CN114702558A

CN114702558A - Novel glucomannose hybrid glycan peptide and preparation method and application thereof

Info

Publication number: CN114702558A
Application number: CN202210358654.6A
Authority: CN
Inventors: 骆俊清; 苟小军
Original assignee: Sichuan Aobang Investment Co ltd
Current assignee: Sichuan Aobang Investment Co ltd
Priority date: 2022-04-06
Filing date: 2022-04-06
Publication date: 2022-07-05
Anticipated expiration: 2042-04-06
Also published as: CN114702558B

Abstract

The invention discloses a novel glucoglyceroheteropolypeptide, a preparation method and application thereof, wherein the novel glucoglyceroheteropolypeptide is a fermentation product of alpha-hemolytic streptococcus 33#, uniform polysaccharide peptide is obtained by separation and purification by adopting a concentration and alcohol precipitation method and an ultrafiltration method, and the structure of the uniform polysaccharide peptide is identified by physical and chemical analysis; according to molar ratio, the monosaccharide composition of the novel glucoglyceroheteropolyglycopeptide is mannose: rhamnose: glucose is 8.22:1:6.46, weight average molecular weight is 10045Da, and molecular weight distribution coefficient is 1.31; the acne removing cosmetic has obvious moisturizing effect, is long in moisturizing time, has an effect of inhibiting oil of propionibacterium acnes, achieves the effects of diminishing inflammation and removing acnes, can balance sebum secretion on the surface of the skin of a human body, enables the repaired skin to be fuller, and reduces residue of acne marks; can avoid the skin surface from loosening and pore blocking.

Description

Novel glucomannose hybrid glycan peptide and preparation method and application thereof

Technical Field

The invention relates to the technical field of biology, in particular to a novel glucoganan polypeptide and a preparation method and application thereof.

Background

Cosmetics are a necessity of people in daily life. The cosmetics on the market at present are divided into moisture retention, whitening, sun protection, acne removal, wrinkle resistance and the like according to functions; the active ingredients in cosmetics can be classified into synthetic and natural sources according to their sources. In recent years, natural active ingredients applied to cosmetics have the advantages of clear effect, small toxic and side effects and the like.

Natural polysaccharides have complex biological activity and function. The polysaccharide contains a large number of hydrophilic groups and shows excellent physicochemical properties such as strong water absorption, emulsibility, high viscosity and good film forming property, and the properties determine that the polysaccharide has the natural advantages of being developed into moisturizing components. Meanwhile, researches show that the polysaccharide can be widely involved in various vital activities of cells to generate various biological functions, such as anti-inflammation, antivirus, anti-tumor, anticoagulation, anti-radiation, anti-aging and the like. Therefore, the application range of natural polysaccharide compounds in the cosmetic field is also increasing. Acne, pimples or comedo is a chronic inflammatory reaction of the pilosebaceous glands of the skin, and polysaccharides with anti-inflammatory activity may remove acne. The acne removing method comprises an oral medicinal preparation and an external medicament for treatment, and although the methods also have certain effects, the method has prominent disadvantages, such as unobvious effect, easy scar left on the face and other toxic and side effects.

alpha-hemolytic Streptococcus (Streptococcus hemolyticus-a-haemolysis) 33# is separated from the oral cavity and throat parts of healthy people, and alpha-Mannatide, also called Mannatide (Mannatide) or polyoxin A, can be obtained by liquid fermentation culture, and is a novel immunopotentiator originated in China, monosaccharide components of the Mannatide are mannose and glucose, the mannose accounts for more than 90% of the total ratio, and the relative molecular mass is 4-6 ten thousand Da. The research shows that the low concentration of the a-mannan peptide can activate the cellular immunity and the humoral immunity of the organism. In the prior art, the strain is not applied to acne-removing cosmetics, and the glucomannan peptide with smaller molecular weight is separated from the fermentation liquor of the strain and is applied to the acne-removing cosmetics.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a novel glucomannatide and a preparation method and application thereof, wherein the novel glucomannatide with smaller molecular weight is separated from alpha-hemolytic Streptococcus (Streptococcus thermophilus-alpha-haemolysis) 33# fermentation liquor and is applied to acne-removing cosmetics, so that the redness and swelling condition on the surface of the skin of a human body is effectively eliminated, sebum secretion on the surface of the skin of the human body can be balanced, the skin can be fuller after being repaired, the residue of acne marks is reduced, and the conditions of looseness and pore blockage on the surface of the skin are avoided.

The purpose of the invention is realized by the following technical scheme: a novel glucan heteropolysaccharide peptide, which has the following sugar chain repeating structural unit:

the novel glucoglyceroheteropolyglycopeptide is a fermentation product of alpha-hemolytic Streptococcus 33# (Streptococcus thermophilus-a-Hemolysis), and the monosaccharide composition of the novel glucoglyceroheteropolyglycopeptide is mannose in terms of molar ratio: rhamnose: glucose 8.22:1:6.46, weight average molecular weight 10045Da, molecular weight distribution coefficient 1.31.

A preparation method of novel glucoglyceroheteropolyglycopeptide comprises the following steps:

s1, performing liquid fermentation by using alpha-hemolytic Streptococcus 33# (Streptococcus thermophilus-a-Hemolysis) as a production bacterium to obtain a fermentation liquid;

s2, centrifuging the fermentation liquor, concentrating the supernatant obtained by centrifuging at the temperature of 60-80 ℃ for 2-6h, adding ethanol with the volume of 3-6 times of that of the concentrated solution for precipitation, centrifuging and collecting the precipitate, removing protein impurities from the precipitate by using a trichloroacetic acid and n-butyl alcohol mixed solution, and performing vacuum freeze drying to obtain a crude product of the polysaccharide peptide;

s3, adding distilled water into the crude polysaccharide peptide product for redissolving, then performing ultrafiltration by using a membrane with the cut-off of 5 kilodaltons, and collecting an ultrafiltration external liquid to obtain a first ultrafiltration external liquid;

s4, performing ultrafiltration on the first ultrafiltration external liquid by using a membrane with the cut-off of 2 ten thousand daltons, and collecting the ultrafiltration external liquid to obtain a second ultrafiltration external liquid;

s5, performing ultrafiltration on the secondary ultrafiltration external liquid by using a membrane with the cut-off of 1 ten thousand daltons, and collecting the concentrated solution in an ultrafiltration bottle to obtain an ultrafiltration concentrated solution;

s6, separating the ultrafiltration concentrated solution by a Superdex75pg gel column to obtain a component of the novel glucoglyceroheteropolyglycopeptide.

In step S1, the pH of the fermentation broth is maintained at 1.5-6.0.

In step S2, the ethanol concentration is 60% to 99.9%.

The novel glucoganaxane peptide is used in anti-acne cosmetics.

The invention has the beneficial effects that: the novel glucoglyceroheteropolysaccharide peptide with smaller molecular weight is separated from the fermentation liquor of alpha-hemolytic Streptococcus 33# (Streptococcus thermophilus-a-Hemolysis), and is applied to the anti-inflammatory acne removing cosmetics, so that the red swelling condition on the skin surface of a human body can be effectively eliminated; the skin moisturizing cream has an obvious skin moisturizing effect, the moisturizing time is long, the moisturizing effect is still good after 4 hours, and sebum secretion on the surface of the skin of a human body can be balanced, so that the skin can be fuller after being repaired, residue of acne marks is reduced, and the conditions of looseness and pore blockage on the surface of the skin are avoided.

Drawings

FIG. 1 is a graph showing the purity of a glucomannan peptide obtained in preparation example 1 by high performance liquid chromatography;

FIG. 2 is an infrared spectrum of a glucan heteropolysaccharide peptide obtained in production example 1;

FIG. 3 is a chromatogram of monosaccharide composition of Glucomannopeptin prepared in preparation example 1;

FIG. 4 is a GC-MS analysis of the total ion flux pattern of a methylated modification product of a Gluglycinan peptide prepared in preparation example 1;

FIG. 5 is 1H NMR of Glucomannopeptin prepared in preparation example 1;

FIG. 6 is a 13C NMR spectrum of Glucomannopeptin prepared in preparation example 1.

Detailed Description

The technical solutions of the present invention are further described in detail below with reference to the accompanying drawings, but the scope of the present invention is not limited to the following.

Examples

A novel glucan heteropolysaccharide peptide, which has the following sugar chain repeating structural unit:

the novel glucoglyceroheteropolyglycopeptide is a fermentation product of alpha-hemolytic Streptococcus 33# (Streptococcus thermophilus-a-Hemolysis), and the monosaccharide composition of the novel glucoglyceroheteropolyglycopeptide is mannose in terms of molar ratio: rhamnose: glucose is 8.22:1:6.46, weight average molecular weight is 10045Da, and molecular weight distribution coefficient is 1.31; the preparation method comprises the following steps:

s1, activating a-hemolytic Streptococcus 33# (Streptococcus thermophilus-a-haemolysis) by using a blood plate, selecting monoclonal to inoculate into a liquid culture medium to prepare a seed solution, transferring 5ml of seed solution to inoculate into 200ml of fermentation culture medium for culture, culturing at 37 ℃ for 20h to obtain fermentation liquor, and performing batch fermentation according to the condition; the pH value of the fermentation liquor is kept between 1.5 and 6.0;

s2, centrifuging the fermentation liquor, concentrating the supernatant obtained by centrifuging at the temperature of 60-80 ℃ for 2-6h, adding ethanol with the volume 4 times of that of the concentrated solution for precipitation, centrifuging to collect the precipitate, washing the precipitate with ethanol for three times, removing protein impurities through a mixed solution of trichloroacetic acid and n-butyl alcohol, and performing vacuum freeze drying to obtain 20.1g of crude polysaccharide peptide; the concentration of the ethanol is 60-99.9%;

s3, adding 10g of the crude polysaccharide peptide product into 500ml of distilled water for redissolving, stirring overnight, centrifuging, taking supernate, performing ultrafiltration by using a membrane with the cut-off of 5 ten thousand daltons, and collecting ultrafiltration external liquid to obtain first ultrafiltration external liquid;

s5, performing ultrafiltration on the secondary ultrafiltration external liquid by using a membrane with the cut-off of 1 ten thousand daltons, collecting the concentrated liquid in an ultrafiltration bottle, and performing vacuum freeze drying on the obtained ultrafiltration concentrated liquid to obtain a crude product of the glucosan hybrid glycan peptide;

s6, dissolving 500mg of the crude product of the glucoglyceroheteropolyglycopeptide in 5mL of deionized water, centrifuging (10000r/min) the supernatant, eluting by using ultrapure water eluent through a Superdex75pg gel column, collecting the eluent, collecting 2mL of the eluent in each tube, controlling the flow rate of the eluent at 0.5mL/min, developing by a phenol sulfate method, detecting the absorbance value by an enzyme-labeling instrument at 490nm, drawing an elution curve, collecting main peak components, combining, and performing vacuum freeze drying to obtain 0.45g of uniform novel glucoglyceroheteropolyglycopeptide.

(1) Structural characterization of the Glucomannan peptides obtained in the examples

The characteristic pattern of the Sephadex peptide on the gel column analyzed by Shodex Ohpak SB-804HQ is shown in FIG. 1, and the chromatographic conditions are as follows: mobile phase: 0.3M Na₂SO₄(ii) a Flow rate: 0.7 mL/min; column temperature: 35 ℃; chromatograph: shimadzu liquid chromatograph; a detector: a difference detector; the relative molecular mass of the glucoglyceroheteropolyglycopeptide was determined to be 10045Da by High Performance Gel Filtration Chromatography (HPGFC).

Taking about 2mg of Glucomannan peptide sample, and determining the infrared spectrum of Glucomannan peptide by potassium desertification tabletting method, the result is shown in FIG. 2, 3408.01cm^-1Is the 0-H stretching vibration absorption peak of glycopeptide saccharide ring, 2932.71cm^-1Is the C-H stretching vibration absorption peak, 1411.73cm^-1And 1050.77cm^-1Respectively C-0 stretching vibration outside the ring and inside the ring. 1654.15cm^-1Is 0-H bending vibration, 911.02cm^-1And 815.74cm^-1D-mannose is shown, which indicates that the glucomannan peptide is a glycopeptide high polymer.

Dissolving about 5mg of glucomannan peptide sample in 0.5mol/L dilute sulfuric acid solution, filling nitrogen to seal a pipe, hydrolyzing at 110 ℃ for 3 hours, neutralizing the hydrolysate, deriving by using methanol-PMP solution, analyzing the derivative product by using a C18 column, and analyzing monosaccharide composition, wherein the molar ratio of the monosaccharide composition is mannose to rhamnose to glucose is 8.22:1: 6.46.

100mg of the glucomannan peptide sample is weighed, methylation modification is carried out, the modified product is completely hydrolyzed by acid and then is analyzed and detected by GC-MS, and the result is shown in figure 4. Based on NIST mass spectrum database matching, the major glycosidic linkages of the glucosan hybrid glycan peptide are linked with: 1 → 4-D-Rha, T-Man, 1 → 4-D-Man, 1 → 3-D-Man, 1 → 2-D-Man, 1 → 6-D-Man, 1 → 3,4-D-Man, 1 → 2,3-D-Man, 1 → 2,4-D-Man, 1 → 3-D-Glc, 1 → 4,6-D-Man, 1 → 3,6-D-Man, 1 → 2,6-D-Man, 1 → 3,4,6-D-Man, 1 → 2,3, 6-D-Glc. According to the peak area size of the total ion current spectrum, deducing 1 → 3-D-Man, 1 → 3,6-D-Man, 1 → 2,3-D-Man, 1 → 2,6-D-Man as a main chain and the rest as side chains or terminal mannose.

Taking 20mg of lyophilized product of Glutamine and heteroglycan peptide, adding D₂O0.5 mL, centrifuged at 10000r/min, the supernatant was transferred to a NMR tube, and one-dimensional NMR spectra were measured at 25 ℃ with a JEOL600M NMR spectrometer, respectively, and the results are shown in FIGS. 5 and 6. FIG. 5 shows the H NMR spectrum of gluconoglycopeptide with chemical shifts at 5.212ppm assigned to the anomeric hydrogen signal of (1 → 4) -a-D-Rha; 5.106ppm anomeric hydrogen signal assigned to (1 → 2,3) -a-D-Man; 4.944ppm anomeric hydrogen signal assigned to (1 → 2,6) -a-D-Man; 4.924ppm anomeric hydrogen signal assigned to T-a-D-Man; 4.902 are assigned to (1 → 2,3,6) - -a-D-Glc; 4.848ppm anomeric hydrogen signal assigned to (1 → 3) -a-D-Glc; 4.845ppm anomeric hydrogen signal assigned to (1 → 4,6) -a-D-Man; 4.818ppm anomeric hydrogen signal assigned to (1 → 3) -a-D-Man; 4.713ppm anomeric hydrogen signal assigned to (1 → 6) - β -D-Man; 4.69ppm anomeric hydrogen signal assigned to (1 → 3,4) - β -D-Man; 4.639ppm anomeric hydrogen signal assigned to (1 → 4) - β -D-Man; 4.619ppm is attributed to the anomeric hydrogen signal of (1 → 3,6) - β -D-Man. FIG. 6 shows the C NMR spectrum of Gluconaglycopeptide with a chemical shift of 102.987ppm of anomeric carbon signal attributed to T-a-D-Man; chemical shift 102.47ppm is attributed to the anomeric carbon signal of (1 → 3,6) - β -D-Man; chemical shift 102.336ppm Allocation to (1 → 3) -a-D-ManA carbon signal; chemical shift 102.19ppm is attributed to the anomeric carbon signal of (1 → 2,6) -a-D-Man; chemical shifts at 100.517ppm were assigned to the anomeric carbon signal of (1 → 6) -a-D-Man; chemical shifts were assigned to the anomeric carbon signal of (1 → 3) -a-D-Glc at 98.162 ppm. The weaker signal at 1.077ppm for chemical shift in FIG. 5 and the signal at 16.747ppm for C6 on (1 → 4) -a-D-Rha for chemical shift in FIG. 6.

(2) Test examples the moisturizing Activity of Glucomacropeptide prepared in the examples

The moisturizing and acne removing gel comprises the following components: glucoglyceroheteropolyglycopeptides, carbomer, propylene glycol, triethanolamine and ultrapure water; the gel comprises the components of, by mass, 5-10% of novel glucoglyceroheteropolyglycopeptide, 1-2.5% of carbomer, 5-10% of propylene glycol, 1-2% of triethanolamine and the balance of ultrapure water. The preparation method of the glucomannan peptide gel cosmetic comprises the following steps:

s1, weighing 10g of glucoglyceroheteropolyglycopeptide, grinding into fine powder, adding into a proper amount of deionized water, stirring to completely dissolve the glucoheteropolyglycopeptide, and uniformly stirring to obtain a glucoheteropolyglycopeptide solution;

s2, adding carbomer into deionized water according to the mass percentage shown in the table 1, stirring to dissolve, and uniformly stirring to obtain a transparent gel matrix;

s3, mixing and stirring the transparent gel matrix and the glucomannan peptide solution uniformly, adding propylene glycol and triethanolamine, and continuously stirring until the mixture is uniform to obtain a mixed solution;

and S4, adding the residual deionized water to the total amount of the mixed solution, and uniformly stirring to obtain the glucomannan peptide gel cosmetic.

Table 1: material proportion of gel cosmetics

The evaluation method of the glucomannan peptide gel cosmetic comprises the following steps:

the determination of the moisture content of the skin stratum corneum proves that the glucogan hybrid glycan peptide has better moisturizing effect on human skin.

The test principle is as follows: when skin is aged, the horny layer of epidermis is thinned, the content of natural moisturizing factors in the horny layer is reduced, the hydration capability of the skin is reduced, the water loss of the skin is obviously increased, cell shrinkage tissues are shrunk, and the skin gradually generates fine wrinkles. Therefore, the degree of skin aging and the efficacy of the anti-aging product can be reflected to some extent by the measurement of skin moisture.

The experimental method comprises the following steps: selecting a test population with normal skin and no cosmetic allergy history, wherein the test population is 22-38 years old, 10 males and 10 females are selected randomly, 5 males and 5 females are selected as experimental groups, and the rest are selected as control groups. Indoor temperature is 25 +/-1 ℃, and humidity is 50% +/-5%; during the test, the face is cleaned, the cleaning time of each person is the same, after the skin is dried, skin with the face and the cheek being 4cm multiplied by 4cm is selected as a test area, and 1.5g of the skin care product is smeared on the skin.

Wherein, the skin care product applied to the experimental group is the glucoglyceroheteropolysaccharide peptide gel cosmetic, and the skin care product applied to the control group is the matrix gel without the glucoglyceroheteropolysaccharide peptide.

And (3) measurement: the test was started 10min after applying the skin care product to each cheek test area.

Measuring the skin water content of the test area by using a digital skin water detector, respectively measuring the water content of the test area after using corresponding skin care products for 1h, 2h, 3h and 4h, respectively testing for 6 times, and taking an average value; the results of the experiment are shown in table 2.

TABLE 2 measurement of skin moisture content

As can be seen from Table 2, the test group had an obvious skin moisturizing effect compared with the control group, and the moisturizing time was longer, and the moisturizing effect was still good after 4 hours.

(3) Test example the inhibition test of Propionibacterium acnes by Glucomannan peptides prepared in the examples

Now, the mass propagation of propionibacterium acnes in hair follicles is widely considered to be a very important factor for inducing whelk and causing whelk deterioration, so that the research on the inhibition effect of propionibacterium acnes is an important method for evaluating the acne removing efficacy of products.

The experimental method comprises the following steps:

(3.1) culture of Propionibacterium acnes

Propionibacterium acnes AB6919 is purchased from China center for type culture species preservation, BHI culture medium is prepared according to the requirements of strain culture conditions, the Propionibacterium acnes strain is inoculated into the BHI culture medium, and is subjected to anaerobic culture for 48h at 37 ℃ in an anaerobic generation bag and is shaken uniformly for later use. Coating a small amount of bacteria beads on a glass slide, fixing and dyeing by adopting a gram dyeing method, and then placing the glass slide under an oil microscope of 100 multiplied by 100 to perform microscopic examination, wherein purple bacteria can be seen under the microscope, and the bacteria beads are slightly bent into a rod or a rod shape and grow without mixed bacteria.

(3.2) dilution of sample solution by multiple

The Propionibacterium acnes-cultured lysis solution was diluted 100-fold (OD600 was about 0.02), and the Glucomannan peptide prepared in example was formulated into 200mg/mL aqueous solution, which was diluted in half into sample solutions of various concentrations of 100, 50.0, 25.0, and 12.5 mg/mL.

(3.3) culture of Strain and drug solution

Adding 90. mu.L of the diluted Propionibacterium acnes-containing solution to each well of a sterile 96-well plate, and then adding 10. mu.L of the diluted sample solution so that the final concentrations of the drugs are 20mg/mL, 10mg/mL, 5mg/mL, 2.5mg/mL, and 1.25 mg/mL; taking 90 mu L of bacterial liquid and 10 mu L of sterile normal saline as negative control; the positive control is clinical common skin anti-inflammatory drugs of minocycline, levofloxacin, azithromycin and erythromycin; transferring the sample into a constant temperature shaking incubator at 37 ℃ and 250r/min for anaerobic culture for 48 h.

(3.4) detection of bacterial growth

And (3) measuring the light absorption value of the mixed solution OD600 of each group of test tube medicines and the propionibacterium acnes by using a microplate reader. The concentration of the drug that has completely no light-absorbing pores is taken as the minimum inhibitory concentration. The bacterial liquid without light absorption is taken for routine gram staining and then is examined by a microscope to prove the growth of the propionibacterium acnes.

(3.5) measurement of minimum inhibitory concentration

The Minimum Inhibitory Concentration (MIC) value of the glucomannan peptide prepared by the plate culture retesting method for detecting Propionibacterium acnes AB6919₅₀Values), the results are shown in table 3.

Table 3: results of MIC testing of samples

Test sample	MIC₅₀Value of
		Glutamine heteroglycan peptides	95μg/mL
Minocycline	9μg/mL
		Levofloxacin	20μg/mL
Azithromycin	130μg/mL
		Erythromycin	140μg/mL

From the test results, the bacteriostatic concentration value of the glucoglyceroheteropolysaccharide peptide is 95 mug/mL, and the glucoglyceroheteropolysaccharide peptide is proved to have a certain function of inhibiting propionibacterium acnes and a certain acne removing function.

(4) Test example the prepared Glucomacropeptide was tested for its anti-acne efficacy

A moisturizing and acne removing gel comprises the following components: glucoglyceroheteropolyglycopeptides, carbomer, propylene glycol, triethanolamine and ultrapure water; the gel comprises the components of 5-10% of glucoglyceroheteropolyglycopeptide, 1-2.5% of carbomer, 5-10% of propylene glycol, 1-2% of triethanolamine and the balance of ultrapure water in percentage by mass. The preparation method of the moisturizing and acne-removing gel comprises the following steps:

s1, weighing the glucoglyceroheteropolyglycopeptide, grinding the glucoglyceroheteropolyglycopeptide into fine powder, adding deionized water into the powder according to the mass percentage shown in the table 4, and stirring the mixture to completely dissolve the glucoheteropolyglycopeptide to obtain a glucoglyceroheteropolyglycopeptide solution;

s2, adding carbomer into a proper amount of deionized water according to the mass percentage shown in the table 4, stirring to dissolve carbomer, and uniformly stirring to obtain a transparent gel matrix;

and S4, adding the rest deionized water to the total amount of the mixed solution, and uniformly stirring to obtain the moisturizing and acne-removing gel.

Table 4: material proportioning

Test method:

40 volunteers with face having acne and inflammation and red swelling are selected, the age is 20-40 years, and the male and female are divided into 2 groups randomly, wherein each group comprises 10 males and 10 females. The anti-inflammatory acne-removing cosmetic prepared by the embodiment is applied to a volunteer every day for 1 time, 1 time and 15min, the cosmetic is tested once a week and continuously tested for 4 weeks, and the volunteer can not replace or use other similar products during the test period. The volunteers were tested with MPA580 skin elasticity tester and VISIA full face analyzer, respectively, and the volume and area of pox on the volunteer's face and the swelling of the volunteer's face were observed, and the results are shown in Table 5.

Table 5: moisturizing and acne removing gel with anti-inflammatory and acne removing effects

Product name	Anti-inflammatory effect on skin	Reduction rate of pox volume	Degree of lactic acid stinging of skin
				Example 1	58％	54％	1.3
Example 2	73％	61％	1.1
				Example 3	85％	75％	0.8

The test results show that the cosmetic added with the glucoraphanin can effectively eliminate the red and swollen condition of the surface of the skin of a human body and balance the sebum secretion of the surface of the skin of the human body, so that the skin can be fuller after being repaired, the residue of acne marks is reduced, and the conditions of looseness and pore blockage on the surface of the skin are avoided.

The foregoing is illustrative of the preferred embodiments of this invention, and it is to be understood that the invention is not limited to the precise form disclosed herein and that various other combinations, modifications, and environments may be resorted to, falling within the scope of the concept as disclosed herein, either as described above or as apparent to those skilled in the relevant art. And that modifications and variations may be effected by those skilled in the art without departing from the spirit and scope of the invention as defined by the appended claims.

Claims

1. A novel glucan heteropolypeptide characterized by having the following repeating structural unit of sugar chain:

2. the novel glucan heteroglycan peptide of claim 1, wherein: the novel glucoglyceroheteropolyglycopeptide is a fermentation product of alpha-hemolytic Streptococcus 33# (Streptococcus thermophilus-a-haemolysis), and the monosaccharide composition of the novel glucoglyceroheteropolyglycopeptide is mannose in terms of molar ratio: rhamnose: glucose 8.22:1:6.46, weight average molecular weight 10045Da, molecular weight distribution coefficient 1.31.

3. A preparation method of novel glucoglyceroheteropolyglycopeptide is characterized by comprising the following steps:

s1, performing liquid fermentation by taking alpha-hemolytic Streptococcus 33# (Streptococcus thermophilus-a-haemolysis) as a production bacterium to obtain fermentation liquor;

s2, concentrating the fermentation liquor, adding ethanol for precipitation, centrifuging and collecting the precipitate to obtain a crude product of the polysaccharide peptide;

s3, adding distilled water into the crude polysaccharide peptide product for redissolving, then carrying out ultrafiltration, and collecting the ultrafiltration external liquid to obtain a first ultrafiltration external liquid;

s4, carrying out ultrafiltration on the first ultrafiltration external liquid again, and collecting the ultrafiltration external liquid to obtain a second ultrafiltration external liquid;

s5, carrying out ultrafiltration on the secondary ultrafiltration external liquid again, and collecting the ultrafiltration bottle internal concentrated liquid to obtain an ultrafiltration concentrated liquid;

s6, carrying out column chromatography on the ultrafiltration concentrated solution, and separating to obtain the component of the novel glucoglyceroheteropolyglycopeptide.

4. The method for producing a novel glucan heteropolyglycopeptide according to claim 3, wherein: in step S1, the pH of the fermentation broth is maintained at 1.5-6.0.

5. The method for producing a novel glucan heteropolyglycopeptide according to claim 3, wherein: in step S2, the ethanol concentration is 60% to 99.9%.

6. The application of the novel glucomannose hybrid glycan peptide in cosmetics is characterized in that: use of the novel glucoganaxane peptide according to any one of claims 1 to 5 in anti-acne cosmetics.