CN110090164A - Cosmetic composition and preparation method thereof comprising coconut candy tunning - Google Patents
Cosmetic composition and preparation method thereof comprising coconut candy tunning Download PDFInfo
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- CN110090164A CN110090164A CN201910086656.2A CN201910086656A CN110090164A CN 110090164 A CN110090164 A CN 110090164A CN 201910086656 A CN201910086656 A CN 201910086656A CN 110090164 A CN110090164 A CN 110090164A
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- 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
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9794—Liliopsida [monocotyledons]
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- 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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/60—Sugars; Derivatives thereof
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- 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
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- 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
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- 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
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
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Abstract
Cosmetic composition and preparation method thereof the present invention relates to the lactobacillus-fermented product comprising coconut candy as effective component, due to the silk polymeric protein of cosmetic the composition of the invention, (HAS3 of hyaluronic acid synthetase -3, Hyaluronic acid synthase-3), 3 (AQP3 of aquaporin, Aquaporins 3) etc. the factor in relation to skin barrier and moisturizing content it is high, to improve skin moisture-keeping effect, and have the effect of improving skin barrier.
Description
Technical field
The present invention relates to the cosmetic compositions and preparation method thereof comprising coconut candy tunning as effective component
(Cosmetic composition comprising fermented coconut sugar extract and
manufacturing method thereof)。
Background technique
Palm sugar or palm sugar (palm sugar), which refer to, boils coconut (coco), palmyra (Palmyra
Palm), the Palmaes such as date (date palm), water coconut (nypa fruticans), Arenga pinnata tree (sugar palm)
The sugar that the sap of coco obtains, coconut candy refer to by the palm sugar of the sap manufacture of coconut (coconut palm) scape.It is known
The main composition of coconut candy includes sucrose (sucrose) 70~79%, glucose (glucose) and fructose (fructose)
Respectively about 3~9% or so [Beck L., 2014, " Coconut sugar:Is it healthier than white
sugar,or just hype”,Theglobe&Mail,Retrieved 30May 2015]。
In the form of crystal (crystal) or particle (granule), blocky (block) or liquid form provide coconut candy, main
It is used for field of food, once attempts that existing refined sugar is replaced to reduce heat used as the coconut candy of natural sugar.In addition,
Also once there is the example using coconut candy in cosmetic field, but the type of its preparation is only limitted to frosted facial mask.
Then, the present inventor makes great efforts to develop the cosmetics for the various dosage forms that coconut candy is utilized, as a result, confirmed coconut palm
The improved function of sub- sugar tunning, so as to complete the present invention.
Existing technical literature
Patent document
Patent document 1: Korean published Patent 10-2012-0095042
Patent document 2: Korean published Patent 10-2015-0058721
Summary of the invention
The object of the present invention is to provide a kind of cosmetic composition and preparation method thereof, the cosmetic combination
Object includes the lactobacillus-fermented product of coconut candy as effective component, to improve skin moisture-keeping and skin texture holding effect.
In order to achieve the above objectives, the present invention provides a kind of cosmetic composition, which, which is used as, has
Imitating ingredient includes by the coconut candy tunning of coconut candy lactobacillus-fermented.
An embodiment according to the present invention, the lactic acid bacteria are the bacterial strains for belonging to Lactobacillus (Lactobacillus).
An embodiment according to the present invention, the coconut candy tunning are prepared by following preparation method, this method packet
It includes: the step of cultivating lactic acid bacteria；The step of lactic acid bacteria strains of the culture are inoculated into coconut sugar juice；And described in culture
The coconut sugar juice of inoculating lactic acid bacterium and the step of fermented.
An embodiment according to the present invention, the cosmetic composition can include coconut of the invention by following
Sugared tunning is prepared as the preparation method of the cosmetic composition of effective component.
An embodiment according to the present invention, the cosmetic composition is by increasing silk polymeric protein, hyaluronic acid conjunction
At enzyme -3 (HAS3, Hyaluronic acid synthase-3), the expression quantity of aquaporin 3 (AQP3, Aquaporins3)
Effect is kept to improve skin moisture-keeping and skin texture.
On the other hand, the present invention provides a kind of preparation method of cosmetic composition, which can
It improves skin moisture-keeping and skin texture keeps effect.
An embodiment according to the present invention, the method are the makeup materials as effective component comprising coconut candy tunning
The preparation method of feed composition, this method comprises: preculture belongs to the step of the lactic acid bacteria of Lactobacillus (Lactobacillus)
Suddenly；The step of lactic acid bacteria strains of the preculture are inoculated into coconut sugar juice；And culture has been inoculated with the lactic acid bacteria
Coconut sugar juice and the step of fermented.
An embodiment according to the present invention, the coconut sugar juice can be containing the coconut candy of 5~15 weight %, but is not
It is defined in this.
An embodiment according to the present invention, the inoculation can be with 1 × 104~1 × 106The pre- training of concentration inoculation of cfu/ml
Feeding lactic acid bacteria, but it is not limited to this.
An embodiment according to the present invention, the fermentation can carry out at 30~40 DEG C, but be not limited to this.According to
One embodiment of the invention, the fermentation can carry out 35~50 hours, but be not limited to this.
An embodiment according to the present invention, the preparation method of the cosmetic composition may include: that preculture belongs to
The step of lactic acid bacteria of Lactobacillus (Lactobacillus)；By the lactic acid bacteria of the preculture with 1 × 105~5 ×
105The concentration of cfu/ml is inoculated into the step of coconut sugar juice of 5 weight %；The coconut sugar juice of the lactic acid bacteria will be inoculated with
Under the conditions of 33~36 DEG C, 120~150rpm the step of shaken cultivation 40~48 hours；And after the centrifuge separation culture
The step of filtering supernatant.
Invention effect
The silk polymeric protein of cosmetic composition comprising coconut candy tunning of the invention, hyaluronic acid synthesis
Enzyme -3 (HAS3, Hyaluronic acid synthase-3), the content of aquaporin 3 (AQP3, Aquaporins 3) are aobvious
It writes and therefore shows that excellent skin moisture-keeping and skin barrier change higher than existing coconut sugar extract or other tunnings
Kind effect.
Detailed description of the invention
Fig. 1 shows the preparation sections of the coconut candy tunning of one embodiment of the invention.
Fig. 2 is silk polymeric protein of the measurement coconut candy lactobacillus-fermented product of the invention according to different coconut candy concentration
(filaggrin) result of expression.
: unprocessed (culture medium for being not added with 10% FBS and addition AA of sample), comparative example 1: 5 weight of coconut candy
Measure % solution, embodiment 1-1: 5 weight % lactobacillus-fermented product of coconut candy, embodiment 1-2: 10 weight % lactic acid bacteria of coconut candy
Tunning, embodiment 1-3: 15 weight % lactobacillus-fermented product of coconut candy,
Fig. 3 is the silk polymeric protein of comparison coconut candy lactobacillus-fermented product of the invention and coconut sugar yeast tunning
(filaggrin) result of expression.
: unprocessed, comparative example 1: coconut sugar juice, comparative example 2: coconut sugar yeast tunning, embodiment 1: according to
The coconut candy lactobacillus-fermented product of embodiment 1-1 preparation.
Fig. 4 is the hyaluronic acid conjunction of comparison coconut candy lactobacillus-fermented product of the invention and coconut sugar yeast tunning
At the result of enzyme (HAS3) expression.
: unprocessed, comparative example 1: coconut sugar juice, comparative example 2: coconut sugar yeast tunning, embodiment 1: according to
The coconut candy lactobacillus-fermented product of embodiment 1-1 preparation.
Fig. 5 is the aquaporin of comparison coconut candy lactobacillus-fermented product of the invention and coconut sugar yeast tunning
The result of 3 (AQP3) expressions.
: unprocessed, comparative example 1: coconut sugar juice, comparative example 2: coconut sugar yeast tunning, embodiment 1: according to
The coconut candy lactobacillus-fermented product of embodiment 1-1 preparation.
Specific embodiment
An embodiment of the invention provides a kind of cosmetic composition, and the cosmetic composition is as effective component
Coconut candy tunning comprising the coconut candy that fermented using lactic acid bacteria.
An embodiment according to the present invention, the lactic acid bacteria, which refers to, ferments to carbohydrate to obtain energy, and generates big
The bacterium for measuring lactic acid (lactic acid), for example, may include Lactobacillus (Lactobacillus), lactococcus
(Lactococcus), Leuconostoc (Leuconostoc), Pediococcus (Pediococcus), Bifidobacterium
(Bifidobacterium) Pseudomonas such as, but it is not limited to this.
An embodiment according to the present invention, the lactic acid bacteria are the bacterial strains for belonging to Lactobacillus (Lactobacillus).
An embodiment according to the present invention, the coconut candy tunning are prepared by following preparation method, this method packet
It includes: the step of cultivating lactic acid bacteria；The step of lactic acid bacteria strains of the culture are inoculated into coconut sugar juice；And described in culture
The coconut sugar juice of inoculating lactic acid bacterium and the step of fermented.
An embodiment according to the present invention, the cosmetic composition can include coconut of the invention by following
Sugared tunning is prepared as the preparation method of the cosmetic composition of effective component.
An embodiment according to the present invention is general with using coconut sugar extract without fermentation procedure, using yeast etc.
The coconut candy tunning that ferments of fermentative microorganism compare, the silk polymeric protein of the coconut candy lactobacillus-fermented product
(filaggrin) content is higher.The silk polymeric protein (filaggrin) is known is present in mammalian epidermal cells
Keratin associated protein plays an important role in epidermis stable state (epidermal homeostasis) adjusting, silk polymeric protein list
Body (monomer) is inserted into lipid film in cuticula, so as to execute skin barrier function.In addition, silk polymeric protein is at angle
The top of matter layer is further processed to discharge free amino acid, thereby assists in moisture-keeping function [referring to Ovaere P, et
al.,2009,“The emerging roles of serine protease cascades in the epidermis”,
Trends Biochem.Sci.34 (9): 453-63 etc.].Therefore, the cosmetic comprising coconut candy tunning of the invention
Composition can be improved the skin texture including improving skin barrier and keep effect and skin moisturization.
Another embodiment according to the present invention provides a kind of change for improving skin moisture-keeping and skin texture holding effect
The preparation method of adornment material compositions, the composition include coconut candy tunning as effective component.
An embodiment according to the present invention, the method are the makeup materials as effective component comprising coconut candy tunning
The preparation method of feed composition, this method comprises: preculture belongs to the step of the lactic acid bacteria of Lactobacillus (Lactobacillus)
Suddenly；The step of lactic acid bacteria strains of the preculture are inoculated into coconut sugar juice；And the culture inoculating lactic acid bacterium
Coconut sugar juice and the step of fermented.
Coconut candy lactobacillus-fermented product of the invention can prepare (Fig. 1) by following process.
First step: the step of preculture lactic acid bacteria
First step is the step in order to make to activate and carry out preculture as the lactic acid bacteria of fermentative microorganism before inoculation
Suddenly.An embodiment according to the present invention, the lactic acid bacteria are lactobacillus genus strains, and the preculture of the lactic acid bacteria can be used
MRS meat soup (MRS broth) culture medium carries out, but is not limited to this, can modify Lactobacillus according to the characteristic of bacterial strain
The usually used culture medium of bacterial strain and condition and use.
Second step: the step of by lactobacillus inoculum to coconut sugar juice
Second step is in order to ferment coconut sugar juice and by the lactobacillus inoculum of the preculture to coconut sugar juice
Step.The coconut sugar juice can be by preparing the dissolution of commercially available coconut candy powder in a solvent.
An embodiment according to the present invention, the coconut sugar juice can contain 1~20 weight %, preferably 5~15 weights
The coconut candy of %, most preferably 4~6 weight % are measured, but is not limited to this.An embodiment according to the present invention, the solvent can
To use distilled water, salt content less than 6% seawater, tap water, the underground water below of pH 8 or thermal water etc., but and it is non-limiting
In this.
An embodiment according to the present invention can be used for the coconut sugar juice of lactobacillus-fermented by the coconut candy
After solution sterilizes 15 minutes at 121 DEG C, it is cooled to the coconut sugar juice lower than 35 DEG C.
The concentration of the lactic acid bacteria of the inoculation can be adjusted, according to the present invention according to content, the condition of culture etc. of coconut candy
An embodiment, the inoculation is the lactic acid bacteria by preculture with 1 × 103~1 × 107Cfu/ml, preferably 1 × 104~1 ×
106Cfu/ml, more preferably 1 × 105~5 × 105The concentration of cfu/ml is inoculated with, but is not limited to this.
Third step: the fermentation step of coconut sugar juice
The step of third step is the culture coconut sugar juice of inoculating lactic acid bacterium and is fermented, can be according to cream
Type and concentration of sour bacterium etc., and consider the growth of bacterium, to select best Culture and fermentation conditions.According to the present invention one is real
Example is applied, the culture can be Liquid Culture.An embodiment according to the present invention, the culture can be in 100~150rpm
Under the conditions of the shaken cultivation that carries out, but be not limited to this.An embodiment according to the present invention, the fermentation can be 30~40
DEG C, preferably carried out at 33~36 DEG C, but be not limited to this.An embodiment according to the present invention, the fermentation can carry out
It 35~50 hours, preferably carries out 40~48 hours, but be not limited to this.
Four steps: filtration step
An embodiment according to the present invention, the preparation process of the coconut candy lactobacillus-fermented product can be with further include:
The step of being filtered before preparing the coconut candy lactobacillus-fermented product.An embodiment according to the present invention, the filtering
It is carried out after can use centrifugation tunning by the method for film filtering supernatant, but is not limited to this, and can benefit
Known method used in filter process used in tunning carries out.
The coconut candy lactobacillus-fermented product or its fraction prepared by the above process can be configured to various forms.This hair
Bright cosmetic composition can have moisturizing lotion, skin softener, toner, astringent, lotion, milk milky lotion,
Moisturizing emulsion, nutritional emulsions, massage cream, nourishing cream, moisturiser, hand lotion, ointment, foundation emulsion, Essence, nutrition Essence,
The various dosage forms such as facial mask, perfumed soap, foam facial cleanser, mildy wash, washing cream, body lotion, shower cream, and liquid can be prepared as
Body, paste, paste, solid, lotion, suspension, microemulsion, microcapsules, particle, ionic liposome, non-ionic vesica point
The various characters such as powder, pulvis, aerosol.
In addition, other than coconut candy tunning or its fraction of the invention, the cosmetic composition can be with
Containing commonly used in cosmetics adjuvant or well known functional component (skin moisture-keeping, improve skin elasticity, whitening, improve wrinkle,
Skin barrier strengthens ingredient etc.).Example as the adjuvant, comprising: concentrating agents, solubilizer, gelling agent, softening agent, filling
Agent, antioxidant, blocking agent, wetting agent, preservative, suspending agent, stabilizer, foaming agent, aromatic, surfactant, fat
Substance, organic solvent, water, ionic or nonionic emulsifier, metal ion chelation agent, chelating agent, vitamin, essential oil, dye
Material, pigment, hydrophily or lipophilic activating agent, lipid vesicle etc., but it is not limited to this.
An embodiment according to the present invention, the cosmetic of the invention as effective component comprising coconut candy tunning
Composition can be in the cosmetic composition containing the substance commonly used in cosmetics preparation, add hair of the invention
1~95 weight % of ferment product, the cosmetic composition for preferably adding 5~90 weight %, but it is not limited to this.When concentration is small
When 1 weight %, it is difficult to skin effect is expected, when concentration is greater than 95 weight %, in the anti-corrosion and use feeling for constituting dosage form
Use in additive may be restricted.
Hereinafter, referring to embodiment the present invention is described in detail.However, following embodiment only illustrates the present invention, of the invention is interior
Appearance is not limited to following embodiment.
<embodiment 1>
The preparation of coconut candy lactobacillus-fermented product of the invention
<1-1>prepares 5 weight % lactobacillus-fermented product of coconut candy
In order to prepare the cosmetic composition comprising coconut candy tunning of the invention, commercially available coconut candy is bought
(Republic of Korea, HEDEN Co., Ltd., www.heden.co.kr), is prepared for coconut candy tunning as follows.Fermentative microorganism
The lactic acid bacteria (KCCM 12203, Lactobacillus sp.) purchased from Korean Culture Center is used.
By the lactic acid bacteria in MRS meat soup (BD, USA) preculture so that initial bacterial count be 1.0 × 105~5.0 ×
105cfu/ml.Distilled water 95g and coconut candy 5g are put into together in conical flask and after making it completely dissolved, gone out at 121 DEG C
It bacterium 15 minutes, then cools to room temperature and is prepared for 5 weight % solution of coconut candy.By the lactobacillus inoculum of the preculture to institute
After stating 5 weight % solution of coconut candy, the shaken cultivation 40~48 hours under the conditions of 33~36 DEG C, 120~150rpm.It will be described
It is centrifugated after ten minutes with the coconut sugar juice of lactobacillus-fermented with 3500rpm, with 0.45um film filtering supernatant and degerming
For testing.
The preparation of 10 weight % lactobacillus-fermented product of<1-2>coconut candy
Commercially available coconut candy (Republic of Korea, HEDEN Co., Ltd., www.heden.co.kr) is bought, is prepared for coconut palm as follows
Sub- sugar tunning.Fermentative microorganism used purchased from Korean Culture Center lactic acid bacteria (KCCM 12203,
Lactobacillus sp.)。
By the lactic acid bacteria in MRS meat soup (BD, USA) preculture so that initial bacterial count be 1.0 × 105~5.0 ×
105cfu/ml.Distilled water 90g and coconut candy 10g are put into together in conical flask and after making it completely dissolved, at 121 DEG C
Sterilizing 15 minutes, then cools to room temperature and is prepared for 10 weight % solution of coconut candy.By the lactobacillus inoculum to the coconut palm
After 10 weight % solution of sub- sugar, the shaken cultivation 40~48 hours under the conditions of 33~36 DEG C, 120~150rpm.Lactic acid will be used
The coconut sugar juice of bacterium fermentation is centrifugated after ten minutes with 3500rpm, and with 0.45um film filtering supernatant and degerming is to be used for
Experiment.
The preparation of 15 weight % lactobacillus-fermented product of<1-3>coconut candy
Commercially available coconut candy (Republic of Korea, HEDEN Co., Ltd., www.heden.co.kr) is bought, is prepared for coconut palm as follows
Sub- sugar tunning.Fermentative microorganism used purchased from Korean Culture Center lactic acid bacteria (KCCM 12203,
Lactobacillus sp.)。
By the lactic acid bacteria in MRS meat soup (BD, USA) preculture so that initial bacterial count be 1.0 × 105~5.0 ×
105cfu/ml.Distilled water 85g and coconut candy 15g are put into together in conical flask and after making it completely dissolved, at 121 DEG C
Sterilizing 15 minutes, then cools to room temperature and is prepared for 15 weight % solution of coconut candy.By the lactobacillus inoculum to the coconut palm
After 15 weight % solution of sub- sugar, the shaken cultivation 40~48 hours under the conditions of 33~36 DEG C, 120~150rpm.By the use
The coconut sugar juice of lactobacillus-fermented is centrifugated after ten minutes with 3500rpm, with 0.45um film filtering supernatant and degerming with
For testing.
It is used for the comparative example tested together with the coconut candy lactobacillus-fermented product of the invention, is to make in the following manner
It is standby.
[comparative example 1] coconut candy (5 weight %) solution
Coconut candy purchased from HEDEN Co., Ltd. (Republic of Korea, www.heden.co.kr) is soluble in water, to make
For 5 weight % solution of coconut candy.Distilled water 95g and coconut candy 5g are put into conical flask together and are made it completely dissolved
Afterwards, it sterilizes 15 minutes at 121 DEG C, then cools to room temperature and be prepared for 5 weight % solution of coconut candy.
[comparative example 2] coconut sugar yeast tunning
Yeast (KCCM 11293, Saccharomyces cerevisae) purchased from Korean Culture Center is connect
Kind is into the coconut sugar juice of the comparative example 1 and is fermented.
Yeast after preculture, is inoculated into Potato dextrose meat soup (potato dextrose broth, BD, USA)
In the 5 weight % solution of coconut candy to sterilize 15 minutes at 121 DEG C, so that initial bacterial count is 1.0 × 105~5.0 ×
105Cfu/ml, and shaken cultivation 40~48 hours 30~33 DEG C, 120~150rpm under the conditions of.Yeast-leavened coconut palm will be used
Sub- sugar juice is centrifugated after ten minutes with 3500rpm, with 0.45um film filtering supernatant and degerming for testing.
<embodiment 2>
Silk polymeric protein (Filaggrin) expression quantity of lactobacillus-fermented product is evaluated according to coconut candy concentration
In order to confirm skin moisture-keeping and barrier of the coconut candy to coconut candy lactobacillus-fermented product of the present invention of various concentration
The influence of strengthening effect, using the sample prepared according to the embodiment<1-1>to<1-3>and comparative example 1 as object, and following institute
Determine the mRNA relative expression levels of the silk polymeric protein (Filaggrin) as skin barrier and moisturizing factor with stating.
As Human keratinocytes strain HaCat purchased from America type culture collection (ATCC,
Rockville, MD, USA), and resisted using 10% fetal calf serum (FBS, ATCC, Rockville, MD, USA) and 1% is added to
The DMEM (Hyclone, Logan, UT, USA) of bacterium-antifungal agent (AA, ATCC, Rockville, MD, USA), in 37 DEG C, 5%
CO2It is cultivated in the incubator of condition.
In order to confirm barrier strengthening effect in Human keratinocytes, by Human keratinocytes with 5 × 105Cell/
After the density in hole is inoculated into 6 porocyte culture plates, using the culture medium for being added to 10% FBS and 1% AA, in 37 DEG C, 5%
CO2It has been cultivated in culture medium 24 hours.
Addition is according to the embodiment<1-1>to<1-3>and each sample of the preparation of comparative example 1 and cultivates 24 hours, recycling
Cell simultaneously adds 1mL tricyclazole (RNA iso, DAKARA, Japan) to separate RNA.Using Nanodrop 2000 (Thermo,
USA it) after quantitative RNA, is reacted 55 minutes at 42 DEG C, reacts 15 minutes at 70 DEG C to synthesize cDNA (Reverse
Transcriptase Mix, ELPIS biotech, South Korea).Using PCR instrument, (Step One Plus, the U.S. apply biology department
System company, the U.S.) reverse transcription polymerase chain reaction (RT-PCR) is carried out, by SYBRGreen (SYBRGreen supermix, beauty
Applied Biosystems, Inc., state, the U.S.) added together with silk polymeric protein (Filaggrin) primer and cDNA, at 94 DEG C into
5 minutes polymerase activations of row, and by 30 seconds at 95 DEG C, 1 minute at 54 DEG C, at 72 DEG C 1 minute polymerization reaction, carry out
40 circulations.The base sequence of primer used in experiment is as shown in table 1 below.Finally by correction beta-actin (β-
Actin) gene analyzes the expression quantity of gene.
Table 1
The base sequence of silk polymeric protein (Filaggrin) primer
As a result, as shown in Fig. 2, the silk for confirming coconut candy lactobacillus-fermented product (embodiment 1-1) of the invention is poly-
Hop protein expression quantity increases compared with untreated fish group (be not added with 10% FBS of sample and be added to the culture medium of AA)
3.83 times, and confirm and control group (comparative example 1) phase for having used the 5 weight % solution of coconut candy without lactobacillus-fermented
Than also substantially increasing.
In addition, the 5 weight % lactobacillus-fermented product (embodiment 1-1) of coconut candy, 10 weight of coconut candy that use in an experiment
It measures between % lactobacillus-fermented product (embodiment 1-2), 15 weight % lactobacillus-fermented product (embodiment 1-3) of coconut candy, no
Silk polymeric protein expression quantity with concentration coconut candy does not show significant difference.
Although will not be shown because of the concentration of coconut candy difference by the coconut candy tunning of one embodiment of the invention preparation
A significant difference for polymeric protein expression quantity is shown, but compared with the coconut sugar juice without fermentation procedure, is shown obvious
Higher silk polymeric protein expression.Refer to color and smell usually as the character of cosmetic material, and with coconut candy
Concentration is higher, and color becomes opaquer and specific scent becomes stronger.Therefore, when in view of being used as cosmetics as described above
Whens character and cost of raw material etc., preferably pass through the coconut candy hydroponics lactic acid bacteria with the coconut candy containing 5 weight % concentration
To prepare coconut candy lactobacillus-fermented product of the invention.
<embodiment 3>
The silk polymeric protein of comparative evaluation coconut candy lactobacillus-fermented product and coconut sugar yeast tunning
(Filaggrin) expression quantity
In order to confirm the skin moisture-keeping and barrier of the coconut candy lactobacillus-fermented product according to the embodiment<1-1>preparation
Strengthening effect, also to the coconut sugar yeast tunning and silk polymeric protein (filaggrin) prepared according to the comparative example 2
MRNA relative expression levels compare.
As Human keratinocytes strain HaCat purchased from America type culture collection (ATCC,
Rockville, MD, USA), and utilize the fetal calf serum (FBS, ATCC, Rockville, MD, USA) and 1% for being added to 10%
The DMEM (Hyclone, Logan, UT, USA) of antibacterial-antifungal agent (AA, ATCC, Rockville, MD, USA), 37 DEG C,
5%CO2It is cultivated in the incubator of condition.
In order to confirm barrier strengthening effect in Human keratinocytes, by Human keratinocytes with 5 × 105Cell/
After the density in hole is inoculated into 6 porocyte culture plates, using the culture medium for being added to 10% FBS and 1% AA, in 37 DEG C, 5%
CO2It has been cultivated in culture medium 24 hours.
Addition is according to the Comparative Examples 1 and 2 and each sample of embodiment<1-1>preparation and cultivates 24 hours, and recycling cell is simultaneously
1mL tricyclazole (RNA iso, DAKARA, Japan) is added to separate RNA.It is quantitative using Nanodrop 2000 (Thermo, USA)
It after RNA, is reacted 55 minutes at 42 DEG C, reacts 15 minutes at 70 DEG C to have synthesized cDNA (Reverse Transcriptase
Mix, ELPIS biotech, South Korea).Using PCR instrument (Step One Plus, Applied biosystems, the U.S.) into
Row reverse transcription polymerase chain reaction (RT-PCR), by SYBRGreen (SYBRGreen supermix, U.S.A. applied biosystem
Company, the U.S.) it is added together with silk polymeric protein (filaggrin) primer and cDNA, 5 minutes polymerases are carried out at 94 DEG C
Activation, and by 30 seconds at 95 DEG C, 1 minute at 54 DEG C, at 72 DEG C 1 minute polymerization reaction, carried out 40 circulations.It is real
The base sequence of primer used in testing is as shown in Table 1 above.Gene is analyzed finally by correction beta-actin gene
Expression quantity.
As a result, as shown in figure 3, confirming the coconut candy lactobacillus-fermented of the invention prepared according to embodiment<1-1>
The silk polymeric protein expression quantity of product (embodiment 1) increases 3.83 times compared with untreated fish group, and confirms and coconut candy
Solution (comparative example 1) is compared with yeast tunning (comparative example 2), shows considerably higher expression quantity.
<embodiment 4>
The hyaluronic acid synthetase of comparative evaluation coconut candy lactobacillus-fermented product and coconut sugar yeast tunning
(HAS3) expression quantity
Effect is strengthened according to the skin moisture-keeping of the coconut candy lactobacillus-fermented product of the embodiment<1-1>preparation in order to confirm
Fruit, by as the hyaluronic acid synthetase -3 of correlation factor (HAS3's (Hyaluronic acid synthase-3, HAS3)
MRNA relative expression levels compare with coconut sugar yeast tunning (comparative example 2).Experimental method is as described below.
As Human keratinocytes strain HaCat purchased from America type culture collection (ATCC,
Rockville, MD, USA), and resisted using 10% fetal calf serum (FBS, ATCC, Rockville, MD, USA) and 1% is added to
The DMEM (Hyclone, Logan, UT, USA) of bacterium-antifungal agent (AA, ATCC, Rockville, MD, USA), in 37 DEG C, 5%
CO2It is cultivated in the incubator of condition.
In order to confirm barrier strengthening effect in Human keratinocytes, by Human keratinocytes with 5 × 105Cell/
After the density in hole is inoculated into 6 porocyte culture plates, using the culture medium for being added to 10% FBS and 1% AA, in 37 DEG C, 5%
CO2It has been cultivated in culture medium 24 hours.
Addition is according to the Comparative Examples 1 and 2 and each sample of embodiment<1-1>preparation and cultivates 24 hours, and recycling cell is simultaneously
1mL tricyclazole (RNA iso, DAKARA, Japan) is added to separate RNA.It is quantitative using Nanodrop 2000 (Thermo, USA)
It after RNA, is reacted 55 minutes at 42 DEG C, reacts 15 minutes at 70 DEG C to have synthesized cDNA (Reverse Transcriptase
Mix, ELPIS biotech, South Korea).Using PCR instrument (Step One Plus, Applied biosystems, the U.S.) into
Row reverse transcription polymerase chain reaction (RT-PCR), by SYBRGreen (SYBRGreen supermix, U.S.A. applied biosystem
Company, the U.S.) it is added together with silk polymeric protein (filaggrin) primer and cDNA, 5 minutes polymerases are carried out at 94 DEG C to swash
It is living, and by 30 seconds at 95 DEG C, 1 minute at 54 DEG C, at 72 DEG C 1 minute polymerization reaction, carried out 40 circulations.Experiment
Used in primer base sequence it is as described in Table 2.The table of gene is analyzed finally by correction beta-actin gene
Up to amount.
Table 2
The base sequence of hyaluronic acid synthetase -3 (HAS3) primer
As a result, as shown in figure 4, confirming the coconut candy lactobacillus-fermented of the invention prepared according to embodiment<1-1>
The HAS3 expression of product (embodiment 1) increases 4.2 times, and confirm compared with negative (-) control group (untreated fish group)
To compared with coconut sugar yeast tunning, considerably higher level (Fig. 4) is also showed that.
<embodiment 5>
Aquaporin 3 (AQP3) table of comparative evaluation coconut candy lactobacillus-fermented product and coconut sugar yeast tunning
Up to amount
Effect is strengthened according to the skin moisture-keeping of the coconut candy lactobacillus-fermented product of the embodiment<1-1>preparation in order to confirm
Fruit, will be as the mRNA relative expression levels of the aquaporin 3 (Aquaporins, AQP3) of correlation factor and coconut sugar yeast
Tunning (comparative example 2) compares.Experimental method is as described below.
As Human keratinocytes strain HaCat purchased from America type culture collection (ATCC,
Rockville, MD, USA), and resisted using 10% fetal calf serum (FBS, ATCC, Rockville, MD, USA) and 1% is added to
The DMEM (Hyclone, Logan, UT, USA) of bacterium-antifungal agent (AA, ATCC, Rockville, MD, USA), in 37 DEG C, 5%
CO2It is cultivated in the incubator of condition.
In order to confirm barrier strengthening effect in Human keratinocytes, by Human keratinocytes with 5 × 105Cell/
After the density in hole is inoculated into 6 porocyte culture plates, using the culture medium for being added to 10% FBS and 1% AA, in 37 DEG C, 5%
CO2It has been cultivated in culture medium 24 hours.
Addition is according to the Comparative Examples 1 and 2 and each sample of embodiment<1-1>preparation and cultivates 24 hours, and recycling cell is simultaneously
1mL tricyclazole (RNA iso, DAKARA, Japan) is added to separate RNA.It is quantitative using Nanodrop 2000 (Thermo, USA)
It after RNA, is reacted 55 minutes at 42 DEG C, reacts 15 minutes at 70 DEG C to have synthesized cDNA (Reverse Transcriptase
Mix, ELPIS biotech, South Korea).Using PCR instrument (Step One Plus, Applied biosystems, the U.S.) into
Row reverse transcription polymerase chain reaction (RT-PCR), by SYBRGreen (SYBRGreen supermix, U.S.A. applied biosystem
Company, the U.S.) it is added together with silk polymeric protein (filaggrin) primer and cDNA, 5 minutes polymerases are carried out at 94 DEG C to swash
It is living, and by 30 seconds at 95 DEG C, 1 minute at 54 DEG C, at 72 DEG C 1 minute polymerization reaction, carried out 40 circulations.Experiment
Used in primer base sequence it is as described in Table 3.The table of gene is analyzed finally by correction beta-actin gene
Up to amount.
Table 3
Aquaporin 3 (AQP3) primer nucleotide sequences
As a result, as shown in figure 5, confirming the coconut candy lactobacillus-fermented of the invention prepared according to embodiment<1-1>
The AQP3 expression of product (embodiment 1) increases 1.7 times, and confirm compared with negative (-) control group (untreated fish group)
To compared with coconut sugar yeast tunning, considerably higher level (Fig. 5) is shown.
Claims (9)
1. a kind of cosmetic composition, wherein
As the coconut candy tunning that effective component includes using lactobacillus-fermented coconut candy.
2. cosmetic composition according to claim 1, which is characterized in that
The genus lactubacillus is in Lactobacillus.
3. cosmetic composition according to claim 1, which is characterized in that
The coconut candy tunning is prepared by following preparation method, this method comprises:
The step of cultivating lactic acid bacteria；
The step of lactic acid bacteria strains of culture are inoculated into coconut sugar juice；And
The step of culture has been inoculated with the coconut sugar juice of the lactic acid bacteria and has been fermented.
4. cosmetic composition according to claim 1, which is characterized in that
The cosmetic composition improves skin moisture-keeping and skin texture keeps effect.
5. a kind of preparation method of the cosmetic composition as effective component comprising coconut candy tunning, wherein
Include:
Preculture belongs to the step of lactic acid bacteria of Lactobacillus；
The step of lactic acid bacteria strains of preculture are inoculated into coconut sugar juice；And
The step of culture has been inoculated with the coconut sugar juice of the lactic acid bacteria and has been fermented.
6. preparation method according to claim 5, which is characterized in that
The coconut sugar juice contains the coconut candy of 5~15 weight %.
7. preparation method according to claim 5, which is characterized in that
By the lactic acid bacteria of preculture with 1 × 104~1 × 106The concentration of cfu/ml is inoculated with.
8. preparation method according to claim 5, which is characterized in that
The fermentation carries out 35~50 hours at 30~40 DEG C.
9. preparation method according to claim 5, which is characterized in that
Include:
Preculture belongs to the step of lactic acid bacteria of Lactobacillus；
By the lactic acid bacteria of preculture with 1 × 105~5 × 105The concentration of cfu/ml is inoculated into the coconut sugar juice of 5 weight %
The step of；
To be inoculated with the coconut sugar juice of the lactic acid bacteria under conditions of 33~36 DEG C, 120~150rpm shaken cultivation 40~
48 hours the step of；And
The step of being centrifugated filtering supernatant after the culture.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JP2002284663A (en) * | 2001-03-27 | 2002-10-03 | Ichimaru Pharcos Co Ltd | Cosmetic composition |
| US20160286820A1 (en) * | 2015-03-30 | 2016-10-06 | Active Micro Technologies, LLC | Fermented coconut antimicrobial product |
| JP2016193857A (en) * | 2015-03-31 | 2016-11-17 | 丸善製薬株式会社 | Skin cosmetic, and food and drink |
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| JP2002284663A (en) * | 2001-03-27 | 2002-10-03 | Ichimaru Pharcos Co Ltd | Cosmetic composition |
| US20160286820A1 (en) * | 2015-03-30 | 2016-10-06 | Active Micro Technologies, LLC | Fermented coconut antimicrobial product |
| JP2016193857A (en) * | 2015-03-31 | 2016-11-17 | 丸善製薬株式会社 | Skin cosmetic, and food and drink |
Non-Patent Citations (2)
| Title |
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| AKIHIKO KOSUGI ET AL.: ""Ethanol and lactic acid production using sap squeezed from old oil palm trunks felled for replanting"", 《JOURNAL OF BIOSCIENCE AND BIOENGINEERING》 * |
| S. CHOOKLIN ET AL.: ""Potential Utilization of Sap from Oil Palm (Elaeis guineensis) for Lactic Acid Production by Lactobacillus casei"", 《JOURNAL OF SUSTAINABLE ENERGY & ENVIRONMENT》 * |
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