CN114702558B - Novel glucomannan peptide and preparation method and application thereof - Google Patents
Novel glucomannan peptide and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a novel glucomannan peptide, a preparation method and application thereof, wherein the novel glucomannan peptide is a fermentation product of alpha-hemolytic streptococcus 33# and is separated and purified by adopting a concentration and alcohol precipitation method and an ultrafiltration method to obtain uniform polysaccharide peptide, and the structure of the uniform polysaccharide peptide is identified through physical and chemical analysis; the monosaccharide composition of the novel glucomannan peptide is mannose: rhamnose: glucose=8.22:1:6.46, with a weight average molecular weight of 10045Da and a molecular weight distribution coefficient of 1.31; the acne-removing cosmetic has obvious moisturizing effect and longer moisturizing time, has the effects of inhibiting propionibacterium acnes and engine oil, achieves the effects of diminishing inflammation and removing acnes, can balance sebum secretion on the surface of human skin, ensures that the repaired skin is plump, and reduces the residue of acne marks; can avoid skin surface loosening and pore blocking.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a novel glucomannan peptide and a preparation method and application thereof.
Background
Cosmetics are a necessity for people's daily life. Cosmetic products in the current market are classified into moisturizing, whitening, sun-screening, acne removing, wrinkle preventing and the like according to functions; the active ingredients in cosmetics can be classified into artificial synthetic and natural sources according to their sources. In recent years, the natural active ingredients applied to cosmetics have the advantages of definite effect, small toxic and side effects and the like.
The natural polysaccharide has complex biological activity and function. Polysaccharides containing a large amount of hydrophilic groups exhibit excellent physicochemical properties such as strong water absorption, emulsifying property, high viscosity and good film forming property, which determine the natural advantage of the polysaccharide for development as a moisturizing ingredient. Meanwhile, the research also shows that the polysaccharide can be widely involved in various vital activities of cells to generate various biological functions, such as anti-inflammatory, antiviral, antitumor, anticoagulation, anti-radiation, anti-aging and the like. Therefore, the application range of natural polysaccharide compounds in the cosmetic field is also becoming wider. "acne", "pimples" or "comedo" is a chronic inflammatory reaction of the pilosebaceous glands of skin, and polysaccharide compounds with anti-inflammatory activity or permission to remove acne. The acne removing method has the advantages of oral medicinal preparation and external medicinal treatment, and the methods have certain effects, but have prominent defects, such as insignificant effects, easy scar on the face and other toxic and side effects.
The alpha-hemolytic streptococcus (Streptococcus hemolyticus-alpha-hemolysis) 33# is separated from the oral cavity and throat parts of healthy people, and can obtain alpha-Mannatide, also called Mannatide or polyantin A (polysaccharide A), which is a novel immunopotentiator initiated in China, wherein the monosaccharide composition of the Mannatide is mannose and glucose, the total ratio of the mannose is more than 90%, and the relative molecular mass is 4-6 ten thousand Da. Studies show that low concentration of a-mannatide can activate cellular immunity and humoral immunity of organism. In the prior art, the glucomannan peptide is not applied to acne-removing cosmetics, and the glucomannan peptide with smaller molecular weight is separated from fermentation liquor of the strain and is used in the acne-removing cosmetics.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, provides a novel glucomannan peptide, a preparation method and application thereof, separates the novel glucomannan peptide with smaller molecular weight from alpha-hemolytic streptococcus (Streptococcus hemolyticus-a-hemolysis) 33# fermentation liquor, and applies the novel glucomannan peptide to acne-removing cosmetics, so that the red swelling condition of the skin surface of a human body is effectively eliminated, and the sebum secretion of the skin surface of the human body can be balanced, so that the skin can be more full after repair, the residue of acne marks is reduced, and the conditions of loose skin surface and pore blocking are avoided.
The aim of the invention is realized by the following technical scheme: a novel glucomannan peptide having the following sugar chain repeating structural units:
the novel glucomannan peptide is a fermentation product of alpha-hemolytic streptococcus 33# (Streptococcus hemolyticus-alpha-Hemolysis), and the monosaccharide composition of the novel glucomannan peptide is mannose: rhamnose: glucose=8.22:1:6.46, with a weight average molecular weight of 10045Da and a molecular weight distribution coefficient of 1.31.
A preparation method of a novel glucomannan peptide comprises the following steps:
s1, carrying out liquid fermentation by taking alpha-hemolytic streptococcus 33# (Streptococcus hemolyticus-alpha-Hemolysis) as a production bacterium to obtain fermentation liquor;
s2, centrifuging the fermentation liquor, concentrating supernatant obtained by centrifugation at 60-80 ℃ for 2-6 hours, adding ethanol precipitation which is 3-6 times of the volume of the concentrate, centrifuging and collecting the precipitate, removing protein impurities from the precipitate by using a trichloroacetic acid and n-butanol mixed solution, and performing vacuum freeze drying to obtain a polysaccharide peptide crude product;
s3, adding distilled water into the polysaccharide peptide crude product for re-dissolution, performing ultrafiltration by using a membrane ultrafiltration with a cutoff of 5 ten thousand daltons, and collecting ultrafiltration external liquid to obtain first ultrafiltration external liquid;
s4, ultrafiltering the first ultrafiltrate by using a membrane with a cutoff of 2 ten thousand daltons, and collecting the ultrafiltrate to obtain a second ultrafiltrate;
s5, ultrafiltering the second ultrafiltrate by using a membrane with a cutoff of 1 kilodaltons, and collecting the concentrated solution in an ultrafiltration bottle to obtain an ultrafiltration concentrated solution;
s6, separating the ultrafiltration concentrated solution by using a Superdex75pg gel column to obtain the novel glucomannan peptide.
In the step S1, the pH value of the fermentation liquor is kept between 1.5 and 6.0.
In the step S2, the concentration of the ethanol is 60% -99.9%.
The novel glucomannan peptide is used in acne-removing cosmetics.
The beneficial effects of the invention are as follows: the novel glucomannan peptide with smaller molecular weight is separated from the fermentation broth of the alpha-hemolytic streptococcus 33# (Streptococcus hemolyticus-a-Hemolysis), and is applied to anti-inflammatory acne-removing cosmetics, so that the red and swollen condition of the skin surface of a human body can be effectively eliminated; the skin moisturizing agent has obvious skin moisturizing effect, has long moisturizing time, has good moisturizing effect after 4 hours, and can balance sebum secretion on the surface of human skin, so that the skin can be plump after repair, the residue of acne marks is reduced, and the situations of loosening and pore blocking on the surface of the skin are avoided.
Drawings
FIG. 1 is a purity chart of a high performance liquid chromatograph of the glucomannan peptide prepared in preparation example 1;
FIG. 2 is an infrared spectrum of the glucomannan peptide prepared in preparation example 1;
FIG. 3 is a chromatogram of the monosaccharide composition of the glucomannan peptide prepared in preparation example 1;
FIG. 4 is a GC-MS analysis total ion flow chart of the methylation-modified product of the glucomannan peptide prepared in preparation example 1;
FIG. 5 is 1H NMR of a glucomannan peptide prepared in preparation example 1;
FIG. 6 is a 13C NMR spectrum of a glucomannan peptide obtained in preparation example 1.
Detailed Description
The technical solution of the present invention will be described in further detail with reference to the accompanying drawings, but the scope of the present invention is not limited to the following description.
Examples
A novel glucomannan peptide having the following sugar chain repeating structural units:
the novel glucomannan peptide is a fermentation product of alpha-hemolytic streptococcus 33# (Streptococcus hemolyticus-alpha-Hemolysis), and the monosaccharide composition of the novel glucomannan peptide is mannose: rhamnose: glucose=8.22:1:6.46, with a weight average molecular weight of 10045Da and a molecular weight distribution coefficient of 1.31; the preparation method comprises the following steps:
s1, activating a-hemolytic streptococcus 33# (Streptococcus hemolyticus-a-Hemolysis) strain by using a blood plate, selecting a monoclonal to inoculate to a liquid culture medium to prepare a seed solution, transferring 5ml of the seed solution to inoculate to 200ml of a fermentation culture medium for culturing, culturing for 20 hours at 37 ℃ to obtain a fermentation liquid, and fermenting in batches 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 supernatant obtained by centrifugation at 60-80 ℃ for 2-6 hours, adding ethanol precipitation with the volume being 4 times of the volume of the concentrate, centrifuging, collecting the precipitate, washing the precipitate with ethanol for three times, removing protein impurities by using a trichloroacetic acid and n-butanol mixed solution, and performing vacuum freeze drying to obtain 20.1g polysaccharide peptide crude product; the concentration of the ethanol is 60% -99.9%;
s3, adding 500ml of distilled water into 10g of the crude polysaccharide peptide product for re-dissolution, stirring overnight, centrifuging, taking supernatant, ultrafiltering with membrane ultrafiltration with interception of 5 ten thousand daltons, and collecting ultrafiltrate to obtain first ultrafiltrate;
s4, ultrafiltering the first ultrafiltrate by using a membrane with a cutoff of 2 ten thousand daltons, and collecting the ultrafiltrate to obtain a second ultrafiltrate;
s5, ultrafiltering the second ultrafiltered external liquid by using a membrane with the interception of 1 ten thousand daltons, collecting concentrated solution in an ultrafiltration bottle, and performing vacuum freeze drying on the obtained ultrafiltered concentrated solution to obtain a glucomannan peptide crude product;
s6, dissolving 500mg of the glucomannan peptide crude product in 5mL of deionized water, eluting supernatant by using ultrapure water eluent through a Superdex75pg gel column, collecting eluent, collecting 2mL of eluent from each tube, controlling the flow rate of the eluent to be 0.5mL/min, performing color development by a phenol sulfate method, detecting absorbance value by an enzyme-labeled instrument 490nm, drawing an elution curve, collecting main peak components, merging, and performing vacuum freeze drying to obtain 0.45g of uniform novel glucomannan peptide.
(1) Structure identification of the glucomannan peptide obtained in the examples
The characteristic spectrum of the glucomannan peptide on the gel column is shown in figure 1 by Shodex OHPak SB-804HQ analysis, and the chromatographic conditions are as follows: mobile phase: 0.3M Na 2 SO 4 The method comprises the steps of carrying out a first treatment on the surface of the Flow rate: 0.7mL/min; column temperature: 35 ℃; chromatograph: island body fluid phase chromatograph; a detector: a differential detector; the relative molecular mass of the glucomannan peptide was determined to be 10045Da by High Performance Gel Filtration Chromatography (HPGFC).
About 2mg of the glucomannan peptide sample is taken, and the infrared spectrum of the glucomannan peptide is measured by adopting a potassium desertification tabletting method, and the result is shown in figure 2 and 3408.01cm -1 Is the telescopic vibration absorption peak of 0-H on the glycopeptide sugar ring, 2932.71cm -1 Is C-H telescopic vibration absorption peak of 1411.73cm -1 And 1050.77cm -1 C-0 stretching vibration outside the ring and inside the ring respectively. 1654.15cm -1 Flexural vibration of 0-H, 911.02cm -1 And 815.74cm -1 D-mannose is indicated, indicating that the glucomannan peptide is a glycopeptide polymer.
About 5mg of glucomannan peptide sample is taken and dissolved in 0.5mol/L dilute sulfuric acid solution, a nitrogen gas is filled for sealing a tube, hydrolysis is carried out for 3 hours at 110 ℃, after the hydrolysis solution is neutralized, methanol-PMP solution is used for derivatization, the derivatization product is analyzed by a C18 column, and the analysis result of monosaccharide composition shows that the molar ratio of the monosaccharide composition is mannose to rhamnose to glucose is 8.22:1:6.46.
100mg of glucomannan peptide sample is weighed, methylation modification is carried out, and analysis and detection are carried out on the modified product by GC-MS after complete acid hydrolysis, and the result is shown in figure 4. Based on NIST mass spectrometry database matching, the main glycosidic linkages of the glucomannan peptide are linked: 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 total ion flow spectrum peak area, 1-3-D-Man, 1-3, 6-D-Man, 1-2, 3-D-Man, 1-2, 6-D-Man are deduced as main chain, and the rest is side chain or terminal mannose.
Adding D into 20mg of glucomannan peptide lyophilized product 2 O0.5 mL was dissolved, centrifuged at 10000r/min, and the supernatant was transferred to a nuclear magnetic resonance tube, and one-dimensional nuclear magnetic resonance spectra were measured at 25℃on a JEOL600M nuclear magnetic resonance spectrometer, respectively, and the results are shown in FIG. 5 and FIG. 6. FIG. 5 shows an H NMR spectrum of a glucomannan peptide with chemical shift assigned to the anomeric hydrogen signal of (1.fwdarw.4) -a-D-Rha at 5.212 ppm; 5.106ppm of the anomeric hydrogen signal attributed to (1→2, 3) -a-D-Man; 4.944ppm is assigned to the (1→2, 6) -a-D-Man anomeric hydrogen signal; 4.924ppm is assigned to the anomeric hydrogen signal of T-a-D-Man; 4.902 is assigned to (1→2,3, 6) - - -a-D-Glc;4.848ppm of an anomeric hydrogen signal ascribed to (1.fwdarw.3) -a-D-Glc; 4.845ppm is assigned to the (1→4, 6) -a-D-Man anomeric hydrogen signal; 4.818ppm is assigned to the (1→3) -a-D-Man anomeric hydrogen signal; 4.713ppm is assigned to the (1→6) - β -D-Man anomeric hydrogen signal; 4.69ppm of the anomeric hydrogen signal assigned to (1→3, 4) - β -D-Man; 4.639ppm is assigned to the (1→4) - β -D-Man anomeric hydrogen signal; 4.619ppm is assigned to the (1→3, 6) - β -D-Man anomeric hydrogen signal. FIG. 6 shows a C NMR spectrum of glucomannan peptide, chemical shift 102.987ppm of the anomeric carbon signal ascribed to T-a-D-Man; chemical shift 102.47ppm is ascribed to an anomeric carbon signal of (1→3, 6) - β -D-Man; chemical shift 102.336ppm is ascribed to an anomeric carbon signal of (1→3) -a-D-Man; chemical shift 102.19ppm is ascribed to an anomeric carbon signal of (1→2, 6) -a-D-Man; a anomeric carbon signal at 100.517ppm attributed to (1→6) -a-D-Man; the chemical shift was ascribed to an anomeric carbon signal of (1.fwdarw.3) -a-D-Glc at 98.162 ppm. The weaker signal in FIG. 5 at 1.077ppm and the signal in FIG. 6 at 16.747ppm attributed to C6 on (1.fwdarw.4) -a-D-Rha.
(2) Test example preparation of the moisturizing Activity of the glucomannan peptide
The gel comprises the following components: glucomannan peptide, carbomer, propylene glycol, triethanolamine and ultrapure water; the gel comprises, by mass, 5-10% of novel glucomannan peptide, 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 type cosmetic comprises the following steps:
s1, weighing 10g of glucomannan peptide, grinding into fine powder, adding into a proper amount of deionized water, stirring to completely dissolve the glucomannan peptide, and uniformly stirring to obtain a glucomannan peptide solution;
s2, adding Xiang Kabo mu of the mass percent shown in the table 1 into deionized water, stirring and dissolving, and uniformly stirring to obtain a transparent gel matrix;
s3, uniformly mixing and stirring the transparent gel matrix and the glucomannan peptide solution, adding propylene glycol and triethanolamine, and continuously stirring until uniform to obtain a mixed solution;
and S4, adding the rest deionized water to the mixed solution to the full amount, and uniformly stirring to obtain the glucomannan peptide gel type cosmetic.
Table 1: material proportion of gel cosmetics
The evaluation method of the glucomannan peptide gel type cosmetic comprises the following steps:
the measurement of the moisture content of the skin horny layer proves that the glucomannan peptide has better moisturizing effect on human skin.
Test principle: the epidermis stratum corneum becomes thinner during skin aging, the content of natural moisturizing factors therein is reduced, the hydration capacity of the skin is reduced, the moisture loss of the skin is obviously increased, and the skin gradually develops fine wrinkles due to shrinkage of cells and tissue atrophy. Thus, the extent of skin aging and the efficacy of the anti-aging product can be reflected to some extent by measurement of skin moisture.
The experimental method comprises the following steps: the group of subjects with normal skin and no cosmetic allergy history was selected, aged 22-38 years, 10 men and 10 women, 5 men and 5 women were randomly selected as experimental groups, and the rest were selected as control groups. Indoor temperature 25+/-1 ℃ and humidity 50% +/-5%; during the test, the face is cleaned, the cleaning time of each person is the same, after the skin is dried, the skin of 4cm multiplied by 4cm of the cheek is selected as a test area, and 1.5g of skin care product is smeared.
The skin care product smeared in the experimental group is the glucomannan peptide gel cosmetic, and the skin care product smeared in the control group is the matrix gel without the glucomannan peptide.
Measurement: in the cheek test areas, the test was started 10min after each application of the skin care product.
Measuring skin moisture content of the test area by using a digital skin moisture detector, respectively measuring the moisture 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 experimental results are shown in table 2.
Table 2 skin moisture content measurement values
As can be seen from table 2, the test group has an obvious skin moisturizing effect compared with the control group, and the moisturizing effect is still good after 4 hours.
(3) Test example prepared glucomannan peptide inhibition test on Propionibacterium acnes
It is now widely believed that the massive proliferation of propionibacterium acnes in hair follicles is a very important factor inducing and deteriorating acne, and therefore, research on the inhibition of propionibacterium acnes is an important method for evaluating the acne-removing efficacy of products.
The experimental method comprises the following steps:
(3.1) cultivation of Propionibacterium acnes
Propionibacterium acnes AB6919 is purchased from China center for type culture collection, a BHI culture medium is prepared according to the requirement of strain culture conditions, the Propionibacterium acnes strain is inoculated into the BHI culture medium, anaerobic culture is carried out for 48 hours at 37 ℃ in an anaerobic generating bag, and shaking is uniform for standby. And (3) a small amount of fungus beads are coated on a glass slide, fixed and dyed by adopting a gram dyeing method, and then the glass slide is placed under a 100 multiplied by 100 oil mirror for microscopic examination, purple bacillus is visible under the mirror, and the glass slide is slightly bent to be in a rod shape or a rod shape and grows in a sterile manner.
(3.2) dilution of the sample solution by a multiple ratio
The propionibacterium acnes cultured lysate was diluted 100-fold (OD 600 of about 0.02), and the glucomannan peptide prepared in the examples was formulated as 200mg/mL aqueous solution, diluted in half to 100, 50.0, 25.0 and 12.5mg/mL different concentrations of sample solutions.
(3.3) cultivation of Strain and medical fluid
90. Mu.L of the diluted propionibacterium acnes-containing solution described above was added to each well of a sterile 96-well plate, followed by 10. Mu.L of the diluted sample solution so that the final drug concentration was 20mg/mL, 10mg/mL, 5mg/mL, 2.5mg/mL and 1.25mg/mL; taking 90 mu L of bacterial liquid and 10 mu L of sterile physiological saline as negative control; the positive control is minocycline, levofloxacin, azithromycin and erythromycin which are commonly used in clinic as skin anti-inflammatory drugs; transferring the sample into a constant temperature shaking incubator at 37 ℃ for anaerobic culture at 250r/min for 48 hours.
(3.4) detection of bacterial growth
The absorbance of the solution OD600 after mixing each group of test tube drug with Propionibacterium acnes was measured with an enzyme-labeled instrument. The drug concentration of the light absorbing wells has been completely absent as the minimum inhibitory concentration. The bacterial liquid without light absorption is taken for routine gram staining and then examined by a microscope to prove whether the propionibacterium acnes grow.
(3.5) determination of minimum inhibitory concentration
The Minimum Inhibitory Concentration (MIC) of the glucomannan peptide prepared by the test case on Propionibacterium acnes AB6919 is detected by adopting a plate culture retest method 50 Values), the results are shown in table 3.
Table 3: sample MIC test results
| Sample to be tested | MIC 50 Value of |
| Glucomannan peptide | 95μg/mL |
| Minocycline | 9μg/mL |
| Levofloxacin | 20μg/mL |
| Azithromycin | 130μg/mL |
| Erythromycin | 140μg/mL |
From the test results, the antibacterial concentration value of the glucomannan peptide is 95 mug/mL, and the glucomannan peptide is proved to have a certain effect of inhibiting propionibacterium acnes and a certain acne removing function.
(4) Test of the acne-removing Effect of the glucomannan peptide prepared in the example
A moisturizing and acne-removing gel comprises the following components: glucomannan peptide, carbomer, propylene glycol, triethanolamine and ultrapure water; the gel comprises, by mass, 5-10% of glucomannan peptide, 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 moisturizing and acne-removing gel comprises the following steps:
s1, weighing glucomannan peptide, grinding into fine powder, adding deionized water into the fine powder according to the mass percentage shown in the table 4, and stirring to completely dissolve the glucomannan peptide to obtain a glucomannan peptide solution;
s2, adding carbomer into a proper amount of deionized water according to the mass percentage shown in the table 4, stirring to dissolve the carbomer, and uniformly stirring to obtain a transparent gel matrix;
s3, uniformly mixing and stirring the transparent gel matrix and the glucomannan peptide solution, adding propylene glycol and triethanolamine, and continuously stirring until uniform to obtain a mixed solution;
and S4, adding the rest deionized water to the mixed solution to a full amount, and uniformly stirring to obtain the moisturizing and acne-removing gel.
Table 4: material proportioning
The test experiment method comprises the following steps:
40 volunteers with acne, inflammation and redness on the face and ages 20-40 were selected, and the volunteers were randomly divided into 2 groups of 10 men and 10 women. The anti-inflammatory acne-removing cosmetic prepared in the embodiment is applied to volunteers for 1 time, 1 time for 15min, tested once a week, continuously tested for 4 weeks, and other similar products cannot be replaced or used together during the test. The volunteers were tested with MPA580 skin elasticity tester and VISIA full face analyzer, respectively, and the volume and area of vaccinia on the faces of the volunteers and the detumescence of the faces of the volunteers were observed, and the results of the tests are shown in Table 5.
Table 5: moisturizing and acne-removing gel with anti-inflammatory and acne-removing effects
| Product name | Skin anti-inflammatory effect | Vaccinia volume reduction rate | Skin lactic acid stinging degree |
| Example 1 | 58％ | 54％ | 1.3 |
| Example 2 | 73％ | 61％ | 1.1 |
| Example 3 | 85％ | 75％ | 0.8 |
According to the test results, the glucomannan peptide-added cosmetic disclosed by the invention can effectively eliminate the red and swollen condition of the skin surface of a human body, and can balance sebum secretion of the skin surface of the human body, so that the skin can be plump after repair, the residue of acne marks is reduced, and meanwhile, the conditions of loosening and pore blocking on the skin surface are avoided.
The foregoing is merely a preferred embodiment of the invention, and it is to be understood that the invention is not limited to the form disclosed herein but is not to be construed as excluding other embodiments, but is capable of numerous other combinations, modifications and environments and is capable of modifications within the scope of the inventive concept, either as taught or as a matter of routine skill or knowledge in the relevant art. And that modifications and variations which do not depart from the spirit and scope of the invention are intended to be within the scope of the appended claims.
Claims (5)
1. A novel glucomannan peptide, characterized in that the novel glucomannan peptide is a fermentation product of a-hemolytic streptococcus Streptococcus hemolyticus-a-hemolysis 33#, and the novel glucomannan peptide has the following sugar chain repeating structural units:
；
the monosaccharide composition of the novel glucomannan peptide is mannose: rhamnose: glucose=8.22:1:6.46, with a weight average molecular weight of 10045Da and a molecular weight distribution coefficient of 1.31.
2. A method for preparing the novel glucomannan peptide according to claim 1, comprising the steps of:
s1, carrying out liquid fermentation by taking a-hemolytic streptococcus Streptococcus hemolyticus-a-hemolysis 33# as a production bacterium to obtain fermentation liquor;
s2, concentrating the fermentation liquor, adding ethanol for precipitation, and centrifugally collecting the precipitation to obtain a polysaccharide peptide crude product;
s3, adding distilled water into the polysaccharide peptide crude product for re-dissolution, performing ultrafiltration, and collecting ultrafiltration external liquid to obtain first ultrafiltration external liquid;
s4, ultrafiltering the first ultrafiltrate again, and collecting the ultrafiltrate to obtain a second ultrafiltrate;
s5, ultrafiltering the second ultrafiltered external liquid again, and collecting concentrated solution in an ultrafiltration bottle to obtain ultrafiltered concentrated solution;
s6, passing the ultrafiltration concentrated solution through a column, and separating to obtain the novel glucomannan peptide.
3. The method for preparing the novel glucomannan peptide according to claim 2, wherein the method comprises the following steps: in the step S1, the pH value of the fermentation liquor is kept between 1.5 and 6.0.
4. The method for preparing the novel glucomannan peptide according to claim 2, wherein the method comprises the following steps: in the step S2, the concentration of the ethanol is 60% -99.9%.
5. The application of the novel glucomannan peptide in the preparation of cosmetics is characterized in that: the novel glucomannan peptide of claim 1 for use in anti-acne cosmetics.
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| EP0255299A2 (en) * | 1986-07-30 | 1988-02-03 | Smithkline Beecham Corporation | Glycopeptide antibiotics |
| US5811403A (en) * | 1996-09-30 | 1998-09-22 | Vanderbilt University | Polysaccharide toxin from Group B β-hemolytic Streptococcus (GBS) having improved purity |
| JPH1180206A (en) * | 1997-08-29 | 1999-03-26 | Hitoshi Ito | Glucomannan separated from agaricus blazei mushroom culture and antitumor agent containing it as active ingredient |
| KR20080008956A (en) * | 2006-07-20 | 2008-01-24 | 박순영 | A NOVEL PHELLINUS STRAIN AND THE METHOD FOR ISOLATING beta;-1,3 GLUCOMANNAN THEREFROM SHOWING IMMUNE ENHANCING ACTIVITY AND ANTITUMOR ACTIVITY |
| JP2009013240A (en) * | 2007-07-03 | 2009-01-22 | Chisso Corp | Polysaccharide, method for producing the same and its application |
| WO2021219530A1 (en) * | 2020-04-28 | 2021-11-04 | Janssen Pharmaceuticals, Inc. | Method for identification and quantification of polysaccharides in complex glycoconjugate compositions |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US11325986B2 (en) * | 2013-11-05 | 2022-05-10 | Dsm Ip Assets B.V. | Heteropolysaccharides |
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| EP0255299A2 (en) * | 1986-07-30 | 1988-02-03 | Smithkline Beecham Corporation | Glycopeptide antibiotics |
| US5811403A (en) * | 1996-09-30 | 1998-09-22 | Vanderbilt University | Polysaccharide toxin from Group B β-hemolytic Streptococcus (GBS) having improved purity |
| JPH1180206A (en) * | 1997-08-29 | 1999-03-26 | Hitoshi Ito | Glucomannan separated from agaricus blazei mushroom culture and antitumor agent containing it as active ingredient |
| KR20080008956A (en) * | 2006-07-20 | 2008-01-24 | 박순영 | A NOVEL PHELLINUS STRAIN AND THE METHOD FOR ISOLATING beta;-1,3 GLUCOMANNAN THEREFROM SHOWING IMMUNE ENHANCING ACTIVITY AND ANTITUMOR ACTIVITY |
| JP2009013240A (en) * | 2007-07-03 | 2009-01-22 | Chisso Corp | Polysaccharide, method for producing the same and its application |
| WO2021219530A1 (en) * | 2020-04-28 | 2021-11-04 | Janssen Pharmaceuticals, Inc. | Method for identification and quantification of polysaccharides in complex glycoconjugate compositions |
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