CN110862862A - Method for simultaneously preparing xanthoceras sorbifolia Bunge oil and xanthoceras sorbifolia Bunge protein by aqueous enzymatic method - Google Patents
Method for simultaneously preparing xanthoceras sorbifolia Bunge oil and xanthoceras sorbifolia Bunge protein by aqueous enzymatic method Download PDFInfo
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/30—Extraction; Separation; Purification by precipitation
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/025—Pretreatment by enzymes or microorganisms, living or dead
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/04—Pretreatment of vegetable raw material
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/001—Refining fats or fatty oils by a combination of two or more of the means hereafter
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/02—Refining fats or fatty oils by chemical reaction
- C11B3/06—Refining fats or fatty oils by chemical reaction with bases
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/10—Refining fats or fatty oils by adsorption
-
- 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
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
Abstract
The invention belongs to the technical field of deep processing of agricultural products, and discloses a method for simultaneously preparing xanthoceras sorbifolia Bunge oil and xanthoceras sorbifolia Bunge protein by an aqueous enzymatic method, which comprises the following steps: obtaining slurry; performing enzymolysis and centrifugation; performing ultrasonic demulsification treatment to obtain free oil; sequentially carrying out hydration degumming, deacidification and decoloration treatment to obtain high-purity xanthoceras sorbifolia oil; adding alkaline protease into the primary crude protein solution to perform alkali extraction twice; obtaining secondary alkali extract and centrifugally separating the secondary alkali extract to obtain secondary protein liquid; and adjusting the pH value, and then carrying out acid precipitation centrifugation and freeze drying to obtain the xanthoceras sorbifolia Bunge protein. The process is simple and safe, and the extracted xanthoceras sorbifolia bunge oil is strong in fragrance, light in color and good in quality; the prepared shinyleaf yellowhorn protein has higher purity, the protein content is higher than 50%, and the shinyleaf yellowhorn protein is suitable for industrial production. The invention not only improves the oil yield, but also has no pollution, high safety and no use of flammable solvent, thereby reducing the possibility of flammability and explosiveness, controlling pollution and ensuring safety; and the cost is low.
Description
Technical Field
The invention belongs to the technical field of deep processing of agricultural products, and particularly relates to a method for simultaneously preparing xanthoceras sorbifolia bunge oil and xanthoceras sorbifolia bunge protein by an aqueous enzymatic method.
Background
Currently, the closest prior art: the shinyleaf yellowhorn belongs to the genus shinyleaf yellowhorn of the family sapindaceae, also called as shinyleaf yellowhorn, shinyleaf yellowhorn and the like, and is a unique precious woody tree species in China. It is not only a kind of fruit and famous woody oil tree species with rich nutrition and delicious taste, but also an important good water and soil conservation tree species capable of improving ecological environment and greening soil and country.
The shinyleaf yellowhorn contains a large amount of grease, the oil content of the seeds is 30.4%, the kernel is rich in nutrition, the oil content is 55% -66%, the content of unsaturated fatty acid in the oil is as high as 94%, wherein the linoleic acid accounts for 36.9%, and the oleic acid accounts for 57.2%; and contains 19 kinds of amino acids, potassium, sodium, calcium, magnesium, iron, zinc and other trace elements and vitamins A, C, E and other vitamins essential for human body. Therefore, the shinyleaf yellowhorn oil is edible oil with high nutritive value. At present, the price of the shinyleaf yellowhorn oil in the international market is over $ 12/kg, and the future is expected to rise.
At present, the main methods for preparing vegetable oil in China are the traditional squeezing method and the traditional extraction method. The traditional squeezing method is simple in process operation and strong in adaptability, but high temperature of more than 200 ℃ generated in the squeezing process causes great damage to unsaturated fatty acid, vitamin K and other important nutrient components in the grease, protein denaturation and the like, and cake dregs can only be used as animal feed and the like; compared with the traditional pressing method, the extraction method is an advanced oil preparation method, the labor intensity is low, the production efficiency is higher, but the organic solvent used for oil extraction is overflowed and volatilized, so that the problems of large adverse effect on human bodies and environment, solvent residue, oil quality reduction and the like cause the development of the oil as edible oil and health-care oil to be limited to a certain extent. Therefore, the development of a new oil preparation process for efficiently and industrially producing high-quality, nutritional and health-care xanthoceras sorbifolia bunge oil is urgently needed.
Meanwhile, the xanthoceras sorbifolia bunge protein has better emulsibility, and can be used as an emulsifier to be added into baked food, frozen food and soup food so as to improve the stability of the product. The shinyleaf yellowhorn protein is rich in amino acid content which is 21.68%, and is protein with better quality. According to WHO recommendation, the amino acid ratio of egg protein is taken as a reference standard, and the evaluation score of the amino acid of the shinyleaf yellowhorn protein is superior to that of peanut protein under the condition of not detecting tryptophan. Therefore, there is a need to develop a method for simultaneously preparing xanthoceras sorbifolia oil and xanthoceras sorbifolia protein.
In summary, the problems of the prior art are as follows:
(1) the traditional squeezing method generates high temperature of more than 200 ℃ in the squeezing process, which causes great damage to unsaturated fatty acid, vitamin K and other important nutrient components in oil and fat, also causes protein denaturation and the like, and the cake meal can only be used as animal feed and the like.
(2) The organic solvent used for oil leaching in the leaching method can be diffused and volatilized, so that the problems of large adverse effect on human bodies and environment, solvent residue, oil quality reduction and the like cause that the development of the oil serving as edible oil and health-care oil is limited to a certain extent.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a method for simultaneously preparing xanthoceras sorbifolia bunge oil and xanthoceras sorbifolia bunge protein by an aqueous enzymatic method.
The invention is realized in such a way that a method for simultaneously preparing xanthoceras sorbifolia Bunge oil and xanthoceras sorbifolia Bunge protein by an aqueous enzymatic method comprises the following steps:
step one, soaking the peeled shinyleaf yellowhorn kernels in clear water for 2 hours, and mixing the soaked shinyleaf yellowhorn kernels with water according to a material-liquid ratio of 2 g: putting 3-9 mL of the mixture into a colloid mill for crushing, crushing shinyleaf yellowhorn kernels to 80-180 meshes, and collecting slurry;
regulating the pH value of the ground slurry to 3.7-6.0, carrying out enzymolysis, heating the enzymolysis liquid to 75-85 ℃ to inactivate enzyme for 15-25 minutes after the enzymolysis reaction is finished, regulating the pH value of the enzymolysis liquid to 4-5, centrifuging the enzymolysis liquid at the speed of 5000r/min for 20 minutes at room temperature, separating to obtain a first layer of clear oil, a second layer of emulsion layer, a third layer of waste liquid and a fourth layer of crude protein in sequence, and absorbing layer by layer to obtain primary xanthoceras sorbifolia Bunge oil, primary emulsion layer liquid and primary crude protein liquid;
performing ultrasonic treatment on the primary emulsion layer to obtain free oil;
step four, mixing the primary xanthoceras sorbifolia Bunge oil obtained in the step two with the free oil obtained in the step three, and sequentially carrying out hydration degumming, deacidification and decoloration treatment to obtain high-purity xanthoceras sorbifolia Bunge oil;
and step five, adjusting the pH value of the primary crude protein liquid to 7.5-8, adding the shinyleaf yellowhorn kernel material liquid in a ratio of 1: 55, stirring the mixture fully, and reacting the mixture for 100 minutes at the temperature of 40 ℃ to obtain primary alkali extract; after the primary alkali extract is naturally cooled to room temperature, centrifuging by using a centrifugal machine to obtain an upper protein solution and a lower precipitate, and separating the upper protein solution and the lower precipitate to obtain a primary protein solution and a primary precipitate;
step six, the obtained primary precipitate is subjected to the treatment of the step five to obtain secondary alkali extract, the secondary alkali extract is centrifuged to obtain an upper layer protein solution and a lower layer precipitate, and the upper layer protein solution and the lower layer precipitate are separated to obtain a secondary protein solution;
and seventhly, uniformly mixing the primary protein liquid and the secondary protein liquid, adjusting the pH value, performing acid precipitation and centrifugation to obtain precipitated starch, and performing freeze drying on the precipitated starch to obtain the shinyleaf yellowhorn protein.
In the second step, the acid for adjusting pH is one or two of hydrochloric acid and citric acid, and the pH of the solution is adjusted and then added to the enzyme solution.
Further, in step two, the enzymatic hydrolysis specifically comprises:
performing enzymolysis for 2 hours at 30-60 ℃ by using 50-500U/g of composite cellulase, then adding 100-500U/g of protease and composite enzyme thereof, and performing enzymolysis for 3 hours at 30-60 ℃;
the cellulase is composite cellulase;
the protease is one or more of papain, neutral protease, trypsin, alkaline protease, pepsin and acidic protease;
the compound enzyme is a compound of protease, xylanase and pectinase.
Further, in the third step, the ultrasonic treatment time is 30 minutes, and the ultrasonic frequency is 25-40k Hz.
Further, in the fourth step, the method for degumming xanthoceras sorbifolia bunge oil comprises the following steps:
(1) inputting the crude oil into a refining tank through a pipeline, heating the crude oil to 75-80 ℃, adding edible salt into warm water at 85 ℃, and melting the edible salt for 20min to form brine, wherein the mass ratio of the salt to the water in the brine is 1: 35;
(2) adding brine into a refining tank, mixing the brine and crude oil according to the mass ratio of 1:7, stirring for 10min under the condition of 60r/min of a stirrer, observing the separation condition of oil and soap, stopping stirring after obvious separation, precipitating for 5h, discharging saponin water until a small amount of oil is discharged, continuously washing twice until all precipitates are discharged, and finishing degumming.
Further, in the fourth step, the deacidification method comprises the following steps: pumping shinyleaf yellowhorn crude oil into an oil refining tank, heating to 65 ℃, adding 12-14% Baume alkali liquor into the oil refining tank, stirring at the speed of 40r/min, continuing to stir for 30min after the alkali liquor is added, observing, starting to heat to 62-65 ℃ when soap grains in emulsion are in a separated state, easily separating soapstock from oil after the final temperature is reached, quickly sinking, standing for 3-7 h, and then washing with water.
Further, the water washing method comprises the following steps: and (3) after standing and settling, discharging soapstock from the bottom of the oil refining tank, adjusting the oil temperature to 70-80 ℃, washing with hot water at the temperature of 5-10 ℃ higher than the oil temperature for 2-4 times, wherein the hot water added each time accounts for 10% -15% of the weight of the oil, continuously stirring at the speed of 45r/min in the washing process, stirring for 35min, standing for 2h, and discharging wastewater.
Further, in the fourth step, the decoloring method comprises: the washed oil contains 0.6 percent of water, the oil is dried firstly, the vacuum residual pressure of a decoloring tank is set to be 4 Kpa-7 Kpa, the oil in an oil refining tank in the step three is sucked into the decoloring tank, the volume of the oil in the decoloring tank accounts for 2/3 of the volume of the tank body of the decoloring tank, the temperature is raised while stirring, the temperature of the oil sucked into the decoloring tank is controlled to be 90-110 ℃, and the water content is reduced to be below 0.2 percent; then absorbing activated clay and activated carbon which are 2 to 5 percent of the weight of the oil into a decoloring tank through vacuum, and maintaining for more than 40min to complete decoloring; and filtering after the decolorization is finished.
Further, in the fifth step, the alkali for adjusting the pH is one or two of sodium hydroxide and potassium hydroxide, and the solution is added into the enzyme solution after the pH of the solution is adjusted.
Further, in step five, the centrifugation specifically comprises:
the centrifugation speed is 4000r/min, and the centrifugation time is 15 min.
In summary, the advantages and positive effects of the invention are:
according to the method for simultaneously preparing the xanthoceras sorbifolia Bunge oil and the xanthoceras sorbifolia Bunge protein by the aqueous enzymatic method, compound enzymolysis is carried out by adopting pectinase and cellulase, and compared with the method for singly using the cellulase or the pectinase for enzymolysis, the method has higher oil extraction rate. The method improves the extraction conditions of the xanthoceras sorbifolia bunge oil, increases the refining process of the xanthoceras sorbifolia bunge oil, reduces energy consumption to the maximum extent on the premise of improving the quality of the xanthoceras sorbifolia bunge finished oil, and improves efficiency. During the water washing process after deacidification, salt with the weight of 5% of water is added at the same time of adding hot water, so that the precipitation is facilitated. In addition, the process is simple and safe, and the extracted xanthoceras sorbifolia bunge oil is strong in fragrance, light in color and good in quality; the prepared shinyleaf yellowhorn protein has higher purity, the protein content is higher than 50%, and the shinyleaf yellowhorn protein is suitable for industrial production.
The invention not only improves the oil yield, but also has no pollution, high safety and no use of flammable solvent, thereby reducing the possibility of flammability and explosiveness, controlling pollution and ensuring safety; and the cost is low.
Drawings
Fig. 1 is a flow chart of a method for simultaneously preparing xanthoceras sorbifolia oil and xanthoceras sorbifolia protein by an aqueous enzymatic method provided by an embodiment of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Aiming at the problems in the prior art, the invention provides a method for simultaneously preparing xanthoceras sorbifolia bunge oil and xanthoceras sorbifolia bunge protein by an aqueous enzymatic method, and the invention is described in detail by combining the attached drawings.
As shown in fig. 1, the method for simultaneously preparing xanthoceras sorbifolia oil and xanthoceras sorbifolia protein by using an aqueous enzymatic method provided by the embodiment of the present invention comprises the following steps:
the method for simultaneously preparing the xanthoceras sorbifolia bunge oil and the xanthoceras sorbifolia bunge protein by the aqueous enzymatic method provided by the embodiment of the invention comprises the following steps:
s101, soaking the peeled shinyleaf yellowhorn kernels in clear water for 2 hours, and mixing the soaked shinyleaf yellowhorn kernels with water according to a material-liquid ratio of 2 g: and 3-9 mL of the mixture is put into a colloid mill for crushing, the shiny-leaved yellowhorn kernels are crushed to 80-180 meshes, and slurry is collected.
S102, adjusting the pH value of the ground slurry to 3.7-6.0, carrying out enzymolysis, after the enzymolysis reaction is finished, heating the enzymolysis liquid to 75-85 ℃ to inactivate enzyme for 15-25 minutes, adjusting the pH value of the enzymolysis liquid to 4-5, centrifuging the enzymolysis liquid at the speed of 5000r/min for 20 minutes at room temperature, separating to obtain a first layer of clear oil, a second layer of emulsion layer, a third layer of waste liquid and a fourth layer of crude protein in sequence, and absorbing layer by layer to obtain primary xanthoceras sorbifolia Bunge oil, primary emulsion layer liquid and primary crude protein liquid.
S103, carrying out ultrasonic treatment on the primary emulsion layer to obtain free oil.
And S104, mixing the primary xanthoceras sorbifolia bunge oil obtained in the step S102 with the free oil obtained in the step S103, and sequentially carrying out hydration degumming, deacidification and decoloration treatment to obtain the high-purity xanthoceras sorbifolia bunge oil.
S105, adjusting the pH value of the primary crude protein liquid to 7.5-8, adding a shinyleaf yellowhorn kernel material liquid ratio of 1: 55, stirring the mixture fully, and reacting the mixture for 100 minutes at the temperature of 40 ℃ to obtain primary alkali extract; and after the primary alkali extract is naturally cooled to room temperature, centrifuging by using a centrifugal machine to obtain an upper protein solution and a lower precipitate, and separating the upper protein solution and the lower precipitate to obtain a primary protein solution and a primary precipitate.
And S106, repeating the treatment of the step S105 on the obtained primary precipitate to obtain a secondary alkali extract, centrifuging the secondary alkali extract to obtain an upper-layer protein solution and a lower-layer precipitate, and separating the upper-layer protein solution and the lower-layer precipitate to obtain a secondary protein solution.
And S107, uniformly mixing the primary protein liquid and the secondary protein liquid, adjusting the pH value, performing acid precipitation and centrifugation to obtain precipitated starch, and performing freeze drying on the precipitated starch to obtain the shinyleaf yellowhorn protein.
In step S102, the acid for adjusting pH provided in the embodiment of the present invention is one or both of hydrochloric acid and citric acid, and the pH of the solution is adjusted and then added to the enzyme solution.
In step S102, the enzymatic hydrolysis provided by the embodiment of the present invention specifically includes:
performing enzymolysis for 2 hours at 30-60 ℃ by using 50-500U/g of composite cellulase, then adding 100-500U/g of protease and composite enzyme thereof, and performing enzymolysis for 3 hours at 30-60 ℃.
The cellulase adopts composite cellulase.
The protease is one or more of papain, neutral protease, trypsin, alkaline protease, pepsin and acidic protease.
The compound enzyme is a compound of protease, xylanase and pectinase.
In step S103, the ultrasonic treatment time provided by the embodiment of the present invention is 30 minutes, and the ultrasonic frequency is 25 to 40 kHz.
In step S104, the method for degumming xanthoceras sorbifolia bunge oil provided by the embodiment of the present invention comprises:
(1) inputting the crude oil into a refining tank through a pipeline, heating the crude oil to 75-80 ℃, adding edible salt into warm water at 85 ℃, and melting the edible salt for 20min to form brine, wherein the mass ratio of the salt in the brine to the water is 1: 35.
(2) Adding brine into a refining tank, mixing the brine and crude oil according to the mass ratio of 1:7, stirring for 10min under the condition of 60r/min of a stirrer, observing the separation condition of oil and soap, stopping stirring after obvious separation, precipitating for 5h, discharging saponin water until a small amount of oil is discharged, continuously washing twice until all precipitates are discharged, and finishing degumming.
In step S104, the deacidification method provided in the embodiment of the present invention includes: pumping shinyleaf yellowhorn crude oil into an oil refining tank, heating to 65 ℃, adding 12-14% Baume alkali liquor into the oil refining tank, stirring at the speed of 40r/min, continuing to stir for 30min after the alkali liquor is added, observing, starting to heat to 62-65 ℃ when soap grains in emulsion are in a separated state, easily separating soapstock from oil after the final temperature is reached, quickly sinking, standing for 3-7 h, and then washing with water.
The water washing method provided by the embodiment of the invention comprises the following steps: and (3) after standing and settling, discharging soapstock from the bottom of the oil refining tank, adjusting the oil temperature to 70-80 ℃, washing with hot water at the temperature of 5-10 ℃ higher than the oil temperature for 2-4 times, wherein the hot water added each time accounts for 10% -15% of the weight of the oil, continuously stirring at the speed of 45r/min in the washing process, stirring for 35min, standing for 2h, and discharging wastewater.
In step S104, the decoloring method provided in the embodiment of the present invention is: the washed oil contains 0.6 percent of water, the oil is dried firstly, the vacuum residual pressure of a decoloring tank is set to be 4 Kpa-7 Kpa, the oil in an oil refining tank in the step three is sucked into the decoloring tank, the volume of the oil in the decoloring tank accounts for 2/3 of the volume of the tank body of the decoloring tank, the temperature is raised while stirring, the temperature of the oil sucked into the decoloring tank is controlled to be 90-110 ℃, and the water content is reduced to be below 0.2 percent; then absorbing activated clay and activated carbon which are 2 to 5 percent of the weight of the oil into a decoloring tank through vacuum, and maintaining for more than 40min to complete decoloring; and filtering after the decolorization is finished.
In step S105, the alkali for adjusting pH provided in the embodiment of the present invention is one or both of sodium hydroxide and potassium hydroxide, and the pH of the solution is adjusted and then added to the enzyme solution.
In step S105, the centrifugation provided in the embodiment of the present invention specifically includes:
the centrifugation speed is 4000r/min, and the centrifugation time is 15 min.
The technical solution and the technical effect of the present invention will be further described with reference to the following embodiments.
Example 1
The method for simultaneously preparing xanthoceras sorbifolia bunge oil and xanthoceras sorbifolia bunge protein by using the aqueous enzymatic method, which is provided by the embodiment 1 of the invention, comprises the following steps:
(1) obtaining slurry: mixing peeled shinyleaf yellowhorn kernels with water according to a material-liquid ratio of 2 g: 3mL of the mixture is put into a colloid mill to be crushed, the shiny-leaved yellowhorn kernels are crushed to 80 meshes, and slurry is collected.
(2) Enzymolysis: and (3) adjusting the pH value of the slurry ground in the step one to 3.7, adding pectinase and cellulase for enzymolysis, and carrying out enzymolysis reaction in a water bath at 45 ℃ for 2.5 hours. After the enzymolysis reaction is finished, heating the enzymolysis liquid to 75 ℃ to inactivate the enzyme for 15 minutes, adjusting the pH of the enzymolysis liquid to 4, and then centrifuging. And (3) carrying out centrifugal separation on the enzymolysis liquid to obtain a first layer of clear oil, a second layer of emulsion layer, a third layer of waste liquid and a fourth layer of crude protein in sequence, and absorbing layer by layer to obtain primary xanthoceras sorbifolia Bunge oil, primary emulsion layer liquid and primary crude protein liquid.
(3) Adding ethanol into the primary emulsion layer liquid, and then carrying out cold bath treatment; the volume ratio of the ethanol to the primary emulsion layer is 1.5:1, the cold bath temperature is-30 ℃, and the cold bath time is 15 min. Introducing high-pressure steam into the enzymolysis liquid after the ethanol cold bath treatment for heat treatment and recovering ethanol, wherein the steam pressure is 15MPa, the steam temperature is 100 ℃, and the steam treatment time is 25 s. And after high-pressure steam treatment, centrifugal separation is carried out to obtain free oil.
(4) And (3) mixing the primary xanthoceras sorbifolia Bunge oil obtained in the step (2) with the free oil obtained in the step (3), and sequentially carrying out hydration degumming, deacidification and decoloration treatment to obtain the high-purity xanthoceras sorbifolia Bunge oil.
(5) Adjusting pH of the primary crude protein liquid to 6.7, adding alkaline protease 1.7% of the weight of xanthoceras sorbifolia bunge kernel, performing alkali extraction to obtain primary alkali extract, centrifuging the primary alkali extract to obtain an upper-layer protein solution and a lower-layer precipitate, and separating the upper-layer protein solution and the lower-layer precipitate to obtain a primary protein liquid and a primary precipitate.
(6) Secondary alkali extraction: and carrying out secondary alkali extraction on the primary precipitate to obtain secondary alkali extraction liquid, centrifuging the secondary alkali extraction liquid to obtain an upper-layer protein solution and a lower-layer precipitate, and separating the upper-layer protein solution and the lower-layer precipitate to obtain a secondary protein solution.
(7) Acid precipitation: and (3) uniformly mixing the primary protein solution and the secondary protein solution, adjusting the pH value, performing acid precipitation and centrifugation to obtain precipitated starch, and performing freeze drying on the precipitated starch to obtain the shinyleaf yellowhorn protein.
Example 2
The method for simultaneously preparing xanthoceras sorbifolia bunge oil and xanthoceras sorbifolia bunge protein by using the aqueous enzymatic method, which is provided by the embodiment 2 of the invention, comprises the following steps:
(1) obtaining slurry: mixing peeled shinyleaf yellowhorn kernels with water according to a material-liquid ratio of 2 g: putting 5mL of the mixture into a colloid mill for crushing, crushing shinyleaf yellowhorn kernels to 120 meshes, and collecting serous fluid.
(2) Enzymolysis: adjusting the pH value of the slurry ground in the step one to 5.0, adding pectinase and cellulase for enzymolysis, and carrying out enzymolysis reaction in a water bath at 50 ℃ for 4 hours. After the enzymolysis reaction is finished, heating the enzymolysis liquid to 80 ℃, inactivating the enzyme for 20 minutes, adjusting the pH of the enzymolysis liquid to 4.5, and centrifuging. And (3) carrying out centrifugal separation on the enzymolysis liquid to obtain a first layer of clear oil, a second layer of emulsion layer, a third layer of waste liquid and a fourth layer of crude protein in sequence, and absorbing layer by layer to obtain primary xanthoceras sorbifolia Bunge oil, primary emulsion layer liquid and primary crude protein liquid.
(3) Adding ethanol into the primary emulsion layer liquid, and then carrying out cold bath treatment; the volume ratio of the ethanol to the primary emulsion layer is 3:1, the cold bath temperature is 0 ℃, and the cold bath time is 20 min. Introducing high-pressure steam into the enzymolysis liquid after the ethanol cold bath treatment for heat treatment and recovering ethanol, wherein the steam pressure is 20MPa, the steam temperature is 110 ℃, and the steam treatment time is 40 s. And after high-pressure steam treatment, centrifugal separation is carried out to obtain free oil.
(4) And (3) mixing the primary xanthoceras sorbifolia Bunge oil obtained in the step (2) with the free oil obtained in the step (3), and sequentially carrying out hydration degumming, deacidification and decoloration treatment to obtain the high-purity xanthoceras sorbifolia Bunge oil.
(5) Adjusting pH of the primary crude protein solution to 7.0, adding alkaline protease 2.2% of the weight of xanthoceras sorbifolia bunge kernel, performing alkali extraction to obtain primary alkali extract, centrifuging the primary alkali extract to obtain an upper-layer protein solution and a lower-layer precipitate, and separating the upper-layer protein solution and the lower-layer precipitate to obtain a primary protein solution and a primary precipitate.
(6) Secondary alkali extraction: and carrying out secondary alkali extraction on the primary precipitate to obtain secondary alkali extraction liquid, centrifuging the secondary alkali extraction liquid to obtain an upper-layer protein solution and a lower-layer precipitate, and separating the upper-layer protein solution and the lower-layer precipitate to obtain a secondary protein solution.
(7) Acid precipitation: and (3) uniformly mixing the primary protein solution and the secondary protein solution, adjusting the pH value, performing acid precipitation and centrifugation to obtain precipitated starch, and performing freeze drying on the precipitated starch to obtain the shinyleaf yellowhorn protein.
Example 3
The method for simultaneously preparing xanthoceras sorbifolia bunge oil and xanthoceras sorbifolia bunge protein by using the aqueous enzymatic method, which is provided by the embodiment 3 of the invention, comprises the following steps:
(1) obtaining slurry: mixing peeled shinyleaf yellowhorn kernels with water according to a material-liquid ratio of 2 g: and (3) putting 9mL of the mixture into a colloid mill for crushing, crushing shinyleaf yellowhorn kernels to 160 meshes, and collecting serous fluid.
(2) Enzymolysis: and (3) adjusting the pH value of the slurry ground in the step one to 6.0, adding pectinase and cellulase for enzymolysis, and carrying out enzymolysis reaction in a water bath at 65 ℃ for 5.5 hours. After the enzymolysis reaction is finished, heating the enzymolysis liquid to 85 ℃ to inactivate the enzyme for 25 minutes, adjusting the pH of the enzymolysis liquid to 5, and then centrifuging. And (3) carrying out centrifugal separation on the enzymolysis liquid to obtain a first layer of clear oil, a second layer of emulsion layer, a third layer of waste liquid and a fourth layer of crude protein in sequence, and absorbing layer by layer to obtain primary xanthoceras sorbifolia Bunge oil, primary emulsion layer liquid and primary crude protein liquid.
(3) Adding ethanol into the primary emulsion layer liquid, and then carrying out cold bath treatment; the volume ratio of the ethanol to the primary emulsion layer is 5.5:1, the cold bath temperature is 30 ℃, and the cold bath time is 25 min. Introducing high-pressure steam into the enzymolysis liquid after the ethanol cold bath treatment for heat treatment and recovering ethanol, wherein the steam pressure is 25MPa, the steam temperature is 120 ℃, and the steam treatment time is 65 s. And after high-pressure steam treatment, centrifugal separation is carried out to obtain free oil.
(4) And (3) mixing the primary xanthoceras sorbifolia Bunge oil obtained in the step (2) with the free oil obtained in the step (3), and sequentially carrying out hydration degumming, deacidification and decoloration treatment to obtain the high-purity xanthoceras sorbifolia Bunge oil.
(5) Adjusting pH of the primary crude protein liquid to 7.4, adding alkaline protease 2.6% of the weight of xanthoceras sorbifolia bunge kernel, performing alkali extraction to obtain primary alkali extract, centrifuging the primary alkali extract to obtain an upper-layer protein solution and a lower-layer precipitate, and separating the upper-layer protein solution and the lower-layer precipitate to obtain a primary protein liquid and a primary precipitate.
(6) Secondary alkali extraction: and carrying out secondary alkali extraction on the primary precipitate to obtain secondary alkali extraction liquid, centrifuging the secondary alkali extraction liquid to obtain an upper-layer protein solution and a lower-layer precipitate, and separating the upper-layer protein solution and the lower-layer precipitate to obtain a secondary protein solution.
(7) Acid precipitation: and (3) uniformly mixing the primary protein solution and the secondary protein solution, adjusting the pH value, performing acid precipitation and centrifugation to obtain precipitated starch, and performing freeze drying on the precipitated starch to obtain the shinyleaf yellowhorn protein.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (10)
1. A method for simultaneously preparing xanthoceras sorbifolia Bunge oil and xanthoceras sorbifolia Bunge protein by an aqueous enzymatic method is characterized by comprising the following steps:
step one, soaking the peeled shinyleaf yellowhorn kernels in clear water for 2 hours, and mixing the soaked shinyleaf yellowhorn kernels with water according to a material-liquid ratio of 2 g: putting 3-9 mL of the mixture into a colloid mill for crushing, crushing shinyleaf yellowhorn kernels to 80-180 meshes, and collecting slurry;
regulating the pH value of the ground slurry to 3.7-6.0, carrying out enzymolysis, heating the enzymolysis liquid to 75-85 ℃ to inactivate enzyme for 15-25 minutes after the enzymolysis reaction is finished, regulating the pH value of the enzymolysis liquid to 4-5, centrifuging the enzymolysis liquid at the speed of 5000r/min for 20 minutes at room temperature, separating to obtain a first layer of clear oil, a second layer of emulsion layer, a third layer of waste liquid and a fourth layer of crude protein in sequence, and absorbing layer by layer to obtain primary xanthoceras sorbifolia Bunge oil, primary emulsion layer liquid and primary crude protein liquid;
performing ultrasonic treatment on the primary emulsion layer to obtain free oil;
step four, mixing the primary xanthoceras sorbifolia Bunge oil obtained in the step two with the free oil obtained in the step three, and sequentially carrying out hydration degumming, deacidification and decoloration treatment to obtain high-purity xanthoceras sorbifolia Bunge oil;
and step five, adjusting the pH value of the primary crude protein liquid to 7.5-8, adding the shinyleaf yellowhorn kernel material liquid in a ratio of 1: 55, stirring the mixture fully, and reacting the mixture for 100 minutes at the temperature of 40 ℃ to obtain primary alkali extract; after the primary alkali extract is naturally cooled to room temperature, centrifuging by using a centrifugal machine to obtain an upper protein solution and a lower precipitate, and separating the upper protein solution and the lower precipitate to obtain a primary protein solution and a primary precipitate;
step six, the obtained primary precipitate is subjected to the treatment of the step five to obtain secondary alkali extract, the secondary alkali extract is centrifuged to obtain an upper layer protein solution and a lower layer precipitate, and the upper layer protein solution and the lower layer precipitate are separated to obtain a secondary protein solution;
and seventhly, uniformly mixing the primary protein liquid and the secondary protein liquid, adjusting the pH value, performing acid precipitation and centrifugation to obtain precipitated starch, and performing freeze drying on the precipitated starch to obtain the shinyleaf yellowhorn protein.
2. The method for simultaneously preparing xanthoceras sorbifolia oil and xanthoceras sorbifolia protein according to claim 1, wherein in the second step, the acid for adjusting pH is one or two of hydrochloric acid and citric acid, and the pH of the solution is adjusted and then added to the enzyme solution.
3. The method for simultaneously preparing xanthoceras sorbifolia bunge oil and xanthoceras sorbifolia bunge protein by using the aqueous enzymatic method as claimed in claim 1, wherein in the second step, the enzymatic hydrolysis specifically comprises the following steps:
performing enzymolysis for 2 hours at 30-60 ℃ by using 50-500U/g of composite cellulase, then adding 100-500U/g of protease and composite enzyme thereof, and performing enzymolysis for 3 hours at 30-60 ℃;
the cellulase is composite cellulase;
the protease is one or more of papain, neutral protease, trypsin, alkaline protease, pepsin and acidic protease;
the compound enzyme is a compound of protease, xylanase and pectinase.
4. The method for simultaneously preparing xanthoceras sorbifolia oil and xanthoceras sorbifolia protein by the aqueous enzymatic method according to claim 1, wherein in the third step, the ultrasonic treatment time is 30 minutes, and the ultrasonic frequency is 25-40k Hz.
5. The method for simultaneously preparing xanthoceras sorbifolia bunge oil and xanthoceras sorbifolia bunge protein by the aqueous enzymatic method according to claim 1, wherein in the fourth step, the degumming method of the xanthoceras sorbifolia bunge oil comprises the following steps:
(1) inputting the crude oil into a refining tank through a pipeline, heating the crude oil to 75-80 ℃, adding edible salt into warm water at 85 ℃, and melting the edible salt for 20min to form brine, wherein the mass ratio of the salt to the water in the brine is 1: 35;
(2) adding brine into a refining tank, mixing the brine and crude oil according to the mass ratio of 1:7, stirring for 10min under the condition of 60r/min of a stirrer, observing the separation condition of oil and soap, stopping stirring after obvious separation, precipitating for 5h, discharging saponin water until a small amount of oil is discharged, continuously washing twice until all precipitates are discharged, and finishing degumming.
6. The method for simultaneously preparing xanthoceras sorbifolia oil and xanthoceras sorbifolia protein by the aqueous enzymatic method as claimed in claim 1, wherein in the fourth step, the deacidification method comprises the following steps: pumping shinyleaf yellowhorn crude oil into an oil refining tank, heating to 65 ℃, adding 12-14% Baume alkali liquor into the oil refining tank, stirring at the speed of 40r/min, continuing to stir for 30min after the alkali liquor is added, observing, starting to heat to 62-65 ℃ when soap grains in emulsion are in a separated state, easily separating soapstock from oil after the final temperature is reached, quickly sinking, standing for 3-7 h, and then washing with water.
7. The method for simultaneously preparing xanthoceras sorbifolia oil and xanthoceras sorbifolia protein by the aqueous enzymatic method according to claim 7, wherein the water washing method comprises the following steps: and (3) after standing and settling, discharging soapstock from the bottom of the oil refining tank, adjusting the oil temperature to 70-80 ℃, washing with hot water at the temperature of 5-10 ℃ higher than the oil temperature for 2-4 times, wherein the hot water added each time accounts for 10% -15% of the weight of the oil, continuously stirring at the speed of 45r/min in the washing process, stirring for 35min, standing for 2h, and discharging wastewater.
8. The method for simultaneously preparing xanthoceras sorbifolia bunge oil and xanthoceras sorbifolia bunge protein by the aqueous enzymatic method according to claim 1, wherein in the fourth step, the decolorization method comprises the following steps: the washed oil contains 0.6 percent of water, the oil is dried firstly, the vacuum residual pressure of a decoloring tank is set to be 4 Kpa-7 Kpa, the oil in an oil refining tank in the step three is sucked into the decoloring tank, the volume of the oil in the decoloring tank accounts for 2/3 of the volume of the tank body of the decoloring tank, the temperature is raised while stirring, the temperature of the oil sucked into the decoloring tank is controlled to be 90-110 ℃, and the water content is reduced to be below 0.2 percent; then absorbing activated clay and activated carbon which are 2 to 5 percent of the weight of the oil into a decoloring tank through vacuum, and maintaining for more than 40min to complete decoloring; and filtering after the decolorization is finished.
9. The method for simultaneously preparing xanthoceras sorbifolia oil and xanthoceras sorbifolia protein according to claim 1, wherein in the fifth step, the alkali for adjusting the pH is one or two of sodium hydroxide and potassium hydroxide, and the pH of the solution is adjusted and then added into the enzyme solution.
10. The method for simultaneously preparing xanthoceras sorbifolia oil and xanthoceras sorbifolia protein by the aqueous enzymatic method as claimed in claim 1, wherein in the fifth step, the centrifugation specifically comprises:
the centrifugation speed is 4000r/min, and the centrifugation time is 15 min.
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| CN111607459A (en) * | 2020-06-04 | 2020-09-01 | 熊红玉 | Method for extracting crude oil and protein from fresh idesia polycarpa fruits |
| CN115820334A (en) * | 2022-12-09 | 2023-03-21 | 吉林省林业科学研究院 | Pilot-scale test process method for preparing shinyleaf yellowhorn oil by ultrasonic composite aqueous enzymatic method |
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