CN115181771A - Method for efficiently preparing feruloyl arabino xylo-oligosaccharide - Google Patents
Method for efficiently preparing feruloyl arabino xylo-oligosaccharide Download PDFInfo
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- CN115181771A CN115181771A CN202210789757.8A CN202210789757A CN115181771A CN 115181771 A CN115181771 A CN 115181771A CN 202210789757 A CN202210789757 A CN 202210789757A CN 115181771 A CN115181771 A CN 115181771A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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Abstract
The invention relates to the field of food processing, in particular to a method for efficiently preparing feruloyl arabino-xylo-oligosaccharides. The method mainly comprises the following steps: the method comprises the steps of taking defatted cereal bran as a raw material, adding amylase for the first time to extrude to remove starch, adding protease for the second time to extrude to remove protein, hydrolyzing xylanase, purifying Amberlite XAD-2 macroporous resin, concentrating and drying to obtain the feruloyl arabo xylooligosaccharide. The method has the advantages of simple process, short time consumption, high yield of the feruloyl arabino xylo-oligosaccharide and the like.
Description
Technical Field
A method for efficiently preparing feruloyl arabino-xylo-oligosaccharide relates to a modification technology of cereal bran by enzyme extrusion, and belongs to the field of food processing.
Background
Cereal bran such as rice bran, wheat bran, etc. contains abundant arabinoxylan, which is polymerized from D-xylopyranose via beta- (1,4) -glycosidic bond, part of xylose in the 2-position or/and 3-position is substituted by alpha-arabinofuranose, and part of arabinose in the 5-position has ferulic acid connected with ester bond. Therefore, the xylanase hydrolyzes the cereal bran to obtain the feruloyl arabino xylo-oligosaccharide. The feruloyl oligosaccharide is recognized as a food ingredient with dual physiological activities of oligosaccharide and ferulic acid by the food and drug administration of the United states, and has higher added value. However, the traditional process usually adopts amylase to hydrolyze bran so as to remove starch and adopts protease water to remove protein, and takes a long time; in addition, the xylanase has low hydrolysis efficiency on the treated bran, so the yield of the feruloyl arabino xylo-oligosaccharide is low. The enzyme extrusion is a novel extrusion technology based on the traditional extrusion, and the essence of the technology is that an extruder is used as an enzyme reactor so as to accelerate the enzymatic reaction. The research team finds that the time for removing starch and protein can be greatly shortened by adding enzyme for extrusion, and the cell wall of the bran is damaged at the same time, so that the hydrolysis efficiency of xylanase on the cereal bran is improved. Based on the discovery, the invention provides a method for efficiently preparing feruloyl arabino xylo-oligosaccharide.
Disclosure of Invention
The invention aims to provide a method for efficiently preparing feruloyl arabino xylo-oligosaccharides, so as to improve the additional value of the cereal bran.
The technical scheme of the invention is as follows: a method for efficiently preparing feruloyl arabino-xylo-oligosaccharides utilizes enzyme-added extrusion to remove starch and protein in cereal bran and destroy cell wall, and then utilizes xylan to hydrolyze bran to obtain feruloyl arabino-xylo-oligosaccharides.
(1) High temperature alpha-amylase extrusion: adjusting the water content of the degreased bran to 30-40%, adding high-temperature alpha-amylase according to the proportion of 720U enzyme per gram of bran, wherein the extrusion temperature is 95 ℃, the outlet temperature is 100 ℃, the screw rotation speed is 50-150 rpm, and the water addition amount is 0.2kg/h;
(2) Adding protease for extrusion: adjusting the water content of the extrudate in the step (1) to 30-40%, adding neutral protease according to the proportion of 1440U enzyme per gram of bran, wherein the extrusion temperature is 50 ℃, the outlet temperature is 60 ℃, the screw rotation speed is 50-150 rpm, the water addition amount is 0.2kg/h, and washing the extrudate by adopting water and 80% ethanol solution after extrusion;
(3) Hydrolysis of xylanase: dispersing the extrudate in a pH 5.0 acetic acid buffer solution, adding xylanase according to the proportion of 2000U enzyme per gram of extrudate, reacting for 3 h at 45 ℃, and centrifuging to obtain an enzymolysis solution;
(4) Purification of feruloyl arabino xylo-oligosaccharide: purifying the feruloyl arabino xylo-oligosaccharide by adopting Amberlite XAD-2 macroporous resin, loading the enzymolysis solution on a column, eluting 2 column volumes by using distilled water, eluting 3 column volumes by using 50% ethanol and eluting 2 column volumes by using 95% ethanol, wherein the flow rate is 2 mL/min, collecting the components eluted by using 50% ethanol, concentrating and drying to obtain the feruloyl arabino xylo-oligosaccharide.
The bran is rice bran, wheat bran or corn bran.
Compared with the prior art, the invention has the following beneficial effects: simple process, short time consumption and high yield of the feruloyl arabino xylo-oligosaccharide.
Drawings
FIG. 1 shows the UV spectrum (A) and the IR spectrum (B) of the arabino-xylo-oligosaccharides obtained from Feruloyl in example 1: a is feruloyl arabino xylo-oligosaccharide obtained by the traditional hydrolysis process, and b is feruloyl arabino xylo-oligosaccharide obtained by the enzyme-added extrusion process.
Detailed Description
Example 1
Adjusting the water content of the defatted rice bran to 40%, adding high-temperature alpha-amylase according to the proportion of 720U enzyme per gram of rice bran, wherein the extrusion temperature is 90 ℃ and the outlet temperature is 100 ℃; adjusting the moisture content of the extrudate to 40%, adding neutral protease according to the proportion of 1440U enzyme per gram of bran, extruding at 50 ℃ and 60 ℃ and extruding under the following other conditions: the screw speed is 50 rpm, and the water addition amount is 0.2 kg/h. After extrusion, the extrudate was washed with water, 80% ethanol solution and then extrudedDispersing the mixture in acetic acid buffer solution with pH 5.0, adding xylanase according to the proportion of 2000U enzyme per gram of extrudate, reacting at 45 ℃ for 3 h, and centrifuging to obtain enzymatic hydrolysate. Purifying the feruloyl arabino xylo-oligosaccharide by adopting Amberlite XAD-2 macroporous resin, loading the enzymolysis solution on a column, eluting 2 column volumes by using distilled water, eluting 3 column volumes by using 50% ethanol and eluting 2 column volumes by using 95% ethanol, wherein the flow rate is 2 mL/min, collecting the components eluted by using 50% ethanol, concentrating and drying to obtain the feruloyl arabino xylo-oligosaccharide. Identifying the structure of the product by using an infrared spectrum chromatograph and an ultraviolet spectrophotometer; and simultaneously measuring the ferulic acid content and the pentose content. In order to embody the effect of the invention, the traditional process is adopted, namely the high temperature alpha-amylase is added for hydrolysis for 40 min at 95 ℃, the neutral protease is added for hydrolysis for 4 h at 50 ℃, and then the same conditions are adopted for xylanase hydrolysis and Amberlite XAD-2 macroporous resin purification to prepare the feruloyl arab xylo-oligosaccharide as a contrast. As shown in FIG. 1, the feruloyl arabino-xylo-oligosaccharides prepared by the two methods have similar ultraviolet spectrum and infrared spectrum, have maximum ultraviolet absorption at 325 nm, and have maximum ultraviolet absorption at 3340, 1650-1900, 1161, 1273, 1517, 1601, 1635, 1040, 990, 850 and 898 cm -1 The like has characteristic absorption peaks, which indicates that the two methods successfully prepare the feruloyl arabino xylo-oligosaccharide and the structures of the two methods are similar. The yield of the feruloyl arabino-xylo-oligosaccharide is 5.78%, and the recovery rates of the ferulic acid and the pentose are 56.38% and 51.49% respectively. However, the yield of the feruloyl arabino xylo-oligosaccharide in the traditional process is 4.32 percent, and the yield of the ferulic acid and the pentose are 40.52 percent and 30.40 percent respectively. Namely, the method greatly improves the yield of the feruloyl arabino xylo-oligosaccharide. Further observing the microstructure of the rice bran with a microscope, it was found that: the traditional enzyme hydrolysis process only removes starch and protein in rice bran, but does not destroy the cell wall structure of the rice bran; the enzymatic extrusion not only removes starch and protein, but also destroys the cell wall structure. Therefore, the hydrolysis efficiency of the xylanase is improved.
Example 2
Adjusting the water content of the defatted rice bran to 30%, adding high-temperature alpha-amylase according to the proportion of 720U enzyme per gram of rice bran, wherein the extrusion temperature is 90 ℃ and the outlet temperature is 100 ℃; adjusting the water content of the extrudate to 30%, adding neutral protease according to the proportion of 1440U enzyme per gram of rice bran, wherein the extrusion temperature is 50 ℃, the outlet temperature is 60 ℃, and the extrusion conditions are as follows: the screw speed is 50 rpm, and the water addition amount is 0.2 kg/h. After extrusion, the extrudate is washed by water and 80% ethanol solution, then the extrudate is dispersed in pH 5.0 acetic acid buffer solution, xylanase is added according to the proportion of 2000U enzyme per gram extrudate, 3 h is reacted at 45 ℃, and enzymolysis solution is obtained by centrifugation. Purifying the feruloyl arabino xylo-oligosaccharide by adopting Amberlite XAD-2 macroporous resin. At this time, the yield of the feruloyl arabino xylo-oligosaccharide was 6.46%, and the recovery rates of ferulic acid and pentose were 57.48% and 57.80%, respectively.
Claims (2)
1. A method for efficiently preparing feruloyl arabino-xylo-oligosaccharide takes defatted bran as a raw material, and comprises the following steps of starch removal, protein removal, xylanase hydrolysis, amberlite XAD-2 macroporous resin purification, concentration and drying, and is characterized in that: extruding by adding high-temperature alpha-amylase to remove starch and adding neutral protease to extrude to remove protein, namely adjusting the moisture content of the bran to 30-40%, adding the high-temperature alpha-amylase according to the proportion of 720U per gram of the bran, wherein the extrusion temperature is 90 ℃ and the outlet temperature is 100 ℃; then adjusting the moisture content of the extrudate to 30-40%, adding neutral protease according to the proportion of 1440U per gram of bran and the extrusion temperature is 50 ℃, and the outlet temperature is 60 ℃; in addition, the screw rotation speed is 50 to 150 rpm, and the water addition amount is 0.2 kg/h.
2. The method for efficiently preparing the feruloyl arabino-xylo-oligosaccharides as claimed in claim 1, wherein: the bran is rice bran, wheat bran or corn bran.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210789757.8A CN115181771A (en) | 2022-07-06 | 2022-07-06 | Method for efficiently preparing feruloyl arabino xylo-oligosaccharide |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210789757.8A CN115181771A (en) | 2022-07-06 | 2022-07-06 | Method for efficiently preparing feruloyl arabino xylo-oligosaccharide |
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| CN115181771A true CN115181771A (en) | 2022-10-14 |
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| CN202210789757.8A Pending CN115181771A (en) | 2022-07-06 | 2022-07-06 | Method for efficiently preparing feruloyl arabino xylo-oligosaccharide |
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- 2022-07-06 CN CN202210789757.8A patent/CN115181771A/en active Pending
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