CN106222159B - Anti-aging co-immobilized enzyme and preparation method thereof - Google Patents

Anti-aging co-immobilized enzyme and preparation method thereof Download PDF

Info

Publication number
CN106222159B
CN106222159B CN201610674530.3A CN201610674530A CN106222159B CN 106222159 B CN106222159 B CN 106222159B CN 201610674530 A CN201610674530 A CN 201610674530A CN 106222159 B CN106222159 B CN 106222159B
Authority
CN
China
Prior art keywords
catalase
glucose oxidase
solution
mixed
aging
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201610674530.3A
Other languages
Chinese (zh)
Other versions
CN106222159A (en
Inventor
桂涛
张洪浩
邓辉
王宝林
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Mianyang Embrace Biotechnology Co Ltd
Original Assignee
Mianyang Embrace Biotechnology Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Mianyang Embrace Biotechnology Co Ltd filed Critical Mianyang Embrace Biotechnology Co Ltd
Priority to CN201610674530.3A priority Critical patent/CN106222159B/en
Publication of CN106222159A publication Critical patent/CN106222159A/en
Application granted granted Critical
Publication of CN106222159B publication Critical patent/CN106222159B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0065—Oxidoreductases (1.) acting on hydrogen peroxide as acceptor (1.11)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
    • C12Y101/03—Oxidoreductases acting on the CH-OH group of donors (1.1) with a oxygen as acceptor (1.1.3)
    • C12Y101/03004—Glucose oxidase (1.1.3.4)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
    • C12Y111/01—Peroxidases (1.11.1)
    • C12Y111/01006—Catalase (1.11.1.6)

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Cosmetics (AREA)
  • Immobilizing And Processing Of Enzymes And Microorganisms (AREA)

Abstract

The invention discloses an anti-aging co-immobilized enzyme which comprises catalase and glucose oxidase, wherein the activity ratio of the catalase to the glucose oxidase is 8: 1-15: 1. the preparation method of the anti-aging co-immobilized enzyme comprises the following preparation steps of dissolving the catalase and the glucose oxidase in a phosphate buffer solution according to the activity ratio to prepare a catalase and glucose oxidase mixed enzyme solution, adding chitosan microspheres into the mixed enzyme solution to prepare a mixed solution, stirring and adsorbing the mixed solution, adding a glutaraldehyde solution with the mass concentration percentage for a crosslinking reaction, standing and balancing to room temperature, finally adding β -D-glucose, electromagnetically stirring, and standing for later use.

Description

Anti-aging co-immobilized enzyme and preparation method thereof
Technical Field
The invention relates to a co-immobilized enzyme and a preparation method thereof, in particular to an anti-aging co-immobilized enzyme and a preparation method thereof.
Background
Free radicals in the human body are mainly divided into endogenous and exogenous free radicals. Endogenous free radicals are mainly derived from the body's own metabolic activities, among which aerobic metabolism is the main one. When the production and elimination of free radicals in vivo are unbalanced, excessive free radicals in vivo can initiate a series of free radical reactions, thereby initiating reactions such as lipid peroxidation and the like. The first free radical in the human body is generated in the aerobic life activity of human: superoxide anion radical (O)2-Cndot.). Superoxide dismutase can scavenge superoxide anion free radical (O)2-Hydrogen peroxide is produced and can penetrate cell membranes, which is a superoxide anion radical (O)2-Cna.) has not been reached. When hydrogen peroxide enters cells, it generates a more oxidative hydroxyl radical (. OH) by Fenton reaction. The hydroxyl radical (. OH) is the most active radical in vivo, and can form lipid peroxide and lipid radical through hydrogen abstraction reaction, molecular rearrangement, double bond conjugation, and combination with oxygen(L.) and through this cycle a chain extension reaction of lipid peroxidation is formed, leading to the constant consumption of lipid molecules and the massive production of lipid peroxides. Lipid peroxidation acts on cell membranes, nucleic acids, proteins and enzymes, causing irreversible damage and turning these substances into oxides or peroxides, which lose their intrinsic action, and the accumulation of these actions contributes to the aging of the body. Thus, a superoxide anion radical (O)2-H) cell damage is mediated by hydrogen peroxide formed by disproportionation. Meanwhile, with age, the accumulation of a large amount of hydrogen peroxide in hair hinders the formation of melanin in hair, resulting in gradual whitening of hair from the inside to the outside.
The exogenous free radicals mainly come from cooking oil smoke, radiation, smoking, alcoholism, sunlight exposure and the like, the skin of a person is in the environment of air, pollution, radiation, ultraviolet rays and the like every day, a large amount of free radicals can be formed in the air, the free radicals can disappear quickly after being generated under normal conditions, and when the free radicals are combined with particles in the air, stable free radicals can be formed to erode the skin and even the body of the person. It is manifested as dark yellow skin, no luster, loose and inelastic skin, and even pigmentation and spots.
Disclosure of Invention
The invention aims to solve the technical problems that people who are often irradiated by computer radiation or ultraviolet rays and are drunk or stay up night can have rough skin and pigmentation, and aims to provide an anti-aging co-immobilized enzyme and a preparation method thereof.
The invention is realized by the following technical scheme:
the anti-aging co-immobilized enzyme comprises catalase and glucose oxidase, wherein the activity ratio of the catalase to the glucose oxidase is 8: 1-15: 1.
Preferably, the activity ratio of the catalase to the glucose oxidase is 9: 1-12: 1.
A preparation method of an anti-aging co-immobilized enzyme comprises the following preparation steps:
a. dissolving catalase and glucose oxidase in a phosphate buffer solution according to the activity ratio to prepare a mixed enzyme solution of catalase and glucose oxidase;
b. b, adding chitosan microspheres into the mixed enzyme solution prepared in the step a to prepare a mixed solution;
c. b, stirring and adsorbing the mixed solution prepared in the step b;
d. adding glutaraldehyde solution to carry out crosslinking reaction after the step c is finished;
e. standing and balancing to room temperature;
f. adding β -D-glucose after the step e is finished, electromagnetically stirring, and standing for later use;
in the step a, the activity ratio of the catalase to the glucose oxidase is 8: 1-15: 1 or 9: 1-12: 1.
Preferably, in the step a, the pH value of the phosphate buffer solution is 3-8, and the concentration is 0.1-0.5 mol/L; in the step c, stirring and adsorbing the mixed solution for 40-55 min at the temperature of 20-50 ℃ and the pH value of 5-7; and d, adding 1.5-3.5 mass percent of glutaraldehyde solution.
Preferably, the specific activity of the catalase is 50000U/mL, and the pH is 6.0; the specific activity of the glucose oxidase is 10000U/g.
Preferably, the volume of the catalase and glucose oxidase mixed enzyme solution prepared in the step a is as follows: the mass of the chitosan microspheres in the step b is 5(mL):2(g) -5 (mL):4 (g).
The anti-aging co-solidifying enzyme acts on the body surface to form a compact anti-oxidation protective layer, which is beneficial to removing hydrogen peroxide generated by oxygen through divalent electron reduction under the action of external conditions such as ultraviolet rays and the like on the body surface, avoiding the generation and removal imbalance of free radicals and initiating a free radical chain reaction to accelerate the aging of human skin, meanwhile, β -D-glucose in the co-solidifying enzyme is decomposed by glucose oxidase to remove excessive oxygen on the body surface and excessive oxygen generated by hydrogen peroxide decomposed by catalase, so that the oxygen content on the body surface reaches dynamic balance, the skin elasticity and white skin are kept, and gluconic acid is generated when the glucose oxidase decomposes β -D-glucose and has the characteristic of chelating copper ions, so that the anti-aging co-solidifying enzyme can help to inhibit the activity of tyrosinase, prevent the generation of melanin and prevent the occurrence of senile plaques.
Compared with the prior art, the invention has the following advantages and beneficial effects:
the invention relates to an anti-aging co-immobilized enzyme and a preparation method thereof, which can remove hydrogen peroxide generated by reduction of oxygen through valence and divalent electrons under the action of external conditions such as ultraviolet rays and the like on the body surface through the synergistic action of catalase and glucose oxidase, decompose β -D-glucose in co-immobilized enzyme to remove excessive oxygen on the body surface and excessive oxygen generated by decomposition of hydrogen peroxide by catalase, ensure that the oxygen content on the body surface reaches dynamic balance, keep the skin elastic and white, and achieve the aim of anti-aging.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples, and the exemplary embodiments and descriptions thereof are only used for explaining the present invention and are not used as limitations of the present invention.
Example 1
A preparation method of an anti-aging co-immobilized enzyme comprises catalase and glucose oxidase, wherein the activity ratio of the catalase to the glucose oxidase is 8: 1.
The preparation method comprises the following preparation steps:
a. dissolving catalase and glucose oxidase in a phosphate buffer solution according to the activity ratio of 8:1 to prepare a mixed enzyme solution of catalase and glucose oxidase, wherein the pH value of the phosphate buffer solution is 3, and the concentration of the phosphate buffer solution is 0.1 mol/L;
b. adding 3g of chitosan microspheres into the mixed enzyme solution prepared in the step a to prepare a mixed solution;
c. b, stirring and adsorbing the mixed solution prepared in the step b for 40min at the temperature of 20 ℃ and under the condition that the pH value is 5;
d. after the step c is finished, adding 1.5 mass percent glutaraldehyde solution, and carrying out crosslinking reaction for 2.5h at the temperature of 5 ℃;
e. standing and balancing to room temperature;
f. and e, adding β -D-glucose 0.8g after the step e is finished, electromagnetically stirring for 30min, and standing for later use.
Wherein the specific activity of the catalase is 50000U/mL, and the pH value is 6.0; the specific activity of the solid glucose oxidase is 10000U/g; the volume of the catalase and glucose oxidase mixed enzyme solution prepared in the step a is as follows: the mass of the chitosan microspheres in the step b is 5(mL):2 (g).
Example 2
A preparation method of an anti-aging co-immobilized enzyme comprises catalase and glucose oxidase, wherein the activity ratio of the catalase to the glucose oxidase is 9: 1.
The preparation method comprises the following preparation steps:
a. dissolving catalase and glucose oxidase into a phosphate buffer solution according to the activity ratio of 9:1 to prepare a mixed enzyme solution of catalase and glucose oxidase, wherein the pH value of the phosphate buffer solution is 5, and the concentration of the phosphate buffer solution is 0.3 mol/L;
b. adding 3g of chitosan microspheres into the mixed enzyme solution prepared in the step a to prepare a mixed solution;
c. b, stirring and adsorbing the mixed solution prepared in the step b for 45min at the temperature of 30 ℃ and the pH value of 6.5;
d. after the step c is finished, adding a glutaraldehyde solution with the mass concentration of 2.5%, and carrying out crosslinking reaction for 2.5h at the temperature of 5 ℃;
e. standing and balancing to room temperature;
f. and e, adding β -D-glucose 0.8g after the step e is finished, electromagnetically stirring for 30min, and standing for later use.
Wherein the specific activity of the catalase is 50000U/mL, and the pH value is 6.0; the specific activity of the solid glucose oxidase is 10000U/g; the volume of the catalase and glucose oxidase mixed enzyme solution prepared in the step a is as follows: the mass of the chitosan microspheres in the step b is 5(mL):3 (g).
Example 3
A preparation method of an anti-aging co-immobilized enzyme comprises catalase and glucose oxidase, wherein the activity ratio of the catalase to the glucose oxidase is 10: 1.
The preparation method comprises the following preparation steps:
a. dissolving catalase and glucose oxidase into a phosphate buffer solution according to the activity ratio of 10:1 to prepare a mixed enzyme solution of catalase and glucose oxidase, wherein the pH value of the phosphate buffer solution is 5, and the concentration of the phosphate buffer solution is 0.3 mol/L;
b. adding 3g of chitosan microspheres into the mixed enzyme solution prepared in the step a to prepare a mixed solution;
c. b, stirring and adsorbing the mixed solution prepared in the step b for 45min at the temperature of 30 ℃ and the pH value of 6.5;
d. after the step c is finished, adding a glutaraldehyde solution with the mass concentration of 2.5%, and carrying out crosslinking reaction for 2.5h at the temperature of 5 ℃;
e. standing and balancing to room temperature;
f. and e, adding β -D-glucose 0.8g after the step e is finished, electromagnetically stirring for 30min, and standing for later use.
Wherein the specific activity of the catalase is 50000U/mL, and the pH value is 6.0; the specific activity of the solid glucose oxidase is 10000U/g; the volume of the catalase and glucose oxidase mixed enzyme solution prepared in the step a is as follows: the mass of the chitosan microspheres in the step b is 5(mL):3 (g).
Example 4
A preparation method of an anti-aging co-immobilized enzyme comprises catalase and glucose oxidase, wherein the activity ratio of the catalase to the glucose oxidase is 12: 1.
The preparation method comprises the following preparation steps:
a. dissolving catalase and glucose oxidase into a phosphate buffer solution according to the activity ratio of 12:1 to prepare a mixed enzyme solution of catalase and glucose oxidase, wherein the pH value of the phosphate buffer solution is 5, and the concentration of the phosphate buffer solution is 0.5 mol/L;
b. adding 3g of chitosan microspheres into the mixed enzyme solution prepared in the step a to prepare a mixed solution;
c. b, stirring and adsorbing the mixed solution prepared in the step b for 45min at the temperature of 30 ℃ and the pH value of 6.5;
d. after the step c is finished, adding a glutaraldehyde solution with the mass concentration of 2.5%, and carrying out crosslinking reaction for 2.5h at the temperature of 5 ℃;
e. standing and balancing to room temperature;
f. and e, adding β -D-glucose 0.8g after the step e is finished, electromagnetically stirring for 30min, and standing for later use.
Wherein the specific activity of the catalase is 50000U/mL, and the pH value is 6.0; the specific activity of the solid glucose oxidase is 10000U/g; the volume of the catalase and glucose oxidase mixed enzyme solution prepared in the step a is as follows: the mass of the chitosan microspheres in the step b is 5(mL):3 (g).
Example 5
A preparation method of an anti-aging co-immobilized enzyme comprises catalase and glucose oxidase, wherein the activity ratio of the catalase to the glucose oxidase is 14: 1.
The preparation method comprises the following preparation steps:
a. dissolving catalase and glucose oxidase into a phosphate buffer solution according to the activity ratio of 14:1 to prepare a mixed enzyme solution of catalase and glucose oxidase, wherein the pH value of the phosphate buffer solution is 5, and the concentration of the phosphate buffer solution is 0.5 mol/L;
b. adding 3g of chitosan microspheres into the mixed enzyme solution prepared in the step a to prepare a mixed solution;
c. b, stirring and adsorbing the mixed solution prepared in the step b for 45min at the temperature of 30 ℃ and the pH value of 6.5;
d. after the step c is finished, adding a glutaraldehyde solution with the mass concentration of 2.5%, and carrying out crosslinking reaction for 2.5h at the temperature of 5 ℃;
e. standing and balancing to room temperature;
f. and e, adding β -D-glucose 0.8g after the step e is finished, electromagnetically stirring for 30min, and standing for later use.
Wherein the specific activity of the catalase is 50000U/mL, and the pH value is 6.0; the specific activity of the solid glucose oxidase is 10000U/g; the volume of the catalase and glucose oxidase mixed enzyme solution prepared in the step a is as follows: the mass of the chitosan microspheres in the step b is 5(mL):3 (g).
Example 6
A preparation method of an anti-aging co-immobilized enzyme comprises catalase and glucose oxidase, wherein the activity ratio of the catalase to the glucose oxidase is 15: 1.
The preparation method comprises the following preparation steps:
a. dissolving catalase and glucose oxidase into a phosphate buffer solution according to the activity ratio of 15:1 to prepare a mixed enzyme solution of catalase and glucose oxidase, wherein the pH value of the phosphate buffer solution is 8, and the concentration of the phosphate buffer solution is 0.5 mol/L;
b. adding 3g of chitosan microspheres into the mixed enzyme solution prepared in the step a to prepare a mixed solution;
c. b, stirring and adsorbing the mixed solution prepared in the step b for 55min at the temperature of 50 ℃ and under the condition that the pH value is 7;
d. after the step c is finished, adding glutaraldehyde solution with the mass concentration percentage of 3.5%, and carrying out crosslinking reaction for 2.5h at the temperature of 5 ℃;
e. standing and balancing to room temperature;
f. and e, adding β -D-glucose 0.8g after the step e is finished, electromagnetically stirring for 30min, and standing for later use.
Wherein the specific activity of the catalase is 50000U/mL, and the pH value is 6.0; the specific activity of the solid glucose oxidase is 10000U/g; the volume of the catalase and glucose oxidase mixed enzyme solution prepared in the step a is as follows: the mass of the chitosan microspheres in the step b is 5(mL):4 (g).
Firstly, performance testing:
the skin smearing mode observation is carried out on 24 persons by using the sample (wherein 16 persons are male, 8 persons are female, and the ages are 28-30 years old), wherein 6 persons are programmers, 6 persons are constructors, 6 persons are literary and professional, 6 persons are sold and classified into A, B, C three groups for comparison experiments, and each group of programmers, the constructors, the literary and professional and the sales are respectively two persons. The application method comprises getting up in the morning, washing face, and smearing before sleeping at night. Building workers are exposed to the sun for a long time and have more physical labor, rough skin and dark yellow skin color; programmers and literary classes mainly radiate for a long time against a computer; sales are mainly due to alcohol consumption. Wherein the number of smokers reaches 12. The co-immobilized enzymes obtained by the proportioning and preparation schemes of the examples are respectively used for three groups of people, and the trial time is two weeks. The three groups of people are tested before trial and after being applicable by a Fisher Keel skin comprehensive index measuring instrument, wherein:
moisture measurement range: 0 to 99.9 percent; the ideal range is as follows: 40-60%;
oil content measurement range: 5 to 50% by weight of the water content; the ideal range is as follows: 15-23%;
the elastic coefficient: 0 to 1; the ideal range is as follows: 0.45 to 0.55.
The test result data are the average values of the group members.
II, testing results:
TABLE 1 group A skin quality test results
Examples Moisture (%) Oil content (%) Elasticity
Before trial 20 8 0.67
1 25 10 0.61
2 31 13 0.58
3 43 16 0.52
4 40 15 0.54
5 37 13 0.55
6 33 13 0.57
Table 2B group skin quality test results
Examples Moisture (%) Oil content (%) Elasticity
Before trial 31 12 0.61
1 37 14 0.59
2 42 17 0.54
3 56 21 0.49
4 53 19 0.50
5 50 16 0.53
6 47 17 0.57
Table 3C group skin quality test results
Examples Moisture (%) Oil content (%) Elasticity
Before trial 29 11 0.60
1 39 15 0.57
2 47 19 0.54
3 53 22 0.50
4 52 20 0.52
5 50 17 0.53
6 46 17 0.56
The anti-aging co-solidifying enzyme acts on the body surface to form a compact anti-oxidation protective layer, namely the catalase is beneficial to removing hydrogen peroxide generated by oxygen reduction through divalent electrons under the external conditions of ultraviolet rays and the like on the body surface, the generation and removal of free radicals are avoided, the aging of the human skin is accelerated by a free radical chain reaction, the anti-aging co-solidifying enzyme is triggered, meanwhile, the glucose oxidase decomposes β -D-glucose in the co-solidifying enzyme to remove the excessive oxygen and the excessive hydrogen peroxide, and the glucose oxidase generates a smooth and balanced oxygen content, so that the skin has a synergistic effect of inhibiting the oxidative decomposition of melanin-glucose oxidase and hydrogen peroxide decomposition-6335, and the skin has a synergistic effect of inhibiting the oxidative decomposition of melanin-glucose oxidase and glucose oxidase, so that the skin has a dark yellow color, no luster, dry roughness and no elasticity, and a small amount of pigment deposition, and spots.
The anti-aging co-immobilized enzyme is mainly suitable for people who are often irradiated by computer radiation and ultraviolet rays, are drunk or stay up all night, has good regulating effect on people who have dry and rough skin and no toughness and pigmentation of fibrous tissues, and has better effect when being matched with cosmetics.
The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are merely exemplary embodiments of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalent substitutions, improvements and the like made within the spirit and principle of the present invention should be included in the scope of the present invention.

Claims (1)

1. The application of the co-immobilized enzyme in preparing the cosmetics for resisting skin aging is characterized in that the preparation method of the co-immobilized enzyme comprises the following steps:
a. dissolving catalase and glucose oxidase into a phosphate buffer solution according to the activity ratio to prepare a mixed enzyme solution of catalase and glucose oxidase;
b. b, adding chitosan microspheres into the mixed enzyme solution prepared in the step a to prepare a mixed solution;
c. b, stirring and adsorbing the mixed solution prepared in the step b;
d. adding glutaraldehyde solution to carry out crosslinking reaction after the step c is finished;
e. standing and balancing to room temperature;
f. adding β -D-glucose after the step e is finished, electromagnetically stirring, and standing for later use;
in the step a, the activity ratio of catalase to glucose oxidase is 10: 1;
in the step a, the pH value of a phosphate buffer solution is 3-8, and the concentration is 0.1-0.5 mol/L; in the step c, stirring and adsorbing the mixed solution for 40-55 min at the temperature of 20-50 ℃ and the pH value of 5-7; in the step d, adding 1.5-3.5% of glutaraldehyde solution by mass concentration;
the volume of the catalase and glucose oxidase mixed enzyme solution prepared in the step a is as follows: in the step b, the mass of the chitosan microspheres is 5mL to 2g to 5mL to 4 g;
the specific activity of the catalase is 50000U/mL, and the pH value is 6.0; the specific activity of the glucose oxidase is 10000U/g.
CN201610674530.3A 2016-08-16 2016-08-16 Anti-aging co-immobilized enzyme and preparation method thereof Active CN106222159B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610674530.3A CN106222159B (en) 2016-08-16 2016-08-16 Anti-aging co-immobilized enzyme and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610674530.3A CN106222159B (en) 2016-08-16 2016-08-16 Anti-aging co-immobilized enzyme and preparation method thereof

Publications (2)

Publication Number Publication Date
CN106222159A CN106222159A (en) 2016-12-14
CN106222159B true CN106222159B (en) 2020-03-20

Family

ID=57552430

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610674530.3A Active CN106222159B (en) 2016-08-16 2016-08-16 Anti-aging co-immobilized enzyme and preparation method thereof

Country Status (1)

Country Link
CN (1) CN106222159B (en)

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101756882A (en) * 2008-11-12 2010-06-30 陈静 Tartaric acid gelatin and preparation method thereof
CN102199592B (en) * 2011-04-02 2013-04-03 重庆大学 Method for preparing mixed immobilized glucose oxidase/catalase microspheres
EP3626310A1 (en) * 2013-05-27 2020-03-25 Rakuto Bio Technologies Ltd. Enzymatic system containing cosmetic compositions

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
Optimal study of Co-immobilized GOD/CAT on Cross-linked Chitosan Microsphere containing L-Lysine;Xiaohua Zhou et al.;《Advanced Materials Research》;20110512;第236-238卷;926-929 *

Also Published As

Publication number Publication date
CN106222159A (en) 2016-12-14

Similar Documents

Publication Publication Date Title
JP2019189596A (en) Application of thermus thermophilus and yeast combined fermentation product
CN112891276A (en) Composition with moisturizing, repairing, anti-aging and anti-saccharification functions and application thereof
CN113855603B (en) Combined fermentation product, skin external preparation containing same, and preparation method and application of combined fermentation product
CN109453051B (en) Whitening and repairing mask and preparation method thereof
CN110664718A (en) Whitening skin care product
JP5838467B2 (en) Deodorants
CN109394640B (en) Enzyme skin care composition and application thereof
CN111135138A (en) Skin conditioner compound containing symbiotic bacteria combined fermentation product, muscle base solution and preparation method thereof
CN106222159B (en) Anti-aging co-immobilized enzyme and preparation method thereof
CN114134088A (en) Choerospondias axillaris fermentation product and preparation method and application thereof
CN111358748B (en) Mask essence containing date palm seed extract and preparation method thereof
CN113456527A (en) NADH microcapsule preparation and preparation method and application thereof
CN116747158A (en) Cosmetic composition capable of resisting wrinkle and compacting through mitochondria and application thereof
CN110227050B (en) Anti-aging repairing slow-release essence and anti-aging repairing mask prepared from same
CN115634163B (en) Whitening skin care essence emulsion and preparation method thereof
CN110680796A (en) Skin care product containing earthworm SOD extract and preparation method thereof
CN115778839A (en) Scalp care agent with hair blacking function and preparation method thereof
CN114081862B (en) Preparation and application of Saccharomyces cerevisiae fermentation product from highland barley wine distiller's yeast in Pan-Himalaya region
CN110368326B (en) Dynamic antioxidant complex, preparation method thereof and cosmetic containing same
KR20130099277A (en) A cosmetic composition comprising hydrolysates of ecklonia cava
CN113952250A (en) Stable anti-photoaging composition and preparation method thereof
CN113876610A (en) Composition capable of enhancing cell energy and improving skin resistance and preparation method and application thereof
CN110882194A (en) Additive-free pink skin-beautifying facial mask liquid and preparation method thereof
CN111437220B (en) Camellia oil fatty acid ester composition and preparation method and application thereof
CN113143818B (en) Whitening and anti-photoaging composition and application thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant