CN113197281A - Oat fiber with lipase and amylase inhibition activity and preparation method and application thereof - Google Patents
Oat fiber with lipase and amylase inhibition activity and preparation method and application thereof Download PDFInfo
- Publication number
- CN113197281A CN113197281A CN202110363167.4A CN202110363167A CN113197281A CN 113197281 A CN113197281 A CN 113197281A CN 202110363167 A CN202110363167 A CN 202110363167A CN 113197281 A CN113197281 A CN 113197281A
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- China
- Prior art keywords
- oat
- lipase
- fiber
- amylase
- water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Abstract
The invention discloses oat fiber with lipase and amylase inhibiting activity, a preparation method and application thereof, and belongs to the field of nutrition and health. The oat fiber rich in lipase inhibitor and amylase inhibitor is prepared by taking the fiber-rich tissue part of oat as a raw material and matching with a specific large-scale radio frequency sterilization mode, an enzymolysis extraction mode and a physical adsorption mode, and has excellent lipase and amylase inhibition activity. Has the characteristics of high purity and good inhibition rate of active ingredients. The oat fiber prepared by the method has obvious inhibition effect on small intestine alpha-amylase and lipase, thereby having the functions of reducing blood sugar and losing weight, being capable of obviously improving intestinal health, controlling blood sugar rise and reducing grease absorption, and providing a new solution for diabetes patients and fat foods.
Description
Technical Field
The invention relates to oat fiber with lipase and amylase inhibition activity, a preparation method and application thereof, and belongs to the field of nutrition and health.
Background
In modern fast-paced life, the daily dietary structure of domestic people usually takes cereals as the main part and vegetables, fruits and meats as the auxiliary parts, especially in recent years, the proportion of dietary fat of residents in China continuously rises, the intake of dietary fiber is obviously reduced, and therefore the number of 'three high population' with hyperglycemia, hypertension and hyperlipidemia rises, and meanwhile, other diseases caused by too little intake of dietary fiber are as follows: the incidence of constipation, intestinal polyps, intestinal cancer and the like is also increasing.
The dietary fiber has positive effect on human health, and has outstanding function in preventing human gastrointestinal tract health. The wheat plants are rich in alpha-amylase inhibitors and lipase inhibitors. Meanwhile, through the certification of the United states Food and Drug Administration (FDA), the water-soluble dietary fiber beta-glucan in the oat is an effective component for reducing blood sugar and blood fat and preventing cardiovascular diseases, can obviously reduce the incidence of the cardiovascular diseases, and can also activate macrophages and improve immunity. The research results of papers published in the international journal 'nutriment' of Germany human nutrition research show that the water-insoluble dietary fiber has positive influence on controlling long-term blood sugar value, is beneficial to preventing type 2 diabetes, and for people with high fasting blood sugar value, the water-insoluble dietary fiber can be taken frequently to improve the blood sugar tolerance; for obese people, the multiple intake of water-insoluble dietary fiber can reduce the level of inflammation in vivo.
However, currently, much attention is paid to extraction rate, structural modification and extraction of beta-glucan for extraction of oat dietary fiber, such as: oxford and the like (influence of ultrasonic modification on physicochemical properties and structures of oat dietary fibers, food science 2020) extract the dietary fibers from the oats, and the oat dietary fibers are modified by adopting an ultrasonic technology, so that the water holding capacity, the water swelling capacity and the oil holding capacity of the oat dietary fibers are obviously improved; liu De Teng and the like (research on an oat bran dietary fiber extraction process, preservation and processing, 2013) research on the extraction rate of dietary fiber extracted from oat bran by an enzyme-alkali combination method and reaches 56.43 percent; the liroc (oat bran extrusion modification process optimization and efficacy research, university of rock river, 2012) optimizes the oat bran extrusion modification process, and the yield of oat beta-glucan is 136.95 mg/g; liu Jing Xue (oat bran dietary fiber extrusion modification process, food industry, 2019) improves the content of soluble dietary fiber in oat bran dietary fiber to 8.8% by optimizing oat fiber extrusion modification process parameters, and improves the content of the soluble dietary fiber by 29.4% compared with the soluble dietary fiber of the original oat bran dietary fiber, and the like.
Cereals are a good source of amylase and lipase inhibitors, such as: liuwen ganoderma and the like (separation and purification and property research of alpha-amylase inhibitor in sticky corn, food industry science and technology, 2009) prepare the alpha-amylase inhibitor from the sticky corn, and the activity of the alpha-amylase inhibitor reaches 165.8U/mg; the yield of the alpha-amylase inhibitor reaches 4.67 percent by improving the extraction process of the white kidney beans, such as the extraction of the alpha-amylase inhibitor in the white kidney beans and the performance research thereof, food science and technology, 2017. Wang Yanfei et al (research on inhibition mechanism of pancreatic lipase inhibitor in rice germ, food technology, 2004) extracted pancreatic lipase inhibitor from rice germ and clarified inhibition mechanism; trypan and the like (research on the pancreatic lipase inhibition effect of tartary buckwheat polysaccharide, Chinese food additive 2015) research on the pancreatic lipase inhibition effect of tartary buckwheat, and elucidation that the pancreatic lipase inhibition rate of tartary buckwheat polysaccharide is 52.71% by changing the pancreatic lipase conformation, and the like. There are few reports of amylase and lipase inhibitors extraction from oats compared to other cereals, and the concern is low, resulting in loss of active ingredients during oat fiber extraction.
The active ingredients in common cereal oats, in addition to the well-known oat beta-glucan, also contain oat fiber with a high activity of lipase inhibitors and amylase inhibitors. However, extracting active ingredients from oats neglects lipase inhibitors and amylase inhibitors of which activity is higher due to the focus on oat beta-glucan in the oat. The water-soluble fiber in the oat fiber accounts for 9.2 percent of the total dietary fiber, and the production process focuses more on oat beta-glucan, so much attention has not been paid to the extraction of lipase inhibitors and amylase inhibitors which are rich in water-insoluble fiber, and the indirect loss of the functionality of the oat dietary fiber, such as the functionality of inhibiting the absorption of saccharides and lipid substances, preventing the increase of blood sugar and reducing the content of cholesterol in the body, is caused.
In conclusion, research and development of oat fiber rich in lipase inhibitors and amylase inhibitors to improve the lipase and amylase inhibition activity are urgent requirements for widening the application direction of oat dietary fiber and the food nutrition and health field.
Disclosure of Invention
The purpose of the invention is as follows: aiming at the problems, the invention develops the oat fiber with lipase and amylase inhibition activity and the preparation method and the application thereof, and solves the technical problem of extracting lipase inhibitors and amylase inhibitors from water-insoluble oat fiber.
The technical scheme is as follows: the technical scheme adopted by the invention is as follows:
one object of the present invention is to provide a method for preparing oat fiber with lipase and amylase inhibition activity, which comprises the following steps:
(1) enzymolysis: dissolving fiber powder of oat in water, adding amylase, protease, lipase, cellulase and anthocyanin enzyme for enzymolysis;
(2) solid-liquid separation: carrying out solid-liquid separation on the enzymolysis liquid after enzymolysis to obtain a solid phase and a liquid phase; decolorizing and extracting active ingredients by enzymolysis, specifically hydrolyzing with anthocyanidin enzyme to remove pigment; residual starch, protein and lipid in the waste water are removed by hydrolyzing amylase, protease and lipase; cellulose in the enzymolysis solution is decomposed by hydrolysis of cellulase, so that the dissolution rate of water-soluble beta-glucan in the enzymolysis solution is improved, and the extraction rates of a water-insoluble amylase inhibitor and a lipase inhibitor are improved;
(3) and respectively drying the solid phase and the liquid phase of the enzymatic hydrolysate to obtain the oat fiber, wherein the oat fiber obtained from the liquid phase is rich in beta-glucan, and the oat fiber obtained from the solid phase is rich in amylase inhibitor and lipase inhibitor, so that the oat fiber has excellent lipase and amylase inhibition activity.
Further, in the step (1), the amylase, the protease, the lipase, the cellulase and the anthocyanidin are added in an amount of 0.5-0.9% (w/w) of the total amount of the fiber powder of oat and water, respectively.
Further, in the step (1), the ratio of the fiber powder of the oat to the water is 0.8-1.2 (w/v).
Further, in the step (1), the enzymolysis conditions are as follows: stirring at 55 deg.C and pH of 5.0-6.0 for 1-2 hr.
Further, in the step (1), the fiber powder is prepared by crushing the fiber-rich tissue part of the oat, removing starch and sterilizing; the method for removing the starch comprises air flow classification, and the sterilization method comprises any one of radio frequency sterilization, ultrasonic sterilization and high-temperature high-pressure sterilization.
Further, the air flow classification condition is 10-15 m/s; the radio frequency sterilization conditions are as follows: distance between polar plates14-16cm, field strength of 0.42 +/-0.03 kV/cm, power density of 2.42 +/-0.03W/g, heat resistance time of 10-15min, and total energy consumption of 2.13X 10 under the condition3-2.16×103J/g。
Furthermore, the solid-liquid separation adopts a static adsorption and NaCl elution mode and adopts D315Resin adsorption, the operating conditions are as follows: adjusting pH to 7.0-8.5, and the protein concentration of the sample liquid is C025-35 g/L, the temperature t is 28-32 ℃, and the rotation speed omega is 150-250 rpm; the concentration of the NaCl solution used for NaCl elution is 0.5-0.8 mol/L. Separating enzymolysis liquid after enzymolysis, and adopting D315Extracting enzymolysis liquid rich in beta-glucan, amylase inhibitor and lipase inhibitor by resin physical adsorption and NaCl elution.
Further, in the step (3), after the solid phase and the liquid phase are respectively dried, the water-insoluble oat fiber and the water-soluble oat fiber are correspondingly obtained. The obtained water-insoluble oat fiber is rich in lipase inhibitor and amylase inhibitor, so that after the water-insoluble oat fiber and the water-soluble oat fiber are mixed, the oat fiber which is rich in lipase inhibitor and amylase inhibitor and has lipase and amylase inhibition activity can be obtained.
Further, in the step (3), the drying manner includes freeze drying, drying under normal pressure/reduced pressure, spray drying, and the like.
It is another object of the present invention to provide an oat fiber having excellent lipase and amylase inhibitory activity.
Another object of the present invention is to provide the use of oat fiber with excellent lipase and amylase inhibitory activity in the fields of food and pharmaceutical products, including but not limited to general food, health food, and formula food for special medical use, and pharmaceutical products, including but not limited to hypoglycemic agents, for achieving or improving the effects of controlling blood sugar, controlling blood lipid, enhancing immunity, etc.
The invention has the beneficial effects that:
(1) the invention provides a lipase inhibitor and an amylase inhibitor which are extracted from water-insoluble fiber in oat by an enzymatic hydrolysis method, and the obtained lipase and amylase have excellent inhibition activity.
(2) The invention adopts physical adsorption and elution methods for purification and enrichment, is nontoxic and harmless, and has little influence on environmental pollution.
(3) The method adopts a large-scale radio frequency-containing method for sterilization, improves the flavor of the raw material for extracting the oat dietary fiber and reduces the sour taste and the bitter taste while achieving the sterilization effect.
(4) The raw materials of the oat dietary fiber prepared by the invention are all food grade, and can be applied to the food fields of common food, health food, formula food with special medical application and the like. It can improve intestinal health, control blood sugar rise, reduce oil absorption, and provide new solution for diabetic and obesity food.
Drawings
FIG. 1 is a flavor radar chart of products obtained in examples 1 to 5 and comparative examples 1 to 2.
FIG. 2 is a graph showing the inhibitory effect of the products obtained in examples 1 to 5 and comparative examples 1 to 2 on α -amylase.
FIG. 3 is a graph showing the lipase inhibitory effects of the products obtained in examples 1 to 5 and comparative examples 1 to 2.
FIG. 4 shows the dietary fiber contents of the products obtained in examples 1 to 5 and comparative examples 1 to 2.
Detailed Description
One object of the present invention is to provide a method for preparing oat fiber with lipase and amylase inhibition activity, which comprises the following steps:
(1) crushing: crushing the fiber-rich tissue part of the oat, and sieving the crushed oat with a 80-mesh sieve to obtain fiber powder;
(2) enzymolysis: dissolving the fiber powder in water, adding amylase, protease, lipase, cellulase and anthocyanin enzyme for enzymolysis, decoloring and extracting active ingredients through enzymolysis, specifically hydrolyzing the anthocyanin enzyme to remove pigments in the fiber powder; residual starch, protein and lipid in the waste water are removed by hydrolyzing amylase, protease and lipase; cellulose in the enzymolysis solution is decomposed by hydrolysis of cellulase, so that the dissolution rates of beta-glucan, an amylase inhibitor and a lipase inhibitor in the enzymolysis solution are improved;
(3) solid-liquid separation: carrying out solid-liquid separation on the enzymolysis liquid after enzymolysis to respectively obtain a liquid phase rich in oat water-soluble fiber and a solid phase rich in oat water-insoluble fiber;
(4) and (3) drying: drying the solid phase and the liquid phase after enzymolysis, and mixing to obtain the high-purity oat fiber rich in beta-glucan, lipase inhibitor and amylase inhibitor and having lipase and amylase inhibition activity.
The following description of the preferred embodiments of the present invention is provided for the purpose of better illustrating the invention and is not intended to limit the invention thereto.
Example 1
A method for preparing oat fiber with lipase and amylase inhibiting activity is provided. The method comprises the following specific steps:
(1) 1000g of the fiber-rich tissue part of oat is pulverized, starch in the oat is removed by an air flow classification method (the air flow classification condition is 15m/s), and then sterilization treatment is carried out by a radio frequency sterilization method. The radio frequency sterilization conditions are as follows: the pole plate spacing RF is 14cm, the field strength is 0.42 +/-0.03 kV/cm, the power density is 2.42 +/-0.03W/g, the heat resistance time is 15min, and the total energy consumption is 2.16 multiplied by 103J/g。
(2) Adding 500g of the powder obtained in the step (1) into 500ml of distilled water, and adding amylase, protease, lipase, cellulase and anthocyanin into the distilled water, wherein the adding amount (mass ratio to substrate) of the enzyme is respectively as follows: 0.9%, 0.9% and 0.5%, adjusting pH to 5.5, temperature to 55 deg.C, and stirring for 1 hr for decolorizing and dissolving active components.
(3) And (3) carrying out solid-liquid separation, specifically extracting lipase inhibitor and amylase inhibitor in the enzymolysis liquid by adopting physical adsorption and elution modes. By using D315Resin adsorption, the adsorption conditions are as follows: adjusting pH to 7.0, and the protein concentration of the sample feed liquid to C030g/L, 30 deg.C and 150 rpm. Then eluted with 0.5mol/L NaCl.
(4) Drying the solid phase separated in the step (3) in a reduced pressure drying mode; and (4) drying the liquid phase separated in the step (3) by adopting a spray drying mode. Respectively obtaining high-purity water-insoluble oat fiber and water-soluble oat fiber rich in amylase inhibitor and lipase inhibitor.
Example 2
The preparation method of the oat fiber with lipase and amylase inhibition activity is the same as that of the oat fiber in example 1, and is different from the preparation method in process parameters, and the preparation method comprises the following specific steps:
(1) 1000g of the fiber-rich tissue part of oat is pulverized, starch in the oat is removed by an air flow classification method (the air flow classification condition is 10m/s), and then sterilization treatment is carried out by a radio frequency sterilization method. The radio frequency sterilization conditions are as follows: the distance RF between the polar plates is 15cm, the field intensity is 0.42 +/-0.03 kV/cm, the power density is 2.42 +/-0.03W/g, the heat resistance time is 10min, and the total energy consumption is 2.17 multiplied by 103J/g。
(2) Adding 500g of the powder obtained in the step (1) into 400ml of distilled water, and adding amylase, protease, lipase, cellulase and anthocyanin into the distilled water, wherein the adding amount (mass ratio to substrate) of the enzyme is respectively as follows: 0.5%, 0.5% and 0.6%, adjusting pH to 5.0, temperature to 53 deg.C, and time to 1.5 hr, stirring continuously, decolorizing and dissolving active ingredients.
(3) And (3) carrying out solid-liquid separation on the enzymolysis liquid, and specifically extracting a lipase inhibitor and an amylase inhibitor in the enzymolysis liquid by adopting physical adsorption and elution modes. By using D315Resin adsorption, the adsorption conditions are as follows: adjusting pH to 7.0, and the protein concentration of the sample liquid to C030g/L, 30 deg.C and 150 rpm. Then eluted with 0.5mol/L NaCl.
(4) Drying the solid phase separated in the step (3) in a reduced pressure drying mode; and (4) drying the liquid phase separated in the step (3) by adopting a spray drying mode. Respectively obtaining high-purity water-insoluble oat fiber and water-soluble oat fiber rich in amylase inhibitor and lipase inhibitor.
Example 3
The preparation method of the oat fiber with lipase and amylase inhibition activity is the same as that of the oat fiber in example 1, and is different from the preparation method in process parameters, and the preparation method comprises the following specific steps:
(1) 1000g of the fiber-rich tissue part of oat is pulverized, starch in the oat is removed by an airflow classification method (the airflow classification condition is 13m/s), and then sterilization treatment is carried out by a radio frequency sterilization method. The radio frequency sterilization conditions are as follows: the pole plate spacing RF is 16cm, the field strength is 0.42 +/-0.03 kV/cm, the power density is 2.42 +/-0.03W/g, the heat resistance time is 13min, and the total energy consumption is 2.18 multiplied by 103J/g。
(2) Adding 500g of the powder obtained in the step (1) into 600ml of distilled water, and adding amylase, protease, lipase, cellulase and anthocyanin into the distilled water, wherein the adding amount (mass ratio to substrate) of the enzyme is respectively as follows: 0.7%, 0.7% and 0.7%, adjusting pH to 6.0, temperature to 50 deg.C, and stirring for 2 hr for decolorizing and dissolving active components.
(3) And (3) carrying out solid-liquid separation on the enzymolysis liquid, and specifically extracting a lipase inhibitor and an amylase inhibitor in the enzymolysis liquid by adopting physical adsorption and elution modes. By using D315Resin adsorption, the adsorption conditions are as follows: adjusting pH to 7.0, and the protein concentration of the sample liquid to C030g/L, 30 deg.C and 150 rpm. Then eluted with 0.5mol/L NaCl.
(4) Drying the solid phase separated in the step (3) in a reduced pressure drying mode; and (4) drying the liquid phase separated in the step (3) by adopting a spray drying mode. Respectively obtaining high-purity water-insoluble oat fiber and water-soluble oat fiber rich in amylase inhibitor and lipase inhibitor.
Example 4
A method for preparing oat fiber with lipase and amylase inhibiting activity is provided. The preparation method is the same as example 1, except that the sterilization mode is ultrasonic sterilization, and the specific steps are as follows:
(1) 1000g of the fiber-rich tissue part of oat is pulverized, starch in the pulverized oat is removed by an airflow classification method (the airflow classification condition is 15m/s), and then the pulverized oat is sterilized by ultrasonic wave. And (3) sterilization conditions: the power is 50W, the amplitude of the bottom of the container is 10.5 mu m, and the time is 15 min.
(2) Adding 500g of the powder obtained in the step (1) into 500ml of distilled water, and adding amylase, protease, lipase, cellulase and anthocyanin into the distilled water, wherein the adding amount (mass ratio to substrate) of the enzyme is respectively as follows: 0.9%, 0.9% and 0.5%, adjusting pH to 5.5, temperature to 55 deg.C, and stirring for 1 hr for decolorizing and dissolving active components.
(3) And (3) carrying out solid-liquid separation on the enzymolysis liquid, and specifically extracting a lipase inhibitor and an amylase inhibitor in the enzymolysis liquid by adopting physical adsorption and elution modes. By using D315Resin adsorption, the adsorption conditions are as follows: adjusting pH to 7.0, and the protein concentration of the sample liquid to C030g/L, 30 deg.C and 150 rpm. Then eluted with 0.5mol/L NaCl.
(4) Drying the solid phase separated in the step (3) in a reduced pressure drying mode; and (4) drying the liquid phase separated in the step (3) by adopting a spray drying mode. Respectively obtaining high-purity water-insoluble oat fiber and water-soluble oat fiber rich in amylase inhibitor and lipase inhibitor.
Example 5
A method for preparing oat fiber with lipase and amylase inhibiting activity is provided. The preparation method is the same as example 1, except that the sterilization mode is high-temperature high-pressure sterilization, and the specific steps are as follows:
(1) 1000g of the fiber-rich tissue part of oat is pulverized, starch in the oat is removed by an airflow classification method (the airflow classification condition is 15m/s), and then the oat is sterilized by a high-temperature and high-pressure sterilization method. And (3) sterilization conditions: the temperature is 120 ℃ and the time is 20 min.
(2) Adding 500g of the powder obtained in the step (1) into 500ml of distilled water, and adding amylase, protease, lipase, cellulase and anthocyanin into the distilled water, wherein the adding amount (mass ratio to substrate) of the enzyme is respectively as follows: 0.9%, 0.9% and 0.5%, adjusting pH to 5.5, temperature to 55 deg.C, and stirring for 1 hr for decolorizing and dissolving active components.
(3) And (3) carrying out solid-liquid separation on the enzymolysis solution, and specifically extracting a lipase inhibitor and an amylase inhibitor in a solid phase of the enzymolysis solution by adopting physical adsorption and elution modes. Adsorbing with D315 resin under the following conditions: adjusting pH to 7.0, and the protein concentration of the sample liquid to C030g/L, 30 deg.C and 150 rpm. Then eluted with 0.5mol/L NaCl.
(4) Drying the solid phase separated in the step (3) in a reduced pressure drying mode; and (4) drying the liquid phase separated in the step (3) by adopting a spray drying mode. Respectively obtaining high-purity water-insoluble oat fiber and water-soluble oat fiber rich in amylase inhibitor and lipase inhibitor.
To better explain the remarkable effects of the present invention, the comparative examples were added as follows:
comparative example 1
A method for preparing oat fiber with lipase and amylase inhibiting activity is provided. The preparation method is the same as example 1, and is different from the method for extracting the lipase inhibitor and the amylase inhibitor without physical adsorption and elution, and the specific steps are as follows:
(1) 1000g of the fiber-rich tissue part of oat is pulverized, starch in the oat is removed by an air flow classification method (the air flow classification condition is 15m/s), and then sterilization treatment is carried out by a radio frequency sterilization method. The radio frequency sterilization conditions are as follows: the plate spacing RF is 14cm, the field strength is 0.42 +/-0.01 kV/cm, the power density is 2.42 +/-0.03W/g, the heat resistance time is 15min, and the total energy consumption is 2.16 multiplied by 103J/g。
(2) Adding 500g of the powder obtained in the step (1) into 500ml of distilled water, and adding amylase, lipase, cellulase and anthocyanin into the distilled water, wherein the addition amount (mass ratio of the enzyme to the substrate) of the enzyme is respectively as follows: 0.9%, 0.9% and 0.5%, adjusting pH to 5.5, temperature to 55 deg.C, and time to 1h, stirring continuously, decolorizing and dissolving active ingredients. Carrying out solid-liquid separation on the enzymolysis liquid by adopting a common precipitation separation method to obtain a solid phase and a liquid phase;
(3) drying the solid phase separated in the step (3) in a reduced pressure drying mode; and (4) drying the liquid phase separated in the step (3) by adopting a spray drying mode. Respectively obtaining water-insoluble oat fiber and water-soluble oat fiber.
Comparative example 2
The preparation method of the oat dietary fiber is the same as that in example 1, the difference is that the enzyme used for enzyme extraction is different, and physical adsorption is changed into a chemical separation method, and the specific steps are as follows:
(1) 1000g of the fiber-rich tissue part of oat is pulverized, starch in the oat is removed by an air flow classification method (the air flow classification condition is 15m/s), and then sterilization treatment is carried out by a radio frequency sterilization method. The radio frequency sterilization conditions are as follows: the plate spacing RF is 14cm, the field strength is 0.42 +/-0.01 kV/cm, the power density is 2.42 +/-0.03W/g, the heat resistance time is 15min, and the total energy consumption is 2.16 multiplied by 103J/g。
(2) Adding the powder obtained in the step (1) into pre-boiled water according to the feed-liquid ratio of 1:8(w/v), boiling for 20min, adding a proper amount of water, cooling to 55 ℃, adding a mixed preparation of amylase and saccharifying enzyme with the mass fraction of 0.5%, preserving heat, stirring and hydrolyzing for 100min, and hydrolyzing starch remained in the bran so as to remove impurities.
(3) And (3) adding NaOH with the mass fraction of 5% into the enzyme water obtained in the step (2), and hydrolyzing for 100min at the temperature of 60 ℃ to fully soften the oat.
(4) Washing the softened oat obtained in the step (3) with tap water to be neutral.
(5) Putting the washed oat obtained in the step (4) into 5% of H according to the feed-liquid ratio of 20:1(w/v)2O2Soaking in the water solution at 50 deg.C for 120min for bleaching, and cleaning with clear water.
(6) And (4) centrifugally dewatering the oat fiber obtained in the step (5), and putting the oat fiber into a drying oven at 70 ℃ until the oat fiber is completely dried.
Characterization and comparison of results for examples and comparative examples:
1. flavour material
The flavor substances of the bottom material are extracted from the sterilized oat fiber powder by adopting an electronic tongue test, and comprise sour taste, astringent taste, bitter taste, delicate flavor, salty taste, bitter aftertaste and astringent aftertaste. The samples include examples 1-5 and comparative examples 1-2, and the method comprises the following steps: 1g of the powder obtained in examples 1-5 and comparative examples 1-2 was put into 100ml of distilled water and stirred for 1min, and then centrifuged to take the supernatant and tested by an electronic tongue under the following experimental conditions: maximum voltage 1.00V, minimum voltage-1.00V, potential step 0.1V, initial pulse 1.00S, pulse width 0.50S. The results are shown in Table 1:
TABLE 1 flavor substances of oat fiber powder extraction base material after sterilization in different sterilization modes
As can be seen from FIG. 1, examples 1 to 3 have a significant improvement effect on various flavors, compared to the oat fiber meal extraction substrate prepared in examples 4 to 5 and comparative examples 1 to 2. Firstly, the unpleasant sour taste, bitter taste, astringent taste, aftertaste and aftertaste are improved obviously, especially the three flavors of sour taste, bitter taste and astringent taste. Secondly, example 1 has a clear advantage in umami taste for both the pleasant salty and umami flavors, while it is relatively more neutral in salty taste. These flavors play an important role in the application of oat fiber powder in food.
2. Inhibitory Effect of alpha-Amylase
The inhibition effect of the products of the specific examples and the comparative examples on the alpha-amylase is determined by adopting a 3, 5-nitrosalicylic acid (DNS) colorimetric method (namely, a Bernfeld method), the dosage of the alpha-amylase in the experiment is 2.5U, the concentration of each sample is 10mg/mL, and the test results are shown in Table 2:
TABLE 2 inhibition of alpha-amylase by different samples
The inhibition effect of the products of the above specific examples and comparative examples on lipase was determined by the olive oil emulsification method, and the test results are shown in table 3:
TABLE 3 inhibition of lipase by different samples
From the data in table 2 and table 3, it can be found that the inhibition rates of the oat dietary fibers prepared in examples 1-5 on alpha-amylase and lipase are obviously improved compared with the inhibition rates of the oat dietary fibers prepared in comparative examples 1-2, especially the inhibition effect obtained in examples 1-3 is the best, and the results again prove the effectiveness of purifying and enriching the alpha-amylase and lipase in the preparation method through radio frequency sterilization, the type of enzyme used in enzymolysis, physical adsorption and elution. Referring to fig. 2 and 3, the increase of the inhibition rate of alpha-amylase and lipase proves that the oat dietary fiber prepared by the method can control the increase of blood sugar and reduce the absorption of oil and fat, and provides a new solution for diabetic and obese food.
3. Purity of oat fiber
The results of the test on the total dietary fiber content in the above specific examples and comparative examples using the method of AOAC 2009.01 are shown in table 4:
TABLE 4 Total dietary fiber content in different samples
As can be seen from the data in Table 4 and FIG. 4, the oat dietary fiber obtained in example 1 has the highest content compared with comparative examples 1-4, and the difference of the preparation process that the content is obviously improved is that D315Resin adsorption and NaCl elution. The purification method has certain reference significance for improving the purity of the extracted cereal dietary fiber.
In conclusion, the invention firstly adopts the airflow classification method to change the proportion of each component in the raw materials and improve the relative content of the dietary fiber of the raw materials.
Secondly, a large-scale radio frequency sterilization mode is adopted, the temperature is controlled to be below 70 ℃, the distance between the polar plates is set to be 14cm, and the sterilization device has obvious effects on reducing sterilization time, reducing energy consumption, reducing the water content of products and improving the sour taste and the bitter taste of the products.
In addition, the residual protein and fat components in the oat soluble dietary fiber can be removed to the maximum extent by adopting an enzymatic method for decolorization and extraction, the yield of the oat soluble dietary fiber is improved, and the enzymatic method for extracting the dietary fiber has mild conditions, small environmental pollution and better development prospect.
Furthermore, D315Compared with the existing common chemical separation method, the resin adsorption and NaCl elution method has the advantages of higher extraction purity, lower cost, simpler operation of the precipitation elution process and less influence on the activity of the enzyme inhibitor.
The key point of the invention lies in that the application of enzymatic extraction is adopted, and the method is effectively combined with physical adsorption and spray drying, the oat dietary fiber can be obtained by simple extraction, and the oat dietary fiber rich in amylase inhibitor and lipase inhibitor prepared by the principle has the characteristics of high purity, high content of water-soluble dietary fiber, high activity of amylase inhibitor and lipase inhibitor, and excellent water holding capacity, water swelling capacity and oil holding capacity. The production process is non-toxic and harmless, has little pollution to the environment and has good industrial application prospect.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (10)
1. A method for preparing oat fiber with lipase and amylase inhibiting activity is characterized by comprising the following steps:
(1) enzymolysis: dissolving fiber powder of oat in water, adding amylase, protease, lipase, cellulase and anthocyanin enzyme for enzymolysis;
(2) solid-liquid separation: carrying out solid-liquid separation on the enzymolysis liquid after enzymolysis to obtain a solid phase and a liquid phase;
(3) and respectively drying the solid phase and the liquid phase of the enzymolysis solution to obtain the oat fiber.
2. The method for preparing oat fiber with lipase and amylase inhibiting activity according to claim 1, wherein in the step (1), amylase, protease, lipase, cellulase and anthocyanidin enzyme are added in an amount of 0.5-0.9% (w/w) of total amount of fiber powder and water of oat, respectively.
3. The method for preparing oat fiber with lipase and amylase inhibiting activity according to claim 1, wherein in the step (1), the ratio of fiber powder of oat to water is 0.8-1.2 (w/v).
4. The method for preparing oat fiber with lipase and amylase inhibiting activity according to claim 1, wherein in the step (1), the enzymolysis condition is as follows: stirring at pH 5.0-6.0 and 50-55 deg.C for 1-2 hr.
5. The method for preparing oat with lipase and amylase inhibiting activity according to claim 1, wherein in the step (1), the fiber powder of oat is prepared by pulverizing fiber-rich tissue part of oat, removing starch and sterilizing; the method for removing the starch comprises air flow classification, and the sterilization method comprises any one of radio frequency sterilization, ultrasonic sterilization and high-temperature high-pressure sterilization.
6. The method of claim 5, wherein the conditions for the air flow fractionation are 10-15 m/s; the radio frequency sterilization conditions are as follows: the space between the polar plates is 14-16cm, the field intensity is 0.42 +/-0.03 kV/cm, the power density is 2.42 +/-0.03W/g, the heat resistance time is 10-15min, and the total energy consumption under the condition is 2.13 multiplied by 103-2.16×103J/g。
7. The method for preparing lipase and amylase with inhibiting activity according to claim 1, wherein in the step (2), the solid-liquid separation is performed by adopting a static adsorption mode and a NaCl elution mode, and the static adsorption mode adopts D315Resin adsorption, the operating conditions are as follows: adjusting pH to 7.0-8.5, and the protein concentration of the sample liquid is C025-35 g/L, the temperature t is 28-32 ℃, and the rotation speed omega is 150-250 rpm; the concentration of the NaCl solution used for NaCl elution is 0.5-0.8 mol/L.
8. The method for preparing oat flour having lipase and amylase inhibitory activity according to claim 1, wherein in the step (3), after the solid phase and the liquid phase are dried respectively, water-insoluble oat fiber and water-soluble oat fiber are obtained; the drying mode comprises any one of freeze drying, normal pressure/reduced pressure drying and spray drying.
9. Oat fiber having lipase and amylase inhibiting activity prepared according to the method of any one of claims 1-8.
10. The use of oat fiber with lipase and amylase inhibiting activity in the fields of food and medicine according to claim 9, wherein the food includes but is not limited to general food, health food, and formula food for special medical use, and the medicine includes but is not limited to hypoglycemic.
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