CN106075412A - A kind of antioxidation SOD compound enzyme capsule and preparation method thereof - Google Patents
A kind of antioxidation SOD compound enzyme capsule and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to a kind of antioxidation SOD compound enzyme capsule; in every 300mg capsule 's content, each constituent content is: SOD 5000～10000 unit of activity; CAT 500～1000 unit of activity; GSH Px 30～50 unit of activity; activity protecting agent 50～100mg; synergetic effect additive 30～50mg, functional oligose 150～200mg.Its preparation method is: first by trehalose, mannitol, lactose is prepared by mixing into activity protecting agent by certain mass ratio, by vitamin C, vitamin E, bata-carotene, GSH, NMN is prepared by mixing into synergetic effect additive according to certain mass ratio, use lauroyl chloride respectively to SOD and CAT again, SOD Yu GSH Px carries out cross-linking covalent modification, and add activity protecting agent, freeze-dried obtain modification SOD compound enzyme lyophilized powder, then by modification SOD compound enzyme lyophilized powder and synergetic effect additive, functional oligose mixed grinding, it is packed into capsule shells, obtain SOD compound enzyme capsule.SOD compound enzyme capsule stability obtained by the present invention is good, body superoxide dismutase (SOD) vigor can be significantly improved by mice and crowd being taken test proof, reduce internal malonaldehyde (MDA) content, there is oxygen-derived free radicals, antioxidation, the health-care effect of slow down aging in purged body.
Description
Technical field
The present invention relates to a kind of health food, be specifically related to a kind of antioxidation SOD compound enzyme capsule.
Background technology
Under the effect of some impairment factors, such as disease, wound, environmental pollution, ionizing radiation, ultraviolet irradiation, chemistry
Medicines etc. are the accumulation of a large amount of reactive oxygen free radical (ROS) in can inducing organism.Active oxygen chemical property is extremely active, has extremely strong
Oxidation susceptibility, easily capture the electronics of the biomolecule such as nucleic acid, aminoacid, cause body molecular level, cellular level and tissue
The various damages of organ level, accelerate the senescence process of body, and induce various disease.
Human body has complete defense system to the removing of reactive oxygen free radical (ROS).Whole defense system includes zymetology
Machine-processed and non-Enzymatic Mechanism.Superoxide dismutase (SOD), catalase (CAT) and paddy is included in zymetology defense system
Light sweet peptide peroxidase (GSH-Px) etc..Non-Enzymatic Mechanism is mainly the free radical scavenger of some small-molecular-weight, have Vc,
VE, beta-carotene, glutathion (GSH), reduced coenzyme Ⅰ (NADH) and NADPH (NADPH) etc..
Superoxide dismutase (SOD) is the ultra-oxygen anion free radical (O generally acknowledged in the world2 -. ) remover,
Ultra-oxygen anion free radical (O under the effect of SOD2 -. ) it is disproportionated into hydrogen peroxide (H2O2) and molecular oxygen (O2), thus generate
Hydrogen peroxide (H2O2) again through the effect point further such as catalase (CAT) or glutathione peroxidase (GSH-Px)
Solving is nontoxic water and oxygen, thus eliminates hydrogen peroxide (H2O2) murder by poisoning to body.SOD is catalyzed O2 -.Dismutation reaction formula is such as
Under:
Formula is it can be seen that organism defence reactive oxygen free radical is only inadequate by SOD, because SOD can only be clear from the reactions above
Except O2 -., the damage of protection activity oxygen-derived free radicals to greatest extent to be reached, thoroughly remove O2 -.And H2O2, really play body thin
The good protection effect of born of the same parents and histoorgan, still needs and wants other antioxidases such as catalase (CAT), peroxidase
And the synergism of glutathione peroxidase (GSH-Px) etc. (POD).Domestic experimentation confirms, at relatively SOD, CAT
During the ability of the lipid peroxidation injury caused by hyperbaric oxygen anti-with GSH-Px, the combined system effect of three kinds of enzymes is than any of which list
The effect of one enzyme to be got well.Also result of study is had to show, the compound enzyme pair containing multiple free radical scavenger with SOD as main component
Single SOD is substantially better than with the protection effect of the free radical resisting damage of the animal test model of type.This prompting, removes freely
Base, prevents its oxidative damage to body, and the effect depending single SOD alone is far from being enough, also needs to other antioxidation simultaneously
Enzyme and the collaborative and mutual aid effect of antioxidant.
At present, domestic market has many containing the antioxidation of SOD composition, defying age, the medicine of anti-disease and health care
Product, wherein have and have much declared patent of invention, as patent No. ZL201010604957.9 disclose a kind of defying age, blood pressure lowering,
Blood fat reducing SOD pharmaceutical composition and preparation method thereof, this patent mainly takes Radix Ginseng powder, Radix Notoginseng powder, Flos Carthami powder, ginkgo powder and Cortex Pini
After extract mixing drying and sterilizing, add SOD mixing, make capsule preparations；Patent No. ZL201110197534.4 discloses SOD
Collagen peptide U.S. face antidotal agent, this patent select natural plants SOD and collagen protein polypeptide, hyaluronic acid, Margarita powder with
And rationally the assembling of ginsenoside extract, there is activating cell function, improve immunity and effect of beautifying and anti-aging；Patent
Number ZL201110254795.5 discloses a kind of SOD complex capsule and preparation method thereof, this patent choose SOD, Radix Ginseng extract,
Margarita powder, Radix Codonopsis extract, Flos Carthami extract, Herb Gynostemmae Pentaphylli extract, spirulina, Cortex Pini Massonianae extract combine with lycium barbarum polysaccharide,
Making enteric capsulation, product has slow down aging, the effect of enhancing human body immunity power；Patent No. ZL200910059831.5 is public
Having opened a kind of fiveleaf gynostemma herb and lophatherum gracile SOD anti-cancer heath drink and preparation method thereof, this patent carries with Herb Gynostemmae Pentaphylli extract, Herba Lophatheri
Take thing, SOD, tea polyphenols are the anticancer health beverage that primary raw material is formulated, sterilizing is made.Product involved by these patent applications
Product are essentially all and form with Chinese medicine extract compatible combination with single SOD, and are inhaled by gastrointestinal digestion with oral way
Receive and enter physical exertion effect, do not account for supplementing other antioxidant reductases, small molecule antioxidant while supplementary SOD
And protector of enzyme activity.Therefore, prior art Shortcomings in terms of SOD activity stability and Antioxidant effectiveness
Part, it is impossible to play and remove interior free yl to greatest extent, the effect of conditioning health.
Summary of the invention
The purpose of the present invention is contemplated to overcome the deficiencies in the prior art, it is provided that a kind of based on high activity, high concentration SOD
Wanting composition and integrate the antioxidation SOD compound enzyme capsule of multiple antioxidase, activity protecting agent and synergetic effect additive, described answers
Synthase capsule contains superoxide dismutase (SOD), catalase (CAT), glutathion peroxidase (GSH-Px), lives
Property protective agent, synergetic effect additive, functional oligose, can the reactive oxygen free radical of excess in purged body to greatest extent, from
And realize effect of antioxidation, slow down aging.
It is an object of the invention to be achieved through the following technical solutions:
This antioxidation SOD compound enzyme capsule, it is characterised in that: containing superoxide dismutase in described compound enzyme capsule
(SOD), catalase (CAT), glutathion peroxidase (GSH-Px), activity protecting agent, synergetic effect additive, functional
Oligosaccharide, in every 300mg capsule 's content, each constituent content is:
SOD 5000～10000 unit of activity,
CAT 500～1000 unit of activity,
GSH-Px 30～50 unit of activity,
Activity protecting agent 50～100mg,
Synergetic effect additive 30～50mg,
Functional oligose 150～200mg.
Described superoxide dismutase (SOD) is pig erythrocyte extract, and unit enzyme activity is 3000～10000U/
mg。
Described catalase (CAT) unit enzyme activity is 2000～5000U/mg.
Described glutathion peroxidase (GSH-Px) unit enzyme activity is 100～500U/mg.
Described activity protecting agent is trehalose, mannitol and the mixture of lactose composition, trehalose, mannitol and lactose
Mass ratio be 2～1:1:0.5.
Described synergetic effect additive is vitamin C, vitamin E, beta-carotene, glutathion (GSH) and nicotiamide monokaryon
The mixture that thuja acid (NMN) forms, vitamin C, vitamin E, beta-carotene, the mass ratio of GSH and NMN are 5～7:1:1:1:
1。
Described oligosaccharide is any one in oligofructose, dextrinosan, oligomeric xylose or chitosan.
It is a further object to provide the preparation method of a kind of antioxidation SOD compound enzyme capsule, it is characterised in that:
Containing superoxide dismutase (SOD), catalase (CAT), glutathion peroxidase in described compound enzyme capsule
(GSH-Px), activity protecting agent, synergetic effect additive, functional oligose, in every 300mg capsule 's content, each constituent content is:
SOD 5000～10000 unit of activity, CAT 500～1000 unit of activity, GSH-Px 30～50 unit of activity, active protection
Agent 50～100mg, synergetic effect additive 30～50mg, functional oligose 150～200mg, comprise the technical steps that:
(1) preparation of activity protecting agent:
Weigh trehalose, mannitol and lactose respectively, be 2～1:1:0.5 according to the mass ratio of trehalose, mannitol and lactose
Ratio mix homogeneously, obtains activity protecting agent, standby；
(2) preparation of synergetic effect additive:
Weigh vitamin C, vitamin E, beta-carotene, GSH and NMN respectively, according to vitamin C, vitamin E, beta-carotene,
The mass ratio of GSH and NMN is the ratio mix homogeneously of 5～7:1:1:1:1, obtains synergetic effect additive, standby；
(3) modification of compound enzyme:
A. weigh the SOD of GSH-Px and proportional quantity 50% by proportioning to be dissolved in deionized water, be configured to total enzyme activity (with SOD enzyme
Live and GSH-Px enzyme work sum meter) composite enzyme solution between 1000～2000U/ml, water-bath is heated to 30
DEG C, with NaOH solution regulatory enzyme liquid pH value to 9.0, the lower dropping dressing agent continuously of stirring also starts timing, maintains enzyme liquid pH value
After 9.0,30 DEG C of insulation reaction 45min, being cooled to 10 DEG C with frozen water, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant
Liquid, through the ultra-filtration membrane device ultrafiltration that molecular cut off is 5000Da, removes unreacted dressing agent, collects and retain concentrated solution,
To modifying complex enzyme liquid I, standby；
B. weigh the SOD of CAT and proportional quantity 50% by proportioning to be dissolved in deionized water, be configured to total enzyme activity and (live with SOD enzyme
And CAT enzyme is lived sum meter) composite enzyme solution between 1000～2000U/ml, water-bath is heated to 30 DEG C, uses
NaOH solution regulatory enzyme liquid pH value is to 7.0, and stirring lower dropping continuously dressing agent also starts timing, maintenance enzyme liquid pH value 7.0,30 DEG C
After insulation reaction 60min, being cooled to 10 DEG C with frozen water, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant, through cutting
The ultra-filtration membrane device ultrafiltration staying molecular weight to be 5000Da, removes unreacted dressing agent, collects and retains concentrated solution, obtains modifying again
Synthase liquid II, standby；
C. merge to modify complex enzyme liquid with modifying complex enzyme liquid II by modification complex enzyme liquid I, add the step weighed by proportional quantity
Suddenly the activity protecting agent prepared by (1), stirring and dissolving mixing postlyophilization, obtain modifying compound enzyme lyophilized powder, standby；
(4) mixture obtains SOD compound enzyme powder:
I.e. synergetic effect additive as prepared by proportional quantity weighs functional oligose and step (2), adds step (3) C made
Standby modification compound enzyme lyophilized powder, is ground, and crosses 100 mesh sieves, obtains SOD compound enzyme powder；
(5) dress powder makes capsule:
Use conventional capsule fill process, the SOD compound enzyme powder of step (4) gained is loaded capsule shells, obtain SOD and be combined
Enzyme capsule.
Described in step (3) a and step (3) b of manufacture method, dressing agent is lauroyl chloride, and addition is that enzymatic solution is total
The 0.5-1.5% of quality.
Described in the step (5) of manufacture method, capsule is enteric coated capsule.
Below in conjunction with the performance of main composition of the present invention, the present invention will be further described:
(1) SOD
SOD is one of most important antioxidase in living organism.The physiological action of SOD is except by disproportionation O2 -., it is allowed to become
H2O and O2, make O2 -.Direct oxidation damaging action be suppressed outside, but also effectively prevent due to O2 -.Start and lead
The chain reaction of free radical that causes and prevent toxicity higher.The generation of OH.Therefore, in SOD compound enzyme system, keep enough
SOD activity and content, for whole antioxidant system defence radical damage play vital effect.
(2) CAT
Catalase (CAT) is removing H in living organism2O2Major Enzymes.The major physiological effect of CAT is catalyzed H exactly2O2,
It is allowed to be decomposed into H2O and O2, on the one hand eliminate H2O2The toxicity of biological active oxygen effect being had, on the other hand also makes H2O2Will not
In with O2 -.Under iron chelate effect, toxic action is higher, damaging effect is bigger in reaction generation.OH, therefore CAT and GSH-Px
Collectively form the second defence line of body defenses oxygen free radical injury.
(3) GSH-Px
Glutathion peroxidase (GSH-Px) is one of enzyme important in body anti-oxidative defense system.GSH-Px's is main
Physiological action is to remove lipid hydroperoxide (ROOH) to be reduced to avirulent ROH, and generation in the tissue that CAT content is little
For CAT, make H2O2It is changed into H2O and O2.Therefore, GSH-Px has removing H simultaneously2O2Work with lipid hydroperoxide (ROOH)
With, constitute anti-oxidative defense system in vivo together with SOD and CAT, have prevention distortion, prevent cell membrane and other biological tissue
Important biomolecule function from peroxide injury.GSH-Px the most constantly reduces, and is the factor of impact aging
One of.
(4) GSH
Glutathion (GSH) is mainly played a role by two ways: one is direct and free radical (R.) reaction, it is allowed to be changed into
Stable molecule (RH)；Two is the hydrogen donor of some macromolecular radical scavenger (such as GSH-Px), for ensureing that it removes H2O2
With ROOH, there is important function.Additionally, GSH also can make alpha-tocopherol regeneration indirectly play a role.
(5) vitamin C, vitamin E
Vitamin C and vitamin E are all very important free radical scavenger, antioxidant, and its major physiological effect not only may be used
Directly to remove O2 -., but also can remove ROOH,.OH、1O2And other free radicals.It addition, some enzyme needs mercapto freely
Base (-SH) could keep activity, and vitamin C can make disulfide group (-S-S) be reduced to sulfydryl (-SH), thus improve relevant enzyme
Activity, plays indirect antioxidant effect, and when finite concentration, vitamin C can also make vitamin E recover prototype, continues
Play its effect removing free radical.Therefore, there is also mutual synergism between vitamin C and vitamin E.
(6) beta-carotene
Bata-carotene is the precursor of vitamin A, is also a kind of antioxidant, has Detoxication, be to safeguard that health is not
The nutrient that can lack, has significant function in anticancer, prevention cardiovascular disease, cataract and antioxidation.Bata-carotene
CCl can be made3 O2 -.It is changed into the CCl of non-free radical3 O2, it is also possible to make free radical (R.) it is changed into stable molecule (RH),
Thus play the damaging action alleviating organic free radical.
(7) NMN
Nicotinamide mononucleotide. (NMN) is played an important role in human body cell energy generates, and it can change into energy i (in vivo) generation
Thank to requisite nicotinamide adenine dinucleotide (NAD), and NAD mediating cell death, antioxidation, oxidative stress, line grain
Body function, many important biological processes such as aging.Experimentation both domestic and external proves, NAD can make have aging resistance merit
The protein " Sirtuin " (deacetylase) of energy is the most active, and when human senility, in body, NAD amount can reduce therewith, and this
Plant change and easily cause the diseases such as metabolism decline and diabetes.NMN is proved in zoopery, has reverse nerve etc.
Partial organ organizes the effect of aging.At present, Japan is carrying out NMN defying age human clinical trial.
(8) trehalose, lactose and mannitol
Trehalose is a kind of safe and reliable natural saccharide, has non-specific protective effect to various bioactivators, can
Using the excellent activity protective agent as pharmaceutical grade protein, enzyme, vaccine and other biological goods.
Lactose and mannitol are all the freeze drying protectants of the biological product such as enzyme, vaccine, make the antioxygens such as SOD, CAT, GSH-Px
Change enzymatic activity is more stable, the holding time extends.Mannitol is also a kind of little molecular radical scavenger simultaneously, plays antioxidation
Synergism.
(9) functional oligose
Oligosaccharide has another name called oligosaccharide, is the little polymer being formed by connecting by glycosidic bond by 2-10 glycosyl.Functional oligose energy
Improve the propagation of microecological environment in human body, beneficially bacillus bifidus and other probiotics, suppression enteral Salmonella and corruption
The growth of bacterium, regulates gastrointestinal function, be eaten for a long time can delaying senility, relieving constipation, antibacterial, anti-cancer, anticancer, alleviate burden of liver, carry
High nutrition absorption rate.Some researchs in recent years show, some functional oligose such as oligofructose, dextrinosan, oligomeric
Galactose etc. not only have the function strengthening immunity and regulating intestinal canal flora, also have enhancing body antioxidative effect.Oligomeric
Sugar is by directly eliminating oxygen-derived free radicals, or by passway of metabolism, activate in body the antioxidase such as SOD, GSH-Px,
Make its antioxidant activity improve, thus reach antioxidation.
This antioxidation SOD compound enzyme capsule proposed according to above technical scheme, has the advantage that
(1) present invention based on high activity, high concentration SOD for main antioxidant enzyme component, and be aided with catalase (CAT)
With glutathion peroxidase (GSH-Px), combine little molecular radical scavenger V simultaneouslyC、VE, beta-carotene, gluathione
Peptide (GSH) and nicotinamide mononucleotide. (NMN), collectively constitute the antioxidation SOD compound enzyme system that a class is unique, in system each
Component can mutually be protected, mutually collaborative and potentiation, and this is the characteristic not available for single antioxidase or single antioxidant.
(2) present invention with low-molecular-weight lauroyl chloride (activated form of lauric acid (Lauric acid, be called for short LA)) for repairing
Decorations agent, respectively SOD Yu CAT, SOD Yu GSH-Px are carried out crosslinking covalent modification, generate covalently bound SOD-LA-CAT with
SOD-LA-GSH-Px compound enzyme system, substantially increases heat resistance and the stability at room temperature of antioxidase, extends
The holding time of antioxidase.
(3) in the SOD compound enzyme system with the connection of same dressing agent lauroyl chloride, due to the existence of CAT and GSH-Px,
Functionally eliminate the H that SOD produces2O2, protect SOD activity, produce good collaborative and mutual aid effect.
(4) product of the present invention is owing to using enteric coated capsule preparation, it is to avoid gastric juice and enzyme of proteolysis are to antioxygens such as SOD
Change the destruction of enzyme, it is ensured that effective ingredient fully absorbing and utilizing at intestinal.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is further detailed explanation, but embodiments of the present invention are also
It is not limited to the scope that this embodiment represents.
Embodiment 1:
The preparation method of this antioxidation SOD compound enzyme capsule, it is characterised in that: containing superoxides in described compound enzyme capsule
Dismutase (SOD), catalase (CAT), glutathion peroxidase (GSH-Px), activity protecting agent, synergetic effect additive,
Oligofructose, in 300g capsule 's content, each constituent content is: SOD 5,500,000 unit of activity, CAT 600,000 unit of activity, GSH-
Px 30,000 unit of activity, activity protecting agent 60g, synergetic effect additive 55g, oligofructose 185g.
The concrete step of preparation process of this antioxidation SOD compound enzyme capsule is as follows:
(1) preparation of activity protecting agent:
It is 2～1:1:0.5 ratios according to mass ratio, weighs trehalose 33g, mannitol 18g, lactose 9 respectively, mix homogeneously, i.e.
Obtain activity protecting agent 60g, standby；
(2) preparation of synergetic effect additive:
According to the ratio that mass ratio is 5～7:1:1:1:1, weigh respectively vitamin C 31g, vitamin E 6g, beta-carotene 6g,
GSH6g, NMN6g, mix homogeneously, obtain synergetic effect additive 55g, standby；
(3) modification of compound enzyme:
A. weigh GSH-Px 30,000 unit of activity, SOD275 ten thousand unit of activity is dissolved in 2L deionized water, is configured to compound enzyme molten
Liquid, is heated to 30 DEG C in water-bath, and with 10%NaOH solution regulatory enzyme liquid pH value to 9.0, stirring lower dropping continuously enzyme is molten
The lauroyl chloride of liquid gross mass 0.8% also starts timing, after maintaining 9.0,30 DEG C of insulation reaction 45min of enzyme liquid pH value, cold with frozen water
But to 10 DEG C, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant, through being the ultrafiltration of 5000Da through molecular cut off
Film device ultrafiltration, removes unreacted dressing agent, collects and retains concentrated solution, obtains modifying complex enzyme liquid I, standby；
B. weigh CAT 600,000 unit of activity, SOD275 ten thousand unit of activity is dissolved in 3L deionized water, is configured to compound enzyme molten
Liquid, is heated to 30 DEG C in water-bath, and with 10%NaOH solution regulatory enzyme liquid pH value to 7.0, stirring lower dropping continuously enzyme is molten
The lauroyl chloride of liquid gross mass 0.6% also starts timing, after maintaining 7.0,30 DEG C of insulation reaction 60min of enzyme liquid pH value, cold with frozen water
But to 10 DEG C, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant, through being the ultrafiltration of 5000Da through molecular cut off
Film device ultrafiltration, removes unreacted dressing agent, collects and retains concentrated solution, obtains modifying complex enzyme liquid II, standby；
C. merge to modify complex enzyme liquid with modifying complex enzyme liquid II by modification complex enzyme liquid I, add the work prepared by step (1)
Property protective agent 60g, stirring and dissolving mixing postlyophilization, obtain modify compound enzyme lyophilized powder, standby；
(4) mixture obtains SOD compound enzyme powder:
Weigh the synergetic effect additive 55g prepared by oligofructose 185g, step (2), add the modification prepared by step (3) C multiple
Synthase lyophilized powder, is ground, and crosses 100 mesh sieves, obtains SOD compound enzyme powder；
(5) dress powder makes capsule preparations:
Use conventional capsule fill process, the SOD compound enzyme powder of step (4) gained is loaded enteric capsule shell, obtains SOD
Compound enzyme capsule preparations 1000, every average net weight 300mg.
Instructions of taking and consumption: oral, adult daily 2 times, each 2.
Embodiment 2:
The preparation method of this antioxidation SOD compound enzyme capsule, it is characterised in that: containing superoxides in described compound enzyme capsule
Dismutase (SOD), catalase (CAT), glutathion peroxidase (GSH-Px), activity protecting agent, synergetic effect additive,
Dextrinosan, in 300g capsule 's content, each constituent content is: SOD 7,000,000 unit of activity, CAT 800,000 unit of activity,
GSH-Px 40,000 unit of activity, activity protecting agent 70g, synergetic effect additive 50g, dextrinosan 180g.
The concrete step of preparation process of this antioxidation SOD compound enzyme capsule is as follows:
(1) preparation of activity protecting agent:
It is 2～1:1:0.5 ratios according to mass ratio, weighs trehalose 31g, mannitol 26g, lactose 13 respectively, mix homogeneously, i.e.
Obtain activity protecting agent 70g, standby；
(2) preparation of synergetic effect additive:
According to the ratio that mass ratio is 5～7:1:1:1:1, weigh respectively vitamin C 30g, vitamin E 5g, beta-carotene 5g,
GSH5g, NMN5g, mix homogeneously, obtain synergetic effect additive 50g, standby；
(3) modification of compound enzyme:
A. weigh GSH-Px 40,000 unit of activity, SOD350 ten thousand unit of activity is dissolved in 2L deionized water, is configured to compound enzyme molten
Liquid, is heated to 30 DEG C in water-bath, and with 10%NaOH solution regulatory enzyme liquid pH value to 9.0, stirring lower dropping continuously enzyme is molten
The lauroyl chloride of liquid gross mass 1.0 % also starts timing, after maintaining 9.0,30 DEG C of insulation reaction 45min of enzyme liquid pH value, uses frozen water
Being cooled to 10 DEG C, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant, through being the super of 5000Da through molecular cut off
Filter membrane device ultrafiltration, removes unreacted dressing agent, collects and retains concentrated solution, obtains modifying complex enzyme liquid I, standby；
B. weigh CAT 800,000 unit of activity, SOD350 ten thousand unit of activity is dissolved in 3L deionized water, is configured to compound enzyme molten
Liquid, is heated to 30 DEG C in water-bath, and with 10%NaOH solution regulatory enzyme liquid pH value to 7.0, stirring lower dropping continuously enzyme is molten
The lauroyl chloride of liquid gross mass 0.8% also starts timing, after maintaining 7.0,30 DEG C of insulation reaction 60min of enzyme liquid pH value, cold with frozen water
But to 10 DEG C, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant, through being the ultrafiltration of 5000Da through molecular cut off
Film device ultrafiltration, removes unreacted dressing agent, collects and retains concentrated solution, obtains modifying complex enzyme liquid II, standby；
C. merge to modify complex enzyme liquid with modifying complex enzyme liquid II by modification complex enzyme liquid I, add the work prepared by step (1)
Property protective agent 70g, stirring and dissolving mixing postlyophilization, obtain modify compound enzyme lyophilized powder, standby；
(4) mixture obtains SOD compound enzyme powder:
Weigh the synergetic effect additive 50g prepared by dextrinosan 180g, step (2), add repairing prepared by step (3) C
Decorations compound enzyme lyophilized powder, is ground, and crosses 100 mesh sieves, obtains SOD compound enzyme powder；
(5) dress powder makes capsule:
Use conventional capsule fill process, the SOD compound enzyme powder of step (4) gained is loaded enteric capsule shell, obtains SOD
Compound enzyme capsule preparations 1000, every average net weight 300mg.
Instructions of taking and consumption: oral, adult daily 2 times, each 2.
Embodiment 3:
The preparation method of this antioxidation SOD compound enzyme capsule, it is characterised in that: containing superoxides in described compound enzyme capsule
Dismutase (SOD), catalase (CAT), glutathion peroxidase (GSH-Px), activity protecting agent, synergetic effect additive,
Oligomeric xylose, in 300g capsule 's content, each constituent content is: SOD 8,000,000 unit of activity, CAT 900,000 unit of activity, GSH-
Px 4.5 ten thousand unit of activity, activity protecting agent 80g, synergetic effect additive 50g, oligomeric xylose 170g.
The concrete step of preparation process of this antioxidation SOD compound enzyme capsule is as follows:
(1) preparation of activity protecting agent:
It is 2～1:1:0.5 ratios according to mass ratio, weighs trehalose 38g, mannitol 28g, lactose 14 respectively, mix homogeneously, i.e.
Obtain activity protecting agent 80g, standby；
(2) preparation of synergetic effect additive:
According to the ratio that mass ratio is 5～7:1:1:1:1, weigh Catergen 8g, vitamin E 5.5g, beta-carotene respectively
5.5g, GSH5.5g, NMN5.5g, mix homogeneously, obtain synergetic effect additive 50g, standby；
(3) modification of compound enzyme:
A. weigh GSH-Px 4.5 ten thousand unit of activity, SOD400 ten thousand unit of activity is dissolved in 3L deionized water, is configured to compound enzyme
Solution, is heated to 30 DEG C in water-bath, with 10%NaOH solution regulatory enzyme liquid pH value to 9.0, drips enzyme under stirring continuously
The lauroyl chloride of solution gross mass 0.9 % also starts timing, after maintaining 9.0,30 DEG C of insulation reaction 45min of enzyme liquid pH value, uses ice
Being water-cooled to 10 DEG C, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant, through being 5000Da's through molecular cut off
Ultra-filtration membrane device ultrafiltration, removes unreacted dressing agent, collects and retains concentrated solution, obtains modifying complex enzyme liquid I, standby；
B. weigh CAT 900,000 unit of activity, SOD400 ten thousand unit of activity is dissolved in 4L deionized water, is configured to compound enzyme molten
Liquid, is heated to 30 DEG C in water-bath, and with 10%NaOH solution regulatory enzyme liquid pH value to 7.0, stirring lower dropping continuously enzyme is molten
The lauroyl chloride of liquid gross mass 0.8% also starts timing, after maintaining 7.0,30 DEG C of insulation reaction 60min of enzyme liquid pH value, cold with frozen water
But to 10 DEG C, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant, through being the ultrafiltration of 5000Da through molecular cut off
Film device ultrafiltration, removes unreacted dressing agent, collects and retains concentrated solution, obtains modifying complex enzyme liquid II, standby；
C. merge to modify complex enzyme liquid with modifying complex enzyme liquid II by modification complex enzyme liquid I, add the work prepared by step (1)
Property protective agent 80g, stirring and dissolving mixing postlyophilization, obtain modify compound enzyme lyophilized powder, standby；
(4) mixture obtains SOD compound enzyme powder:
Weigh the synergetic effect additive 50g prepared by oligomeric xylose 170g, step (2), add the modification prepared by step (3) C multiple
Synthase lyophilized powder, is ground, and crosses 100 mesh sieves, obtains SOD compound enzyme powder；
(5) dress powder makes capsule preparations:
Use conventional capsule fill process, the SOD compound enzyme powder of step (4) gained is loaded enteric capsule shell, obtains SOD
Compound enzyme capsule preparations 1000, every average net weight 300mg.
Instructions of taking and consumption: oral, adult daily 2 times, each 2.
Embodiment 4:
The preparation method of this antioxidation SOD compound enzyme capsule, it is characterised in that: containing superoxides in described compound enzyme capsule
Dismutase (SOD), catalase (CAT), glutathion peroxidase (GSH-Px), activity protecting agent, synergetic effect additive,
Chitosan, in 300g capsule 's content, each constituent content is: SOD 9,500,000 unit of activity, CAT 950,000 unit of activity, GSH-Px
50000 unit of activity, activity protecting agent 90g, synergetic effect additive 55g, chitosan 155g.
The concrete step of preparation process of this antioxidation SOD compound enzyme capsule is as follows:
(1) preparation of activity protecting agent:
It is 2～1:1:0.5 ratios according to mass ratio, weighs trehalose 45g, mannitol 30g, lactose 15 respectively, mix homogeneously, i.e.
Obtain activity protecting agent 90g, standby；
(2) preparation of synergetic effect additive:
According to the ratio that mass ratio is 5～7:1:1:1:1, weigh vitamin C 34.2g, vitamin E 5.2g, beta-carotene respectively
5.2g, GSH5.2g, NMN5.2g, mix homogeneously, obtain synergetic effect additive 55g, standby；
(3) modification of compound enzyme:
A. weigh GSH-Px 50,000 unit of activity, SOD475 ten thousand unit of activity is dissolved in 3L deionized water, is configured to compound enzyme molten
Liquid, is heated to 30 DEG C in water-bath, and with 10%NaOH solution regulatory enzyme liquid pH value to 9.0, stirring lower dropping continuously enzyme is molten
The lauroyl chloride of liquid gross mass 1.0 % also starts timing, after maintaining 9.0,30 DEG C of insulation reaction 45min of enzyme liquid pH value, uses frozen water
Being cooled to 10 DEG C, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant, through being the super of 5000Da through molecular cut off
Filter membrane device ultrafiltration, removes unreacted dressing agent, collects and retains concentrated solution, obtains modifying complex enzyme liquid I, standby；
B. weigh CAT 950,000 unit of activity, SOD475 ten thousand unit of activity is dissolved in 4.5L deionized water, is configured to compound enzyme molten
Liquid, is heated to 30 DEG C in water-bath, and with 10%NaOH solution regulatory enzyme liquid pH value to 7.0, stirring lower dropping continuously enzyme is molten
The lauroyl chloride of liquid gross mass 0.8% also starts timing, after maintaining 7.0,30 DEG C of insulation reaction 60min of enzyme liquid pH value, cold with frozen water
But to 10 DEG C, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant, through being the ultrafiltration of 5000Da through molecular cut off
Film device ultrafiltration, removes unreacted dressing agent, collects and retains concentrated solution, obtains modifying complex enzyme liquid II, standby；
C. merge to modify complex enzyme liquid with modifying complex enzyme liquid II by modification complex enzyme liquid I, add the work prepared by step (1)
Property protective agent 90g, stirring and dissolving mixing postlyophilization, obtain modify compound enzyme lyophilized powder, standby；
(4) mixture obtains SOD compound enzyme powder:
Weigh the synergetic effect additive 55g prepared by chitosan 155g, step (2), add the modification prepared by step (3) C and be combined
Enzyme lyophilized powder, is ground, and crosses 100 mesh sieves, obtains SOD compound enzyme powder；
(5) dress powder makes capsule preparations:
Use conventional capsule fill process, the SOD compound enzyme powder of step (4) gained is loaded enteric capsule shell, obtains SOD
Compound enzyme capsule preparations 1000, every average net weight 300mg.
Instructions of taking and consumption: oral, adult daily 2 times, each 2.
Comparative example 1:
Difference with embodiment 1 is: do not modify SOD, CAT and GSH-Px.
Formula: in 300g capsule 's content, each constituent content is: SOD 5,500,000 unit of activity, CAT 600,000 unit of activity,
GSH-Px 30,000 unit of activity, activity protecting agent 60g, synergetic effect additive 55g, oligofructose 185g.
Concrete step of preparation process is as follows:
(1) preparation of activity protecting agent:
It is 2～1:1:0.5 ratios according to mass ratio, weighs trehalose 33g, mannitol 18g, lactose 9 respectively, mix homogeneously, i.e.
Obtain activity protecting agent 60g, standby；
(2) preparation of synergetic effect additive:
According to the ratio that mass ratio is 5～7:1:1:1:1, weigh respectively vitamin C 31g, vitamin E 6g, beta-carotene 6g,
GSH6g, NMN6g, mix homogeneously, obtain synergetic effect additive 55g, standby；
(3) mixture obtains SOD compound enzyme powder:
SOD 5,500,000 unit of activity, CAT 600,000 unit of activity, GSH-Px 30,000 unit of activity, oligofructose is weighed by proportioning
185g, the activity protecting agent 60g prepared by addition step (1), the synergetic effect additive 55g prepared by step (2), be ground,
Cross 100 mesh sieves, obtain SOD compound enzyme powder；
(4) dress powder makes capsule:
Use conventional capsule fill process, the SOD compound enzyme powder of step (3) gained is loaded enteric capsule shell, obtains SOD
Compound enzyme capsule preparations 1000, every average net weight 300mg.
Comparative example 2:
Difference with embodiment 2 is: do not add CAT and GSH-Px in formula.
Formula: in 300g capsule 's content, each constituent content is: SOD 7,000,000 unit of activity, activity protecting agent 70g, association
With synergist 50g, dextrinosan 180g.
Concrete step of preparation process is as follows:
(1) preparation of activity protecting agent:
It is 2～1:1:0.5 ratios according to mass ratio, weighs trehalose 31g, mannitol 26g, lactose 13 respectively, mix homogeneously, i.e.
Obtain activity protecting agent 70g, standby；
(2) preparation of synergetic effect additive:
According to the ratio that mass ratio is 5～7:1:1:1:1, weigh respectively vitamin C 30g, vitamin E 5g, beta-carotene 5g,
GSH5g, NMN5g, mix homogeneously, obtain synergetic effect additive 50g, standby；
(3) modification of SOD:
A. weigh SOD700 ten thousand unit of activity to be dissolved in 5L deionized water, be configured to SOD enzymatic solution, heat temperature raising in water-bath
To 30 DEG C, with 10%NaOH solution regulatory enzyme liquid pH value to 9.0, the lower lauroyl chloride dripping enzyme liquid gross mass 1.0 % continuously of stirring
And start timing, after maintaining 9.0,30 DEG C of insulation reaction 45min of enzyme liquid pH value, it is cooled to 10 DEG C with frozen water, 6000rpm low temperature (4
～8 DEG C) centrifugal 8min, collect supernatant, through through the ultra-filtration membrane device ultrafiltration that molecular cut off is 5000Da, removing unreacted
Dressing agent, collects and retains concentrated solution, obtain modification SOD enzyme liquid, standby；
B. adding the activity protecting agent 70g prepared by step (1) in modification SOD enzyme liquid, stirring and dissolving mixes postlyophilization,
Obtain modification SOD enzyme lyophilized powder, standby；
(4) mixture obtains SOD powder:
Weigh the synergetic effect additive 50g prepared by dextrinosan 180g, step (2), add repairing prepared by step (3) b
Decorations SOD enzyme lyophilized powder, is ground, and crosses 100 mesh sieves, obtains SOD powder；
(5) dress powder makes capsule:
Use conventional capsule fill process, the SOD powder of step (4) gained is loaded enteric capsule shell, obtains SOD capsule system
Agent 1000, every average net weight 300mg.
Comparative example 3:
Difference with embodiment 2 is: replace synergetic effect additive with the edible corn starch of equivalent in formula.
Formula: in 300g capsule 's content, each constituent content is: SOD 7,000,000 unit of activity, CAT 800,000 unit of activity,
GSH-Px 40,000 unit of activity, activity protecting agent 70g, edible corn starch 50g, dextrinosan 180g.
Concrete step of preparation process is as follows:
(1) preparation of activity protecting agent:
It is 2～1:1:0.5 ratios according to mass ratio, weighs trehalose 31g, mannitol 26g, lactose 13 respectively, mix homogeneously, i.e.
Obtain activity protecting agent 70g, standby；
(2) modification of compound enzyme:
A. weigh GSH-Px 40,000 unit of activity, SOD350 ten thousand unit of activity is dissolved in 2L deionized water, is configured to compound enzyme molten
Liquid, is heated to 30 DEG C in water-bath, and with 10%NaOH solution regulatory enzyme liquid pH value to 9.0, stirring lower dropping continuously enzyme is molten
The lauroyl chloride of liquid gross mass 1.0 % also starts timing, after maintaining 9.0,30 DEG C of insulation reaction 45min of enzyme liquid pH value, uses frozen water
Being cooled to 10 DEG C, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant, through being the super of 5000Da through molecular cut off
Filter membrane device ultrafiltration, removes unreacted dressing agent, collects and retains concentrated solution, obtains modifying complex enzyme liquid I, standby；
B. weigh CAT 800,000 unit of activity, SOD350 ten thousand unit of activity is dissolved in 3L deionized water, is configured to compound enzyme molten
Liquid, is heated to 30 DEG C in water-bath, and with 10%NaOH solution regulatory enzyme liquid pH value to 7.0, stirring lower dropping continuously enzyme is molten
The lauroyl chloride of liquid gross mass 0.8% also starts timing, after maintaining 7.0,30 DEG C of insulation reaction 60min of enzyme liquid pH value, cold with frozen water
But to 10 DEG C, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant, through being the ultrafiltration of 5000Da through molecular cut off
Film device ultrafiltration, removes unreacted dressing agent, collects and retains concentrated solution, obtains modifying complex enzyme liquid II, standby；
C. merge to modify complex enzyme liquid with modifying complex enzyme liquid II by modification complex enzyme liquid I, add the work prepared by step (1)
Property protective agent 70g, stirring and dissolving mixing postlyophilization, obtain modify compound enzyme lyophilized powder, standby；
(3) mixture obtains SOD compound enzyme powder:
Weigh edible corn starch 50g, dextrinosan 180g, add the modification compound enzyme lyophilizing prepared by step (2) C
Powder, is ground, and crosses 100 mesh sieves, obtains SOD compound enzyme powder；
(4) dress powder makes capsule:
Use conventional capsule fill process, the SOD compound enzyme powder of step (3) gained is loaded enteric capsule shell, obtains SOD
Compound enzyme capsule preparations 1000, every average net weight 300mg.
The finished capsule product obtaining above-described embodiment and comparative example carries out stability and the product function effect of antioxidase
Test evaluation.
The stability test evaluation of antioxidase in SOD compound enzyme capsule:
The heat stability of SOD: the capsule 's content that Example 1 and comparative example 1 obtain respectively, is configured to same concentration solution,
It is respectively placed under different temperatures insulation 2 hours, then detection SOD vigor, and calculates phase on the basis of the vigor of enzyme at 25 DEG C
To vigor (relative activity=enzyme of enzyme is at initial (benchmark) vigor of the vigor/enzyme in certain moment), the results are shown in Table 1:
As known from Table 1, the SOD vigor in embodiment 1 and comparative example 1 all rises with temperature and declines, but the SOD of embodiment 1 is relatively
The SOD relative activity of comparative example 1 is high.Illustrate that modified by lauroyl chloride SOD is significantly stronger than unmodified SOD to the tolerance of heat.
The heat stability of CAT Yu GSH-Px: the capsule 's content that Example 1 and comparative example 1 obtain respectively, is configured to same
One strength solution, is respectively placed in 50 DEG C of water-baths insulation 2 hours, and timing sampling mensuration CAT Yu GSH-Px lives at regular intervals
Power, and calculate relative activity on the basis of (when 0) enzyme activity of recording before insulation, the results are shown in Table 2:
As known from Table 2, under the conditions of 50 DEG C, the prolongation the most in time of CAT Yu the GSH-Px vigor of embodiment 1 and comparative example 1 and
Rapid decrease, the vigor of CAT with GSH-Px processing 1.5 hours comparative examples 1 completely loses, and CAT and GSH-of embodiment 1
Px vigor still remnants have 27% and 48%；.Illustrate that the thermostability of CAT and GSH-Px through modified by lauroyl chloride is than unmodified
CAT and GSH-Px is strong.
The stability that antioxidase is at room temperature deposited: the capsule that embodiment 1 and comparative example 1 obtain is put and deposits at room temperature
Put 1 year, the vigor of SOD, CAT and the GSH-Px in its content of sampling determination the most respectively, and to place
Calculate relative activity on the basis of the enzyme activity recorded for 1 day, the results are shown in Table 3:
As can be seen from Table 3, after compound enzyme capsule at room temperature places 1 year, the SOD vigor of embodiment 1 is stablized constant, and contrasts
The SOD vigor of example 1 declines nearly 1/3；The CAT vigor of embodiment 1 and comparative example 1 is the most more stable in 1-3 month, 12 months
The CAT vigor of the latter two all declines quickly, but the CAT vigor of embodiment 1 is apparently higher than comparative example 1；The GSH-Px of embodiment 1
Vigor is more stable in 1-6 month, and after 12 months, its relative activity still has 66%, and the GSH-Px vigor of comparative example 13 months
Rapid decrease later, after 12 months, its relative activity only has 20%.The antioxidase the passing through modification antioxidation compared with unmodified is described
Enzyme good stability at room temperature, long shelf-life.
The anti-oxidation efficacy test evaluation of SOD compound enzyme capsule:
Natural aged mouse gavages SOD compound enzyme capsule of the present invention and tests the impact of internal Antioxidant Indexes:
The male mice in kunming 66 of 12 monthly ages, body weight 40～50g is chosen in this test, is randomly divided into 6 groups, respectively matched group
1, matched group 2, matched group 3, test I group, test II group and test III group, often group 11.Matched group 1 gavages distilled water, comparison
Group 2 gavages the capsule obtained by comparative example 2, and matched group 3 gavages the capsule obtained by comparative example 3, tests I group and gavages embodiment 1
Obtained by capsule, test II group gavage embodiment 2 obtained by capsule, test III group gavage embodiment 3 obtained by capsule.
Dissolve capsule 's content with distilled water, all in gavage mode by the dosed administration of 200mg/kg.bw every day, every day 1 time, give continuously
Medicine (or distilled water) 30 days, matched group 1 gavages the distilled water of equivalent.After experiment terminates, coeliac artery takes blood, uses Nanjing to build up
Bioengineering Research Institute provide test kit and carry out serum superoxides discrimination respectively according to the method for test kit description introduction
Change enzyme (SOD) vigor (hydroxylamine assay), glutathion peroxidase (GSH-Px) vigor (DTNB method) and malonaldehyde (MDA) to contain
The detection of amount (TBA method).
Result such as table 4, table 5, table 6:
As can be seen from Table 4, under same dose, the SOD vigor being administered each dosage group is all remarkably higher than non-administration matched group 1
(P ＜ 0.05 or P < 0.01), each test group (test I group, test II group and test III group) SOD vigor pole be significantly higher than to
Medicine matched group 1(P ＜ 0.01.), and it is significantly higher than comparative example administration group (matched group 2 and matched group 3).Show in product of the present invention
The combination of each antioxidant content serves the effect of Synergistic, and it is evident in efficacy is better than single antioxidase or other antioxidation
Being used alone of agent, illustrates that SOD compound enzyme capsule of the present invention has the work being obviously enhanced nature aged mouse serum activity of SOD
With.
As can be seen from Table 5, compare with non-administration matched group 1, matched group 2, matched group 3 and the GSH-Px of test I group
Vigor is all without significant difference (P ＜ 0.05), but the GSH-Px vigor of test II group and test III group is significantly higher than non-administration pair
According to group 1 (P ＜ 0.05).Illustrate that SOD compound enzyme capsule of the present invention can significantly improve nature aged mouse GSH-Px in serum vigor.
In table 6 it can be seen that the MDA content being administered each dosage group be substantially less than non-administration matched group 1 (P ＜ 0.05 or P <
0.01), and each test group (test I group, test II group and test III group) MDA content pole be substantially less than non-administration matched group 1
(P ＜ 0.01.)；Test group of the present invention (test compared with comparative example administration group (matched group 2 and matched group 3), under same dose
I group, test II group and test III group) all can significantly reduce MDA content (P < 0.05), SOD compound enzyme capsule energy of the present invention is described
Significantly reduce nature aged mouse Content of MDA, there is antioxidative effect.
The test that internal Antioxidant Indexes is affected by human oral's SOD compound enzyme capsule:
Select on a voluntary basis the age 40-60 year, certain community in urban areas middle-aged and elderly people that physical condition is good be tested
Object, each 20 people of matched group 1, matched group 2, matched group 3 and test-meal group, wherein matched group 1 does not make any process, and matched group 2 takes
With the capsule obtained by comparative example 2, matched group 3 takes the capsule obtained by comparative example 3, obtained by test-meal group takes embodiment 2
Capsule.Instructions of taking and dosage is: take 2 times for each person every day, early, evening each the most once, each 2 (300mg/ grain), take continuously
With 30 days, during test, experimenter did not change original life and dietary habit, normal diet.After experiment terminates, 12 is little on an empty stomach
Time vein haemospasia.SOD vitality test uses xanthine oxidase (hydroxylamine assay), and MDA assay uses thiobarbituricacidα-
(TBA) method, SOD and MDA testing cassete is built up Bioengineering Research Institute by Nanjing and provides.
Result such as table 7, table 8:
As seen from Table 7, the SOD vigor that matched group 1 test-meal is forward and backward is without marked difference (P ＜ 0.05), matched group 2 and matched group 3
The forward and backward SOD vigor of test-meal all has marked difference (P ＜ 0.05)；SOD vigor after matched group 2 test-meal compares also with matched group 1
Have and significantly increase (P ＜ 0.01)；After test group test-meal with compare SOD vigor before test-meal and have pole to significantly improve (P < 0.01), with
Matched group 1 and matched group 3 compare SOD vigor has pole to significantly improve (P < 0.01).Illustrate that SOD compound enzyme capsule of the present invention has aobvious
Write the effect improving middle-aged and elderly people serum activity of SOD.
As seen from Table 8, before test-meal, test group compares MDA content all without significant difference (p ＜ 0.05), examination with each matched group
After food, test group MDA content significantly reduces (p ＜ 0.05) with comparing before test-meal, compares with matched group, MDA after test group test-meal
Content significantly reduces (p ＜ 0.05).Illustrate that SOD compound enzyme capsule of the present invention can significantly reduce middle-aged and elderly people Content of MDA,
The decline of MDA content the most just imply that interior free yl is swept off, and the anti-oxidation function of body is increased dramatically.
Claims (10)
1. an antioxidation SOD compound enzyme capsule, it is characterised in that: containing superoxide dismutase in described compound enzyme capsule
(SOD), catalase (CAT), glutathion peroxidase (GSH-Px), activity protecting agent, synergetic effect additive and function
Property oligosaccharide, in every 300mg capsule 's content, each constituent content is:
SOD 5000～10000 unit of activity,
CAT 500～1000 unit of activity,
GSH-Px 30～50 unit of activity,
Activity protecting agent 50～100mg,
Synergetic effect additive 30～50mg,
Functional oligose 150～200mg.
A kind of antioxidation SOD compound enzyme capsule the most according to claim 1, it is characterised in that: described SOD is that Sanguis sus domestica is red
Cell extract, unit enzyme activity is 3000～10000U/mg.
A kind of antioxidation SOD compound enzyme capsule the most according to claim 1, it is characterised in that: described catalase
(CAT) unit enzyme activity is 2000～5000U/mg.
A kind of antioxidation SOD compound enzyme capsule the most according to claim 1, it is characterised in that: described glutathion mistake
Oxide enzyme (GSH-Px) unit enzyme activity is 100～500U/mg.
A kind of antioxidation SOD compound enzyme capsule the most according to claim 1, it is characterised in that: described activity protecting agent
For the mixture of trehalose, mannitol and lactose composition, the mass ratio of trehalose, mannitol and lactose is 2～1:1:0.5.
A kind of antioxidation SOD compound enzyme capsule the most according to claim 1, it is characterised in that: described synergetic effect additive
The mixture formed for vitamin C, vitamin E, beta-carotene, glutathion (GSH) and nicotinamide mononucleotide. (NMN), dimension
Raw element C, vitamin E, beta-carotene, the mass ratio of GSH and NMN are 5～7:1:1:1:1.
A kind of antioxidation SOD compound enzyme capsule the most according to claim 1, it is characterised in that: described oligosaccharide is low
Any one in Polyfructose., dextrinosan, oligomeric xylose or chitosan.
8. the preparation method of the antioxidation SOD compound enzyme capsule described in an any one of claim 1-7, it is characterised in that: main
Include following processing step:
(1) preparation of activity protecting agent:
Weigh trehalose, mannitol and lactose respectively, be 2～1:1:0.5 according to the mass ratio of trehalose, mannitol and lactose
Ratio mix homogeneously, obtains activity protecting agent, standby；
(2) preparation of synergetic effect additive:
Weigh vitamin C, vitamin E, beta-carotene, GSH and NMN respectively, according to vitamin C, vitamin E, beta-carotene,
The mass ratio of GSH and NMN is the ratio mix homogeneously of 5～7:1:1:1:1, obtains synergetic effect additive, standby；
(3) modification of compound enzyme:
A. weigh the SOD of GSH-Px and proportional quantity 50% by proportioning to be dissolved in deionized water, be configured to total enzyme activity (with SOD enzyme
Live and GSH-Px enzyme work sum meter) composite enzyme solution between 1000～2000U/ml, water-bath is heated to 30
DEG C, with NaOH solution regulatory enzyme liquid pH value to 9.0, the lower dropping dressing agent continuously of stirring also starts timing, maintains enzyme liquid pH value
After 9.0,30 DEG C of insulation reaction 45min, being cooled to 10 DEG C with frozen water, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant
Liquid, through the ultra-filtration membrane device ultrafiltration that molecular cut off is 5000Da, removes unreacted dressing agent, collects and retain concentrated solution,
To modifying complex enzyme liquid I, standby；
B. weigh the SOD of CAT and proportional quantity 50% by proportioning to be dissolved in deionized water, be configured to total enzyme activity and (live with SOD enzyme
And CAT enzyme is lived sum meter) composite enzyme solution between 1000～2000U/ml, water-bath is heated to 30 DEG C, uses
NaOH solution regulatory enzyme liquid pH value is to 7.0, and stirring lower dropping continuously dressing agent also starts timing, maintenance enzyme liquid pH value 7.0,30 DEG C
After insulation reaction 60min, being cooled to 10 DEG C with frozen water, 6000rpm low temperature (4～8 DEG C) is centrifuged 8min, collects supernatant, through cutting
The ultra-filtration membrane device ultrafiltration staying molecular weight to be 5000Da, removes unreacted dressing agent, collects and retains concentrated solution, obtains modifying again
Synthase liquid II, standby；
C. merge to modify complex enzyme liquid with modifying complex enzyme liquid II by modification complex enzyme liquid I, add the step weighed by proportional quantity
Suddenly the activity protecting agent prepared by (1), stirring and dissolving mixing postlyophilization, obtain modifying compound enzyme lyophilized powder, standby；
(4) mixture obtains SOD compound enzyme powder:
I.e. synergetic effect additive as prepared by proportional quantity weighs functional oligose and step (2), adds step (3) C made
Standby modification compound enzyme lyophilized powder, is ground, and crosses 100 mesh sieves, obtains SOD compound enzyme powder；
(5) dress powder makes capsule:
Use conventional capsule fill process, the SOD compound enzyme powder of step (4) gained is loaded capsule shells, obtain SOD and be combined
Enzyme capsule.
The preparation method of a kind of antioxidation SOD compound enzyme capsule the most according to claim 8, it is characterised in that: step (3)
Dressing agent described in a and step (3) b is lauroyl chloride, and addition is the 0.5-1.5% of enzymatic solution gross mass.
The preparation method of a kind of antioxidation SOD compound enzyme capsule the most according to claim 8, it is characterised in that: step
(5) described capsule is enteric coated capsule.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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| CN108378226A (en) * | 2018-03-07 | 2018-08-10 | 余姚辉农农业科技有限公司 | A kind of complex enzyme formulation and its application in feed |
| CN113604462A (en) * | 2021-09-13 | 2021-11-05 | 清华大学 | Metal organic framework material-enzyme compound and preparation method and application thereof |
| CN114344455A (en) * | 2022-01-19 | 2022-04-15 | 宝莱福健康科技研究(中山)有限公司 | Preparation method and application of anti-aging composition |
| CN117126822A (en) * | 2023-10-27 | 2023-11-28 | 山东爱维德生物科技有限公司 | Adamantine modified SOD enzyme for arborescent aloe SOD composite pure dew and application thereof |
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| CN117126822A (en) * | 2023-10-27 | 2023-11-28 | 山东爱维德生物科技有限公司 | Adamantine modified SOD enzyme for arborescent aloe SOD composite pure dew and application thereof |
| CN117126822B (en) * | 2023-10-27 | 2023-12-22 | 山东爱维德生物科技有限公司 | Adamantine modified SOD enzyme for arborescent aloe SOD composite pure dew and application thereof |
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