Efficient preparation method of wheat bran feruloyl Arabic xylooligosaccharide
Through extrusion and puffing, ultrafine crushing, high-pressure cooking and microwave-assisted multi-enzyme synergistic technology, the problem of low extraction efficiency of Arabic xylose in wheat bran is solved, and high-efficiency extraction of high content of ferulic acid is achieved Arabic xylooligosaccharides ensure the maintenance of its functional characteristics.
Patent Information
- Application Number
- CN202411400126.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-09
- Publication Date
- 2024-12-13
AI Technical Summary
The extraction efficiency of feruloyl Arabin oligosaccharide in wheat bran is low and there is a lack of efficient extraction methods.
Using physical means such as extrusion puffing, ultrafine crushing, high-pressure cooking, microwave and other physical means combined with multi-enzyme synergistic enzymes, breaking the glycosidic bonds of feruloyl arabinoxilan by pretreatment, primary and secondary ultrafine crushing, extrusion Pressure puffing, high-pressure cooking, microwave-assisted multi-enzyme coenzyme decomposition and separation and purification steps to extract feruloyl Arabin-oligomeroxylose.
The efficient extraction of feruloyl Arabin xylosol is achieved, the extraction efficiency is improved, and the high content of disaccharides and trisaccharides is obtained. The enzymatic process will not destroy its structure, which is conducive to the development of functional characteristics.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of oligosaccharide preparation, and particularly relates to a method for efficiently preparing wheat bran feruloyl arabinoxylo-oligosaccharide. Background Art
[0002] As an internationally popular oligosaccharide, xylooligosaccharide has many functional properties. For example, it can regulate intestinal flora and help the growth of beneficial intestinal flora; it is beneficial for people with diabetes, obesity and other "three highs" to eat; it has antioxidant and immunity-enhancing effects, so it is widely used in food, feed and pharmaceutical industries. Feruloyl arabinoxo-oligosaccharide is a dual-functional xylooligosaccharide with a main chain composed of xylose and a side chain connected to feruloyl with antioxidant activity. It can be used as an ingredient for baked goods, beverages, pasta and other foods to improve the functional and nutritional properties of food, and has broad application prospects.
[0003] During the processing of cereals (wheat), a large amount of byproduct wheat bran is produced. Wheat bran is rich in phenolic acids and non-digestible polysaccharides, but the low utilization rate of wheat bran leads to its low added value. The output of wheat bran is close to 20% of the wheat processing volume. my country's annual wheat bran output reaches more than 20 million tons, which is a very rich raw material for the preparation of oligoxylose. The mass content of hemicellulose in wheat bran is about 30%, second only to the 33% content in corn cobs used for industrial production of oligoxylose. The hemicellulose composition and structure of wheat bran and corn cobs are different. Wheat bran is rich in feruloyl arabinoxylan, but there is a lack of corresponding extraction methods, or the extraction efficiency is relatively low. It is necessary to develop an efficient method for extracting feruloyl arabinoxylan from wheat bran. Summary of the invention
[0004] Aiming at the problem of low extraction efficiency of feruloyl arabinoxylo-oligosaccharides from wheat bran in the prior art, the present invention provides a method for efficiently preparing feruloyl arabinoxylo-oligosaccharides from wheat bran, and the glycosidic bonds of feruloyl arabinoxylo-oligosaccharides from wheat bran are broken by physical means such as extrusion puffing, ultrafine grinding, high-pressure cooking, microwave, etc., combined with multi-enzyme synergistic enzymolysis, thereby establishing a green and efficient preparation technology for feruloyl arabinoxylo-oligosaccharides.
[0005] The present invention is achieved through the following technical solutions: An efficient preparation method of wheat bran feruloyl arabinoxylo-oligosaccharide comprises the following steps: (1) Pretreatment: adding water and high temperature amylase to the wheat bran for primary enzymolysis, adding alkaline protease for secondary enzymolysis after enzymolysis, inactivating the enzyme after enzymolysis, filtering, and washing with precipitated water to obtain wheat bran free of starch and protein; (2) Initial ultrafine grinding: The wheat bran from which starch and protein have been removed is dried to a constant weight and then subjected to initial ultrafine grinding; (3) Extrusion and puffing to break the bran wall: add water to the ultrafinely crushed wheat bran to condition it to 30-45wt%, extrusion and puffing to break the bran wall, and cut into segments; (4) Secondary ultrafine grinding: the wheat bran cut in step (3) is dried to a constant weight and subjected to secondary ultrafine grinding; (5) High-pressure cooking and breaking the bran wall: After the second ultrafine grinding, add water and mix well to form a paste, and then treat it under high pressure at 120-125℃ for 20-25min to obtain wheat bran paste; (6) Microwave-assisted multi-enzyme synergistic enzymolysis: The wheat bran paste was treated with microwaves, and then water, ligninase, cellulase and endo-β-1,4-xylanase were added. The enzymolysis was stirred at 30-50 rpm for 12-16 h. After the enzymolysis was completed, the enzyme was inactivated to obtain a wheat bran suspension. (7) Separation and purification: After centrifugation of the wheat bran suspension, the supernatant was collected, filtered to remove impurities, adsorbed on a macroporous adsorption resin Amberlite XAD-2 at a flow rate of 1-1.5 mL / min, and eluted with 50% ethanol at a flow rate of 0.5-1 mL / min; (8) Drying: The eluate from step (7) is vacuum concentrated to an extract, washed with water until there is no ethanol smell, and freeze-dried to obtain feruloyl arabinoxylo-oligosaccharide.
[0006] Furthermore, in step (1), the mass ratio of wheat bran to water is 1-2:8-9; the amount of high temperature amylase added is 0.4-0.6% of the mass of wheat bran and water; the amount of alkaline protease added is 0.7-0.9% of the mass of wheat bran and water. Furthermore, in step (1), the conditions for the first enzymatic hydrolysis are 90-95°C for 3-4 hours and then cooled to 60-65°C; the conditions for the second enzymatic hydrolysis are 60-65°C for 3-4 hours.
[0007] Furthermore, the drying conditions in step (2) and step (4) are 80-90°C.
[0008] Furthermore, in step (3), the extrusion puffing wall breaking condition is 105-110°C.
[0009] Furthermore, the wheat bran after the initial ultrafine grinding in step (2) is passed through a 100-mesh sieve; and the wheat bran after the secondary ultrafine grinding in step (4) is passed through a 120-mesh sieve.
[0010] Furthermore, in step (5), the mass ratio of the wheat bran to water after the secondary ultrafine grinding is 7-8:5-6.
[0011] Furthermore, in step (6), the microwave power is 150-200 W, the treatment is carried out for 25-30 min, and then after cooling to 25-30° C., water having a volume 4 to 6 times that of the wheat bran paste is added; the amount of ligninase added is 0.02 to 0.04% of the mass of the wheat bran paste; the amount of cellulase added is 0.04 to 0.06% of the mass of the wheat bran paste; and the amount of β-1,4-xylanase added is 0.06 to 0.08% of the mass of the wheat bran paste.
[0012] Furthermore, the enzymatic hydrolysis conditions in step (6) are stirring enzymatic hydrolysis at 30-50 rpm for 12-16 hours; the eluate in step (8) is vacuum concentrated at 50-60°C, and the freeze-drying conditions are -40--50°C.
[0013] Furthermore, the enzyme inactivation treatment is carried out at 95-110° C. for 8-15 min.
[0014] Beneficial Effects (1) The present invention uses physical means such as extrusion puffing, ultrafine grinding, high-pressure cooking, microwave, etc., combined with multi-enzyme synergistic enzymolysis, to break the glycosidic bonds of wheat bran feruloyl arabinoxylan, thereby establishing a green and efficient preparation technology for feruloyl arabinoxylo-oligosaccharides. The extraction efficiency of feruloyl arabinoxylo-oligosaccharides is high, and the content of disaccharides and trisaccharides is high; (2) The present invention uses three enzymes, ligninase, cellulase and endo-β-1,4-xylanase, to treat the wheat bran paste in a coordinated microwave treatment. The enzymatic hydrolysis efficiency is high and the structure of the feruloyl arabinoxylo-oligosaccharides in the wheat bran will not be destroyed, which is conducive to the performance of its functional properties. DETAILED DESCRIPTION
[0015] The present invention will be further described below in conjunction with the embodiments. However, the scope of the present invention is not limited to the following embodiments. It will be appreciated by those skilled in the art that various changes and modifications may be made to the present invention without departing from the spirit and scope of the present invention. Example 1 (1) Pretreatment: The wheat bran was crushed and passed through an 80-mesh sieve. 85 parts of water and 0.5% of the mass of wheat bran and water were added to 15 parts of wheat bran for primary enzymolysis. The enzymolysis was carried out at 90-95°C for 3.5 hours. After the enzymolysis, the temperature was lowered to 60-65°C. Alkaline protease was added to the wheat bran and water at a mass of 0.8% for secondary enzymolysis. The enzymolysis was carried out at 60-65°C for 3.5 hours. After the enzymolysis was completed, the temperature was raised to 100°C and kept warm for 10 minutes to inactivate the enzyme. The enzymolysis solution was filtered with an 80-mesh non-woven gauze. The precipitate was washed with water 5 times to obtain wheat bran with de-starch and de-proteinized. (2) Initial ultrafine grinding: The wheat bran from which starch and protein have been removed is dried at 85°C for 8 h until constant weight is reached, and then initially ultrafine ground. After grinding, the ground wheat bran is passed through a 100-mesh sieve. (3) Extrusion and puffing to break the bran wall: After ultrafine grinding, add water to condition the bran to 40wt%, extrusion and puffing to break the bran wall at 105-110℃, and cut into small pieces of about 1 cm; (4) Secondary ultrafine grinding: The wheat bran cut into segments in step (3) is dried at 85° C. for 8 h to a constant weight, firstly crushed to 60 mesh using a rotary vane mill, then subjected to secondary ultrafine grinding, and then passed through a 120-mesh sieve after grinding; (5) High-pressure cooking and breaking the bran wall: 75 parts of the second ultrafinely crushed wheat bran are added with 60 parts of water and mixed into a paste, and then high-pressure cooking is performed at 120-125°C for 20 minutes to obtain wheat bran paste; (6) Microwave-assisted multi-enzyme synergistic enzymolysis: The wheat bran paste was subjected to microwave treatment at a microwave power of 180 W for 25 min. Then, water 5 times the volume of the wheat bran paste, 0.03% ligninase by weight of the wheat bran paste, 0.05% cellulase by weight of the wheat bran paste, and 0.07% endo-β-1,4-xylanase by weight of the wheat bran paste were added. The enzymolysis was carried out under stirring at 40 rpm for 15 h. After the enzymolysis was completed, the temperature was raised to 100 °C and kept warm for 10 min to inactivate the enzymes to obtain a wheat bran suspension. (7) Separation and purification: The wheat bran suspension was centrifuged at 4000 rpm for 18 min, the supernatant was collected and filtered with non-woven fabric to remove impurities, and then adsorbed by macroporous adsorption resin Amberlite XAD-2 at a flow rate of 1.2 mL / min, and eluted with 50% ethanol at a flow rate of 0.8 mL / min; (8) Drying: The eluate from step (7) is vacuum concentrated at 55°C to obtain an extract, washed with water until there is no ethanol smell, and freeze-dried at -45°C to obtain feruloyl arabinoxylo-oligosaccharide.
[0016] Example 2 (1) Pretreatment: The wheat bran was crushed and passed through an 80-mesh sieve. 90 parts of water and 0.6% of the mass of wheat bran and water were added to 20 parts of wheat bran for primary enzymolysis. The enzymolysis was carried out at 90-95°C for 4 hours. After the enzymolysis, the temperature was lowered to 60-65°C. Alkaline protease was added to the wheat bran and water at a mass of 0.7% for secondary enzymolysis. The enzymolysis was carried out at 60-65°C for 4 hours. After the enzymolysis was completed, the temperature was raised to 100°C and kept warm for 10 minutes to inactivate the enzyme. The enzymolysis solution was filtered with an 80-mesh non-woven gauze. The precipitate was washed with water 5 times to obtain wheat bran with de-starch and de-proteinized. (2) Initial ultrafine grinding: The wheat bran from which starch and protein have been removed is dried at 90°C for 8 h until constant weight is reached, and then initially ultrafine ground. After grinding, the ground wheat bran is passed through a 100-mesh sieve. (3) Extrusion and puffing to break the bran wall: After ultrafine grinding, add water to condition the bran to 45wt%, extrusion and puffing to break the bran wall at 105-110℃, and cut into small pieces of about 1cm; (4) Secondary ultrafine grinding: The wheat bran cut into segments in step (3) is dried at 90° C. for 8 h to a constant weight, firstly ground into 60 mesh using a rotary vane mill, then subjected to secondary ultrafine grinding, and then passed through a 120-mesh sieve after grinding; (5) High-pressure cooking and breaking the bran wall: 80 parts of the second ultrafinely crushed wheat bran are added with 60 parts of water and mixed into a paste, and then high-pressure cooking is performed at 120-125°C for 25 minutes to obtain wheat bran paste; (6) Microwave-assisted multi-enzyme synergistic enzymolysis: The wheat bran paste was subjected to microwave treatment at a microwave power of 150 W for 30 min. Then, water 6 times the volume of the wheat bran paste, 0.02% ligninase by weight of the wheat bran paste, 0.06% cellulase by weight of the wheat bran paste, and 0.08% endo-β-1,4-xylanase by weight of the wheat bran paste were added. The enzymolysis was carried out by stirring at 40 rpm for 16 h. After the enzymolysis was completed, the temperature was raised to 100 °C and kept warm for 10 min to inactivate the enzymes to obtain a wheat bran suspension. (7) Separation and purification: The wheat bran suspension was centrifuged at 4000 rpm for 20 min, the supernatant was collected and filtered with non-woven fabric to remove impurities, and then adsorbed by macroporous adsorption resin Amberlite XAD-2 at a flow rate of 1.5 mL / min, and eluted with 50% ethanol at a flow rate of 1 mL / min; (8) Drying: The eluate from step (7) is vacuum concentrated at 60°C to obtain an extract, washed with water until there is no ethanol smell, and freeze-dried at -50°C to obtain feruloyl arabinoxylo-oligosaccharide.
[0017] Example 3 (1) Pretreatment: The wheat bran was crushed and passed through an 80-mesh sieve. 85 parts of water and 0.4% of the mass of wheat bran and water were added to 15 parts of wheat bran for primary enzymolysis. The enzymolysis was carried out at 90-95°C for 4 hours. After the enzymolysis, the temperature was lowered to 60-65°C. Alkaline protease was added to the wheat bran and water at 0.9% of the mass of water for secondary enzymolysis. The enzymolysis was carried out at 60-65°C for 4 hours. After the enzymolysis was completed, the temperature was raised to 100°C and kept warm for 10 minutes to inactivate the enzyme. The enzymolysis solution was filtered with a 60-80 mesh non-woven gauze. The obtained precipitate was washed with water 5 times to obtain wheat bran with de-starch and de-proteinized. (2) Initial ultrafine grinding: The wheat bran from which starch and protein have been removed is dried at 90°C for 8 h until constant weight is reached, and then initially ultrafine ground. After grinding, the ground wheat bran is passed through a 100-mesh sieve. (3) Extrusion and puffing to break the bran wall: After ultrafine grinding, add water to condition the bran to 30wt%, extrusion and puffing to break the bran wall at 105-110℃, and cut into small pieces of about 1cm; (4) Secondary ultrafine grinding: The wheat bran cut into segments in step (3) is dried at 90° C. for 8 h to a constant weight, firstly ground into 60 mesh using a rotary vane mill, then subjected to secondary ultrafine grinding, and then passed through a 120-mesh sieve after grinding; (5) High-pressure cooking and breaking the bran wall: 70 parts of the second ultrafinely crushed wheat bran are added with 50 parts of water and mixed into a paste, and then high-pressure cooking is performed at 120-125°C for 20 minutes to obtain wheat bran paste; (6) Microwave-assisted multi-enzyme synergistic enzymolysis: The wheat bran paste was subjected to microwave treatment at a microwave power of 200 W for 25 min. Then, water 4 times the volume of the wheat bran paste, 0.02% ligninase by weight of the wheat bran paste, 0.06% cellulase by weight of the wheat bran paste, and 0.06% endo-β-1,4-xylanase by weight of the wheat bran paste were added. The enzymolysis was stirred at 50 rpm for 16 h. After the enzymolysis was completed, the temperature was raised to 100 °C and kept warm for 10 min to inactivate the enzymes to obtain a wheat bran suspension. (7) Separation and purification: The wheat bran suspension was centrifuged at 4000 rpm for 20 min, the supernatant was collected and filtered with non-woven fabric to remove impurities, and then adsorbed by macroporous adsorption resin Amberlite XAD-2 at a flow rate of 1 mL / min, and eluted with 50% ethanol at a flow rate of 0.5 mL / min; (8) Drying: The eluate from step (7) is vacuum concentrated at 60°C to obtain an extract, washed with water until there is no ethanol smell, and freeze-dried at -50°C to obtain feruloyl arabinoxylo-oligosaccharide.
[0018] Comparative Example 1 Different from Example 1, step (6) of Comparative Example 1 does not involve microwave treatment, and the remaining conditions are the same as those of Example 1.
[0019] Comparative Example 2 Different from Example 1, no ligninase and cellulase were added in step (6) of Comparative Example 2, and the other conditions were the same as those of Example 1.
[0020] Comparative Example 3 Different from Example 1, no endo-β-1,4-xylanase was added in step (6) of Comparative Example 3, and the other conditions were the same as those of Example 1.
[0021] The content and yield of feruloyl arabinoxooligosaccharides in the finished products obtained in Examples 1 to 3 and Comparative Examples 1 to 3, as well as the content of xylobiose and xylotriose in xylose were tested, and the results are shown in Table 1 below: Table 1 As can be seen from Table 1, the three enzymes of ligninase, cellulase and endo-β-1,4-xylanase in the present invention cooperate with microwave treatment to treat wheat bran paste. The three enzymes cooperate with microwave action to have high enzymatic hydrolysis efficiency and will not destroy the structure of wheat bran feruloyl arabinoxylo-oligosaccharides, which is beneficial to the performance of its functional properties.
Claims
1. An efficient preparation method of wheat bran feruloyl arabinoxylo-oligosaccharide, characterized in that: The following steps are involved: (1) Pretreatment: adding water and high temperature amylase to the wheat bran for primary enzymolysis, adding alkaline protease for secondary enzymolysis after enzymolysis, inactivating the enzyme after enzymolysis, filtering, and washing with precipitated water to obtain wheat bran free of starch and protein; (2) Initial ultrafine grinding: The wheat bran from which starch and protein have been removed is dried to a constant weight and then subjected to initial ultrafine grinding; (3) Extrusion and puffing to break the bran wall: add water to the ultrafinely crushed wheat bran to condition it to 30-45wt%, extrusion and puffing to break the bran wall, and cut into segments; (4) Secondary ultrafine grinding: the wheat bran cut in step (3) is dried to a constant weight and subjected to secondary ultrafine grinding; (5) High-pressure cooking and breaking the bran wall: After the second ultrafine grinding, add water and mix well to form a paste, and then treat it under high pressure at 120-125°C for 20-25 minutes to obtain wheat bran paste; (6) Microwave-assisted multi-enzyme synergistic enzymolysis: The wheat bran paste was treated with microwaves, and then water, ligninase, cellulase and endo-β-1,4-xylanase were added. The enzymolysis was stirred at 30-50 rpm for 12-16 h. After the enzymolysis was completed, the enzyme was inactivated to obtain a wheat bran suspension. (7) Separation and purification: After centrifugation of the wheat bran suspension, the supernatant was collected, filtered to remove impurities, adsorbed on a macroporous adsorption resin Amberlite XAD-2 at a flow rate of 1-1.5 mL / min, and eluted with 50% ethanol at a flow rate of 0.5-1 mL / min; (8) Drying: The eluate from step (7) is vacuum concentrated to an extract, washed with water until there is no ethanol smell, and freeze-dried to obtain feruloyl arabinoxylo-oligosaccharide.
2. The efficient preparation method of wheat bran feruloyl arabinoxylo-oligosaccharide according to claim 1, characterized in that: In step (1), the mass ratio of wheat bran to water is 1-2:8-9; the amount of high temperature amylase added is 0.4-0.6% of the mass of the wheat bran and water; and the amount of alkaline protease added is 0.7-0.9% of the mass of the wheat bran and water.
3. The efficient preparation method of wheat bran feruloyl arabinoxylo-oligosaccharide according to claim 1, characterized in that: In step (1), the conditions for the first enzymatic hydrolysis are 90-95°C for 3-4 hours and then cooled to 60-65°C; the conditions for the second enzymatic hydrolysis are 60-65°C for 3-4 hours.
4. The efficient preparation method of wheat bran feruloyl arabinoxylo-oligosaccharide according to claim 1, characterized in that: The drying conditions in step (2) and step (4) are 80-90°C.
5. The efficient preparation method of wheat bran feruloyl arabinoxylo-oligosaccharide according to claim 1, characterized in that: The extrusion and puffing wall breaking conditions in step (3) are 105-110°C.
6. The efficient preparation method of wheat bran feruloyl arabinoxylo-oligosaccharide according to claim 1, characterized in that: The wheat bran after the initial ultrafine grinding in step (2) is passed through a 100-mesh sieve; and the wheat bran after the secondary ultrafine grinding in step (4) is passed through a 120-mesh sieve.
7. The efficient preparation method of wheat bran feruloyl arabinoxylo-oligosaccharide according to claim 1, characterized in that: The mass ratio of the wheat bran and water after the secondary ultrafine grinding in step (5) is 7-8:5-6.
8. The efficient preparation method of wheat bran feruloyl arabinoxylo-oligosaccharide according to claim 1, characterized in that: In step (6), the microwave power is 150-200 W and the treatment is carried out for 25-30 min. Then, after cooling to 25-30° C., water having a volume 4 to 6 times that of the wheat bran paste is added. The amount of ligninase added is 0.02 to 0.04% of the mass of the wheat bran paste. The amount of cellulase added is 0.04 to 0.06% of the mass of the wheat bran paste. The amount of β-1,4-xylanase added is 0.06 to 0.08% of the mass of the wheat bran paste.
9. The efficient preparation method of wheat bran feruloyl arabinoxylo-oligosaccharide according to claim 1, characterized in that: The enzymatic hydrolysis conditions in step (6) are stirring enzymatic hydrolysis at 30-50 rpm for 12-16 hours; the eluate in step (8) is vacuum concentrated at 50-60°C, and the freeze-drying conditions are -40--50°C.
10. The efficient preparation method of wheat bran feruloyl arabinoxylo-oligosaccharide according to claim 1, characterized in that: The enzyme inactivation treatment conditions are 95~110℃ for 8~15min.