CN114271483A - Preparation method of nattokinase embedded substance - Google Patents
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- Enzymes And Modification Thereof (AREA)
Abstract
The invention provides a preparation method of a nattokinase embedded substance, which comprises the following steps: the method comprises the steps of firstly adopting crushed and puffed grains as a core material carrier to adsorb nattokinase, then compounding sodium alginate with pectin, xanthan gum, gellan gum, propolis and the like as a first layer of coating wall material, and adopting zein as a second layer of coating wall material to prepare the double-layer embedded nattokinase microcapsule, wherein the nattokinase microcapsule has high embedding rate and storage rate. The preparation method of the nattokinase microcapsule has the advantages of simple process, short time consumption and low loss, can greatly improve the utilization rate of raw materials, and reduces the loss of nattokinase in the preparation process.
Description
Technical Field
The invention relates to the technical field of embedding and slow release of protein, in particular to a preparation method of a nattokinase embedded substance.
Background
The nattokinase is alkaline serine proteinase which can effectively dissolve artificial thrombus, has strong fibrinolytic activity under proper conditions, has more obvious thrombolytic effect compared with the current medicament urokinase for treating thrombus clinically and the like, and has no toxic or side effect. In recent years, nattokinase has been receiving wide attention due to its strong thrombolytic effect. According to research, the thrombolysis effect of nattokinase is mainly achieved by directly dissolving fibrin into small molecular peptides, amino acids and the like through the nattokinase, stimulating vascular endothelial cells to generate tissue type plasminogen activator (t-AP), activating in vivo prourokinase to be converted into urokinase, degrading and inactivating inhibitor (PAI-1) of the plasminogen activator and directly or indirectly regulating and controlling the four aspects of fibrinolysis. Natto kinase not only has the effect of dissolving thrombus, but also has the effects of resisting hypertension, reducing blood fat, resisting platelet and improving blood circulation, plays an important role in maintaining thyroid health, and is proved to be a substitute treatment scheme for asthma. Therefore, the natto kinase is promising as a novel thrombolytic drug or special diet development. However, the nattokinase has certain limitation in the application process. The stability of nattokinase is easily influenced by temperature, pH, metal ions, small molecular compounds, organic matters and the like. The nattokinase is stable when the pH value is 5.0-12.0, and can be quickly inactivated when the pH value is lower than 3. Under normal conditions, the pH value of gastric acid of a human body is between 0.9 and 1.8, gastric juice is diluted after meals, the pH value can rise to about 3.5, and if the nattokinase is directly eaten by oral administration, the nattokinase is influenced by strong acid in the stomach to cause the spatial structure of the nattokinase to be damaged, so that more pepsin binding sites are exposed, and the thrombolytic activity is lost. In addition, the digestion in the stomach of a human body generally takes two hours, which requires that certain measures are taken for the nattokinase to protect the nattokinase from strong acid in the stomach.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides the preparation method of the nattokinase embedded substance, which has the advantages of simple process, short time consumption and low loss, can greatly improve the utilization rate of raw materials and reduce the loss of nattokinase in the preparation process, and the obtained nattokinase microcapsule has high embedding rate and storage rate.
The technical scheme provided by the invention is as follows: a preparation method of nattokinase embedded substance comprises the following steps:
(1) preparing natto kinase liquid: placing the bacillus natto in a liquid LB culture medium for liquid fermentation, freezing and centrifuging the fermentation liquor, obtaining supernatant fluid which is nattokinase crude enzyme liquid, and ultrafiltering the nattokinase crude enzyme liquid through an ultrafiltration membrane to obtain nattokinase liquid;
(2) preparing a low-expansion-degree cereal raw material: taking a grain raw material, adjusting the water content of the grain according to the water content of the material being 10-20%, uniformly stirring, pouring the mixture into a double-screw extrusion puffing machine for extrusion puffing, drying the puffed grain and crushing for later use;
(3) the puffed grain is loaded with nattokinase: adding the nattokinase solution into the crushed puffed grains according to the mass ratio of the crushed puffed grains to the nattokinase solution of 0.1-2: 1, and uniformly stirring to obtain a nattokinase-loaded puffed grain mixture;
(4) preparing microcapsules: adopting secondary coating, adding the coating material into the mixture obtained in the step (3) according to the mass ratio of the first layer of coating material to the mixture obtained in the step (3) of 1: 1-5 by adopting a composite sodium alginate gel solution with the mass concentration of 0.1-5% as a first layer of coating material, uniformly stirring, drying and crushing; and then carrying out second-layer coating with a zein ethanol solution according to the mass ratio of 1-10: 1, and drying to obtain the nattokinase microcapsule.
Further, the preparation method of the natto kinase liquid in the step (1) comprises the following steps: placing the bacillus natto in a liquid LB culture medium for liquid fermentation, freezing and centrifuging fermentation liquor at 4-15 ℃ at 6000-12000 r/min for 10-30 min, wherein the supernatant is a crude nattokinase solution, passing the crude nattokinase solution through 1 ten thousand ultrafiltration membranes, passing the trapped fluid through 3 ten thousand ultrafiltration membranes, and obtaining a permeate liquid which is the nattokinase solution to be coated.
Further, the molecular weight of the nattokinase in the nattokinase solution obtained in the step (1) is 27000-28000.
Further, the preparation method of the low-puffing-degree cereal raw material in the step (2) comprises the following steps: the method comprises the steps of adopting grain raw materials, adding water according to the water content of 10-20% of the materials to adjust the water content of the grains, uniformly stirring, pouring into a double-screw extrusion puffing machine for extrusion puffing, setting the feeding speed of the extruder to be 14-18 r/min, the screw rotating speed to be 100-120 r/min, the processing temperature to be 150-180 ℃, drying the puffed grains and then crushing for later use.
Further, the grain raw materials are selected from one or two of rice, corn, millet and other grain raw materials according to a mass ratio of 1-10: 1-10 or three of the following components in a mass ratio of 1-10: 1-10: 1 to 10 parts by weight.
Further, the compound sodium alginate gel substance in the step (4) is prepared by matching sodium alginate with any one or two or three of pectin, xanthan gum, propolis and gellan gum; sodium alginate and any one of pectin, xanthan gum, propolis and gellan gum in a mass ratio of 1-10: 1-10, or sodium alginate and any two of the two substances in a mass ratio of 1-10: 1-10, or sodium alginate and any three of the substances in a mass ratio of 1-10: 1-10 are prepared into a 0.1-5% composite sodium alginate gel solution.
Further, when the second coating is performed in the step (4), adding the zein ethanol solution into the mixture after the first coating, and uniformly mixing, or spraying the zein ethanol solution into the mixture after the first coating, and uniformly spraying.
Furthermore, the zein ethanol solution is obtained by dissolving zein in a solvent with volume fraction of 60-95% ethanol according to the mass concentration of 0.1-5%.
The invention firstly adopts puffed grains to load nattokinase, then uses sodium alginate and pectin, xanthan gum, gellan gum, propolis and other materials to compound as a first layer of coating wall material, and uses zein as a second layer of coating wall material to carry out double-layer embedding to obtain the nattokinase microcapsule. The puffed grains are loose and porous, have higher loading capacity and can adsorb the natto kinase liquid to a greater extent. The sodium alginate can form a protective film outside the puffed grain loaded with the nattokinase, and has a certain protective effect on the nattokinase. Because sodium alginate has porous property, and the protective effect of natto kinase is limited by coating with a certain glue substance alone, and the penetration of gastric juice cannot be completely and effectively limited, other materials are adopted for secondary coating. Zein has good film forming property, is insoluble in water but easily soluble in 60-95% ethanol solution, and is insoluble in acidic solution. Therefore, the zein is adopted to carry out the second coating on the nattokinase, and the nattokinase microcapsule with high embedding rate and storage rate is obtained. After the microcapsule is subjected to simulated stomach digestion for 2 hours, the nattokinase can still keep the enzyme activity of 90 percent or more.
Detailed Description
The present invention will be further described with reference to the following specific examples.
Example 1
Placing the bacillus natto in a liquid LB culture medium for liquid fermentation, freezing and centrifuging the fermentation liquor at 6000r/min for 20min at 4 ℃, wherein the supernatant is the crude nattokinase solution, then passing the crude nattokinase solution through 1 ten thousand ultrafiltration membranes, passing the trapped fluid through 3 ten thousand ultrafiltration membranes, and the permeate is the nattokinase solution to be coated.
Selecting corn as a puffing raw material, adding a certain amount of water according to the following formula, adjusting the water content to 16%, putting the corn into a double-screw extrusion puffing machine for extrusion puffing, setting the feeding speed of the extruder to be 14r/min, the screw rotating speed to be 100r/min, the processing temperature to be 150 ℃, drying and crushing the puffed corn for later use.
M- - -the total mass (g) of the added corn, rice and millet;
A0-final material moisture content (%);
A1、A2、A3-original moisture content (%) of corn, rice, millet;
C1、C2、C3ratio of corn, rice and millet (%)
Adding the nattokinase solution into the crushed puffed grains according to the mass ratio of the crushed puffed grains to the nattokinase solution of 1:1, and uniformly stirring to obtain a nattokinase-loaded puffed grain mixture; mixing the mixture with a composite sodium alginate gel solution with the mass concentration of 0.5% according to the mass ratio of 1:1, wherein the composite sodium alginate gel solution is prepared according to the mass ratio of 7:3 of sodium alginate to xanthan gum, and stirring uniformly; and then mixing the obtained product with a zein ethanol solution according to the mass ratio of 1:1, uniformly stirring, wherein the zein ethanol solution is prepared by dissolving zein in ethanol with the volume fraction of 60% according to the mass concentration of 2.5%, and carrying out vacuum freeze drying and crushing to obtain the nattokinase microcapsule.
Example 2
Placing the bacillus natto in a liquid LB culture medium for liquid fermentation, freezing and centrifuging fermentation liquor at 15 ℃ at 10000r/min for 15min, wherein the supernatant is crude nattokinase, then passing the crude nattokinase through 1 ten thousand ultrafiltration membranes, passing the trapped fluid through 3 ten thousand ultrafiltration membranes, and obtaining the permeate as the nattokinase to be coated.
Selecting corn and millet as puffing raw materials, adding a certain amount of water according to a formula with a material ratio of 8:1, adjusting the water content to 16%, putting the mixture into a double-screw extrusion puffing machine for extrusion puffing, setting the feeding speed of the extruder to be 16r/min, the screw rotating speed to be 110r/min and the processing temperature to be 160 ℃, drying and crushing the puffed grains for later use.
Adding the nattokinase solution into the crushed puffed grains according to the mass ratio of the crushed puffed grains to the nattokinase solution of 1.5:1, and uniformly stirring to obtain a nattokinase-loaded puffed grain mixture; mixing the mixture with a composite sodium alginate gel solution with the mass concentration of 1% according to the mass ratio of 1:2, wherein the mass ratio of the composite sodium alginate gel substances in the composite sodium alginate gel solution is sodium alginate to gellan gum to propolis is 5:2:3, and uniformly stirring; and then spraying the zein ethanol solution on the nattokinase-puffed grain-composite sodium alginate gel substance which is continuously stirred according to the mass ratio of 1:3 with the zein ethanol solution, wherein the zein ethanol solution is formed by dissolving zein in ethanol with the volume fraction of 60% according to the mass concentration of 2%, and carrying out pre-dehumidification cold air drying and screening to obtain the nattokinase microcapsule.
Example 3
Placing the bacillus natto in a liquid LB culture medium for liquid fermentation, freezing and centrifuging the fermentation liquor at 4 ℃ at 8000r/min for 20min, wherein the supernatant is the crude nattokinase solution, then passing the crude nattokinase solution through 1 ten thousand ultrafiltration membranes, passing the trapped fluid through 3 ten thousand ultrafiltration membranes, and the permeate is the nattokinase solution to be coated.
Selecting corn, millet and rice as puffing raw materials, adding a certain amount of water according to a material ratio of 6:2:2, adjusting the water content to 16%, putting into a double-screw extrusion puffing machine for extrusion puffing, setting the feeding speed of the extruder to be 18r/min, the screw rotation speed to be 120r/min, the processing temperature to be 180 ℃, drying and crushing the puffed grains for later use.
Adding the nattokinase solution into the crushed puffed grains according to the mass ratio of the crushed puffed grains to the nattokinase solution of 1.5:1, and uniformly stirring to obtain a nattokinase-loaded puffed grain mixture; mixing the mixture with a composite sodium alginate gel solution with the mass concentration of 2% according to the mass ratio of 1:1.5, wherein the mass ratio of the composite sodium alginate gel substances in the composite sodium alginate gel solution is that the mass ratio of sodium alginate to pectin to xanthan gum is 6:2:2, and uniformly stirring; and then spraying the zein ethanol solution on the nattokinase-puffed grain-composite sodium alginate gel substance which is continuously stirred according to the mass ratio of 1:2.5, wherein the zein ethanol solution is prepared by dissolving zein in ethanol with the volume fraction of 65% according to the mass concentration of 3%, drying by a fluidized bed and screening to obtain the nattokinase microcapsule.
The nattokinase microcapsule prepared in the embodiment is used as a sample, and the protection effect of the nattokinase embedding treatment on the nattokinase activity is detected by simulating gastric digestion: preparing simulated gastric electrolyte (containing pepsin), adjusting pH to 2, sampling 0.5g, adding 2ml of prepared gastric juice, keeping the mixture in a shaking table at 37 ℃ and 120r/min in a dark place for 2h, and measuring the enzymatic activity of the nattokinase stored in the microcapsules by adopting a fibrin plate method.
As a result: after the microcapsule is subjected to simulated digestion in stomach for 1h, the nattokinase can keep the enzyme activity of 90% or more. Compared with other nattokinase microcapsules, the microcapsule has obvious advantages.
The above description is only a detailed description of specific embodiments of the present invention, and should not be taken as limiting the invention, and any modifications, equivalents, improvements, etc. made on the design concept of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A preparation method of a nattokinase embedded substance is characterized by comprising the following steps:
(1) preparing natto kinase liquid: placing the bacillus natto in a liquid LB culture medium for liquid fermentation, freezing and centrifuging the fermentation liquor, obtaining supernatant fluid which is nattokinase crude enzyme liquid, and ultrafiltering the nattokinase crude enzyme liquid through an ultrafiltration membrane to obtain nattokinase liquid;
(2) preparing a low-expansion-degree cereal raw material: taking grain raw materials, adjusting the water content of the grains according to the water content of the materials being 10-20%, uniformly stirring, pouring the materials into a double-screw extrusion puffing machine for extrusion puffing, drying the puffed grains and crushing the puffed grains for later use;
(3) the puffed grain is loaded with nattokinase: adding the nattokinase solution into the crushed puffed grains according to the mass ratio of the crushed puffed grains to the nattokinase solution of 0.1-2: 1, and uniformly stirring to obtain a nattokinase-loaded puffed grain mixture;
(4) preparing microcapsules: adopting secondary coating, adding the coating material into the mixture obtained in the step (3) according to the mass ratio of the first layer of coating material to the mixture obtained in the step (3) of 1: 1-5 by adopting a composite sodium alginate gel solution with the mass concentration of 0.1-5% as a first layer of coating material, uniformly stirring, drying and crushing; and then carrying out second-layer coating with a zein ethanol solution according to the mass ratio of 1-10: 1, and drying to obtain the nattokinase microcapsule.
2. The method for preparing nattokinase-embedded material according to claim 1, wherein the nattokinase solution prepared in the step (1) is prepared by the following steps: placing the bacillus natto in a liquid LB culture medium for liquid fermentation, freezing and centrifuging fermentation liquor at 4-15 ℃ at 6000-12000 r/min for 10-30 min, wherein the supernatant is a crude nattokinase solution, passing the crude nattokinase solution through 1 ten thousand ultrafiltration membranes, passing the trapped fluid through 3 ten thousand ultrafiltration membranes, and obtaining a permeate liquid which is the nattokinase solution to be coated.
3. The method for preparing nattokinase-embedded material according to claim 2, wherein the nattokinase-embedded material comprises the following steps: the molecular weight of the nattokinase solution obtained in the step (1) is 27000-28000.
4. The method for preparing nattokinase-embedded product according to claim 1, wherein the low swelling degree cereal raw material of the step (2) is prepared by the following steps: the method comprises the steps of adopting grain raw materials, adding water according to the water content of 10-20% of the materials to adjust the water content of the grains, uniformly stirring, pouring into a double-screw extrusion puffing machine for extrusion puffing, setting the feeding speed of the extruder to be 14-18 r/min, the screw rotating speed to be 100-120 r/min, the processing temperature to be 150-180 ℃, drying the puffed grains and then crushing for later use.
5. The method for preparing nattokinase-embedded material according to claim 4, wherein the nattokinase-embedded material comprises the following steps: the grain raw materials are selected from one or two of rice, corn and millet according to a mass ratio of 1-10: 1-10 or three of the rice, the corn and the millet are mixed according to a mass ratio of 1-10: 1-10.
6. The method for preparing nattokinase-embedded material according to claim 1, comprising the steps of: the composite sodium alginate gel solution in the step (4) is prepared by matching sodium alginate with any one or two or three of pectin, xanthan gum, propolis and gellan gum; sodium alginate and any one of pectin, xanthan gum, propolis and gellan gum in a mass ratio of 1-10: 1-10, or sodium alginate and any two of the two substances in a mass ratio of 1-10: 1-10, or sodium alginate and any three of the substances in a mass ratio of 1-10: 1-10 are prepared into a 0.1-5% composite sodium alginate gel solution.
7. The method for preparing nattokinase-embedded material according to claim 1, comprising the steps of: and (4) when the second coating is carried out in the step (4), adding the zein ethanol solution into the mixture after the first coating, and uniformly mixing, or spraying the zein ethanol solution into the mixture after the first coating, and uniformly spraying.
8. The method for preparing nattokinase-embedded material according to claim 7, comprising the steps of: the zein ethanol solution is obtained by dissolving zein in a solvent with volume fraction of 60-95% ethanol according to the mass concentration of 0.1-5%.
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Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103053988A (en) * | 2012-12-20 | 2013-04-24 | 徐州工程学院 | Method for preparing microcapsule thermal reaction type shrimp flavor material by utilizing porous starch |
| CN103340408A (en) * | 2013-07-26 | 2013-10-09 | 湖南农业大学 | Rice antioxidant peptide microcapsule and preparation method thereof |
| CN105087539A (en) * | 2015-09-18 | 2015-11-25 | 黑龙江省科学院微生物研究所 | Preparation method of nattokinase microcapsule |
| CN106310243A (en) * | 2016-10-13 | 2017-01-11 | 武汉骏安生物科技有限公司 | Natto kinase capsule and preparation method thereof |
| CN106668844A (en) * | 2016-10-08 | 2017-05-17 | 中国农业科学院油料作物研究所 | Natto kinase microcapsule and preparation method thereof |
| CN109007807A (en) * | 2018-08-13 | 2018-12-18 | 上海应用技术大学 | A kind of microcapsules fruit ferment powder and preparation method thereof |
| CN109722427A (en) * | 2019-03-06 | 2019-05-07 | 武汉轻工大学 | A kind of preparation method of Nattokinase freeze-dried powder |
| CN111748502A (en) * | 2020-07-31 | 2020-10-09 | 黄河三角洲京博化工研究院有限公司 | Strain for high-yield nattokinase and method for preparing nattokinase product by using strain |
| CN111990642A (en) * | 2020-07-03 | 2020-11-27 | 蒋胜 | Anti-fatigue microcapsule with slow release effect, preparation method and application |
| CN112295514A (en) * | 2020-10-29 | 2021-02-02 | 湖北工业大学 | Preparation method and application of core-shell structure microcapsule particles |
-
2021
- 2021-12-16 CN CN202111540638.0A patent/CN114271483A/en active Pending
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103053988A (en) * | 2012-12-20 | 2013-04-24 | 徐州工程学院 | Method for preparing microcapsule thermal reaction type shrimp flavor material by utilizing porous starch |
| CN103340408A (en) * | 2013-07-26 | 2013-10-09 | 湖南农业大学 | Rice antioxidant peptide microcapsule and preparation method thereof |
| CN105087539A (en) * | 2015-09-18 | 2015-11-25 | 黑龙江省科学院微生物研究所 | Preparation method of nattokinase microcapsule |
| CN106668844A (en) * | 2016-10-08 | 2017-05-17 | 中国农业科学院油料作物研究所 | Natto kinase microcapsule and preparation method thereof |
| CN106310243A (en) * | 2016-10-13 | 2017-01-11 | 武汉骏安生物科技有限公司 | Natto kinase capsule and preparation method thereof |
| CN109007807A (en) * | 2018-08-13 | 2018-12-18 | 上海应用技术大学 | A kind of microcapsules fruit ferment powder and preparation method thereof |
| CN109722427A (en) * | 2019-03-06 | 2019-05-07 | 武汉轻工大学 | A kind of preparation method of Nattokinase freeze-dried powder |
| CN111990642A (en) * | 2020-07-03 | 2020-11-27 | 蒋胜 | Anti-fatigue microcapsule with slow release effect, preparation method and application |
| CN111748502A (en) * | 2020-07-31 | 2020-10-09 | 黄河三角洲京博化工研究院有限公司 | Strain for high-yield nattokinase and method for preparing nattokinase product by using strain |
| CN112295514A (en) * | 2020-10-29 | 2021-02-02 | 湖北工业大学 | Preparation method and application of core-shell structure microcapsule particles |
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