CN105853342B - Purposes of the polyphenol extract as tyrosinase inhibitor a kind of in spirulina - Google Patents
Purposes of the polyphenol extract as tyrosinase inhibitor a kind of in spirulina Download PDFInfo
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- CN105853342B CN105853342B CN201610326240.XA CN201610326240A CN105853342B CN 105853342 B CN105853342 B CN 105853342B CN 201610326240 A CN201610326240 A CN 201610326240A CN 105853342 B CN105853342 B CN 105853342B
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- spirulina
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- tyrosinase
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- 235000016425 Arthrospira platensis Nutrition 0.000 title claims abstract description 89
- 240000002900 Arthrospira platensis Species 0.000 title claims abstract description 89
- 229940082787 spirulina Drugs 0.000 title claims abstract description 89
- 239000000284 extract Substances 0.000 title claims abstract description 87
- 235000013824 polyphenols Nutrition 0.000 title claims abstract description 79
- 150000008442 polyphenolic compounds Chemical class 0.000 title claims abstract description 76
- 101710147108 Tyrosinase inhibitor Proteins 0.000 title claims abstract description 16
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000000470 constituent Substances 0.000 claims abstract description 13
- 235000019441 ethanol Nutrition 0.000 claims abstract description 7
- 230000002829 reductive effect Effects 0.000 claims abstract description 5
- 238000004108 freeze drying Methods 0.000 claims abstract description 4
- 239000007787 solid Substances 0.000 claims abstract description 4
- 238000002360 preparation method Methods 0.000 claims abstract description 3
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 claims description 28
- 239000000243 solution Substances 0.000 claims description 15
- 229940074391 gallic acid Drugs 0.000 claims description 13
- 235000004515 gallic acid Nutrition 0.000 claims description 13
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 claims description 12
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- 238000006243 chemical reaction Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
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- 229910021642 ultra pure water Inorganic materials 0.000 claims description 6
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- 239000003795 chemical substances by application Substances 0.000 claims description 5
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- 208000037259 Amyloid Plaque Diseases 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 230000001629 suppression Effects 0.000 claims 1
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- 108060008724 Tyrosinase Proteins 0.000 abstract description 33
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- 239000000758 substrate Substances 0.000 description 11
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- WTDRDQBEARUVNC-UHFFFAOYSA-N L-Dopa Natural products OC(=O)C(N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-UHFFFAOYSA-N 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
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- 239000002994 raw material Substances 0.000 description 5
- 230000002087 whitening effect Effects 0.000 description 5
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 241000192700 Cyanobacteria Species 0.000 description 3
- 235000008100 Ginkgo biloba Nutrition 0.000 description 3
- 244000194101 Ginkgo biloba Species 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
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- XKZQKPRCPNGNFR-UHFFFAOYSA-N 2-(3-hydroxyphenyl)phenol Chemical compound OC1=CC=CC(C=2C(=CC=CC=2)O)=C1 XKZQKPRCPNGNFR-UHFFFAOYSA-N 0.000 description 2
- 241001474374 Blennius Species 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 235000011201 Ginkgo Nutrition 0.000 description 2
- 241001256169 Gracilaria asiatica Species 0.000 description 2
- 241000631636 Ishige Species 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 241000206572 Rhodophyta Species 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 230000002596 correlated effect Effects 0.000 description 2
- 230000000875 corresponding effect Effects 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000002532 enzyme inhibitor Substances 0.000 description 2
- 229940125532 enzyme inhibitor Drugs 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 229960004502 levodopa Drugs 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- -1 polyphenol compound Chemical class 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- MPDGHEJMBKOTSU-YKLVYJNSSA-N 18beta-glycyrrhetic acid Chemical compound C([C@H]1C2=CC(=O)[C@H]34)[C@@](C)(C(O)=O)CC[C@]1(C)CC[C@@]2(C)[C@]4(C)CC[C@@H]1[C@]3(C)CC[C@H](O)C1(C)C MPDGHEJMBKOTSU-YKLVYJNSSA-N 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 description 1
- 240000004670 Glycyrrhiza echinata Species 0.000 description 1
- 235000001453 Glycyrrhiza echinata Nutrition 0.000 description 1
- 235000006200 Glycyrrhiza glabra Nutrition 0.000 description 1
- 235000017382 Glycyrrhiza lepidota Nutrition 0.000 description 1
- 241000199919 Phaeophyceae Species 0.000 description 1
- 108010053210 Phycocyanin Proteins 0.000 description 1
- 102000018120 Recombinases Human genes 0.000 description 1
- 108010091086 Recombinases Proteins 0.000 description 1
- 241000195474 Sargassum Species 0.000 description 1
- 241000593522 Sargassum thunbergii Species 0.000 description 1
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- BULOCEWDRJUMEL-UHFFFAOYSA-N benzene formaldehyde Chemical class C=O.C1=CC=CC=C1.C=O BULOCEWDRJUMEL-UHFFFAOYSA-N 0.000 description 1
- 150000001558 benzoic acid derivatives Chemical class 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000003139 biocide Substances 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
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- 230000036772 blood pressure Effects 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 229910001431 copper ion Inorganic materials 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- MHUWZNTUIIFHAS-CLFAGFIQSA-N dioleoyl phosphatidic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OCC(COP(O)(O)=O)OC(=O)CCCCCCC\C=C/CCCCCCCC MHUWZNTUIIFHAS-CLFAGFIQSA-N 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000000640 hydroxylating effect Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- BEJNERDRQOWKJM-UHFFFAOYSA-N kojic acid Chemical class OCC1=CC(=O)C(O)=CO1 BEJNERDRQOWKJM-UHFFFAOYSA-N 0.000 description 1
- 229960004705 kojic acid Drugs 0.000 description 1
- WZNJWVWKTVETCG-UHFFFAOYSA-N kojic acid Natural products OC(=O)C(N)CN1C=CC(=O)C(O)=C1 WZNJWVWKTVETCG-UHFFFAOYSA-N 0.000 description 1
- 229940010454 licorice Drugs 0.000 description 1
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- 230000009467 reduction Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 235000003724 spirulina extract Nutrition 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
Abstract
Purposes the invention discloses the polyphenol extract in a kind of spirulina as tyrosinase inhibitor, using spirulina polyphenol extract as one of active constituent or unique active constituent is used to prepare tyrosinase inhibitor；The preparation method of the spirulina polyphenol extract: using volumetric concentration to extract spirulina for 80% ethyl alcohol, obtains spirulina polyphenolic extract, then spirulina polyphenolic extract is concentrated under reduced pressure in 40 DEG C, solid spirulina polyphenol extract is obtained after freeze-drying.Inhibition of the polyphenol that the present invention extracts for the first time from spirulina to tyrosinase activity, it will be applied to the fields such as skin-whitening, medical treatment, preserving fruit and vegetable utilizing for it and reliable theoretical foundation is provided, and also will provide new way for the tyrosinase inhibitor of comprehensive utilization spirulina resource and research safety, economy.
Description
Technical field
The present invention relates to the polyphenol extracts in the technical field of biological agent more particularly to a kind of spirulina as junket ammonia
The purposes of sour enzyme inhibitor.
Background technique
Seeweed polyphenol is that a kind of mainly separation and Extraction has from cyanobacteria, green alga, red algae and the big marine algae of brown alga four
The general name of the polyphenol compound of the extensive bioactivity such as antibacterial, anti-oxidant, antitumor, antiviral and blood pressure lowering.Iwai research
After brown algae polyphenols are taken in by mouse as a kind of natural phlorose enzyme inhibitor, mouse blood sugar level is controlled
System, can effectively prevent the generation of diabetes；Lu Hongyu et al. extracts 4 kinds by extraction and hyperfiltration technique from full edge sargassum
Polyphenol extract, and test the oxidation resistance of 4 kinds of extracts and the influence to sw579 cell Proliferation, as the result is shown brown alga
The total antioxidant capacity and molecular size range of polyphenol extract are negatively correlated, and the inhibiting effect and concentration to tumour cell are positively correlated；
The researchs such as Wei Yuxi discovery sargassum thunbergii polyphenol can interfere the activity of coagulation factor and reduce Ca inside and outside blood platelet2+Transport, it is anticoagulant
Blood significant effect.Studies have shown that seeweed polyphenol is in food and medicine field, there are researching values.
Tyrosinase energy hydroxylating single phenol substrate and oxidation diphenol substrate, are that double-core copper ion is contained in a kind of activated centre
Oxidoreducing enzyme.Melanin, which is built up, will lead to fruits and vegetables brown stain, skin splash and melanoma and is formed, and tyrosinase activity
Size is closely related with Melanin productions in organism.Typical tyrosinase inhibitor has kojic acid, polyphenol compound, benzene
Formaldehyde and benzoate analog derivative etc. have been widely used in studying fruit and vegetable fresh-keeping agent, white-making ingredient and Biocidal
Agent.Spirulina is a kind of ancient filamentous cyanobacteria, is rich in phycocyanin, spirulina polyphenol, micromolecular polysaccharide isoreactivity ingredient.I
State's spirulina is resourceful, is spirulina raw material big producer, but China's spirulina is substantially carried out simple utilization, and active constituent
Exploitation Depth is inadequate.
In view of this, the present inventor's research and devising a kind of polyphenol extract in spirulina as tyrosinase inhibition
Thus the purposes of agent, this case generate.
Summary of the invention
Purposes the purpose of the present invention is to provide the polyphenol extract in a kind of spirulina as tyrosinase inhibitor,
Spirulina polyphenol is extracted from spirulina raw material, and tyrosinase inhibition test is carried out to it, to be spirulina money
The development and utilization in source provide experimental basis, while developing a kind of natural economic, safety tyrosinase inhibitor.
To achieve the goals above, the technical scheme adopted by the invention to solve the technical problem is that:
Purposes of the polyphenol extract as tyrosinase inhibitor in a kind of spirulina, with spirulina polyphenol extract work
Tyrosinase inhibitor is used to prepare for one of active constituent or unique active constituent.
As the preferred embodiment of embodiment, using spirulina polyphenol extract as one of active constituent or unique activity at
Divide for anti-senile plaque or inhibit the antistaling agent of food brown stain.
As the preferred embodiment of embodiment, the preparation method of the spirulina polyphenol extract: use volumetric concentration for
80% ethyl alcohol extracts spirulina, obtains spirulina polyphenolic extract, then spirulina polyphenolic extract is concentrated under reduced pressure in 40 DEG C,
Solid spirulina polyphenol extract is obtained after freeze-drying.
As the preferred embodiment of embodiment, the measuring method of spirulina polyphenol content: spectrophotometry is used, with forint phenol
Reagent develops the color to the polyphenol in extract, determines polyphenol content: ultrapure water dissolves 0.110g gallic acid, using 100mL capacity
Bottle constant volume, obtains gallic acid Standard Stock solutions；Be diluted to 10 respectively with galla turcica gallic acid Standard Stock solutions again, 20,
30,40 and 50 μ gmL-1Serial solution, blank control is done with ultrapure water, 1.0mL dilution is taken to be placed in colorimetric cylinder respectively,
5.0mL forint phenol solution is added, reacts 6min, 4.0mL 7.5%Na is added2CO3Solution mixes, after avoid light place 1h, in wave
Light absorption value is measured under long 765nm, makes standard curve；Spirulina polyphenol content is according to standard curve colorimetric estimation.
Inhibition of the polyphenol that the present invention extracts for the first time from spirulina to tyrosinase activity will be applied to skin beauty for it
The fields such as white, medical, preserving fruit and vegetable utilizing provide reliable theoretical foundation, also will for comprehensive utilization spirulina resource and research safety,
Economic tyrosinase inhibitor provides new way.
Detailed description of the invention
Fig. 1 is the standard curve of gallic acid of the present invention；
Fig. 2 is inhibiting effect of the spirulina polyphenol extract of the present invention to tyrosinase diphenol enzyme activity；
Fig. 3 is measurement of the spirulina polyphenol extract of the present invention to tyrosinase diphenolase inhibiting mechanism；Wherein, straight line 1-
The concentration of spirulina polyphenol corresponding to 5 is respectively 0,0.67,1.33,2.00 and 2.67mgmL-1；
Fig. 4 is the measurement that spirulina polyphenol extract of the present invention inhibits type and inhibition constant to tyrosinase diphenolase；
Wherein, the concentration of spirulina polyphenol corresponding to straight line 1-5 is respectively 0,0.67,1.33,2.00 and 2.67mgmL-1。
Specific embodiment
1 materials and methods
1.1 material
1.1.1 reagent and drug spirulina powder are provided by refreshing health care product Co., Ltd, Fujian Province six；Forint phenol reagent, does not have
Gallate-based, tyrosinase are purchased from Sigma company；Natrium carbonicum calcinatum is purchased from Shantou Xilong Chemical Factory；Ethyl alcohol, levodopa etc.
Experimental material is purchased from Shanghai traditional Chinese medicines chemical reagent Co., Ltd.
1.1.2 instrument ultraviolet specrophotometer, Australian GBC instrument company；Thermostat water bath, Jintan City, Jiangsu Province
Hong Ke instrument plant；FE20K acidometer, Shanghai plum Teller-support benefit Instrument Ltd.；Vacuum freeze drier, Zhengzhou tomorrow
Experimental instruments and equipment limited.
1.2 method
1.2.1 the extraction of spirulina polyphenol extracts phenolic substances using 80% ethyl alcohol using spirulina powder as raw material, then will
Spirulina polyphenolic extract is concentrated under reduced pressure in 40 DEG C, and solid spirulina polyphenol extract is obtained after freeze-drying.
1.2.2 the measurement of spirulina polyphenol content uses spectrophotometry, with forint phenol reagent to more in extract
Phenol colour developing, determines polyphenol content.Ultrapure water dissolves 0.110g gallic acid, and 100mL volumetric flask constant volume obtains the storage of gallic acid standard
Standby solution.It is diluted to 10,20,30,40 and 50 μ gmL respectively with galla turcica gallic acid Standard Stock solutions again-1Series it is molten
Liquid does blank control with ultrapure water, 1.0mL dilution is taken to be placed in colorimetric cylinder respectively, and 5.0mL forint phenol solution, reaction is added
4.0mL 7.5%Na is added in 6min2CO3Solution mixes, and after avoid light place 1h, light absorption value is measured at wavelength 765nm, makes
Standard curve.Spirulina polyphenol content is according to standard curve colorimetric estimation.1.2.3 spirulina polyphenol extract is to tyrosinase two
Inhibitory effect of the inhibiting effect extract of phenolase to enzyme: by immobilized substrate concentration and enzyme concentration, it is dense to measure different effect object
Relative surplus enzyme activity under degree.Spirulina polyphenol extract is dissolved in dimethyl sulfoxide (DMSO), and the effect of various concentration is made
Object solution.0.5mmol·L-1L-DOPA substrate solution and 2.89 μ gmL-1Enzyme solution uses 50mmolL-16.8 phosphoric acid of pH is slow
Fliud flushing is prepared.In 3mL reaction system, 0.1mL effector solution is first added, adds 2.8mL in advance in 30 DEG C of waters bath with thermostatic control
The substrate solution of 10min is eventually adding 0.1mL enzyme solution, quickly mixes.Measurement wavelength is set as 475nm, measures reaction system
Growth straight line of the OD value with the reaction time, extinction coefficient epsilon=3700Lmol-1·cm-1, by enzyme activity, calculate half and press down
Rate IC processed50Value.
Inhibiting mechanism of the extract to enzyme: by immobilized substrate concentration, change enzyme amount, measure under different effect object concentration
Enzyme activity.In 0.5mmolL containing 2.8mL-1In the live body system of L-DOPA substrate, measurement enzyme amount is 1.15,1.73,2.31,
2.89 and 3.46 μ gmL-1When, various concentration spirulina polyphenol extract acts on the reaction rate of lower enzyme, judges extract to enzyme
The mechanism of action.
Inhibition type, inhibition constant of the extract to enzyme: by fixed enzyme concentration, change the amount of substrate, measure different effects
Answer the enzyme activity under object concentration.Fixed enzyme amount is 2.89 μ gmL-1, it is respectively 0,0.67 in spirulina polyphenol extract concentration,
1.33,2.00 and 2.67mgmL-1Live body system in, measurement L-DOPA concentration be 0.3,0.4,0.5,0.6 and 0.7mmol
L-1When enzyme reaction speed.By Lineweave-Burk double-reciprocal plot, calculates and compare kinetic parameter KmAnd Vm, judgement
Inhibit type.
2 results and analysis
Total phenol content in 2.1 spirulina polyphenol extracts
Folin-Ciocalteus reaction normal curve such as Fig. 1 of gallic acid.Gallic acid mass concentration is in 0~50 μ
g·mL-1When, light absorption value A765There is good linear relationship with concentration of standard solution.The linear regression of gallic acid standard working curve
Equation is Y=0.00123X-0.0086, coefficient R2=0.9996 (Y is light absorption value, and X is gallic acid concentration).According to not having
Light absorption value of the standard working curve formula and spirulina solution of gallate-based under 765nm wavelength, obtains in spirulina extract
Polyphenol content be 35.05mgg-1。
Influence of the 2.2 spirulina polyphenol extracts to tyrosinase vigor
Fixed L-DOPA concentration and enzyme concentration measure various concentration spirulina polyphenol extract to Inhibition of Tyrosinase Activity.By
Fig. 2 illustrates spirulina polyphenol extract it is found that the reaction rate of tyrosinase declines with the increase of spirulina polyphenol concentration
There are inhibiting effect, rate IC half-suppressed to tyrosinase vigor50For 6.23mgmL-1。
Inhibiting mechanism of the 2.3 spirulina polyphenol extracts to tyrosinase
Immobilized substrate L-DOPA concentration changes enzyme amount in live body system, measures the relationship of enzyme amount and enzyme activity.It can by Fig. 3
Know, with the increase of spirulina polyphenol extract concentration, straight slope is successively reduced, and straight-line intersection is origin, illustrates spiral
Algae polyphenol extract is reversible to the process of inhibition of tyrosinase, and the addition of spirulina polyphenol extract does not have in reduction system effectively
Enzyme amount, but the enzyme activity of tyrosinase is suppressed, and catalytic efficiency reduces.
Inhibition type and inhibition constant of the 2.4 spirulina polyphenol extracts to tyrosinase
Fixed enzyme concentration, changes the amount of substrate, measures influence of the various concentration spirulina polyphenol extract to enzyme activity.Such as
Shown in Fig. 4 (a), when acting on tyrosinase with extract, it is bis- that the reaction rate and DOPA concentration of enzyme make Lineweaver-Burk
Inverse figure, obtains one group of straight line that intersection point is located at the second quadrant.This shows the raising with spirulina polyphenol extract concentration,
KmValue becomes larger, VmValue becomes smaller, and inhibiting type is mixing competitive type.With the slope and vertical axis intercept of straight line in Fig. 4 (a) to spiral
Algae polyphenol extract makees Fig. 4 (b), Fig. 4 (c), obtains inhibition constant KIAnd KISRespectively 1.94mgmL-1And 6.83mgmL-1。
The comparison of 2.5 marine algae extracts and Chinese medicine whitening composition to tyrosinase inhibitory action
Cyanobacteria (spirulina), green alga (marginal tuber), red algae (gracilaria asiatica) and brown alga (okamura ishige algae) and Chinese medicine (ginkgo leaf,
Radix Glycyrrhizae) extract is shown in Table 1 to the enzyme kinetics parameter of tyrosinase.As shown in Table 1, the extract of six kinds of separate sources is to junket ammonia
The process of inhibition of sour enzyme is all reversible.By comparing rate IC half-suppressed50Value, can obtain six kinds of extracts to the inhibitory effect of tyrosinase
Successively are as follows: okamura ishige algae extract > marginal tuber extract > licorice > spirulina polyphenol extract > ginkgo biloba p.e >
Gracilaria asiatica extract.
1 natural extract of table compares tyrosinase diphenolase inhibitory effect
Table 1Comparison of inhitory effect of natural extracts on
diphenolase of tyrosinase
3 discuss
Melanin deposition will lead to skin splash and fruits and vegetables brown stain, and production quantity is related with the activity of tyrosinase.From day
Separate and extract safe and efficient tyrosinase inhibitor in right plant and be applied to exploitation preserving fruit and vegetable utilizing, cosmetics whitening etc.
Product is always research hotspot, but less about the research report that seaweed active constituent inhibits tyrosinase.
Multiple nutritional components and bioactive substance are rich in spirulina, because there is removing toxic substances, enhance immune, antibacterial, antioxygen
Change, it is antitumor and other effects and by favor.Research spirulina-active ingredient becomes for development function food, cosmetics and drug
New hot spot.Development of the China Jing Guo recent decades has become spirulina raw material big producer, the world, but the spirulina former material in China
For material based on simply utilizing, product finishing stage is inadequate.
Herein using spirulina powder as raw material, polyphenols is extracted from spirulina powder using ethyl alcohol, studies it to junket
The depression effect of propylhomoserin enzyme activity, and compare the inhibitory effect of seaweed active constituent and Chinese medicine whitening composition.From result of study
It is found that spirulina polyphenol content is lower, polyphenol content 35.05mg in the spirulina polyphenol extract that 80% ethyl alcohol extracts
g-1；Spirulina polyphenol extract has inhibiting effect to tyrosinase, to the rate (IC half-suppressed of diphenolase50) it is 6.29mgmL-1.Extract is reversible to the inhibition of tyrosinase diphenolase, shows that spirulina polyphenol extract can reduce enzymatic efficiency, without
Reduce effective enzyme amount.It is mixed type, inhibition constant K that spirulina polyphenol extract, which inhibits type to enzyme,I=1.94mgmL-1、KIS
=6.83mgmL-1, show spirulina polyphenol extract can simultaneously with resolvase and enzyme-substrate complex ining conjunction with, and with dissociate
The binding ability of enzyme is stronger；Compare the kinetic parameter of four kinds of marine algae extracts and Chinese medicine whitening composition it is found that marine algae extract
It inhibits tyrosinase activity, and the inhibitory effect of extracting section object is better than Chinese medical extract whitening agent (Radix Glycyrrhizae, ginkgo in the market
Leaf extract), this provides possibility to the inhibiting effect of tyrosinase to continue to study other active constituents in spirulina.
Inhibition of the spirulina polyphenol extract to tyrosinase activity will be applied to skin-whitening, medical treatment, fruits and vegetables guarantor for it
Fresh equal fields provide reliable theoretical foundation, also by the tyrosinase to comprehensively utilize spirulina resource and research safety, economy
Inhibitor provides new way.
The above is only the preferred embodiment of the present invention, the range implemented of the present invention that therefore, it cannot be limited according to, i.e., according to
Equivalent changes and modifications made by the invention patent range and description, should still be within the scope of the present invention.
Claims (3)
1. the polyphenol extract in a kind of spirulina is preparing the purposes in tyrosinase inhibitor, it is characterised in that: with spiral
Algae polyphenol extract is used to prepare tyrosinase inhibitor as one of active constituent or unique active constituent；The spirulina
The preparation method of polyphenol extract: using volumetric concentration to extract spirulina for 80% ethyl alcohol, obtain spirulina polyphenolic extract,
Spirulina polyphenolic extract is concentrated under reduced pressure in 40 DEG C again, solid spirulina polyphenol extract is obtained after freeze-drying.
2. the polyphenol extract in a kind of spirulina as described in claim 1 is preparing the purposes in tyrosinase inhibitor,
It is characterized by: using spirulina polyphenol extract as one of active constituent or unique active constituent is used for anti-senile plaque or suppression
The antistaling agent of food brown stain processed.
3. the polyphenol extract in a kind of spirulina as described in claim 1 is preparing the purposes in tyrosinase inhibitor,
It is characterized by: the measuring method of spirulina polyphenol content: spectrophotometry is used, with forint phenol reagent to more in extract
Phenol colour developing, determines polyphenol content: ultrapure water dissolves 0.110g gallic acid and obtains galla turcica acidity scale using 100mL volumetric flask constant volume
Quasi- stock solution；It is diluted to 10,20,30,40 and 50 μ gmL respectively with gallic acid Standard Stock solutions again-1Series it is molten
Liquid does blank control with ultrapure water, 1.0mL dilution is taken to be placed in colorimetric cylinder respectively, and 5.0mL forint phenol solution, reaction is added
4.0mL 7.5%Na is added in 6min2CO3Solution mixes, and after avoid light place 1h, light absorption value is measured at wavelength 765nm, makes
Standard curve；Spirulina polyphenol content is according to standard curve colorimetric estimation.
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| CN101450052A (en) * | 2007-11-30 | 2009-06-10 | 中国科学院上海药物研究所 | Use of black mulberry extract in preparing tyrosinase inhibitor |
| CN102342398A (en) * | 2011-06-28 | 2012-02-08 | 文渊 | Radioresistant xylitol |
| CN103800275A (en) * | 2014-02-08 | 2014-05-21 | 李美凤 | Whitening anti-aging cosmetic and preparation method thereof |
| CN104983658A (en) * | 2015-06-25 | 2015-10-21 | 天峨县平昌生态农业有限公司 | Freckle-removing essence and preparing method thereof |
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|---|---|---|---|---|
| CN101450052A (en) * | 2007-11-30 | 2009-06-10 | 中国科学院上海药物研究所 | Use of black mulberry extract in preparing tyrosinase inhibitor |
| CN102342398A (en) * | 2011-06-28 | 2012-02-08 | 文渊 | Radioresistant xylitol |
| CN103800275A (en) * | 2014-02-08 | 2014-05-21 | 李美凤 | Whitening anti-aging cosmetic and preparation method thereof |
| CN104983658A (en) * | 2015-06-25 | 2015-10-21 | 天峨县平昌生态农业有限公司 | Freckle-removing essence and preparing method thereof |
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