CN115029204A - Wine with SOD activity and preparation method thereof - Google Patents
Wine with SOD activity and preparation method thereof Download PDFInfo
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- CN115029204A CN115029204A CN202210836960.6A CN202210836960A CN115029204A CN 115029204 A CN115029204 A CN 115029204A CN 202210836960 A CN202210836960 A CN 202210836960A CN 115029204 A CN115029204 A CN 115029204A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/04—Preparation of other alcoholic beverages by mixing, e.g. for preparation of liqueurs
- C12G3/05—Preparation of other alcoholic beverages by mixing, e.g. for preparation of liqueurs with health-improving ingredients, e.g. flavonoids, flavones, polyphenols or polysaccharides
- C12G3/055—Preparation of other alcoholic beverages by mixing, e.g. for preparation of liqueurs with health-improving ingredients, e.g. flavonoids, flavones, polyphenols or polysaccharides extracted from plants
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Nutrition Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Botany (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a wine with SOD activity and a preparation method thereof, wherein the wine comprises SOD-containing plant extract, modified albumin, white spirit and a cross-linking agent; wherein the modified albumin-forming nanoparticles coat the SOD-containing plant extract. The SOD in the plant extract is coated by albumin and then is fused with the white spirit, so that the characteristics and typicality of the traditional white spirit are kept, the taste of the traditional white spirit is kept, no peculiar smell is generated, and the palatability is good. Meanwhile, SOD can keep activity in the white spirit and reach the intestines through the stomach of a human body for absorption, thereby efficiently removing excessive free radicals in the body and balancing the function of the human body; albumin is simultaneously absorbed as an important nutrient in the human body. The preparation process utilizes the difference of physicochemical properties of albumin in water and ethanol of white spirit to form albumin nanoparticles to coat SOD in the plant extract, and has the advantages of mild reaction, simple operation, few preparation steps and high reaction speed.
Description
Technical Field
The invention belongs to the technical field of functional white spirit, and particularly relates to wine with SOD activity and a preparation method thereof.
Background
White spirit is an indispensable part in Chinese traditional culture; from the dietary point of view, the proper amount of white spirit is helpful for promoting digestion and stimulating appetite. And the white spirit is fermented and brewed by grains, so the white spirit has the nutrient components contained in the grains, and the drinking of the white spirit can supplement various nutrient substances required by human bodies: protein, fat, saccharide, vitamins, inorganic salt and water. The wine can also generate a large amount of heat, can ensure normal life activities of human bodies, and can play a role in dispelling cold. In addition, drinking can protect the heart and blood vessels and keep people away from cardiovascular diseases.
Excess free radicals in the human body cause cellular damage and pigmentation, and human aging is a consequence of free radical accumulation and impaired clearance. Superoxide dismutase (SOD), a natural free radical scavenger, has very high stability and efficiency against oxidizing agents and can be used to prevent oxidative damage.
Therefore, the combination of the white spirit and SOD can efficiently remove the surplus free radicals in the body while drinking the spirit, and the balance of the human body function is one of the ways of realizing the white spirit functionalization and changing the concept of drinking and hurting the body of people; moreover, the white spirit is combined with SOD, so that the damage of alcohol to human bodies can be reduced when the white spirit is drunk excessively, and the white spirit containing SOD can be used in formal occasions such as business banquet, parties and the like.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention has the first object of providing the wine with SOD activity, wherein SOD components are derived from plant extracts, have higher SOD activity, are fused with the white wine, can stably exist in the white wine, keep the characteristics and typicality of the traditional white wine, keep the mouthfeel of the traditional white wine, have no peculiar smell and have good palatability; has the effects of repairing and protecting human body cells, efficiently eliminating excess free radicals in the body and balancing human body functions.
The second purpose of the invention is to provide a preparation method of the wine with SOD activity.
One of the purposes of the invention can be achieved by adopting the following technical scheme:
a wine with SOD activity comprises the following components in parts by weight: 2.2-20 parts of SOD-containing plant extract, 1-5 parts of modified albumin, 500 parts of white spirit and 0.01-0.5 part of cross-linking agent; wherein the modified albumin-forming nanoparticles coat the SOD-containing plant extract.
Further, the SOD-containing plant extract is one or a combination of more than two of a opuntia ficus-indica extract, a ampelopsis grossedentata extract, a chrysanthemum extract and a ginseng extract.
Further, the SOD-containing plant extract comprises the following components in parts by weight:
1-5 parts of opuntia ficus-indica extract, 1-5 parts of ampelopsis grossedentata extract, 1-5 parts of chrysanthemum extract and 1-5 parts of ginseng extract.
Further, the white spirit is brewed by pure grains; the mass fraction of alcohol in the white spirit is 38-60%.
Further, the modified albumin is formed by connecting an amino group at the surface end of albumin with mPEG (methoxy polyethylene glycol), wherein the albumin is one or a composition of more than two of human serum albumin, bovine serum albumin and ovalbumin; the mPEG is methoxy polyethylene glycol aldehyde; the molecular weight of the polyethylene glycol propionaldehyde is 350-750 g/mol.
Further, the preparation method of the modified albumin comprises the following steps:
dissolving the albumin and the mPEG with the mass ratio of 1:1-3 into water, adding a reducing agent according to the mass ratio of 1:2-4 of the mPEG to the reducing agent, slowly stirring until the mixture is uniformly mixed, reacting for 6-24h at 4-15 ℃, finishing the reaction, and freeze-drying to obtain the modified albumin.
Further, the cross-linking agent is glucose.
The second purpose of the invention can be achieved by adopting the following technical scheme:
a method for preparing wine with SOD activity comprises the following steps:
step S1, dissolving the modified albumin and the SOD-containing plant extract into water, and uniformly stirring to obtain a mixed solution;
step S2, carrying out high-pressure homogenization treatment on the mixed solution through microjet to obtain a homogenized solution;
and step S3, adding white spirit and glucose into the homogenized liquid, stirring uniformly, and standing to obtain the wine with SOD activity.
Further, the working pressure is controlled to be 50 to 100MPa during the high-pressure homogenization treatment in the step S2, and the time of the homogenization treatment is 5 to 10 min.
Further, the standing in the step S3 is carried out for 2-12h at the temperature of 40-50 ℃.
Compared with the prior art, the invention has the beneficial effects that:
1. the wine with SOD activity is based on the traditional fermented white wine, and the albumin is used for coating SOD in the plant extract and then is fused with the white wine, so that the wine body keeps the characteristics and typicality of the traditional white wine, keeps the taste of the traditional white wine, has no peculiar smell and has good palatability. The albumin is utilized to form albumin nano particles in the white spirit to coat SOD in the plant extract, so that the SOD can keep activity in the white spirit, and can reach the intestinal part for absorption without being decomposed through the stomach of a human body, thereby efficiently removing the excess free radicals in the body and balancing the function of the human body. Albumin is simultaneously absorbed as an important nutrient in the human body.
2. The preparation method of the wine with SOD activity, which is disclosed by the invention, utilizes the physical and chemical property difference of albumin in water and ethanol of white spirit to form albumin nanoparticles to coat SOD in plant extracts, can be realized without additional reagents and reaction processes, and is simple to operate, few in preparation steps, high in reaction speed and free of special equipment.
Drawings
Fig. 1 is an SEM image of nanoparticles of modified albumin coated SOD containing plant extracts prepared in example.
Detailed Description
The technical solution of the present invention will be clearly and completely described below with reference to the specific embodiments. It is to be understood that the described embodiments are merely some embodiments of the invention, and not all embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
SOD is a kind of metal enzyme, generally obtained from animal blood, plant or by microbial fermentation and genetic engineering method, in the course of using, must avoid the destruction of amino acid structure and pepsin enzymolysis when directly eating that high temperature and organic reagent cause, therefore need special storage and edible method.
A wine with SOD activity comprises the following components in parts by weight: 2.2-20 parts of SOD-containing plant extract, 1-5 parts of modified albumin, 500 parts of white spirit and 0.01-0.5 part of cross-linking agent; wherein the modified albumin-forming nanoparticles coat the SOD-containing plant extract.
The SOD of the invention is derived from plant extracts, and the SOD is obtained by the plant extracts, so that the effects of other active substances in the plant extracts can be utilized to enhance the functions of human bodies. In view of the fact that white spirit contains a large amount of ethanol, the spatial structure of amino acid in SOD and coordination of copper ions and zinc ions realize the action of active enzyme of SOD, and ethanol serving as an organic reagent is easy to damage the spatial structure of the amino acid, so that the SOD activity is reduced and even inactivated. Therefore, the SOD component in the plant extract is coated by the nano-particles formed by the modified albumin, so that the coated SOD can exist in the white spirit for a long time and keeps activity. And after the modified albumin is directly drunk, the modified albumin resists the decomposition of pepsin or the SOD successfully passes through the stomach to reach the small intestine and is released through the decomposition of the modified albumin, and the SOD is absorbed by the small intestine to realize the retention of activity and play a role in eliminating surplus free radicals in the body and balancing the functions of a human body.
In one embodiment, the SOD-containing plant extract is one or a combination of two or more of opuntia ficus-indica extract, ampelopsis grossedentata extract, chrysanthemum extract, and ginseng extract.
The plant extract contains SOD active components, wherein the Opuntia Ficus lndica extract contains SOD components, and is obtained by extracting stem, leaf, root, fruit, and outer skin of Opuntia Ficus lndica plant.
Ampelopsis grossedentata is a common medicinal material for minority people in Yao, Dong, Tujia and the like in China. The Ampelopsis grossedentata extract is rich in flavonoids, polyphenols and the like, and has antioxidant and anti-inflammatory effects; and has effects in scavenging free radicals, and delaying skin aging.
Taking stems and leaves of Ampelopsis grossedentata (Ampelopsis grossedentata) of Ampelopsis as raw materials, extracting with solvent, filtering, and spray drying to obtain Ampelopsis grossedentata extract; wherein the solvent is water, ethanol or ethanol water solution; the ratio of material to liquid is 1: 10-30 parts of; the extraction temperature is 40-100 ℃, and the extraction time is 1-h.
The flos Chrysanthemi extract is prepared from dry cephalanthus of flos Chrysanthemi of Compositae, and contains flavonoids and phenols as main active ingredients. The chrysanthemum extract chrysanthemum can be obtained by ultrasonic extraction: ultrasonically treating dry capitula of flos Chrysanthemi in 5-10 times of 50-80% ethanol solution at 25-40 deg.C for 30-120min, extracting for 2-3 times, concentrating the extractive solution, and vacuum drying to obtain flos Chrysanthemi extract.
Ginseng is known as the king of all herbs in China, is a marked medicinal material in traditional Chinese medicines, and is widely applied to medicines, health care products and cosmetics. Modern medical research proves that the traditional Chinese medicine composition has good protection effect on the central nervous system, the cardiovascular and cerebrovascular system, the respiratory system, the blood and hematopoietic system, the endocrine system and the reproductive system. The main effective component contained in the ginseng is ginsenoside, and other components such as volatile oil, polysaccharide, amino acid, trace elements and the like are also contained; the ginseng extract can be obtained by a general extraction method.
The modified albumin is mainly characterized in that functional groups are connected to albumin, so that the solubility and the stability of the albumin in white spirit are improved. The cross-linking agent can make the amino part of lysine residue and the guanidyl part of arginine on the side chain of albumin undergo condensation reaction with aldehyde group, so that the nano-particles formed by solidifying albumin are stable.
The components of the invention can keep the characteristics and typicality of the traditional white spirit, keep the taste of the traditional white spirit, have no peculiar smell and have good palatability. Wherein, the SOD can be absorbed into blood rapidly to promote blood circulation by using alcohol as carrier; promoting alcohol metabolism of liver, and relieving hangover; removing surplus free radicals in human blood and reducing blood viscosity; SOD can accelerate alcohol metabolism in liver, timely discharge alcohol metabolite out of body, relieve alcohol damage to liver, and repair and activate cells; the white spirit is rich in SOD active enzyme and various amino acids and mineral substances necessary for human bodies, and has the functions of promoting and bidirectionally regulating the immune system of the human bodies; SOD has super-strong oxidation resistance, can inhibit synthesis and release of inflammatory factors such as prostaglandin, leukotriene and the like, penetrates the barrier of prostatic lipid membrane by virtue of high penetrability of white spirit, removes toxin, expands blood vessels, promotes blood circulation, removes meridian obstruction, removes blood stasis, and improves local microcirculation disturbance; SOD can protect female endocrine system, regulate female physiological cycle and menstrual period irregularity, and postpone climacteric period.
In one embodiment, the SOD-containing plant extract comprises the following composition in parts by weight:
1-5 parts of opuntia ficus-indica extract, 1-5 parts of ampelopsis grossedentata extract, 1-5 parts of chrysanthemum extract and 1-5 parts of ginseng extract.
In one embodiment, the white spirit is brewed by pure grains; the mass fraction of alcohol in the white spirit is 38-60%. The white spirit brewed by pure grains contains nutrient substances in grains, the mass fraction of the white spirit is 38-60%, the SOD coated by albumin can exist stably, the white spirit basically contains the component proportion of the white spirit sold on the market, and the white spirit with SOD activity can be prepared by taking the white spirit sold on the market as a raw material.
In one embodiment, the modified albumin is formed by connecting an amino group at the surface end of albumin with mPEG, and the albumin is one or a combination of more than two of human serum albumin, bovine serum albumin and ovalbumin; the mPEG is methoxy polyethylene glycol aldehyde; the molecular weight of the polyethylene glycol propionaldehyde is 350-750 g/mol.
The albumin contains N-terminal amino, so that albumin can be modified and modified by taking the amino as a connecting functional group to react with other substances, and the human serum albumin, the bovine serum albumin and the ovalbumin are commonly used albumins in the field and have high nutritional value. The mPEG is a derivative of polyethylene glycol, the albumin is modified by methoxy polyethylene glycol aldehyde, aldehyde groups in the methoxy polyethylene glycol aldehyde are used as reaction groups to react with amino groups of the albumin, and the active aldehyde groups couple the polyethylene glycol to the amino groups of protein molecules, so that the albumin is modified.
The choice of mPEG firstly has a repeating unit with an ethanol structure, so that in white spirit, the solubility and the dispersibility of the modified albumin in the white spirit can be enhanced, and the modified albumin can be uniformly dispersed in the white spirit without agglomeration or precipitation after being formed into nanoparticles. On the other hand, the chain structure of mPEG is on the surface of the modified albumin, so that the contact of ethanol molecules in the white spirit to the modified albumin nanoparticles can be reduced, and the nanoparticles are prevented from being damaged. In addition, when the white spirit reaches the stomach after drinking, the mPEG can also prevent the damage of gastric acid or pepsin to albumin, so that the albumin nanoparticles can continuously coat and protect the SOD.
In one embodiment, the modified albumin is prepared by the following steps:
dissolving the albumin and the mPEG with the mass ratio of 1:1-3 into water, adding a reducing agent according to the mass ratio of 1:2-4 of the mPEG to the reducing agent, slowly stirring until the mixture is uniformly mixed, reacting for 6-24h at 4-15 ℃, finishing the reaction, and freeze-drying to obtain the modified albumin.
The modified albumin is prepared by modifying albumin by mPEG reduction alkylation technology, wherein a reducing agent is helpful for realizing stable alkylation modification. Preferably, the reducing agent is sodium cyanoborohydride.
In one embodiment, the cross-linking agent is glucose. The cross-linking agent is used to make the albumin-forming nanoparticles more robust. The commonly used cross-linking agent is aldehyde, but the aldehyde has substances to human bodies, so the application takes glucose as the cross-linking agent, and the using amount of the cross-linking agent is not high, so that the taste and the mouthfeel of the white spirit can not be changed by adding a small amount of glucose, and the pungency of the white spirit can be reduced to a certain extent, so that the white spirit is more soft and soft.
The invention also provides a preparation method of the wine with SOD activity, which comprises the following steps:
step S1, dissolving the modified albumin and the SOD-containing plant extract into water, and uniformly stirring to obtain a mixed solution;
step S2, carrying out high-pressure homogenization treatment on the mixed solution through microjet to obtain a homogenized solution;
and step S3, adding white spirit and glucose into the homogenized solution, stirring uniformly, and standing to obtain the wine with SOD activity.
Step S2 is to use micro-jet high pressure homogenization treatment to mix the modified albumin and the plant extract containing SOD uniformly and disperse the SOD component in the plant extract containing SOD uniformly to realize the full contact with the modified albumin. And the albumin has viscosity and colloid, so that the modified albumin can be close to the SOD due to the binding property of the modified albumin to copper and zinc ions in the SOD in the process of contacting the SOD, and the SOD is coated when the modified albumin forms nanoparticles.
Step S3, adding liquor and glucose into the homogeneous liquid, wherein the liquor contains 38-60% by mass of ethanol, which belongs to organic reagent, so that the modified albumin is denatured and aggregated in the organic reagent ethanol, and the glucose is used as a cross-linking agent to enable amino parts or other active groups of residues on side chains of the albumin to perform condensation reaction with aldehyde groups, so that the albumin is solidified into nanoparticles, and the SOD is coated and protected in the process.
In one embodiment, the working pressure during the high-pressure homogenization treatment in step S2 is controlled to be 50 to 100MPa, and the time for the homogenization treatment is 5 to 10 min. The high-pressure homogenization of the microjet can ensure that the modified albumin and the SOD-containing plant extract are fully and uniformly mixed, and on the other hand, the microjet is utilized to form micro particles in the mixed solution, so that when the white spirit is added, the modified albumin can be fully cured to form nano particles with uniform particle sizes, and the structure is stable; the coated SOD is uniform and the activity is stable.
In one embodiment, the standing in step S3 is at 40-50 deg.C for 2-12 h.
Example 1:
wine with SOD activity
Dissolving 0.1mol of ovalbumin and 0.1mol of methoxy polyethylene glycol aldehyde with the molecular weight of 350g/mol into water, then adding 0.2mol of sodium cyanoborohydride, slowly stirring until the mixture is uniformly mixed, reacting for 24 hours at 4 ℃, finishing the reaction, dialyzing the reaction solution, and freeze-drying to obtain the modified albumin. Dissolving 1 part of opuntia ficus-indica extract, 3 parts of ampelopsis grossedentata extract, 5 parts of chrysanthemum extract and 1 part of ginseng extract in 10 parts of water, adding 3 parts of modified albumin, uniformly stirring to obtain a mixed solution, then treating the mixed solution at 50MPa for 10min by using microjet to obtain a homogeneous solution, adding the homogeneous solution and 0.01 part of glucose into 500 parts of white spirit with the mass fraction of 38% of ethanol, uniformly stirring, standing at 40 ℃ for 12h, and naturally cooling to room temperature to obtain the wine with SOD activity.
Example 2:
wine with SOD activity
Dissolving 0.1mol of ovalbumin and 0.2mol of methoxy polyethylene glycol aldehyde with the molecular weight of 550g/mol into water, then adding 0.6mol of sodium cyanoborohydride, slowly stirring until the mixture is uniformly mixed, reacting for 15h at 10 ℃, finishing the reaction, dialyzing the reaction solution, and freeze-drying to obtain the modified albumin. Dissolving 3 parts of opuntia ficus-indica extract, 1 part of ampelopsis grossedentata extract, 3 parts of chrysanthemum extract and 5 parts of ginseng extract in 10 parts of water, adding 1 part of modified albumin, uniformly stirring to obtain a mixed solution, then treating the mixed solution for 7min at 75MPa by using microjet to obtain a homogeneous solution, adding the homogeneous solution and 0.25 part of glucose into 500 parts of white spirit with the mass fraction of 45% of ethanol, uniformly stirring, standing for 7h at 45 ℃, and naturally cooling to room temperature to obtain the wine with SOD activity.
Example 3:
wine with SOD activity
Dissolving 0.1mol of ovalbumin and 0.3mol of methoxy polyethylene glycol aldehyde with the molecular weight of 750g/mol into water, then adding 0.9mol of sodium cyanoborohydride, slowly stirring until the mixture is uniformly mixed, reacting for 6 hours at 15 ℃, finishing the reaction, dialyzing the reaction solution, and freeze-drying to obtain the modified albumin. Dissolving 5 parts of Opuntia ficus-indica extract, 5 parts of Ampelopsis grossedentata extract, 1 part of chrysanthemum extract and 3 parts of ginseng extract in 10 parts of water, adding 5 parts of modified albumin, uniformly stirring to obtain a mixed solution, then treating the mixed solution for 5min at 100MPa by using microjet to obtain a homogeneous solution, adding the homogeneous solution and 0.5 part of glucose into 500 parts of white spirit with the mass fraction of ethanol of 60%, uniformly stirring, standing for 2h at 50 ℃, and naturally cooling to room temperature to obtain the wine with SOD activity.
Example 4:
wine with SOD activity
Dissolving 0.1mol of bovine serum albumin and 0.15mol of methoxy polyethylene glycol aldehyde with the molecular weight of 550g/mol into water, then adding 0.15mol of sodium cyanoborohydride, slowly stirring until the mixture is uniformly mixed, reacting for 18h at 5 ℃, finishing the reaction, dialyzing the reaction solution, and freeze-drying to obtain the modified albumin. Dissolving 2 parts of Opuntia ficus-indica extract, 4 parts of Ampelopsis grossedentata extract, 5 parts of chrysanthemum extract and 2 parts of ginseng extract in 10 parts of water, adding 4 parts of modified albumin, uniformly stirring to obtain a mixed solution, treating the mixed solution for 7min at 75MPa by using microjet to obtain a homogeneous solution, adding the homogeneous solution and 0.25 part of glucose into 500 parts of white spirit with the mass fraction of ethanol of 45%, uniformly stirring, standing for 7h at 45 ℃, and naturally cooling to room temperature to obtain the wine with SOD activity.
Example 5:
wine with SOD activity
Dissolving 0.1mol of human serum albumin and 0.25mol of methoxy polyethylene glycol aldehyde with the molecular weight of 400g/mol into water, then adding 0.63mol of sodium cyanoborohydride, slowly stirring until the mixture is uniformly mixed, reacting for 12 hours at 10 ℃, finishing the reaction, dialyzing the reaction solution, and freeze-drying to obtain the modified albumin. Dissolving 4 parts of opuntia ficus-indica extract, 1 part of ampelopsis grossedentata extract, 3 parts of chrysanthemum extract and 5 parts of ginseng extract in 10 parts of water, adding 4 parts of modified albumin, uniformly stirring to obtain a mixed solution, then treating the mixed solution for 10min at 50MPa by using microjet to obtain a homogeneous solution, adding the homogeneous solution and 0.1 part of glucose into 500 parts of white spirit with the mass fraction of 50% of ethanol, uniformly stirring, standing at 45 ℃ for 7h, and naturally cooling to room temperature to obtain the wine with SOD activity.
Comparative example 1
Compared with the example 2, 3 parts of opuntia ficus-indica extract, 1 part of ampelopsis grossedentata extract, 3 parts of chrysanthemum extract and 5 parts of ginseng extract are dissolved in 10 parts of water without using albumin, the mixture is uniformly stirred to obtain a mixed solution, then the mixed solution is treated for 7min under 75MPa by using microjet to obtain a homogeneous solution, the homogeneous solution and 0.25 part of glucose are added into 500 parts of white spirit with the mass fraction of ethanol of 45%, the mixture is uniformly stirred, then the mixture is kept stand at 45 ℃ for 7h, and then the temperature is naturally reduced to the room temperature to obtain the wine.
Comparative example 2
In contrast to example 2, no microfluidization was used for homogenization and sonication was used for treatment. Dissolving 0.1mol of ovalbumin and 0.2mol of methoxy polyethylene glycol aldehyde with the molecular weight of 550g/mol into water, then adding 0.6mol of sodium cyanoborohydride, slowly stirring until the mixture is uniformly mixed, reacting for 15h at 10 ℃, finishing the reaction, dialyzing the reaction solution, and freeze-drying to obtain the modified albumin. Dissolving 3 parts of opuntia ficus-indica extract, 1 part of ampelopsis grossedentata extract, 3 parts of chrysanthemum extract and 5 parts of ginseng extract in 10 parts of water, adding 1 part of modified albumin, uniformly stirring to obtain a mixed solution, carrying out ultrasonic treatment on the mixed solution for 7min to obtain a homogeneous solution, adding the homogeneous solution and 0.25 part of glucose into 500 parts of white spirit with the mass fraction of ethanol of 45%, uniformly stirring, standing at 45 ℃ for 7h, and naturally cooling to room temperature to obtain the wine with SOD activity.
Comparative example 3
Compared to example 2, albumin was not modified. Dissolving 3 parts of opuntia ficus-indica extract, 1 part of ampelopsis grossedentata extract, 3 parts of chrysanthemum extract and 5 parts of ginseng extract in 10 parts of water, adding 1 part of ovalbumin, uniformly stirring to obtain a mixed solution, carrying out ultrasonic treatment on the mixed solution for 7min to obtain a homogeneous solution, adding the homogeneous solution and 0.25 part of glucose into 500 parts of white spirit with the mass fraction of ethanol of 45%, uniformly stirring, standing at 45 ℃ for 7h, and naturally cooling to room temperature to obtain the wine with SOD activity.
Comparative example 4
Compared with the example 2, 0.1mol of ovalbumin and 0.2mol of methoxy polyethylene glycol aldehyde with the molecular weight of 550g/mol are dissolved in water without using SOD-containing plant extracts, then 0.6mol of sodium cyanoborohydride is added, the mixture is slowly stirred until the mixture is uniformly mixed and reacts for 15 hours at 10 ℃, the reaction is finished, and the modified albumin is obtained after the reaction liquid is dialyzed and freeze-dried. Adding 1 part of modified albumin into 10 parts of water, uniformly stirring to obtain a mixed solution, treating the mixed solution for 7min under 75MPa by using microjet to obtain a homogeneous solution, adding the homogeneous solution and 0.25 part of glucose into 500 parts of white spirit with the mass fraction of 45% of ethanol, uniformly stirring, standing for 7h at 45 ℃, and naturally cooling to room temperature to obtain the wine with SOD activity.
Test examples
1. SEM characterization
After removing water and ethanol from the wine having SOD activity prepared in example 2, freeze-drying the wine to remove water and ethanol, the remaining solid was modified albumin to coat the nanoparticles containing SOD plant extract, and the nanoparticles were characterized by electron microscopy, the results are shown in fig. 1. As can be seen from FIG. 1, the nanoparticles formed were rough in surface, relatively uniform in shape, nearly spherical, and between 200 and 300nm in diameter.
2. In vitro antioxidant Activity test
The wine having SOD activity of example 2 and the products of comparative examples 1 to 4 were tested for superoxide anion radical scavenging performance, p-OH scavenging ability.
Determination of superoxide anion scavenging ability: the superoxide anion scavenging ability is measured by pyrogallol autoxidation. Taking 1mL of newly prepared white spirit of example 2 and comparative examples 1-4, adding 4.5mL of 50mmol/L Tris-HCl (Tris: trihydroxyaminomethane) buffer solution with pH of 8.2 and 3.2mL of distilled water, uniformly mixing, keeping the temperature at 27 ℃ for 10min, immediately taking out, adding 0.3mL of preheated 3mmol/L pyrogallol solution at 25 ℃, taking distilled water as a reference solution, measuring absorbance at 320nm every 30s, measuring for 5min, and calculating the increase of absorbance per minute in a linear range. The blank control solution was replaced with 1mL of distilled water. The clearance was calculated according to the formula:
clearance (%) (. DELTA.Ae. DELTA.As)/. DELTA.Ae
Wherein Ae is the increase value of absorbance per minute of the blank control group; as is the increase in absorbance per minute for the sample set. After leaving the samples of example 2 and comparative examples 1 to 4 for 60 days, the superoxide anion scavenging ability was re-measured, and the results are shown in Table 1:
TABLE 1 superoxide anion radical scavenging capacity
As can be seen from Table 1, example 2 shows better superoxide anion radical scavenging ability than comparative examples 1-4, wherein comparative example 1 is not added with albumin and can not carry out coating protection on plant extracts containing SOD, so although the white spirit of comparative example 1 is added with SOD, the SOD can also have 67.14% superoxide anion scavenging ability when being added, but with the preservation of the plant extracts containing SOD in the white spirit, the activity of SOD is slowly inactivated by ethanol in the white spirit, the superoxide anion scavenging ability is reduced to only 32% of the original value after the preservation for 60 days, and the effect is greatly reduced. Comparative example 2 no homogenization treatment was performed using microjet during the preparation process, so the stability of albumin-coated SOD in the prepared wine having SOD activity was not strong, and the activity of SOD was lost after long-term storage for 60 days. The albumin of comparative example 3 is not modified, so that the albumin after forming nanoparticles directly contacts with ethanol in the white spirit without hindrance, thereby accelerating the damage of the albumin, and the SOD activity is also affected. In comparative example 4, no plant extract was added, and thus SOD components were substantially not contained, and thus superoxide anion radical scavenging ability was substantially not exhibited.
In vitro simulated gastric digestion tolerance experiment
Pepsin was suspended to a final concentration of 3g/L in PBS buffer at pH 3.0 as simulated gastric juice, the same volume of the white spirit having SOD activity of example 2 and the samples of comparative examples 1-4 were added, respectively, after shaking at constant temperature of 37 ℃ for 1 hour, the pH was adjusted to 7, and the change in superoxide anion radical scavenging ability before and after simulated gastric digestion tolerance was compared, and the results are shown in table 2.
TABLE 2 simulation of superoxide anion radical scavenging changes following gastric digestion
As can be seen from Table 2, the white spirit of example 2 still has a superoxide anion scavenging ability of 97.62% of the original performance of SOD after simulated gastric digestion, while the white spirit of comparative example 1 is not coated with albumin, and is directly added into the white spirit containing SOD plant extracts, and after simulated gastric digestion, the superoxide anion scavenging ability of SOD is only 12.32% of the original performance, which indicates that most of the SOD active ingredients of comparative example 1 are consumed or decomposed and lose the superoxide anion scavenging ability after simulated gastric digestion. Comparative examples 2 and 3 illustrate the stability and simulated gastric digestion tolerance of microfluidics treatment and albumin modified by mPEG. In contrast, comparative example 4, which contains no SOD component, shows a weak ability to scavenge superoxide anions even after simulated gastric digestion, and thus shows little change.
The OH scavenging ability was tested as follows:
adding 7mmol/L FeSO into the reaction system 4 And 7mmol/L salicylic acid-ethanol solution each 1mL, then 1mL sample solution, and finally 1mL 6mmol/L H 2 O 2 Starting reaction, keeping the temperature at 37 ℃ for 0.5h, taking distilled water as reference solution, and measuring the light absorption value A at 510nm 0 . Adding 1mL of distilled water into the blank control solution to replace the sample solution, and measuring the absorbance A 1 Adding 1mL of distilled water into the hydrolysate to replace H 2 O 2 The absorbance was measured as A 2 . Clearance was calculated according to the formula: has a p-OH clearance of (1- (A) 0 -A 2 )/A 1 ) x 100%. After the samples of example 2 and comparative examples 1 to 4 were left alone for 60 days, the superoxide anion scavenging ability was re-measured, and the results are shown in Table 3:
TABLE 3 test results for OH scavenging ability
As can be seen from Table 3, example 2 and comparative examples 1 to 4 have similar OH scavenging ability to superoxide anion radical scavenging results.
Therefore, as can be seen from tables 1 to 3, the SOD coated plant extract containing SOD as a white spirit additive can maintain better activity after long-term coexistence of SOD and white spirit, and can remove free radicals with high efficiency.
In conclusion, the wine with SOD activity is based on the traditional fermented white spirit, and the albumin is used for coating SOD in the plant extract and then is fused with the white spirit, so that the wine body keeps the characteristics and typicality of the traditional white spirit, keeps the taste of the traditional white spirit, has no peculiar smell and has good palatability. The albumin is utilized to form albumin nano particles in the white spirit to coat SOD in the plant extract, so that the SOD can keep activity in the white spirit, and can reach the intestinal part for absorption without being decomposed through the stomach of a human body, thereby efficiently removing the excess free radicals in the body and balancing the function of the human body. Albumin is simultaneously absorbed as an important nutrient in the human body. The preparation method has the advantages of simple operation, few preparation steps, high reaction speed and no need of special equipment.
The above embodiments are only preferred embodiments of the present invention, and the scope of the present invention should not be limited thereby, and any insubstantial changes and substitutions made by those skilled in the art based on the present invention are intended to be covered by the claims.
Claims (10)
1. The wine with SOD activity is characterized by comprising the following components in parts by weight: 2.2-20 parts of SOD-containing plant extract, 1-5 parts of modified albumin, 500 parts of white spirit and 0.01-0.5 part of cross-linking agent; wherein the modified albumin-forming nanoparticles coat the SOD-containing plant extract.
2. The wine with SOD activity according to claim 1, wherein the SOD-containing plant extract is one or more of Opuntia ficus-indica extract, Ampelopsis grossedentata extract, Chrysanthemum extract, and Panax ginseng extract.
3. The wine with SOD activity as claimed in claim 2, wherein the SOD-containing plant extract comprises the following components in parts by weight:
1-5 parts of opuntia ficus-indica extract, 1-5 parts of ampelopsis grossedentata extract, 1-5 parts of chrysanthemum extract and 1-5 parts of ginseng extract.
4. The wine with SOD activity according to claim 1, wherein the white wine is brewed from pure grain; the mass fraction of alcohol in the white spirit is 38-60%.
5. The wine with SOD activity according to claim 1, wherein the modified albumin is albumin with surface amino groups linked to mPEG, and the albumin is one or a combination of two or more of human serum albumin, bovine serum albumin and ovalbumin; the mPEG is methoxy polyethylene glycol aldehyde; the molecular weight of the polyethylene glycol propionaldehyde is 350-750 g/mol.
6. The wine with SOD activity as claimed in claim 5, wherein the modified albumin is prepared by the following steps:
dissolving the albumin and the mPEG with the mass ratio of 1:1-3 into water, adding a reducing agent according to the mass ratio of 1:2-4 of the mPEG to the reducing agent, slowly stirring until the mixture is uniformly mixed, reacting for 6-24h at 4-15 ℃, finishing the reaction, and freeze-drying to obtain the modified albumin.
7. Wine with SOD activity as claimed in claim 1, wherein the cross-linking agent is glucose.
8. The preparation method of the wine with SOD activity is characterized by comprising the following steps:
step S1, dissolving the modified albumin and the SOD-containing plant extract into water, and uniformly stirring to obtain a mixed solution;
step S2, carrying out high-pressure homogenization treatment on the mixed solution through microjet to obtain a homogenized solution;
and step S3, adding white spirit and glucose into the homogenized liquid, stirring uniformly, and standing to obtain the wine with SOD activity.
9. The method for preparing a wine having SOD activity according to claim 8,
and S2, controlling the working pressure to be 50-100 MPa during the high-pressure homogenization treatment, and controlling the time of the homogenization treatment to be 5-10 min.
10. The method for preparing a wine having SOD activity according to claim 8,
and step S3, standing for 2-12h at 40-50 ℃.
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