics, probiotics and probiotic polypeptides, the technical solution includes: probiotic strains are fermented to obtain probiotic fermentation liquid, which is stored in a probiotic fermentation tank, and centrifuged to obtain a probiotic concentrate and a probiotic clear liquid, which are respectively stored in a probiotic concentrate tank and a probiotic clear liquid tank, wherein the probiotic concentrate is freeze-dried and packaged to obtain a probiotic product; the probiotic clear liquid after centrifugation enters a silicon carbide membrane processing system through a variable frequency delivery pump I, and is separated to obtain a silicon carbide membrane concentrated liquid A and a silicon carbide membrane permeate A, the silicon carbide membrane concentrated liquid A is delivered to the probiotic concentrate tank by a variable frequency delivery pump II, and the silicon carbide membrane permeate A is delivered to the probiotic concentrate tank by a variable frequency delivery pump II. The frequency conversion pump III enters the ceramic membrane treatment system, and after separation, ceramic membrane concentrate B and ceramic membrane permeate B are obtained. The ceramic membrane concentrate B enters the ceramic membrane concentrate transfer tank through the frequency conversion pump IV, and is then transported to the probiotic concentrate tank through the frequency conversion pump V; the ceramic membrane permeate B enters the nanofiltration membrane treatment system through the frequency conversion pump VI, and is separated to obtain nanofiltration membrane concentrate C and nanofiltration membrane clear liquid C. The nanofiltration membrane concentrate C enters the nanofiltration membrane concentrate storage tank through the frequency conversion pump VII, and is transported and packaged by the metering pump to obtain the probiotic polypeptide product. The nanofiltration membrane permeate C is transported to the biochemical treatment system for treatment through the frequency conversion pump VIII.

[0007] Among them, the specifications of the probiotic fermentation tank are 1-3 m 3 ; Among them, the specifications of the probiotic concentrate tank are 1-3 m 3 ; Among them, the specifications of the probiotic liquid tank are 1-3 m 3 ; Wherein, the flow rate of the variable frequency delivery pump 1 is 500-1500 L / h; Among them, the silicon carbide membrane processing device uses a silicon carbide membrane tube with a pore size of 500-1000 nm. Its main purpose is to intercept probiotic strains with a pore size larger than 500-1000 nm and transport them into the probiotic concentrate tank; Among them, the flow rate of variable frequency delivery pump II is 300-1000 L / h; Among them, the flow rate of variable frequency delivery pump III is 300-1000 L / h; Among them, the ceramic membrane treatment device uses a ceramic membrane with a pore size of 50-100 nm. Its main purpose is to further intercept probiotics and macromolecular substances. The ceramic membrane concentrate is used as the raw material for probiotics; Among them, the flow rate of variable frequency delivery pump IV is 300-1000 L / h; Among them, the specifications of the ceramic membrane concentrate transfer tank are 1-2 m 3 ; Among them, the flow rate of variable frequency delivery pump V is 300-500 L / h; Among them, the flow rate of variable frequency delivery pump VI is 300-500 L / h; Among them, the nanofiltration membrane treatment device uses a nanofiltration membrane with a pore size of 1-2 nm. Its main purpose is to intercept the probiotic polypeptide solution. Small molecular amino acids and inorganic salts produced during the fermentation process pass through the membrane; Among them, the flow rate of variable frequency delivery pump VII is 300-500 L / h; Among them, the flow rate of variable frequency delivery pump VIII is 300-500 L / h; Among them, the flow rate of the variable frequency metering pump is 50-150 L / h.

[0008] Compared with the existing technology, the present invention has significant advantages: first, the process integrates physicochemical and biochemical treatment processes, and has the advantages of high efficiency, stability, economy, environmental protection, small footprint, etc., and has strong promotion characteristics, and can be widely used in the treatment of various probiotics such as Bacillus, Bifidobacterium, and Lactobacillus; second, the process uses the membrane integrated treatment technology of the composite silicon carbide membrane treatment system, the ceramic membrane treatment system and the nanofiltration membrane treatment system to grade and purify probiotics and probiotic polypeptides, realizes the recycling of probiotics and probiotic polypeptides in the probiotic centrifugal clear liquid, realizes the production of probiotics and probiotic polypeptide products, and has important economic value; finally, the process effectively reduces the organic matter content in the nanofiltration membrane clear liquid, and reduces the load of the subsequent biochemical treatment system. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 The present invention provides a membrane-integrated processing process for probiotics and probiotic polypeptides. DETAILED DESCRIPTION

[0010] Comparative Example 1: Probiotic (Bifidobacterium) strains were fermented by conventional methods to obtain probiotic fermentation liquid (i.e., Bifidobacterium fermentation liquid), which was stored in a 2 m 3 The probiotic fermentation tank was centrifuged to obtain the probiotic concentrate (i.e., bifidobacterium concentrate) and the probiotic clear solution (i.e., bifidobacterium clear solution), which were then added to the 2 m 3 Probiotic concentrate tank and 2 m 3 The probiotic concentrate is stored in a tank, where the probiotic product is obtained through conventional freeze-drying and packaging processes; the probiotic concentrate is transported to the biochemical treatment system for treatment via a 1500L / h variable frequency delivery pump VIII. Example

[0011] Probiotic (Bifidobacterium) strains were fermented by conventional methods to obtain probiotic fermentation liquid (i.e., Bifidobacterium fermentation liquid), which was stored in 1 m 3The probiotic fermentation tank was centrifuged to obtain the probiotic concentrate (i.e., bifidobacterium concentrate) and the probiotic clear solution (i.e., bifidobacterium clear solution), which were then added to 1 m 3 Probiotic concentrate tank and 1 m 3 The probiotic clear liquid tank is used for storage, wherein the probiotic concentrate is subjected to conventional freeze-drying and packaging processes to obtain a probiotic product; the probiotic clear liquid enters the silicon carbide membrane treatment system with a pore size of 500 nm through a 500 L / h variable frequency delivery pump I, and the probiotics and organic solutions larger than 500 nm are retained to form a silicon carbide membrane concentrate, which is delivered to the probiotic concentrate tank through a 300 L / h variable frequency delivery pump II, while the silicon carbide membrane permeate with a pore size smaller than 500 nm enters the ceramic membrane treatment system with a pore size of 50 nm through a 300 L / h variable frequency delivery pump III, and the probiotics and macromolecular organic solutions larger than 50 nm are retained by the ceramic membrane to form a ceramic membrane concentrate, which is delivered to a 1 m 3 The ceramic membrane concentrate transfer tank is connected to the probiotic concentrate tank through liquid level control and is transported to the probiotic concentrate tank by a 300 L / h delivery pump V. The probiotic polypeptide solution with a pore size of less than 50 nm enters the nanofiltration membrane treatment system with a pore size of 1 nm through a 300 L / h variable frequency delivery pump VI. The probiotic polypeptide molecules with a pore size greater than 1 nm are retained by the nanofiltration membrane. The formed nanofiltration membrane concentrate enters the nanofiltration membrane concentrate storage tank through a 300 L / h variable frequency delivery pump VII, and is transported and packaged by a 50 L / h variable frequency metering pump to obtain a probiotic polypeptide product. The nanofiltration membrane permeate with a pore size less than 1 nm is transported to the biochemical treatment system for treatment through a 300 L / h variable frequency delivery pump VIII. Example

[0012] Probiotic (Bifidobacterium) strains were fermented by conventional methods to obtain probiotic fermentation liquid (i.e. Bifidobacterium fermentation liquid), which was stored in a 3 m 3 The probiotic fermentation tank was centrifuged to obtain the probiotic concentrate (i.e., bifidobacterium concentrate) and the probiotic clear solution (i.e., bifidobacterium clear solution), which were then respectively placed in a 3 m 3 Probiotic concentrate tank and 3 m 3The probiotic clear liquid tank is used for storage, wherein the probiotic concentrate is subjected to conventional freeze-drying and packaging processes to obtain a probiotic product; the probiotic clear liquid enters the silicon carbide membrane treatment system with a pore size of 1000 nm through a 1500 L / h variable frequency delivery pump I, and the probiotics and organic solutions larger than 1000 nm are retained to form a silicon carbide membrane concentrate, which is delivered to the probiotic concentrate tank through a 1000 L / h variable frequency delivery pump II, while the silicon carbide membrane permeate with a size smaller than 1000 nm enters the ceramic membrane treatment system with a pore size of 100 nm through an 800 L / h variable frequency delivery pump III, and the probiotics and macromolecular organic solutions larger than 100 nm are retained by the ceramic membrane to form a ceramic membrane concentrate, which is delivered to a 1.5 m 3 The ceramic membrane concentrate transfer tank is connected to the probiotic concentrate tank through liquid level control and is transported to the probiotic concentrate tank by a 500 L / h delivery pump V. The probiotic polypeptide solution with a pore size of less than 100 nm enters the nanofiltration membrane treatment system with a pore size of 2 nm through a 500 L / h variable frequency delivery pump VI. The probiotic polypeptide molecules with a pore size greater than 2 nm are retained by the nanofiltration membrane. The formed nanofiltration membrane concentrate enters the nanofiltration membrane concentrate storage tank through a 500 L / h variable frequency delivery pump VII, and is transported and packaged by a 150 L / h variable frequency metering pump to obtain a probiotic polypeptide product. The nanofiltration membrane permeate with a pore size less than 2 nm is transported to the biochemical treatment system for treatment through a 500 L / h variable frequency delivery pump VIII. Example

[0013] Probiotic (Bifidobacterium) strains were fermented by conventional methods to obtain probiotic fermentation liquid (i.e., Bifidobacterium fermentation liquid), which was stored in a 2 m 3 The probiotic fermentation tank was centrifuged to obtain the probiotic concentrate (i.e., bifidobacterium concentrate) and the probiotic clear solution (i.e., bifidobacterium clear solution), which were then added to the 2 m 3 Probiotic concentrate tank and 2 m 3 The probiotic clear liquid tank is used for storage, wherein the probiotic concentrate is subjected to conventional freeze-drying and packaging processes to obtain a probiotic product; the probiotic clear liquid enters the silicon carbide membrane treatment system with a pore size of 800 nm through a 1000 L / h variable frequency delivery pump I, and the probiotics and organic solutions larger than 800 nm are retained to form a silicon carbide membrane concentrate, which is delivered to the probiotic concentrate tank through a 1000 L / h variable frequency delivery pump II, while the silicon carbide membrane permeate with a pore size smaller than 800 nm enters the ceramic membrane treatment system with a pore size of 80 nm through an 800 L / h variable frequency delivery pump III, and the probiotics and macromolecular organic solutions larger than 80 nm are retained by the ceramic membrane to form a ceramic membrane concentrate, which is delivered to a 1.5 m 3The ceramic membrane concentrate transfer tank is controlled by liquid level and transported to the probiotic concentrate tank through a 500 L / h delivery pump V. The probiotic polypeptide solution with a pore size of less than 100 nm enters the nanofiltration membrane treatment system with a pore size of 1.5 nm through a 500 L / h variable frequency delivery pump VI. The probiotic polypeptide molecules with a pore size greater than 1.5 nm are retained by the nanofiltration membrane. The formed nanofiltration membrane concentrate enters the nanofiltration membrane concentrate storage tank through a 400 L / h variable frequency delivery pump VII, and is transported and packaged by an 80 L / h variable frequency metering pump to obtain a probiotic polypeptide product. The nanofiltration membrane permeate with a pore size less than 1.5 nm is transported to the biochemical treatment system for treatment through a 400 L / h variable frequency delivery pump VIII.

[0014] Compared with the conventional probiotic production process (Control Example 1), the present invention realizes the graded production of probiotics and probiotic polypeptides, the yield of probiotics increases by 4.1%-5.2%, the proportion of probiotic polypeptide yield to probiotics is 1.3%-2.1%, and the load of subsequent biochemical treatment is greatly reduced. The COD value of the wastewater entering the end biochemical treatment is reduced by about 81.5%-87.9%.

Claims

1. A membrane-integrated treatment process for probiotics, probiotics, and probiotic polypeptides, characterized by: Probiotics are fermented by conventional methods to obtain probiotic fermentation liquid, which is stored in 1-3 m 3 The probiotics fermentation tank is centrifuged to obtain the probiotics concentrate and the probiotics clear solution, which are then fed into 1-3 m 3 Probiotic concentrate tank and 1-3 m 3 The probiotic clear liquid tank is used for storage, and the probiotic concentrate is subjected to conventional freeze-drying and packaging processes to obtain a probiotic product; the probiotic clear liquid enters a silicon carbide membrane treatment system with a certain pore size through a 500-1500 L / h variable frequency delivery pump I, and the probiotics and organic solutions larger than the pore size of the silicon carbide membrane are retained to form a silicon carbide membrane concentrate, which is delivered to the probiotic concentrate tank through a 500-1500 L / h variable frequency delivery pump II, while the silicon carbide membrane permeate smaller than the pore size of the silicon carbide membrane enters a ceramic membrane treatment system with a certain pore size through a 500-1500 L / h variable frequency delivery pump III, and the probiotics and macromolecular organic solutions larger than the pore size of the ceramic membrane are retained by the ceramic membrane to form a ceramic membrane concentrate, which is delivered to a 1-2 m 3 The ceramic membrane concentrate transfer tank is controlled by the liquid level and is transported to the probiotic concentrate tank by the 300-500 L / h delivery pump V. The probiotic polypeptide solution smaller than the pore size of the ceramic membrane enters the nanofiltration membrane treatment system with a certain pore size through the 300-500 L / h variable frequency delivery pump VI. The probiotic polypeptide molecules larger than the pore size of the nanofiltration membrane are retained by the nanofiltration membrane. The formed nanofiltration membrane concentrate enters the nanofiltration membrane concentrate storage tank through the 300-500 L / h variable frequency delivery pump VII, and is transported and packaged by the 50-150 L / h variable frequency metering pump to obtain the probiotic polypeptide product. The nanofiltration membrane permeate smaller than the pore size of the nanofiltration membrane is transported to the biochemical treatment system for treatment by the 300-500 L / h variable frequency delivery pump VIII.

2. A membrane-integrated treatment process for probiotics, probiotics, and probiotic polypeptides according to claim 1, characterized in that: In the silicon carbide film processing device, the pore size of the silicon carbide film is in the range of 500-1000 nm.

3. A membrane-integrated treatment process for probiotics, probiotics, and probiotic polypeptides according to claim 1, characterized in that: The ceramic membrane processing device has a ceramic membrane pore size range of 50-100 nm.

4. A membrane-integrated treatment process for probiotics, probiotics, and probiotic polypeptides according to claim 1, characterized in that: The nanofiltration membrane treatment device has a pore size range of 1-2 nm.

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