CN114874356B - Method for extracting arabinoxylan from wheat bran as raw material, arabinoxylan and application - Google Patents
Method for extracting arabinoxylan from wheat bran as raw material, arabinoxylan and application Download PDFInfo
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- CN114874356B CN114874356B CN202210496576.6A CN202210496576A CN114874356B CN 114874356 B CN114874356 B CN 114874356B CN 202210496576 A CN202210496576 A CN 202210496576A CN 114874356 B CN114874356 B CN 114874356B
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0057—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Xylans, i.e. xylosaccharide, e.g. arabinoxylan, arabinofuronan, pentosans; (beta-1,3)(beta-1,4)-D-Xylans, e.g. rhodymenans; Hemicellulose; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
- A23L33/25—Synthetic polymers, e.g. vinylic or acrylic polymers
- A23L33/26—Polyol polyesters, e.g. sucrose polyesters; Synthetic sugar polymers, e.g. polydextrose
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention discloses a method for extracting arabinoxylan by taking wheat bran as a raw material, which comprises the following steps: cleaning wheat bran for multiple times, drying, crushing and sieving for later use; extraction pretreatment: carrying out weak base treatment; extruding and puffing; extraction treatment: extracting with water, centrifuging, and collecting supernatant; adjusting the pH value, centrifuging again, and collecting supernatant; vacuum concentrating the collected supernatant, adding ethanol for precipitation, centrifuging, and collecting precipitate; after redissolving, dialyzing and removing salt components in the solution; vacuum concentrating, and freeze drying to obtain arabinoxylan. The method reduces pollution while improving extraction efficiency, can fully utilize wheat bran resources, realizes the added value of the wheat bran resources, and changes waste into valuable; the extraction method is simple, the cost is low, the pollution is small, and the arabinoxylan prepared by the method has higher level of soluble dietary fiber and good application prospect.
Description
Technical Field
The invention belongs to the technical field of agricultural and sideline product deep processing, and particularly relates to a method for extracting arabinoxylan from wheat bran, the arabinoxylan and application of the arabinoxylan.
Background
Dietary fiber refers to a plant-derived food component that cannot be decomposed by human digestive enzymes, and is mainly non-starch polysaccharides and lignin. Earlier researches show that the soluble dietary fiber has good relieving effect on hypertension, hyperlipidemia, type II diabetes, obesity and the like, and can effectively enhance gastrointestinal tract immunity and improve intestinal health. With the improvement of living standard of people, the proportion of three-high foods and fine foods in daily diet of people is continuously increased, so that dietary fiber nutrients are seriously lost in dietary structure. At the same time, the prevalence of diabetes and other cardiovascular diseases in the country has also increased year by year. According to the latest data released in 2021 by the international diabetes union (IDF), the number of diabetic patients (20-79 years old) in China reaches 1.409 hundred million, and the number of type II diabetic patients accounts for about 90%. Meanwhile, the early-stage diabetes population in adults in China is as high as 50.1%, and 5-10% of patients develop diabetes every year. Therefore, the enhanced addition of dietary fiber in food is important and has become a great demand for the nation and people.
Wheat is one of main crops in China, and the annual yield of bran which is a processing byproduct is up to 1.3 hundred million tons. Arabinoxylan (AX), a major soluble dietary fiber in plant cells as a major hemicellulose component (about 35% of the dry cell weight), is abundantly present in wheat bran. However, a large part of AX in nature is not effectively utilized, resulting in a large waste of resources. Therefore, the wheat bran is used as the raw material to efficiently extract the AX, and the AX is used as the dietary fiber raw material to be added into related foods and medicines in an enhanced manner, so that the daily dietary fiber intake level of people can be improved. In view of the important health role of AX, the national food safety risk assessment center has already publicly requested an opinion on new food raw material arabinoxylan in 3 months and 14 days 2022, which is the new food raw material for which an opinion was first requested in 2022. As one of the functional sugars, the arabinoxylan has various health-care functions and is expected to be applied to functional foods for aiding digestion, controlling blood sugar, improving immunity, losing weight and the like.
At present, the extraction methods of AX are mainly divided into three main categories: hot water extraction, alkaline extraction and enzymatic extraction. Because AX is closely combined with components such as protein, starch, cellulose and the like in bran through covalent bonds such as ester bonds, ether bonds and the like, the content of free AX components is low, and hot water leaching can only dissolve out part of free AX components, the extraction yield is generally low and the energy consumption is high. In most basic researches, researchers generally extract AX in small batches by using weak base, and the weak base can effectively destroy covalent bonds such as intermolecular ester bonds and the like, so that the bound AX is effectively released, and the extraction efficiency can be remarkably improved. In some studies, xylanase, cellulase and ultrasonic and microwave treatment are also used for auxiliary extraction of AX, but the defects of high energy consumption and incapability of continuous industrial production are also existed. Therefore, the development of the high-efficiency extraction and preparation process of AX has important significance for the development of novel dietary fiber raw materials and the value-added application of agricultural and sideline products.
Through searching, the following patent publications related to the patent application of the invention are found:
1. a method for preparing araboxylan (CN 108191996A) comprises processing byproduct of corn drum skin of corn starch as raw material, treating with ultrasonic wave and microwave-assisted alkaline method, centrifuging, adjusting, centrifuging, concentrating under reduced pressure, precipitating with ethanol, centrifuging, and lyophilizing to obtain araboxylan of corn drum skin. The invention adopts a physical auxiliary method of ultrasonic wave and microwave on the basis of an alkaline method, fully utilizes the high penetrating power and high heating efficiency of the microwave and the cavitation action of the ultrasonic wave, greatly improves the yield of polysaccharide, shortens the production period, and realizes waste utilization because the raw material of the invention is a byproduct from starch processing and greatly increases economic benefit. The arabinoxylan prepared by the invention has a certain in vitro antioxidant activity and is expected to be developed into a food additive with better antioxidant activity.
The arabinoxylan prepared by the above publications is extracted by an alkaline method, so that the pollution is serious, and the basic analytical structure of the arabinoxylan can be damaged. Meanwhile, the ultrasonic wave and the microwave have higher requirements on cooperative treatment and serious energy consumption. In addition, the preparation method of AX is complicated in process steps and limited in application range.
2. A preparation method of arabinoxylan and its product (CN 111574640A) specifically comprise the following steps: (1) removing starch from the wheat bran; (2) obtaining a polysaccharide solution by adopting an alkali extraction method; (3) deproteinizing the polysaccharide solution; (4) Carrying out enzyme treatment and dialysis on the deproteinized polysaccharide solution; (5) carrying out enzymolysis on the dialyzed polysaccharide solution; (6) Respectively carrying out alcohol precipitation grading on the polysaccharide solution which is not subjected to enzymolysis and the polysaccharide solution which is subjected to enzymolysis to obtain araboxylan with different molecular weights; (7) And (3) carrying out secondary enzyme treatment and secondary dialysis on the arabinoxylan with different molecular weights. The preparation method is simple and feasible, can improve the utilization rate of the raw material wheat bran, can obtain different fractions of arabinoxylan, has different structures, has different degrees of functional effects of reducing blood sugar, resisting oxidation and the like, and has high medical value.
The arabinoxylan prepared by the above publications is extracted by an alkaline method, has a large influence on the environment, and is expensive due to the adoption of a method of multiple enzyme treatments. In addition, the processing method of grading alcohol precipitation has poor repeatability, and the quality stability of products cannot be ensured.
3. A method for extracting arabinoxylan and protein fiber from wheat bran (CN 104264262B) mainly comprises the following steps: primarily crushing wheat bran, pretreating with ethanol, centrifuging to obtain insoluble substances, primarily extruding the insoluble substances by an extruder, then performing superfine crushing, inactivating enzyme by using a complex enzyme and gravity action, separating supernate from precipitate to obtain a mixed solution rich in arabinoxylan, concentrating the supernate, precipitating with an organic solvent, concentrating and drying to obtain a product rich in arabinoxylan, and performing vacuum drying on the precipitate to obtain a protein fiber product. The method provided by the invention can extract the arabinoxylan with high added value from the grain processing byproduct bran, and the byproduct in the extraction process can be used as animal feed, thereby increasing the added value of the product and improving the economic benefit of grain processing enterprises.
The arabinoxylan prepared by the above publications adopts enzymatic extraction, has high extraction cost, low arabinoxylan yield, high energy consumption for ultramicro-pulverization treatment, and limited application range. The wheat bran is crushed to a certain degree, and the fine treatment cannot well improve the yield of the product, so the efficiency is poor.
By contrast, the present patent application is intrinsically different from the above patent publications.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a method for extracting arabinoxylan by taking wheat bran as a raw material, the arabinoxylan and application.
The technical scheme adopted by the invention for solving the technical problem is as follows:
a method for extracting arabinoxylan from wheat bran comprises the following steps:
s1: cleaning wheat bran for multiple times, drying, crushing and sieving for later use;
s2: extraction pretreatment: performing weak base treatment on the wheat bran obtained through the S1 treatment to obtain treated wheat bran;
s3: extruding and puffing the wheat bran obtained through the S2 treatment to obtain pretreated wheat bran;
s4: extraction treatment: extracting the wheat bran obtained by the S3 treatment by using a water extraction method, centrifuging, and collecting supernatant;
s5: adjusting the pH of the supernatant collected in the step S4, centrifuging again, and collecting the supernatant;
s6: concentrating the supernatant collected in S5 in vacuum, adding ethanol for precipitation until the precipitate is complete, centrifuging, and collecting the precipitate;
s7: redissolving the precipitate collected in S6, and removing salt components in the precipitate after dialysis;
s8: and (3) carrying out vacuum concentration on the product obtained by the S7 treatment, and then carrying out freeze drying to obtain the arabinoxylan.
Further, the weak base treatment in step S2 is: spraying NaCO accounting for 2-5% of the wheat bran 3 Solution of NaCO 3 The concentration of the solution is 0.5-1M, and the solution is treated at 35 ℃ for 12-24h. Preferably, the weak base is present at a concentration of 1M, preferably for a period of 24h.
Further, in the step S3, the extrusion and the expansion are carried out under the conditions that the temperature is 60-160 ℃, the moisture content is 40-60%, and the rotating speed of a screw is 50-100rpm. The preferred temperature is from 100 to 160 ℃. The moisture content is 40-60%, preferably 40%. The screw speed is 50-100rpm. Preferably, the screw speed is 100rpm.
Further, when the water extraction method is used for extraction in the step S4, the mass ratio of wheat bran to water is 1:20, the temperature is 100 ℃, and the extraction time is 24 hours; the rotation speed of the centrifuge is 4000-5000rpm. Preferably, the rotation speed of the centrifuge is 4500rpm.
Further, the pH value in the step S5 is 4, and the rotation speed of the centrifugation is 4000-5000rpm. Preferably, the speed of centrifugation is 4500rpm.
Further, in the step S6, heating is carried out during vacuum concentration, the temperature after heating is 60 ℃, the concentration is carried out until the volume is 1/3 of that of the original dialysate, the mass concentration of the ethanol is 90-98%, the ethanol is stopped to be added when the mass concentration of the ethanol in the solution is 70-80%, then the solution is kept stand for 8-12 hours, and the rotation speed of centrifugation is 4000-5000rpm. Preferably, the rotation speed of the centrifuge is 4500rpm.
Further, the temperature of the precipitate in the step S7 is 60 ℃, and the redissolution time is 4-6 hours, preferably 6 hours; the dialysis is carried out by using a dialysis bag, the interception size of the dialysis bag is 8000-14000Da, water is adopted for dialysis to be used as dialysate for cleaning, the dialysis is carried out for three days, and the dialysate is changed every 3-5 hours until the conductivity of the dialysate is similar to or equal to that of pure water.
Further, in the step S8, the temperature is increased during vacuum concentration, the temperature after heating is 60 ℃, and the concentration is carried out until the volume is 1/2 of that of the original dialysate.
The arabinoxylan is prepared by the method for extracting the arabinoxylan by taking the wheat bran as the raw material.
Furthermore, the total sugar content of the arabinoxylan reaches 75%, the molecular weight of the arabinoxylan is 640kDa, the monosaccharide consists of arabinose and xylose, and the molar percentage of the arabinose and xylose is 46.09% and 53.91%;
use of arabinoxylans as described above in the preparation of functional food products.
The beneficial effects obtained by the invention are as follows:
1. the method is a method for extracting the arabinoxylan by taking wheat bran as a raw material, and aims at solving the problems that the yield of the water extraction method commonly used for the arabinoxylan is low, the structure of the arabinoxylan can be damaged by alkaline extraction, the pollution is serious, and the price of an enzyme extraction method is high, the release of the arabinoxylan in the wheat bran is promoted by adopting alkaline treatment and an extrusion puffing technology, the pollution is reduced while the extraction efficiency is improved, the wheat bran resource can be fully utilized, the additional value is realized, and the waste is changed into valuable; the extraction method is simple, the cost is low, the pollution is small, and the arabinoxylan prepared by the method has higher level of soluble dietary fiber and good application prospect.
2. The invention provides an extraction scheme of the arabinoxylan and measures related physicochemical properties. The extraction method can better destroy the surface structure of the bran, is beneficial to the dissolution of the araboxylan, increases the content of soluble dietary fiber, reduces the content of insoluble dietary fiber and greatly improves the yield of the dietary fiber.
3. Aiming at the technical defects of the extraction of the existing arabinoxylan, the invention develops an efficient preparation process of extracting AX by hot water by combining weak alkali spraying with extrusion and puffing pretreatment. The specific principle of the method is as follows: increasing pores on the surface of the bran through weak base treatment, and promoting a solvent to enter the bran; the high-temperature and high-pressure conditions of extrusion puffing promote the covalently bound AX in the bran to be converted into a free state, so that the AX can be dissolved out by a water extraction method, and the extraction rate of the AX is high. In addition, the weak base is only added into AX in a small amount of spraying manner in the process, so that alkali waste liquid is hardly generated, and the extrusion and expansion process is simple and convenient to treat, and is particularly suitable for continuous industrial production.
4. Compared with the common alkaline extraction method, the method of the invention can greatly reduce the generated organic waste liquid, adopts a hydrothermal leaching mode and only uses 2 percent of NaCO 3 Compared with the enzyme extraction method, the method does not use expensive enzyme preparation, reduces the cost and simultaneously reserves the effective components of dietary fibers in the wheat bran.
5. In the method, when weak base spraying treatment, extrusion and expansion pretreatment and hot water extraction are simultaneously used for synergistic extraction, the yield of the obtained arabinoxylan is obviously higher than that of single treatment or the effect of simultaneous treatment of two factors. According to the invention, when only hot water extraction is adopted, the yield of the arabinoxylan is only 1.59%, the yield reaches 4.23% after extrusion and expansion pretreatment is adopted, and when NaCO3 spraying treatment and extrusion and expansion pretreatment are simultaneously adopted, the yield of the arabinoxylan is 6 times that of the arabinoxylan extracted only by hot water and 2.5 times that of the arabinoxylan extracted only by extrusion and expansion pretreatment.
Drawings
FIG. 1 is an appearance diagram of an arabinoxylan extract according to the present invention;
FIG. 2 is a glucose standard curve according to the present invention;
FIG. 3 is a chart of molecular weight determination of arabinoxylans according to the present invention.
FIG. 4 is a graph showing the effect of arabinoxylan on starch digestion characteristics according to the present invention;
FIG. 5 is a graph showing the effect of arabinoxylan on the digestion profile of bread in accordance with the present invention;
FIG. 6 is a graph showing the variation of the glycolytic properties of arabinoxylan in bread processing according to the present invention.
Detailed Description
The present invention will be further described in detail with reference to examples for better understanding, but the scope of the present invention is not limited to the examples.
The raw materials used in the invention are all conventional commercial products unless otherwise specified, the methods used in the invention are all conventional in the field, and the quality of each substance used in the invention is conventional quality.
A method for extracting arabinoxylan from wheat bran comprises the following steps:
s1: cleaning wheat bran for multiple times, drying, crushing and sieving for later use;
s2: extraction pretreatment: performing weak base treatment on the wheat bran obtained through the S1 treatment to obtain treated wheat bran;
s3: extruding and puffing the wheat bran obtained through the S2 treatment to obtain pretreated wheat bran;
s4: extraction treatment: extracting the wheat bran obtained by the S3 treatment by using a water extraction method, centrifuging, and collecting supernatant;
s5: adjusting the pH of the supernatant collected in the step S4, centrifuging again, and collecting the supernatant;
s6: concentrating the supernatant collected in S5 in vacuum, adding ethanol for precipitation to be complete, centrifuging, and collecting the precipitate;
s7: redissolving the precipitate collected in S6, and removing salt components in the precipitate after dialysis;
s8: and (4) carrying out vacuum concentration on the product obtained by the S7 treatment, and then carrying out freeze drying to obtain the arabinoxylan.
Preferably, the weak base treatment in step S2 is: spraying NaCO accounting for 2-5% of the wheat bran 3 Solution of NaCO 3 The concentration of the solution is 0.5-1M, and the solution is treated at 35 ℃ for 12-24h. The concentration of weak base is preferably 1M, and the time is preferably 24h.
Preferably, the extrusion puffing is carried out in the step S3 under the conditions that the temperature is 60-160 ℃, the moisture content is 40-60%, and the screw rotation speed is 50-100rpm. The preferred temperature is from 100 to 160 ℃. The moisture content is 40-60%, preferably 40%. The screw speed is 50-100rpm. Preferably the screw speed is 100rpm.
Preferably, when the water extraction method is used for extraction in the step S4, the mass ratio of wheat bran to water is 1:20, the temperature is 100 ℃, and the extraction time is 24 hours; the rotation speed of the centrifuge is 4000-5000rpm. Preferably, the speed of centrifugation is 4500rpm.
Preferably, the pH value in the step S5 is 4, and the rotation speed of the centrifugation is 4000-5000rpm. Preferably, the speed of centrifugation is 4500rpm.
Preferably, the heating is carried out during vacuum concentration in the step S6, the temperature after the heating is 60 ℃, the concentration is carried out until the volume is 1/3 of that of the original dialysate, the mass concentration of the ethanol is 90-98%, the ethanol is stopped to be added when the mass concentration of the ethanol in the solution is 70-80%, then the standing is carried out for 8-12 hours, and the rotation speed of the centrifugation is 4000-5000rpm. Preferably, the speed of centrifugation is 4500rpm.
Preferably, the temperature for redissolving the precipitate in the step S7 is 60 ℃, and the redissolution time is 4-6 hours, preferably 6 hours; the dialysis is carried out by using a dialysis bag, the interception size of the dialysis bag is 8000-14000Da, water is adopted for dialysis to be used as dialysate for cleaning, the dialysis is carried out for three days, and the dialysate is changed every 3-5 hours until the conductivity of the dialysate is similar to or equal to that of pure water.
Preferably, the step S8 is carried out by heating in vacuum concentration, wherein the temperature after heating is 60 ℃, and the concentration is carried out until the volume is 1/2 of the original dialysate.
The arabinoxylan is prepared by the method for extracting the arabinoxylan by taking the wheat bran as the raw material.
Preferably, the total sugar content of the arabinoxylan reaches 75%, the molecular weight of the arabinoxylan is 640kDa, the monosaccharide consists of arabinose and xylose, and the molar percentages of the arabinose and xylose are 46.09% and 53.91%;
use of arabinoxylans as described above in the preparation of functional food products.
Specifically, the preparation and detection are as follows:
example 1
A method for extracting arabinoxylan from wheat bran comprises the following steps:
s1: cleaning wheat bran for multiple times, drying for 3 hours at 105 ℃, crushing and sieving with a 60-mesh sieve for later use;
s2: extraction pretreatment: spraying 1M NaCO with the quality of 2% of wheat bran to the wheat bran obtained by the S1 treatment 3 Treating the solution at 35 ℃ for 24h;
s3: extruding and puffing the wheat bran obtained by the S2 under the conditions that the temperature is 100-160 ℃, the moisture content is 40% and the screw rotation speed is 100rpm to obtain pretreated wheat bran;
s4: extraction treatment: the ratio of the wheat bran obtained by the S3 treatment to water is 1:20, extracting at 100 ℃ for 24h at the rotating speed of 4500rpm, and collecting supernatant;
s5: adjusting the pH of the supernatant collected in the step S4 to be =4, centrifuging again at the rotating speed of 4500rpm, and collecting the supernatant;
s6: concentrating the supernatant collected in S5 at 60 ℃ in vacuum to evaporate the volume of the supernatant to 1/3 of the original volume, adding 98% ethanol until the mass concentration of the ethanol in the solution is 75%, standing for 6 hours, centrifuging at 4500rpm after flocculent precipitates are completely precipitated, and collecting the precipitates;
s7: adding the precipitate collected in the step S6 into distilled water at 60 ℃, stirring for redissolving, filling into a dialysis bag with the interception amount of 8000-14000Da after complete redissolution, wherein the filling amount is about 1/4 of that of the dialysis bag, dialyzing for 3 days, changing the distilled water every 3 hours, and finishing when the electric conductivity of the intercepted solution is similar to that of pure water;
s8: and (3) concentrating the product obtained by the S6 treatment in vacuum at the temperature of 60 ℃, evaporating the volume of the supernatant to 1/2 of the original volume, and freeze-drying to obtain the arabinoxylan. See fig. 1.
Example 2
A method for extracting arabinoxylan from wheat bran comprises the following steps:
s1: cleaning wheat bran for multiple times, drying for 3 hours at 105 ℃, crushing and sieving with a 60-mesh sieve for later use;
s2: extraction pretreatment: spraying 0.5M NaCO with the quality of 2% of wheat bran on the wheat bran obtained by the S1 treatment 3 Treating the solution at 35 deg.C for 24h
S3: extruding and puffing the wheat bran obtained by the S2 under the conditions that the temperature is 60-120 ℃, the moisture content is 40% and the screw rotation speed is 100rpm to obtain pretreated wheat bran;
s4: extraction treatment: the ratio of the wheat bran obtained by the S3 treatment to water is 1:20, extracting at 100 ℃ for 24h at the rotating speed of 4500rpm, and collecting supernatant;
s5: adjusting the pH of the supernatant collected in the step S4 to be =4, centrifuging again at the rotating speed of 4500rpm, and collecting the supernatant;
s6: concentrating the supernatant collected in the step S5 at 60 ℃ in vacuum, evaporating the volume of the supernatant to 1/3 of the original volume, adding 98% ethanol until the mass concentration of the ethanol in the solution is 75%, standing for 6 hours, centrifuging at 4500rpm after flocculent precipitates are completely precipitated, and collecting precipitates;
s7: adding distilled water into the precipitate collected in the step S6, stirring and redissolving at 60 ℃, loading into a dialysis bag with the interception amount of 8000-14000Da after complete redissolution, wherein the loading amount is about 1/4 of that of the dialysis bag, dialyzing for 3 days, changing the distilled water every 3 hours, and finishing when the electric conductivity of the intercepted liquid is similar to that of pure water;
s8: and (3) concentrating the product obtained by the S6 treatment in vacuum at the temperature of 60 ℃, evaporating the volume of the supernatant to 1/2 of the original volume, and freeze-drying to obtain the arabinoxylan. See fig. 1.
Example 3
A method for extracting arabinoxylan from wheat bran comprises the following steps:
s1: cleaning wheat bran for multiple times, drying for 3 hours at 105 ℃, crushing and sieving with a 60-mesh sieve for later use;
s2: extraction pretreatment: spraying 1M NaCO with the quality of 2% of wheat bran to the wheat bran obtained by the S1 treatment 3 Treating the solution at 35 ℃ for 24 hours;
s3: extruding and puffing the wheat bran obtained by the S2 under the conditions that the temperature is 100-160 ℃, the moisture content is 60% and the screw rotation speed is 100rpm to obtain pretreated wheat bran;
s4: extraction treatment: the ratio of the wheat bran obtained by the S3 treatment to water is 1:20, extracting at 100 ℃ for 24h at the rotating speed of 4500rpm, and collecting supernate;
s5: adjusting the pH of the supernatant collected in the step S4 to be =4, centrifuging again at the rotating speed of 4500rpm, and collecting the supernatant;
s6: concentrating the supernatant collected in the step S5 at 60 ℃ in vacuum, evaporating the volume of the supernatant to 1/3 of the original volume, adding 98% ethanol until the mass concentration of the ethanol in the solution is 75%, standing for 6 hours, centrifuging at 4500rpm after flocculent precipitates are completely precipitated, and collecting precipitates;
s7: adding the precipitate collected in the step S6 into distilled water at 60 ℃, stirring for redissolving, filling into a dialysis bag with the interception amount of 8000-14000Da after complete redissolution, wherein the filling amount is about 1/4 of that of the dialysis bag, dialyzing for 3 days, changing the distilled water every 3 hours, and finishing when the electric conductivity of the intercepted solution is similar to that of pure water;
s8: and (3) concentrating the product obtained by the S6 treatment in vacuum at the temperature of 60 ℃, evaporating the volume of the supernatant to 1/2 of the original volume, and freeze-drying to obtain the arabinoxylan. See fig. 1.
The correlation test of the present invention is as follows:
1. purification and physicochemical analysis of arabinoxylan
1. Determination of Total sugar
TABLE 1
| Standard | Distilled water | |||
| 1 | 0.0mL | 2.0mL | 0μg/ |
|
| 2 | 0.4mL | 1.6mL | 40μg/ |
|
| 3 | 0.8mL | 1.2mL | 80μg/ |
|
| 4 | 1.2mL | 0.8mL | 120μg/ |
|
| 5 | 1.6mL | 0.4mL | 160μg/ |
|
| 6 | 2.0mL | 0.0mL | 200μg/mL |
A0.2 mg/mL standard solution was prepared, and a standard curve was prepared as shown in Table 1. 10mg of sample are weighed and 1mL of 72% H is added 2 SO 4 And stirring for 30min at room temperature until the sample is dissolved. 2mL of the diluted sample was taken, and 50. Mu.L of 80% phenol solution was added to each sample tube, and 50. Mu.L of distilled water was added to the blank tube. 3.0mL of concentrated sulfuric acid was added to each tube. Mixing, and cooling at room temperature. The measurement was carried out with an ultraviolet-visible spectrophotometer at a wavelength of 490 nm. The results show that the AX total sugar content reached 75% (fig. 2).
2. Determination of molecular weight
The molecular weight and distribution of the polysaccharide samples were determined by High Performance Liquid Chromatography (HPLC) equipped with a differential detector. And calibrating the molecular weight of the sample by using the dextrans with different molecular weights as standard substances. 2mg of polysaccharide sample was dissolved in 1mL of 0.1mol/mL sodium nitrate (NaNO) 3 ) Preparing into 2mg/mL solution, filtering with 0.22 μm polyethersulfone filter membrane after the sample is completely dissolved, collectingThe filtrate is ready for use. The chromatographic conditions are chromatographic columns: ultrahydrogel Column Linear,10 μm,7.8mm × 300mm; mobile phase: 0.1mg/mL sodium nitrate; flow rate: 0.6mL/min; temperature of the column oven: at 40 ℃. The analysis showed that the crude sugar consisted mainly of a fraction with a molecular weight of 640kDa with a peak time of 14min (FIG. 3).
3. Monosaccharide composition determination
The method for identifying the monosaccharide composition and the proportion of each monosaccharide in a sample by using an ion chromatograph comprises the following steps:
weighing 1-2 mg of polysaccharide sample into an ampoule bottle, adding 1-2ml of 2mol/L trifluoroacetic acid (TFA), sealing by an alcohol lamp, hydrolyzing in an oil bath at 120 ℃ for 3 hours, carrying out reduced pressure rotary evaporation to remove redundant TFA, adding 0.5-2 ml of anhydrous methanol, carrying out reduced pressure rotary drying, repeating for 5 times, completely dissolving the product into 2ml of ultrapure water, mixing uniformly, filtering by a 0.22 mu water system filter membrane, and then loading. Each standard monosaccharide (rhamnose, arabinose, xylose, mannose, galactose, glucose, galacturonic acid, glucuronic acid) was dissolved and then analyzed with a GC under the same conditions. The analysis results showed that the arabinoxylan monosaccharide composition was arabinose and xylose, and the mole percentage was 46.09% and 53.91% (table 2).
TABLE 2 monosaccharide composition of arabinoxylans according to the invention
2. Comparison of the method of the present invention with the conventional extraction method of arabinoxylan
TABLE 3 comparative Effect of the extraction method of arabinoxylan according to the present invention compared to the conventional method
As can be seen from Table 3, the method of the invention produces much less organic waste liquid than the conventional alkaline extraction method, and the invention adopts a hydrothermal leaching method, only 2% of NaCO is used 3 Compared with the enzyme extraction method, the method does not use expensive enzyme preparation, reduces the cost and simultaneously reserves the effective components of dietary fibers in the wheat bran.
3. The method has the synergistic effects of weak base spraying treatment, extrusion and puffing pretreatment and hot water extraction
Comparative example 1
The procedure of comparative example 1 was the same as that of example 1 except for the differences shown in Table 4.
TABLE 4 synergistic effect of spraying weak base, extrusion-puffing pretreatment and hot water extraction in the method of the present invention
Comparative example 2
The procedure of comparative example 2 was the same as that of example 2 except for the differences shown in Table 5.
TABLE 5 synergistic effect of spraying with weak base, extrusion-puffing pretreatment, and hot water extraction in the method of the present invention
Comparative example 3
The procedure of comparative example 3 was the same as that of example 3 except for the differences shown in Table 6.
TABLE 6 synergistic Effect of the weak base spraying treatment, the extrusion-puffing pretreatment and the hot water extraction in the method of the present invention
As can be seen from tables 1, 2 and 3, when weak alkali spraying treatment, extrusion and expansion pretreatment and hot water extraction are simultaneously used for synergistic extraction, the yield of the obtained arabinoxylan is obviously higher than that of single treatment or the effect of the two factors when the two are simultaneously treated. Meanwhile, as can be seen from tables 1, 2 and 3, when only hot water extraction is adopted, the yield of the arabinoxylan is only 1.59%, and after extrusion and puffing pretreatment, the yield reaches 4.23%, and when NaCO3 spraying treatment and extrusion and puffing pretreatment are adopted, the yield of the arabinoxylan is 6 times that of the arabinoxylan only extracted by hot water and 2.5 times that of the arabinoxylan only extracted by extrusion and puffing pretreatment.
4. AX can inhibit the retrogradation of starch during baking and processing of bread, and AX has significant inhibitory effect on bread digestion (1) differential scanning calorimetry
Sample (150 mg) was first mixed with5mL of 5g/L pepsin was mixed in 0.05MHCl and incubated at 37 ℃ for 30 minutes with shaking by adding 8 glass beads. Thereafter, 5mL of 0.25M sodium acetate buffer was added, followed by 2.5mL of the enzyme mixture (3.0 g of pancreatin was dispersed in 20mL of water, stirred for 15 minutes, and then centrifuged at 1049g for 10 minutes to collect the pancreatin supernatant. Then 60mg of amyloglucosidase was dissolved in 1.7mL of water and added to 15.0mL of the pancreatin supernatant to give the desired enzyme mixture). Hydrolysis was carried out in a shaking water bath at 37 ℃ for 0, 20, 40, 60, 90, 120, 180 and 240 minutes, respectively, and then immediately the sample was diluted in anhydrous ethanol to inactivate the enzyme. After centrifugation at 9000g for 2 minutes, the glucose content of the supernatant was measured with a D-glucose assay kit (glucose oxidase/peroxidase, GOPOD Specification K-GLUK) from Megazyme International Ireland Ltd. Glucose contents in the gel system at 20 and 120min of hydrolysis are marked as G 20 And G 120 According to which the starch components are defined as (RDS), slowly Digestible Starch (SDS) and Resistant Starch (RS) using the following formula:
fast digestible starch (RDS) = (G) 20 -G 0 )×0.9÷TS×100
Slowly Digestible Starch (SDS) = (G) 120 -G 20 )×0.9÷TS×100
Resistant Starch (RS) = (TS-RDS-SDS) ÷ TS × 100
Wherein TS is the Total Starch (TS) content of the gel composite measured
The results of the experiment are shown in fig. 4 and 5. As can be seen in FIG. 4, AX inhibits the retrogradation of starch during baking and processing of bread.
As can be seen from fig. 5, AX has a significant bread digestion inhibitory effect, and AX having a higher molecular weight has a stronger bread digestion inhibitory effect.
(2) Glycolytic properties
Preparing a culture medium: (1) 400mg of Na2CO3, 900mg of NaCl, (NH 4) 2SO4 900mg, 900mg of K2HPO4, 50mg of CaCl2, 50mg of MgSO4, 1mg of FeSO 4. 7H2O 20mg, 20mg of MnSO 4. H2O, 1mg of Resazurin, and CoCl. 6H2O 2mg were weighed into a beaker, 995mL of pure water was added thereto, the mixture was dissolved by ultrasound, the pH was adjusted to 6.8 with 5M HCl, the mixture was boiled by heating, the mixture was placed in a 1L brown bottle, bubbles were removed by ultrasound for 30min, and N2 was added thereto and sterilized. (2) In a 10mL centrifuge tube, 2mg of pantothenic acid cap, 2mg of nicotinamide, 6 mg of vitamin B, 2mg of vitamin B, 12mg of vitamin B, 0.1mg of p-aminobenzoyl acid and 100mg of L-cysteine hydrochloride were weighed, and 4mL of distilled water was added. (3) Vc 5mg, folic acid 5mg and vitamin B12.5 mg are weighed into a beaker, and the volume is adjusted to 100mL by a volumetric flask. Adding 1mL of (3) into (2), filtering with 0.22 μm sterile filter membrane to obtain filtrate (4), opening the anaerobic incubator, adding the required substances, adjusting to anaerobic state, and mixing sterilized (1) and (4).
The sample processing method comprises the following steps: 0.2g of porcine colonic digest was weighed out in an anaerobic chamber and 0.2mL of 1% sugar solution was added. Only pig colonic digests and anaerobic medium were added to the blank. Sealing the centrifuge tube with sealing film, placing into anaerobic bag, performing shake culture at 37 deg.C, taking out corresponding samples after 0h, 12h, 24h and 48h, centrifuging at 11000rad/min for 20min, and measuring the supernatant for pH value and total sugar concentration by the following specific measurement method.
1) Determination of the pH value
The results of the measurement using a pH meter are shown in FIG. 6 a.
2) Determination of the Total sugar concentration
TABLE 1
| Standard | Distilled water | |||
| 1 | 0.0mL | 2.0mL | 0μg/ |
|
| 2 | 0.4mL | 1.6mL | 40μg/ |
|
| 3 | 0.8mL | 1.2mL | 80μg/ |
|
| 4 | 1.2mL | 0.8mL | 120μg/ |
|
| 5 | 1.6mL | 0.4mL | 160μg/ |
|
| 6 | 2.0mL | 0.0mL | 200μg/mL |
A0.2 mg/mL standard solution was prepared, and a standard curve was prepared as shown in Table 1. 10mg of sample was weighed and 1mL of 72% H was added 2 SO 4 And stirred at room temperature for 30min until the sample is dissolved. 2mL of the diluted sample was taken, and 50. Mu.L of 80% phenol solution was added to each sample tube, and 50. Mu.L of distilled water was added to the blank tube. 3.0mL of concentrated sulfuric acid was added to each tube. Mixing, and cooling at room temperature. The measurement was carried out with an ultraviolet-visible spectrophotometer at a wavelength of 490 nm. The results are shown in FIG. 6 b.
As can be seen from fig. 6, compared to direct consumption of AX, AX added to bread slowed the glycolysis rate in the intestine, was more beneficial for the microbial utilization in the distal colon, and the prebiotic effect was enhanced (fig. 6).
Although the embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, and therefore the scope of the invention is not limited to the embodiments disclosed.
Claims (9)
1. A method for extracting arabinoxylan from wheat bran as a raw material is characterized by comprising the following steps: the method comprises the following steps:
s1: cleaning wheat bran for multiple times, drying, crushing and sieving for later use;
s2: extraction pretreatment: performing weak base treatment on the wheat bran obtained through the S1 treatment to obtain treated wheat bran;
s3: extruding and puffing the wheat bran obtained through the S2 treatment to obtain pretreated wheat bran;
s4: extraction treatment: extracting the wheat bran obtained by the S3 treatment by using a water extraction method, centrifuging, and collecting supernatant;
s5: adjusting the pH value of the supernatant collected in the step S4, centrifuging again, and collecting the supernatant;
s6: concentrating the supernatant collected in S5 in vacuum, adding ethanol for precipitation until the precipitate is complete, centrifuging, and collecting the precipitate;
s7: redissolving the precipitate collected in S6, and removing salt components in the precipitate after dialysis;
s8: performing vacuum concentration on the product obtained by the S7 treatment, and then performing freeze drying to obtain the arabinoxylan;
the weak base treatment in the step S2 comprises the following steps: spraying Na accounting for 2-5% of the wheat bran 2 CO 3 Solution, na 2 CO 3 The concentration of the solution is 0.5-1M, and the solution is treated at 35 ℃ for 12-24h;
in the step S3, the extrusion and the expansion are carried out under the conditions that the temperature is 60-160 ℃, the water content is 40-60 percent, and the rotating speed of a screw is 50-100rpm.
2. The method for extracting arabinoxylan from wheat bran as a raw material according to claim 1, wherein the method comprises the following steps: when the water extraction method is used for extraction in the step S4, the mass ratio of the wheat bran to the water is 1:20, the temperature is 100 ℃, and the extraction time is 24 hours; the rotation speed of the centrifuge is 4000-5000rpm.
3. The method for extracting arabinoxylan from wheat bran as a raw material according to claim 1, wherein the method comprises the following steps: in the step S5, the pH value is 4, and the rotation speed of the centrifugation is 4000-5000rpm.
4. The method for extracting arabinoxylan from wheat bran as a raw material according to claim 1, wherein the method comprises the following steps: and S6, heating in vacuum concentration, wherein the temperature after heating is 60 ℃, concentrating until the volume is 1/3 of that of the original dialysate, the mass concentration of ethanol is 90-98%, adding ethanol until the mass concentration of the ethanol in the solution is 70-80%, stopping adding the ethanol, standing for 6-12 hours, and the rotation speed of centrifugation is 4000-5000rpm.
5. The method for extracting arabinoxylan from wheat bran as a raw material according to claim 1, wherein the method comprises the following steps: the temperature of the precipitate in the step S7 is 60 ℃, and the redissolution time is 4-6 hours; the dialysis is carried out by using a dialysis bag, the interception size of the dialysis bag is 8000-14000Da, water is adopted for dialysis to be used as dialysate for cleaning, the dialysis is carried out for three days, and the dialysate is changed every 3-5 hours until the conductivity of the dialysate is similar to or equal to that of pure water.
6. The method for extracting arabinoxylan from wheat bran as a raw material according to any one of claims 1 to 5, wherein: and S8, heating during vacuum concentration, wherein the temperature after heating is 60 ℃, and concentrating to 1/2 of the volume of the original dialysate.
7. The arabinoxylan produced by the method for extracting arabinoxylan using wheat bran as a raw material according to any one of claims 1 to 6.
8. Arabinoxylan according to claim 7, characterized in that: the total sugar content of the arabinoxylan reaches 75%, the molecular weight of the arabinoxylan is 640kDa, the monosaccharide consists of arabinose and xylose, and the molar percentage of the arabinose and xylose is 46.09% and 53.91%.
9. Use of arabinoxylans as claimed in claim 7 or 8 in the preparation of functional food products.
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| CN103497258A (en) * | 2013-10-11 | 2014-01-08 | 黑龙江八一农垦大学 | Method for extracting and purifying active polysaccharides of wheat bran |
| CN103626886A (en) * | 2013-11-20 | 2014-03-12 | 中国农业科学院农产品加工研究所 | Method for extracting cereal seed coat active polysaccharide through assisted treatment of hot-pressing steam spray explosion method |
| CN105517450A (en) * | 2013-06-21 | 2016-04-20 | 杜邦营养生物科学有限公司 | Methods and compositions to improve the nutritional value of lignocellulosic biomass |
| CN106962947A (en) * | 2017-03-28 | 2017-07-21 | 贵阳学院 | Combined method extracts soluble dietary fiber in potato residues |
| CN111574640A (en) * | 2020-06-18 | 2020-08-25 | 华中农业大学 | Preparation method of arabinoxylan and product |
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| CN105517450A (en) * | 2013-06-21 | 2016-04-20 | 杜邦营养生物科学有限公司 | Methods and compositions to improve the nutritional value of lignocellulosic biomass |
| CN103497258A (en) * | 2013-10-11 | 2014-01-08 | 黑龙江八一农垦大学 | Method for extracting and purifying active polysaccharides of wheat bran |
| CN103626886A (en) * | 2013-11-20 | 2014-03-12 | 中国农业科学院农产品加工研究所 | Method for extracting cereal seed coat active polysaccharide through assisted treatment of hot-pressing steam spray explosion method |
| CN106962947A (en) * | 2017-03-28 | 2017-07-21 | 贵阳学院 | Combined method extracts soluble dietary fiber in potato residues |
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