Breast milk oligosaccharide extraction and concentration device

Abstract

The utility model discloses a breast milk oligosaccharide extraction and concentration device which comprises an ultrasonic stirrer for carrying out ultrasonic treatment on a mixed solution of breast milk oligosaccharide fermentation liquor and lysozyme, and a microwave extractor for carrying out microwave treatment on a mixed solution of a solution pumped by the ultrasonic stirrer and ethanol, the first centrifugal machine is used for carrying out centrifugal separation treatment on the mixed liquid subjected to microwave treatment, the first storage tank is used for storing clear liquid separated by the first centrifugal machine, the ultrafiltration membrane system is used for carrying out ultrafiltration treatment on the clear liquid, and the nanofiltration membrane system is used for carrying out nanofiltration treatment on filtrate of the ultrafiltration membrane system; the activated carbon adsorption system is used for carrying out decoloration and impurity removal treatment on permeate liquid subjected to nanofiltration; the crystallization system is used for carrying out crystallization treatment on filtrate subjected to decoloration and impurity removal; and the second centrifugal machine is used for carrying out centrifugal treatment on materials subjected to crystallization treatment. The extraction and concentration device can improve the purity and yield of the product extracted from the breast milk oligosaccharide fermentation liquor.

Description

Technical Field

[0001] The utility model relates to a breast milk oligosaccharide extraction and concentration device, belonging to the technical field of extraction and concentration. Background Art

[0002] Human milk oligosaccharides (HMOs) are complex mixed oligosaccharides naturally present in breast milk and are the third largest nutrient in human breast milk after lactose and lipids. HMOs are mainly present in breast milk in free form and are a type of oligosaccharide composed of 3 to 10 monosaccharide units linked by glycosidic bonds to form straight or branched chains. The biosynthetic pathway based on bioengineered bacteria such as Escherichia coli and yeast has the advantages of low production cost and low environmental pollution. It is a synthetic method actively used in today's commercial products. At present, the biosynthetic pathway of bioengineered bacteria is mostly used to prepare human milk oligosaccharides, but the extraction process of biological fermentation broth containing human milk oligosaccharides is limited, and the purity and yield of the human milk oligosaccharides finally extracted are low. Utility Model Content

[0003] The utility model aims to provide a human milk oligosaccharide extraction and concentration device, which can improve the purity and yield of the product extracted from the human milk oligosaccharide fermentation liquid.

[0004] In order to achieve the above purpose, the technical solution adopted by the utility model is:

[0005] A human milk oligosaccharide extraction and concentration device, comprising:

[0006] An ultrasonic stirrer is used to perform ultrasonic treatment on a mixture of human milk oligosaccharide fermentation liquid and lysozyme;

[0007] A microwave extractor is connected to the ultrasonic stirrer and is used to perform microwave treatment on a mixture of the solution and ethanol pumped into the ultrasonic stirrer;

[0008] A first centrifuge, connected to the microwave extractor, is used to perform centrifugal separation on the mixed liquid after microwave treatment;

[0009] A first storage tank, connected to the first centrifuge, for storing the clear liquid separated by the first centrifuge;

[0010] An ultrafiltration membrane system, connected to the first storage tank, for ultrafiltration treatment of the clear liquid;

[0011] The nanofiltration membrane system is connected to the permeation side of the ultrafiltration membrane system and is used to perform nanofiltration treatment on the filtrate of the ultrafiltration membrane system;

[0012] The activated carbon adsorption system is connected to the permeate side of the nanofiltration membrane system and is used to decolorize and remove impurities from the permeate after nanofiltration;

[0013] The crystallization system is connected to the activated carbon adsorption system and is used to perform crystallization treatment on the filtrate after decolorization and impurity removal;

[0014] The second centrifuge is connected to the crystallization system and is used to centrifuge the material after the crystallization treatment to obtain high-purity human milk oligosaccharide crystals and crystallization mother liquor.

[0015] Preferably, it also includes:

[0016] A second storage tank, connected to the second centrifuge, for storing the crystallization mother liquor separated by the second centrifuge;

[0017] The spray drying system is connected to the second storage tank and is used to spray dry the crystallization mother liquor to obtain low-concentration human milk oligosaccharide dry powder.

[0018] Preferably, the membrane in the ultrafiltration membrane system is a ceramic membrane with multiple channels.

[0019] Preferably, the molecular weight cut-off of the ceramic membrane is 0.1-25KD.

[0020] Preferably, the molecular weight cut-off of the nanofiltration membrane in the nanofiltration membrane system is 200-500Da.

[0021] Preferably, the crystallization system is a multiple-effect evaporation crystallization system.

[0022] The beneficial effects of the utility model are:

[0023] Ultrasound-assisted lysozyme is used to break the bacterial cell wall to release intracellular human milk oligosaccharides, and microwave-assisted ethanol extraction is then used to promote the exudation of human milk oligosaccharides from the cells, which can effectively increase the product yield. Centrifugation, ultrafiltration, nanofiltration, and activated carbon decolorization and impurity removal are used to improve product purity. The mother liquor after crystallization is spray-dried to obtain low-concentration human milk oligosaccharide dry powder to maximize utilization. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a flow chart of the utility model;

[0025] Figure 2 This is a device diagram of the utility model.

[0026] The main reference numerals in the figure have the following meanings:

[0027] 1. Ultrasonic agitator, 2. Microwave extractor, 3. First centrifuge, 4. First storage tank, 5. Ultrafiltration membrane system, 6. Nanofiltration membrane system, 7. Activated carbon adsorption system, 8. Crystallization system, 9. Second centrifuge, 10. Second storage tank, 11. Spray drying system, 12. Pump. DETAILED DESCRIPTION

[0028] The utility model is specifically introduced below in conjunction with the accompanying drawings and embodiments.

[0029] like Figure 1-2 As shown, this embodiment is a human milk oligosaccharide extraction and concentration device, including an ultrasonic agitator 1, a pump 12 (boosting pump or centrifugal pump, the same below), a microwave extractor 2, a pump 12, a first centrifuge 3, a first storage tank 4, a pump 12, an ultrafiltration membrane system 5, a nanofiltration membrane system 6, an activated carbon adsorption system 7, a pump 12, a crystallization system 8, a second centrifuge 9, a second storage tank 10, a pump 12 and a spray drying system 11, which are sequentially connected through pipelines.

[0030] Among them, the ultrasonic agitator 1 is used for mixing and ultrasonic treatment of human milk oligosaccharide fermentation liquid and lysozyme. The human milk oligosaccharide fermentation liquid and lysozyme are added into the ultrasonic agitator 1 and mixed evenly under the action of ultrasound and stirring, so that the bacterial cell wall is broken to release the human milk oligosaccharides in the cell; the mixed liquid is then pumped into the microwave extractor 2 through the machine pump 12, and ethanol is added to the microwave extractor 2 at the same time. The ethanol and the liquid pumped into the ultrasonic agitator 1 are further mixed evenly under the action of microwaves and stirring through the microwave extractor 2 to promote the infiltration of human milk oligosaccharides from the cell; the liquid in the microwave extractor 2 is then pumped into the first centrifuge 3 through the machine pump 12 for centrifugal separation to remove impurities such as bacteria, and the human milk oligosaccharides separated in the first centrifuge 3 The clear liquid flows to the first storage tank 4 for storage, and is pumped into the ultrafiltration membrane system 5 by the machine pump 12 to remove residual bacteria, suspended matter and other impurities to obtain ultrafiltration clear liquid; the ultrafiltration clear liquid further passes through the nanofiltration membrane system 6 to remove small molecular impurities, thereby obtaining nanofiltration membrane permeate; the nanofiltration membrane permeate is decolorized and impurity-removed by the activated carbon in the activated carbon adsorption system 7, thereby obtaining activated carbon filtrate with less impurities, and the activated carbon filtrate is pumped into the crystallization system 8 for evaporation and crystallization, and is further separated by the second centrifuge 9 to obtain high-purity human milk oligosaccharide crystals and crystallization mother liquor; the crystallization mother liquor flows into the second storage tank 10 for storage and is sent to the spray drying system 11 by the machine pump 12 to further remove water to obtain low-concentration human milk oligosaccharide dry powder.

[0031] Further, the membrane in the ultrafiltration membrane system 5 is a low molecular weight ceramic membrane, the molecular weight cutoff of the ceramic membrane is 0.1-25KD, preferably 15KD, the ceramic membrane has a diameter of 52mm, a length of 1.5m, 61 channels and a channel diameter of 4mm, and a single membrane tube filtration area of ​​1.29m2; the operating pressure of the ultrafiltration membrane system 5 is 0.6-1MPa, the membrane surface flow rate is 4m / s, and the concentration multiple is more than 10 times. The molecular weight cutoff of the nanofiltration membrane in the nanofiltration membrane system 6 is 200-500Da, preferably 400Da, the system operating pressure is 2.0-3.0MPa, and the concentration multiple is more than 10 times.

[0032] Example 1

[0033] Step 1: add human milk oligosaccharide fermentation liquid and lysozyme (final lysozyme concentration is 0.2 mg / ml) into ultrasonic agitator 1, and perform ultrasonic treatment for 30 minutes under stirring (ultrasonic power 200 W, working 20 seconds, interval 10 seconds).

[0034] Step 2, the feed liquid in the ultrasonic stirrer 1 is pumped into the microwave extractor 2 and ethanol is added (ethanol concentration is 70%, and the volume ratio of ethanol to feed liquid is 10:1), and then microwave treatment is performed (microwave power is 1000w, treatment time is 5min) after mixing evenly.

[0035] Step 3: pump the feed liquid in the microwave extractor 2 into the first centrifuge 3 for centrifugal separation to remove impurities such as bacteria and collect the supernatant; then pump the supernatant into the ultrafiltration membrane system 5 to remove residual bacteria, suspended matter and other impurities to obtain ultrafiltration clear liquid;

[0036] Among them, the operating pressure of ultrafiltration membrane system 5 is 0.6MPa, the membrane surface flow rate is 4m / s, and the concentration multiple is 20 times; the membrane used is a ceramic membrane with a corresponding molecular weight cutoff of 15KD. The ceramic membrane has a diameter of 52mm and a length of 1.5m. It has 61 channels with a channel diameter of 4mm. The filtration area of ​​a single membrane tube is 1.29m 2 .

[0037] Step 4: The ultrafiltration clear liquid enters the nanofiltration membrane system 6 to remove small molecular impurities, thereby obtaining a nanofiltration membrane permeate; the permeate is then decolorized and impurity-removed by the activated carbon adsorption system 7, thereby obtaining an activated carbon filtrate with less impurities;

[0038] The molecular weight cutoff of the nanofiltration membrane in the nanofiltration membrane system 6 is 400Da, the system operating pressure is 2.0MPa, and the concentration multiple is 20 times.

[0039] In step 5, the activated carbon filtrate is pumped into the crystallization system 8 for evaporation and crystallization, and then centrifuged to obtain high-purity human milk oligosaccharide crystals and crystallization mother liquor; the crystallization mother liquor flows into the second storage tank 10 for storage and is pumped into the spray drying system 11 for further dehydration to obtain low-concentration human milk oligosaccharide dry powder.

[0040] Comparative Example 1

[0041] The difference from Example 1 is that no microwave treatment was performed after the addition of ethanol.

[0042] Step 1: add human milk oligosaccharide fermentation liquid and lysozyme (final lysozyme concentration is 0.2 mg / ml) into ultrasonic agitator 1, and perform ultrasonic treatment for 30 minutes under stirring (ultrasonic power 200 W, working 20 seconds, interval 10 seconds).

[0043] Step 2: pump the feed liquid in ultrasonic stirrer 1 into a conventional stirrer and add ethanol (ethanol concentration 70%, and the volume ratio of ethanol to feed liquid is 10:1), and stir.

[0044] In step 3, the feed liquid in the conventional agitator is pumped into the first centrifuge 3 for centrifugal separation to remove impurities such as bacteria and collect the supernatant; the supernatant is then pumped into the ultrafiltration membrane system 5 to remove residual bacteria, suspended matter and other impurities to obtain ultrafiltration clear liquid;

[0045] Among them, the operating pressure of ultrafiltration membrane system 5 is 0.6MPa, the membrane surface flow rate is 4m / s, and the concentration multiple is 20 times; the membrane used is a ceramic membrane with a corresponding molecular weight cutoff of 15KD. The ceramic membrane has a diameter of 52mm and a length of 1.5m. It has 61 channels with a channel diameter of 4mm. The filtration area of ​​a single membrane tube is 1.29m 2 .

[0046] Step 4: The ultrafiltration clear liquid enters the nanofiltration membrane system 6 to remove small molecular impurities, thereby obtaining a nanofiltration membrane permeate; the permeate is then decolorized and impurity-removed by the activated carbon adsorption system 7, thereby obtaining an activated carbon filtrate with less impurities;

[0047] The molecular weight cutoff of the nanofiltration membrane in the nanofiltration membrane system 6 is 400Da, the system operating pressure is 2.0MPa, and the concentration multiple is 20 times.

[0048] In step 5, the activated carbon filtrate is pumped into the crystallization system 8 for evaporation and crystallization, and then centrifuged to obtain high-purity human milk oligosaccharide crystals and crystallization mother liquor; the crystallization mother liquor flows into the second storage tank 10 for storage and is pumped into the spray drying system 11 for further dehydration to obtain low-concentration human milk oligosaccharide dry powder.

[0049] Comparative Example 2

[0050] The difference from Example 1 is that no nanofiltration treatment was performed.

[0051] Step 1: add human milk oligosaccharide fermentation liquid and lysozyme (final lysozyme concentration is 0.2 mg / ml) into ultrasonic agitator 1, and perform ultrasonic treatment for 30 minutes under stirring (ultrasonic power 200 W, working 20 seconds, interval 10 seconds).

[0052] Step 2, the feed liquid in the ultrasonic stirrer 1 is pumped into the microwave extractor 2 and ethanol is added (ethanol concentration is 70%, and the volume ratio of ethanol to feed liquid is 10:1), and then microwave treatment is performed (microwave power is 1000w, treatment time is 5min) after mixing evenly.

[0053] Step 3: pump the feed liquid in the microwave extractor 2 into the first centrifuge 3 for centrifugal separation to remove impurities such as bacteria and collect the supernatant; then pump the supernatant into the ultrafiltration membrane system 5 to remove residual bacteria, suspended matter and other impurities to obtain ultrafiltration clear liquid;

[0054] Among them, the operating pressure of ultrafiltration membrane system 5 is 0.6MPa, the membrane surface flow rate is 4m / s, and the concentration multiple is 20 times; the membrane used is a ceramic membrane with a corresponding molecular weight cutoff of 15KD. The ceramic membrane has a diameter of 52mm and a length of 1.5m. It has 61 channels with a channel diameter of 4mm. The filtration area of ​​a single membrane tube is 1.29m 2 .

[0055] In step 4, the ultrafiltration clear liquid is decolorized and impurities removed by an activated carbon adsorption system 7, thereby obtaining an activated carbon filtrate.

[0056] In step 5, the activated carbon filtrate is pumped into the crystallization system 8 for evaporation and crystallization, and then centrifuged to obtain high-purity human milk oligosaccharide crystals and crystallization mother liquor; the crystallization mother liquor flows into the second storage tank 10 for storage and is pumped into the spray drying system 11 for further dehydration to obtain low-concentration human milk oligosaccharide dry powder.

[0057] The yield of human milk oligosaccharide crystals obtained in Example 1 and Comparative Examples 1-2 was calculated and the product purity was tested. The results are shown in Table 1.

[0058] Table 1 Yield and purity of human milk oligosaccharide crystals obtained in Example 1 and Comparative Examples 1-2

[0059] Human milk oligosaccharide crystal yield Purity of human milk oligosaccharide crystals Example 1 76-78％ 95.3-96.5％ Comparative Example 1 71-73％ 94.2-95.7％ Comparative Example 2 73-75％ 90.2-91.3％

[0060] It can be seen from Table 1 that after the human milk oligosaccharide fermentation liquid is treated by the extraction and concentration device provided by the utility model, the yield and purity of human milk oligosaccharide crystals can be improved to a certain extent.

[0061] The above is only a preferred implementation of the utility model patent. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the utility model patent. These improvements and modifications should also be regarded as the protection scope of the utility model patent.

Claims

1. A human milk oligosaccharide extraction and concentration device, characterized in that: include: An ultrasonic stirrer is used to perform ultrasonic treatment on a mixture of human milk oligosaccharide fermentation liquid and lysozyme; A microwave extractor is connected to the ultrasonic stirrer and is used to perform microwave treatment on a mixture of the solution and ethanol pumped into the ultrasonic stirrer; A first centrifuge, connected to the microwave extractor, is used to perform centrifugal separation on the mixed liquid after microwave treatment; A first storage tank, connected to the first centrifuge, for storing the clear liquid separated by the first centrifuge; An ultrafiltration membrane system, connected to the first storage tank, for ultrafiltration treatment of the clear liquid; The nanofiltration membrane system is connected to the permeation side of the ultrafiltration membrane system and is used to perform nanofiltration treatment on the filtrate of the ultrafiltration membrane system; The activated carbon adsorption system is connected to the permeate side of the nanofiltration membrane system and is used to decolorize and remove impurities from the permeate after nanofiltration; The crystallization system is connected to the activated carbon adsorption system and is used to perform crystallization treatment on the filtrate after decolorization and impurity removal; The second centrifuge is connected to the crystallization system and is used to centrifuge the material after the crystallization treatment to obtain high-purity human milk oligosaccharide crystals and crystallization mother liquor.

2. A human milk oligosaccharide extraction and concentration device according to claim 1, characterized in that: Also includes: A second storage tank, connected to the second centrifuge, for storing the crystallization mother liquor separated by the second centrifuge; The spray drying system is connected to the second storage tank and is used to spray dry the crystallization mother liquor to obtain low-concentration human milk oligosaccharide dry powder.

3. A human milk oligosaccharide extraction and concentration device according to claim 1, characterized in that: The membrane in the ultrafiltration membrane system is a ceramic membrane with multiple channels.

4. A human milk oligosaccharide extraction and concentration device according to claim 3, characterized in that: The molecular weight cut-off of the ceramic membrane is 0.1-25KD.

5. The human milk oligosaccharide extraction and concentration device according to claim 1, characterized in that: The molecular weight cut-off of the nanofiltration membrane in the nanofiltration membrane system is 200-500Da.

6. A human milk oligosaccharide extraction and concentration device according to claim 1, characterized in that: The crystallization system is a multiple-effect evaporation crystallization system.

Images