CN105420317A - Preparation method of mannatide - Google Patents

Preparation method of mannatide Download PDF

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CN105420317A
CN105420317A CN201510888970.4A CN201510888970A CN105420317A CN 105420317 A CN105420317 A CN 105420317A CN 201510888970 A CN201510888970 A CN 201510888970A CN 105420317 A CN105420317 A CN 105420317A
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streptococel
preparation
fermentation
substratum
amino acid
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毛跟年
贾莹
张诗韵
许牡丹
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Shaanxi University of Science and Technology
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Shaanxi University of Science and Technology
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Abstract

The invention provides a preparation method of mannatide. The preparation method comprises the following steps: preparing konjac polypeptide liquid from konjac flying powder through aspergillus terricola and bacillus licheniformis co-fermentation; then processing the konjac polypeptide liquid by virtue of ultra-filtration, and collecting a filtrate with relative molecular mass ranging from 10KD to 100KD; and conducting gel column chromatography on the filtrate, so that the purified mannatide is obtained. The preparation method disclosed by the invention fills the technical blank of preparing the mannatide from the konjac flying powder through the combination of a fermentation method, an ultra-filtration technology and a chromatographic technology; and meanwhile, a new way is created for the production of a high added-value product through the development and utilization of the konjac flying powder; therefore, the preparation method is of great significance to the fields of food, medicine and the like.

Description

A kind of preparation method of Streptococel
Technical field
The invention belongs to fermentable and medicinal peptide field, relate to a kind of preparation method of glycopeptide, be specifically related to a kind of preparation method of Streptococel.
Background technology
Streptococel former name is polyactin, is a kind of biological immunopotentiator that China initiates.It is the mixture together formed by the Streptococel molecular combinations of different chain length, there is not small molecules carbohydrate and total free aminoacids, and its primary structure is that polypeptide chain is connected with mannosans.Modern pharmacology and clinical study show: Streptococel has obvious biological activity, and it can regulate body immunologic derangement, significantly strengthens autoimmunity.Existing be morely used for the treatment of immunologic hypofunction or dysimmunity disease, if the assisting therapy of repeated cold, recurrent respiratory tract infection, oligoleukocythemia and tumour is to alleviate the untoward reaction etc. of Radiotherapy chemotherapy, and achieving good curative effect, clinical application is more and more extensive.
Fry starch of konjak is in the konjaku powder course of processing, the fine powder that the quality descended slowly and lightly around is light, particle is little.Containing abundant nutritive ingredient in fry starch of konjak, protein content 23.8%, water-soluble sugar (seminose and glucose) 8.95%, it is the good source containing high protein, low mannosans, but because fry starch of konjak exists special egg fishy smell and stink, main as animal-feed at present, utilization ratio is low.
At present, domestic Streptococel of preparing mainly extracts from strain cultured solution, originates more single.Such as, the Chinese patent that number of patent application is 03117579.1, denomination of invention is Streptococel and preparation technology and purposes is that separation and Extraction is refining from the tunning of Alpha-hemolytic streptococcus forms Streptococel.The Chinese patent that the Chinese patent that number of patent application is 200710093102.2, denomination of invention is the preparation method of high-purity mannatide and number of patent application are 200710093103.7, denomination of invention is the preparation method of Streptococel, all with No. H1S-33, Alpha-hemolytic streptococcus for bacterial strain, carry out liquid fermenting and prepare Streptococel.Not seeing with fry starch of konjak is at present the method that Streptococel prepared by raw material.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of Streptococel, the method take fry starch of konjak as raw material, obtain Streptococel by fermentation, the method can widen the source of Streptococel, again for the exploitation of fry starch of konjak provide a new way.
For achieving the above object, the technical solution used in the present invention is:
A preparation method for Streptococel, comprises the following steps:
1) ferment: in every 100mL substratum, add fry starch of konjak 1 ~ 3g, adopt aspergillus terricola and Bacillus licheniformis symbiotic fermentation, after having fermented, the centrifugal rear removal thalline of fermentation liquor, get supernatant liquor, add 1 ~ 5 times wherein to the purified water of supernatant volume, then use trichoroacetic acid(TCA) adjust ph to 3.8 ~ 4.2, remove precipitation after leaving standstill, the supernatant liquor obtained after removing precipitation is konjaku glycopeptide liquid;
2) ultrafiltration: be the ultra-filtration membrane of 100KD and 10KD by molecular weight cut-off successively by konjaku glycopeptide liquid, collects molecular weight at the ultrafiltrated of 10 ~ 100KD section, obtains Streptococel crude product;
3) with gel filtration chromatography, separation and purification is carried out to Streptococel crude product, namely obtain the Streptococel of purifying.
Described step 1) in zymotechnique be: the pH value of substratum is 7.0 ~ 11.0, and the bacterial classification ratio of aspergillus terricola and Bacillus licheniformis is 1:(1 ~ 3), bacterial concentration is 5 × 10 7~ 5 × 10 9individual/mL, inoculum size is 4 ~ 12%, and fermentation time is 24 ~ 48h, and leavening temperature is 25 ~ 38 DEG C.
Described step 1) in substratum be add peptone 1g, extractum carnis 1.5g, NaNO in every 100mL distilled water 30.1 ~ 0.4g, K 2hPO 40.05 ~ 0.2g, KCl0.02 ~ 0.1g, MgSO 4.7H 2o0.02 ~ 0.1g and FeSO 4obtain after 0.002 ~ 0.01g.
Described step 1) in fermented liquid at 4 DEG C to remove thalline after the centrifugal 10 ~ 30min of the rotating speed of 8000 ~ 12000r/min.
Described step 3) in use chromatography column specification be (1.0 ~ 20) cm × (50 ~ 100) cm, interior fill is SephadexG-75.
Described step 3) in gel filtration chromatography condition be: applied sample amount is 3 ~ 300mL, sample concentration is 100 ~ 200mg/mL, eluent is distilled water, elution flow rate is 0.5 ~ 80mL/min, often 3 ~ 6mL collected by pipe, at 280nm and 460nm place mensuration light absorption value, collection merging has sugar simultaneously, the component of peptide absorption peak concentrates, and obtains the Streptococel of purifying after vacuum lyophilization.
Described vacuum lyophilization is dry 12 ~ 24h under-110 ~-60 DEG C and 60 ~ 800Pa.
Seminose contained in obtained Streptococel and the mass ratio of glucose are 1.7:(1 ~ 4).
Necessary amino acid whose quality contained in obtained Streptococel is 29.61% ~ 31.26% of amino acid total mass.
Amino acid contained in obtained Streptococel comprises Threonine, Serine, aspartic acid, L-glutamic acid, glycine, L-Ala, halfcystine, tyrosine, α-amino-isovaleric acid, methionine(Met), Isoleucine, leucine, phenylalanine, Methionin, Histidine, arginine and proline(Pro).
Relative to prior art, beneficial effect of the present invention is:
The preparation method of Streptococel provided by the invention, adds fry starch of konjak in the medium, obtains konjaku glycopeptide liquid by aspergillus terricola and Bacillus licheniformis symbiotic fermentation; Then Streptococel crude product is obtained through ultrafiltration; The Streptococel of purifying is obtained again through gel filtration chromatography separation and purification.The present invention is that raw material combining with fermentation method, ultrafiltration and chromatography isolation technique prepare Streptococel with fry starch of konjak, has excavated the utility value of fry starch of konjak, has widened the preparation method of Streptococel.Use method of the present invention, Streptococel can be obtained, confirm that fry starch of konjak is a kind of ideal source of Streptococel, having filled up take fry starch of konjak as the technological gap that Streptococel prepared by raw material, the source of Streptococel can be widened, simultaneously also for fry starch of konjak exploitation, produce high value added product and open a new road, significant in food, medicine and other fields.
Further, the monose of the Streptococel that the present invention obtains, aminoacid component are analyzed, prove that monose contained in obtained Streptococel is mainly seminose and glucose, and the mass ratio of seminose and glucose is 1.7:(1 ~ 4).And found in the Streptococel that the present invention obtains containing Threonine, Serine, aspartic acid, L-glutamic acid, glycine, L-Ala, halfcystine, tyrosine, α-amino-isovaleric acid, methionine(Met), Isoleucine, leucine, phenylalanine, Methionin, Histidine, arginine and proline(Pro) totally 17 seed amino acids by detection, wherein must account for 29.61% ~ 31.26% of total amino acid content by amino acid, there are very high using value and market outlook.
Accompanying drawing explanation
Fig. 1 is the amino acid collection of illustrative plates of mannosans poly saccharide peptide standard product;
Fig. 2 is the amino acid collection of illustrative plates of the Streptococel that the present invention obtains;
Fig. 3 is the high-efficient liquid phase chromatogram of seminose, glucose hybrid standard product;
Fig. 4 is the high-efficient liquid phase chromatogram of the monose hydrolyzate of the Streptococel that the present invention obtains.
Embodiment
Technical scheme of the present invention is described in detail below in conjunction with embodiment.Should be understood that following examples are only not used in for illustration of the present invention to limit the scope of the invention.Without departing from the spirit and substance of the case in the present invention, the amendment make step of the present invention or condition or alternative, all belongs to protection scope of the present invention.
The invention provides a kind of preparation method of Streptococel, take fry starch of konjak as raw material, is total to fermentation method prepares konjaku glycopeptide liquid by aspergillus terricola and Bacillus licheniformis; Then adopt the method for ultrafiltration to process konjaku glycopeptide liquid, collect the filtered solution of relative molecular mass at 10 ~ 100KD; Filtered solution is separated through gel filtration chromatography, the Streptococel of purifying can be obtained.It take fry starch of konjak as the technological gap that raw material combining with fermentation method, ultrafiltration and chromatographic technique prepare Streptococel that the present invention has filled up, also be the exploitation of fry starch of konjak simultaneously, produce high value added product and open a Tiao Xin road, significant in food, medicine and other fields.
Below in conjunction with specific embodiment, the present invention is further elaborated.
Embodiment 1
1) ferment: 1g fry starch of konjak is joined and sends out in 100mL ferment substratum, aspergillus terricola and Bacillus licheniformis is adopted to carry out symbiotic fermentation, after having fermented, fermented liquid removes thalline through the centrifugal 30min of 8000r/min at 4 DEG C, supernatant liquor adds 3 times of purified water, pH to 3.8 adjusted by trichoroacetic acid(TCA), and go precipitation after leaving standstill, supernatant liquor is konjaku glycopeptide liquid.
Wherein, the consisting of of substratum: peptone 1g, extractum carnis 1.5g, NaNO 30.2g, K 2hPO 40.2g, KCl0.08g, MgSO 4.7H 2o0.04g, FeSO 40.004g, distilled water 100mL.The pH value of substratum is 7, and the bacterial classification ratio of dwell native mould and Bacillus licheniformis is 1:1, and bacterial concentration is diluted to 5 × 10 7individual/mL, inoculum size is 10% (V/V, inoculum size refers to the volume fraction of the bacterium liquid accessed in the substratum of unit volume), and leavening temperature is 30 DEG C, and fermentation time is 30h.
2) ultrafiltration: be the ultra-filtration membrane of 100KD and 10KD by molecular weight cut-off successively by konjaku glycopeptide liquid, collects molecular weight distribution in the ultrafiltrated of 10 ~ 100KD section.
3) specification is adopted to be that the SephadexG-75 gel chromatography column of 1cm × 50cm is to the further separation and purification of ultrafiltrated.Gel column separation condition is: applied sample amount 3mL, and sample concentration is 150mg/mL, and eluent is distilled water, and elution flow rate is 0.5mL/min.Often 5mL collected by pipe.Collect and merge the component at 280nm and 460nm absorption peak place, at-60 DEG C, vacuum lyophilization 15h under 60Pa after concentrated, the Streptococel of purifying can be obtained.
Embodiment 2
1) ferment: 1.5g fry starch of konjak is joined in 100mL fermention medium, aspergillus terricola and Bacillus licheniformis is adopted to carry out symbiotic fermentation, after having fermented, fermented liquid removes thalline through the centrifugal 20min of 10000r/min at 4 DEG C, supernatant liquor adds 5 times of purified water, pH to 4 adjusted by trichoroacetic acid(TCA), and go precipitation after leaving standstill, supernatant liquor is konjaku glycopeptide liquid.
Wherein, the consisting of of substratum: peptone 1g, extractum carnis 1.5g, NaNO 30.1g, K 2hPO 40.15g, KCl0.1g, MgSO 4.7H 2o0.06g, FeSO 40.01g, distilled water 100mL.The pH value of substratum is 9, and the bacterial classification ratio of dwell native mould and Bacillus licheniformis is 1:2, and bacterial concentration is diluted to 1 × 10 8individual/mL, inoculum size is 12% (V/V), and leavening temperature is 38 DEG C, and fermentation time is 24h.
2) ultrafiltration: be the ultra-filtration membrane of 100KD and 10KD by molecular weight cut-off successively by konjaku glycopeptide liquid, collects molecular weight distribution in the ultrafiltrated of 10 ~ 100KD section.
3) specification is adopted to be that the SephadexG-75 gel chromatography column of 5cm × 60cm is to the further separation and purification of ultrafiltrated.Gel column separation condition is: applied sample amount 10mL, and sample concentration is 100mg/mL, and eluent is distilled water, and elution flow rate is 5mL/min.Often 3mL collected by pipe.Collect and merge the component at 280nm and 460nm absorption peak place, at-70 DEG C, vacuum lyophilization 12h under 110Pa after concentrated, the Streptococel of purifying can be obtained.
Embodiment 3
1) ferment: 2.5g fry starch of konjak is joined in 100mL fermention medium, aspergillus terricola and Bacillus licheniformis is adopted to carry out symbiotic fermentation, after having fermented, fermented liquid removes thalline through the centrifugal 10min of 12000r/min at 4 DEG C, supernatant liquor adds 4 times of purified water, pH to 3.9 adjusted by trichoroacetic acid(TCA), and go precipitation after leaving standstill, supernatant liquor is konjaku glycopeptide liquid.
Wherein, the consisting of of substratum: peptone 1g, extractum carnis 1.5g, NaNO 30.4g, K 2hPO 40.05g, KCl0.02g, MgSO 4.7H 2o0.1g, FeSO 40.002g, distilled water 100mL.The pH value of substratum is 10, and the bacterial classification ratio of dwell native mould and Bacillus licheniformis is 1:3, and bacterial concentration is diluted to 5 × 10 9individual/mL, inoculum size is 4% (V/V), and leavening temperature is 25 DEG C, and fermentation time is 48h.
2) ultrafiltration: be the ultra-filtration membrane of 100KD and 10KD by molecular weight cut-off successively by konjaku glycopeptide liquid, collects molecular weight distribution in the ultrafiltrated of 10 ~ 100KD section.
3) specification is adopted to be that the SephadexG-75 gel chromatography column of 10cm × 80cm is to the further separation and purification of ultrafiltrated.Gel column separation condition is: applied sample amount 50mL, and sample concentration is 200mg/mL, and eluent is distilled water, and elution flow rate is 10mL/min.Often 4mL collected by pipe.Collect and merge the component at 280nm and 460nm absorption peak place, at-80 DEG C, vacuum lyophilization 20h under 500Pa after concentrated, the Streptococel of purifying can be obtained.
Embodiment 4
1) ferment: 2g fry starch of konjak is joined in 100mL fermention medium, aspergillus terricola and Bacillus licheniformis is adopted to carry out symbiotic fermentation, after having fermented, fermented liquid removes thalline through the centrifugal 25min of 9000r/min at 4 DEG C, supernatant liquor adds 2 times of purified water, pH to 4.1 adjusted by trichoroacetic acid(TCA), and go precipitation after leaving standstill, supernatant liquor is konjaku glycopeptide liquid.
Wherein, the consisting of of substratum: peptone 1g, extractum carnis 1.5g, NaNO 30.3g, K 2hPO 40.1g, KCl0.04g, MgSO 4.7H 2o0.02g, FeSO 40.008g, distilled water 100mL.The pH value of substratum is 11, and the bacterial classification ratio of dwell native mould and Bacillus licheniformis is 1:2.5, and bacterial concentration is diluted to 5 × 10 8individual/mL, inoculum size is 6% (V/V), and leavening temperature is 35 DEG C, and fermentation time is 36h.
2) ultrafiltration: be the ultra-filtration membrane of 100KD and 10KD by molecular weight cut-off successively by konjaku glycopeptide liquid, collects molecular weight distribution in the ultrafiltrated of 10 ~ 100KD section.
3) specification is adopted to be that the SephadexG-75 gel chromatography column of 15cm × 80cm is to the further separation and purification of ultrafiltrated.Gel column separation condition is: applied sample amount 120mL, and sample concentration is 180mg/mL, and eluent is distilled water, and elution flow rate is 30mL/min.Often 6mL collected by pipe.Collect and merge the component at 280nm and 460nm absorption peak place, at-90 DEG C, vacuum lyophilization 18h under 600Pa after concentrated, the Streptococel of purifying can be obtained.
Embodiment 5
1) ferment: 3g fry starch of konjak is joined in 100mL fermention medium, aspergillus terricola and Bacillus licheniformis is adopted to carry out symbiotic fermentation, after having fermented, fermented liquid removes thalline through the centrifugal 15min of 11000r/min at 4 DEG C, supernatant liquor adds 1 times of purified water, pH to 4.2 adjusted by trichoroacetic acid(TCA), and go precipitation after leaving standstill, supernatant liquor is konjaku glycopeptide liquid.
Wherein, the consisting of of substratum: peptone 1g, extractum carnis 1.5g, NaNO 30.25g, K 2hPO 40.12g, KCl0.06g, MgSO 4.7H 2o0.08g, FeSO 40.006g, distilled water 100mL.The pH value of substratum is 8, and the bacterial classification ratio of dwell native mould and Bacillus licheniformis is 1:1.5, and bacterial concentration is diluted to 9 × 10 8individual/mL, inoculum size is 8% (V/V), and leavening temperature is 28 DEG C, and fermentation time is 44h.
2) ultrafiltration: be the ultra-filtration membrane of 100KD and 10KD by molecular weight cut-off successively by konjaku glycopeptide liquid, collects molecular weight distribution in the ultrafiltrated of 10 ~ 100KD section.
3) specification is adopted to be that the SephadexG-75 gel chromatography column of 20cm × 100cm is to the further separation and purification of ultrafiltrated.Gel column separation condition is: applied sample amount 300mL, and sample concentration is 160mg/mL, and eluent is distilled water, and elution flow rate is 80mL/min.Often 5mL collected by pipe.Collect and merge the component at 280nm and 460nm absorption peak place, at-110 DEG C, vacuum lyophilization 24h under 800Pa after concentrated, the Streptococel of purifying can be obtained.
Analyzing and testing is carried out to the monosaccharide components, aminoacid component etc. of Streptococel prepared by the present invention below.
1. qualitative analysis-amino acid collection of illustrative plates measures
Adopt high performance liquid chromatography to carry out qualitative analysis-amino acid collection of illustrative plates to Streptococel prepared by the present invention to measure.Chromatographic column C 18(250mm × 4.6mm, 5 μm); Moving phase: A phase is acetonitrile-water (4:1), B phase is 0.1moL/L sodium-acetate-acetonitrile (93:7, pH6.4); Sampling volume: 10uL; Volumetric flow rate: 1.0mL/min; Determined wavelength: 254nm; Column temperature: 25 DEG C.
The amino acid collection of illustrative plates of Streptococel amino acid collection of illustrative plates contrast mannosans poly saccharide peptide standard product the present invention obtained can carry out qualitative analysis to it.As shown in Figure 1, the Streptococel amino acid collection of illustrative plates that the present invention records as shown in Figure 2 for the amino acid collection of illustrative plates of mannosans poly saccharide peptide standard product.Being labeled as wherein in Fig. 1: 1.Ser; 2.Gly; 3.Tyr; 4.Cys; 5.Val; 6.Met; 7.Phe; 8.Ile; 9.Lev; 10.Lys.Being labeled as in Fig. 2: 1.Ser; 2.Gly; 3.Cys; 4.Val; 5.Met; 6.Phe; 7.Ile; 8.Lev; 9.Lys.
As can be seen from Fig. 1 and Fig. 2, between 35 ~ 43min, amino acid collection of illustrative plates and the mannosans poly saccharide peptide standard product of Streptococel prepared by the present invention are basically identical on appearance time and peak shape, and similarity is higher.This illustrates that the amino acid classes of the amino acid classes that Streptococel prepared by the present invention contains and mannosans poly saccharide peptide standard product is basically identical.
2. monosaccharide components analysis
The monosaccharide components of high performance liquid chromatography to the Streptococel that the present invention obtains is adopted to analyze.Chromatographic column C 18(250mm × 4.6mm, 5 μm); Moving phase: redistilled water-acetonitrile (76:24); Sampling volume: 10uL; Volumetric flow rate: 1.0mL/min; Determined wavelength: 250nm; Column temperature: 35 DEG C.
The mixed standard solution of accurate formulation seminose, glucose, the concentration of glucose and seminose is all 0.1mg/mL.Being diluted to concentration is respectively 0.02mg/mL, 0.04mg/mL, 0.06mg/mL, 0.08mg/mL and 0.10mg/mL, and carries out derivatize mark, absorbs 10 μ L sample introductions respectively, measures its peak area, can draw monose ratio by typical curve regression equation.
Seminose, glucose hybrid standard product high-efficient liquid phase chromatogram are as shown in Figure 3.Streptococel monose hydrolyzate high-efficient liquid phase chromatogram of the present invention as shown in Figure 4.
Peak area reference standard curve is gone out from the sample in Fig. 3, Fig. 4, monose mainly seminose and glucose contained in the Streptococel that the present invention obtains, and the content of seminose and glucose (quality) is than being 1.7:(1 ~ 4).
3. aminoacid component detects
Adopt the aminoacid component of amino acid automatic tester to the Streptococel that the present invention obtains to detect, find that it contains Threonine, Serine, aspartic acid, L-glutamic acid, glycine, L-Ala, halfcystine, tyrosine, α-amino-isovaleric acid, methionine(Met), Isoleucine, leucine, phenylalanine, Methionin, Histidine, arginine, proline(Pro) totally 17 seed amino acids.Wherein must account for 29.61% ~ 31.26% of amino acid total mass by aminoacids content.

Claims (10)

1. a preparation method for Streptococel, is characterized in that, comprises the following steps:
1) ferment: in every 100mL substratum, add fry starch of konjak 1 ~ 3g, adopt aspergillus terricola and Bacillus licheniformis symbiotic fermentation, after having fermented, the centrifugal rear removal thalline of fermentation liquor, get supernatant liquor, add 1 ~ 5 times wherein to the purified water of supernatant volume, then use trichoroacetic acid(TCA) adjust ph to 3.8 ~ 4.2, remove precipitation after leaving standstill, the supernatant liquor obtained after removing precipitation is konjaku glycopeptide liquid;
2) ultrafiltration: be the ultra-filtration membrane of 100KD and 10KD by molecular weight cut-off successively by konjaku glycopeptide liquid, collects molecular weight at the ultrafiltrated of 10 ~ 100KD section, obtains Streptococel crude product;
3) with gel filtration chromatography, separation and purification is carried out to Streptococel crude product, namely obtain the Streptococel of purifying.
2. the preparation method of Streptococel according to claim 1, it is characterized in that: described step 1) in zymotechnique be: the pH value of substratum is 7.0 ~ 11.0, the bacterial classification ratio of aspergillus terricola and Bacillus licheniformis is 1:(1 ~ 3), bacterial concentration is 5 × 10 7~ 5 × 10 9individual/mL, inoculum size is 4 ~ 12%, and fermentation time is 24 ~ 48h, and leavening temperature is 25 ~ 38 DEG C.
3. the preparation method of Streptococel according to claim 1, is characterized in that: described step 1) in substratum be add peptone 1g, extractum carnis 1.5g, NaNO in every 100mL distilled water 30.1 ~ 0.4g, K 2hPO 40.05 ~ 0.2g, KCl0.02 ~ 0.1g, MgSO 4 .7H 2o0.02 ~ 0.1g and FeSO 4obtain after 0.002 ~ 0.01g.
4. according to the preparation method of the Streptococel in claim 1-3 described in any one, it is characterized in that: described step 1) in fermented liquid at 4 DEG C to remove thalline after the centrifugal 10 ~ 30min of the rotating speed of 8000 ~ 12000r/min.
5. according to the preparation method of the Streptococel in claim 1-3 described in any one, it is characterized in that: described step 3) in the chromatography column specification that uses for (1.0 ~ 20) cm × (50 ~ 100) cm, interior fill is SephadexG-75.
6. according to the preparation method of the Streptococel in claim 1-3 described in any one, it is characterized in that: described step 3) in gel filtration chromatography condition be: applied sample amount is 3 ~ 300mL, sample concentration is 100 ~ 200mg/mL, eluent is distilled water, elution flow rate is 0.5 ~ 80mL/min, often 3 ~ 6mL collected by pipe, light absorption value is measured at 280nm and 460nm place, collection merging has sugar simultaneously, the component of peptide absorption peak concentrates, and obtains the Streptococel of purifying after vacuum lyophilization.
7. the preparation method of Streptococel according to claim 6, is characterized in that: described vacuum lyophilization is dry 12 ~ 24h under-110 ~-60 DEG C and 60 ~ 800Pa.
8. according to the preparation method of the Streptococel in claim 1-3 described in any one, it is characterized in that: seminose contained in obtained Streptococel and the mass ratio of glucose are 1.7:(1 ~ 4).
9. the preparation method of Streptococel according to claim 8, is characterized in that: necessary amino acid whose quality contained in obtained Streptococel is 29.61% ~ 31.26% of amino acid total mass.
10. the preparation method of Streptococel according to claim 8, is characterized in that: amino acid contained in obtained Streptococel comprises Threonine, Serine, aspartic acid, L-glutamic acid, glycine, L-Ala, halfcystine, tyrosine, α-amino-isovaleric acid, methionine(Met), Isoleucine, leucine, phenylalanine, Methionin, Histidine, arginine and proline(Pro).
CN201510888970.4A 2015-12-07 2015-12-07 Preparation method of mannatide Pending CN105420317A (en)

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CN107011419A (en) * 2017-04-21 2017-08-04 南京中医药大学 Active glycopeptide of Radix Isatidis with antiviral activity and preparation method and application

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CN106117384A (en) * 2016-06-27 2016-11-16 天津汇滨生物科技有限公司 A kind of preparation method of Candida mannan Mn
CN107011419A (en) * 2017-04-21 2017-08-04 南京中医药大学 Active glycopeptide of Radix Isatidis with antiviral activity and preparation method and application
CN107011419B (en) * 2017-04-21 2020-11-10 南京中医药大学 Isatis root active glycopeptide with antiviral activity and preparation method and application thereof

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Application publication date: 20160323