CN104262501A - Method for extracting grifola frondosa fermentation broth polysaccharide by virtue of dual-membrane process - Google Patents
Method for extracting grifola frondosa fermentation broth polysaccharide by virtue of dual-membrane process Download PDFInfo
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- CN104262501A CN104262501A CN201410516379.1A CN201410516379A CN104262501A CN 104262501 A CN104262501 A CN 104262501A CN 201410516379 A CN201410516379 A CN 201410516379A CN 104262501 A CN104262501 A CN 104262501A
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Abstract
The invention discloses a method for extracting grifola frondosa fermentation broth polysaccharide by virtue of a dual-membrane process. According to the method for extracting the grifola frondosa fermentation broth polysaccharide by virtue of the dual-membrane process, the grifola frondosa fermentation broth polysaccharide is obtained by processing and separating grifola frondosa fermentation broth through a micro-filtration membrane and an ultra-filtration membrane under a specific condition. Due to adoption of the method, the traditional 'water-extraction and ethanol-precipitation' extraction mode of grifola frondosa fermentation broth polysaccharide is changed, the purification effect of grifola frondosa fermentation broth polysaccharide is improved, the amount of ethanol in the production is reduced and the economic efficiency is increased; and meanwhile, by adopting the method, the emission of filtrate wastewater can be reduced and social benefits are significantly increased.
Description
Technical field
The invention belongs to microbial technology field, relate to a kind of method extracting fungi exocellular polysaccharide, more particularly, is a kind of novel method extracting polyose from Polyporus frondosus fermenting liquor based on membrane technique.
Background technology
The saccharan that polyose from Polyporus frondosus fermenting liquor is made up of other monose such as D-Glucose, D-wood sugar, D-Fucose, D-MANNOSEs, being the protein-polysaccharide material with physiologically active using modern biotechnology Hydrolysis kinetics from frondosa fermentation filtrate, is the principle active component of Grifola frondosa.Experiment proves, polyose from Polyporus frondosus fermenting liquor has restraining effect to various virus, as virus of AIDS (HIV.1), hsv, cytomegalovirus, influenza virus etc.Polyose from Polyporus frondosus fermenting liquor can improve immune function of human body, leukocyte increasing, and the treatment of polyose from Polyporus frondosus fermenting liquor to hepatitis has significant effect.
It is carry out repeatedly alcohol alcohol precipitation after the frondosa fermentation filtrate after being separated is concentrated to certain multiple that traditional polyose from Polyporus frondosus fermenting liquor is produced, and removing monose wherein, improves polysaccharide content.But in alcohol precipitation process due to monose often along with polysaccharide simultaneously by under alcohol precipitation, therefore monose clearance lower, needs repeatedly alcohol precipitation, and expend alcohol and manually more, production cost is higher; Meanwhile, traditional hot condensing mode, consumes a large amount of steam, has considerable influence to production cost and quality product.
Summary of the invention
The object of the invention is the deficiency in order to solve above-mentioned polyose from Polyporus frondosus fermenting liquor separation purification method, thus devise a kind of novel environment-friendly process based on membrane technique separation and purification polyose from Polyporus frondosus fermenting liquor, this technique had both improve the purification effect of polyose from Polyporus frondosus fermenting liquor, added economic benefit; Additionally reduce the discharge of waste water, add social benefit.
To achieve these goals, a kind of bi-membrane method of the present invention extracts the method for polyose from Polyporus frondosus fermenting liquor, and the method comprises the steps:
(1) Grifola gigantea (Pers.) Piat. Fermented liquid is passed through microfiltration membrane, obtain micro-filtration dialyzate and mycelium, collect micro-filtration dialyzate; Wherein the aperture of microfiltration membrane is at 0.05-1 micron, and service temperature is 10-70 degree, operating pressure: entrance pressure is 2-8bar, goes out pressure for 0-5bar;
(2) the micro-filtration dialyzate in step (1) is passed through ultra-filtration membrane, obtain ultrafiltration dialysis liquid and concentrated solution, collect concentrated solution; Wherein, ultrafiltration selective retention molecular weight is the ultra-filtration membrane of 800-1000Da, and service temperature is 38-50 degree, operating pressure: entrance pressure is 1-8 bar, goes out pressure for 0.5-3bar;
(3) be that 30-70% obtains thickened pulp by ultrafiltration and concentration liquid in step (2) being concentrated to solid content under 40-80 degrees celsius;
(4) thickened pulp step (3) obtained, at 60-85 degree Celsius, is dried under the condition of more than vacuum tightness-0.08MPa, and drying to dry thing moisture content is mass percent 3-8%, to obtain final product.The dry thing of gained is polyose from Polyporus frondosus fermenting liquor.
Preferred micro-filtration employing aperture is the microfiltration membrane of 0.05-0.5 micron, and most preferably micro-filtration employing aperture is the microfiltration membrane of 0.05 micron, and micro-filtration adopts the service temperature of 50-60 degree, and entrance pressure is 2-4bar, goes out pressure for 1-2 bar.Preferred ultrafiltration adopts molecular weight cut-off to be the ultra-filtration membrane of 900-1000Da, and ultrafiltration adopts the service temperature of 40-45 degree, and entrance pressure is 6-8 bar, goes out pressure for 1-3 bar.
Micro-filtration of the present invention film used is ceramic membrane or organic membrane.Ultrafiltration mould material used can select the one in PAN, PVDF, PC, PES, PP, PE composite membrane.
The polyose from Polyporus frondosus fermenting liquor that Grifola gigantea (Pers.) Piat. Fermented liquid extracts is because of bio-active products in fact, comparatively responsive to temperature, if temperature is more than 60 degree, often raises 5 degree, polyose from Polyporus frondosus fermenting liquor index decline 10%.If be no more than within the scope of 60 degree, have chosen the operation that 20,30,40,50,60 degree of 5 temperature carry out membrane sepn respectively, found that, its physical and chemical index of polyose from Polyporus frondosus fermenting liquor obtained at these 5 temperature operations is more or less the same, but membrane flux and maintain the energy consumption that this temperature produces and differ widely under differing temps, and under low temperature, membrane flux is less, and water coolant is larger needed for maintenance low temperature, temperature raises, and membrane flux increases, and operating process does not need water coolant or needs few water coolant.To sum up, preferred film filtration temperature is 40-60 degree, wherein preferred microfiltration membrane 50-60 degree, ultra-filtration membrane 40-45 degree.
The present invention adopts Grifola gigantea (Pers.) Piat. Fermented liquid to pass through microfiltration membrane, obtains micro-filtration dialyzate and the mycelium of fermented liquid, and mycelium directly cryodrying pulverizing can make fermentation Grifolas frondosa germ powder; Micro-filtration dialyzate is by ultra-filtration membrane, and obtain ultrafiltration dialysis liquid and concentrated solution, ultrafiltration dialysis liquid can be used as fermentation batches water; Significantly production cost can be reduced after two film process.
Production method of the present invention has following beneficial effect compared with the prior art:
1. micro-filtration is except mycelium raising mycelia yield, and in operating process, mechanical power is destroyed, and high degree saves adenosine in mycelium, N.F,USP MANNITOL composition, and supporting cost is low, without the need to the device such as sheet frame, whizzer.
2. the filtering accuracy of ultra-filtration membrane is high, and monose clearance is high, improves polyose from Polyporus frondosus fermenting liquor quality product; Meanwhile, containing monose in ultrafiltration dialysis liquid, can be used as fermentation batches water, decrease sewage load.
3. carry out pre-concentration with ultra-filtration membrane, realize cold concentration, reduce steam consumption, reduce production cost.
4. the alcohol-free consumption of production process of the present invention, greatly reduces production cost and expenditure of time, and meanwhile, the present invention realizes clean automatic production, greatly improves Working environment.
5. membrane technique is at home and abroad constantly promoted widely in recent years; film cost is also in continuous decline; membrane separation and purification technology is applied in the leaching process of polyose from Polyporus frondosus fermenting liquor and will produces significant economic benefit; application membrane technique can not only improve the quality of products, and can also reduce the discharge of waste water, protection of the environment; realize Automated condtrol and cleaner production; reduce production cost, the Working environment of workman can also be improved, increase work efficiency.
Adopt the polysaccharide yield that obtains of present method up to 5.32g/L, polysaccharide content 27.9%, monose 2.21%.Polysaccharide determination method phenol-sulfuric acid process, monose measuring method DNS method measures.Compared with traditional water extraction, the extraction yield of polyose from Polyporus frondosus fermenting liquor improves at least 1.0 percentage points, and polysaccharide content improves more than 10 percentage points, and contents of monosaccharides reduces at least 3 percentage points, and extraction time shortens more than 3 times, and cost significantly reduces; Extract whole process and avoid heat, thus decrease the destruction of high temperature to polysaccharide; Instrument operation of equipment is convenient, and level of automation is higher, is convenient to scale operation.
Embodiment
In order to understand the present invention, below by embodiment, the invention will be further described, but do not limit the present invention.
embodiment 1:
Process for extracting polyose from Polyporus frondosus fermenting liquor of the present invention carries out in the steps below:
1. Grifola gigantea (Pers.) Piat. Fermented liquid source: administrative center provides Grifola frondosa strain by Chinese microorganism strain preservation, bacterial strain is transferred to slant medium 26-28 degree cellar culture about 10 days, obtains the Grifola frondosa strain of activation; The Grifola frondosa strain activated is inoculated in seed culture medium, and the concussion of 26-28 degree is cultivated and is obtained Grifola frondosa primary seed solution in 4-5 days; Primary seed solution is accessed secondary seed medium, and 26-28 degree cultivates about 4 days; Secondary seed medium is accessed fermention medium, and 26-28 degree cultivates about 10 days, and fermentation ends is Grifola gigantea (Pers.) Piat. Fermented liquid.
2. micro-filtration filters: filtered by micro-filtration by above-mentioned Grifola gigantea (Pers.) Piat. Fermented liquid 7500Kg (fermented liquid solid content is 3.7%), obtain the wet mycelium (water content is 76%) of micro-filtration dialyzate 6370Kg and 1130Kg of Grifola frondosa, the transmittance of dialyzate is 83%, and wet mycelium directly cryodrying can pulverize obtained fermentation Grifolas frondosa germ powder.The ceramic membrane of micro-filtration filters mould material used to be aperture be 50nm, concrete technology condition is: service temperature is 50-60 degree, and entrance pressure is 3.0bar, and go out pressure for 1.0bar, pressure difference is 2.0bar.
3. ultra-filtration filters: the micro-filtration dialyzate 6370Kg of above-mentioned gained is obtained ultrafiltration dialysis liquid 5640Kg and 730kg ultrafiltration and concentration liquid by ultra-filtration filters, the transmittance of dialyzate is that 95%, 5640Kg dialyzate can be used as fermentation batches water.The ultra-filtration membrane of ultrafiltration mould material used to be molecular weight cut-off the be PVDF of 1000DaD, processing condition are: service temperature is 40 degree, and entrance pressure is 7bar, and go out pressure for 2bar, pressure difference is 5bar.
4. cryoconcentration: be 0.09MPa in vacuum tightness by the ultrafiltration and concentration liquid 730kg of above-mentioned ultrafiltration gained, thickening temperature is concentrate under 60 degree, Grifola frondosa concentrated solution is concentrated into the polyose from Polyporus frondosus fermenting liquor thickened pulp 49.5Kg that solid content is 57%.
5. low-temperature vacuum drying: the 49.5Kg polyose from Polyporus frondosus fermenting liquor thickened pulp obtained by above-mentioned cryoconcentration enters vacuum drying oven dries, and obtains 29.2kg polyose from Polyporus frondosus fermenting liquor crude product (moisture content 4.2%).Concrete technology condition is: drying temperature 75-85 degree, and vacuum tightness is-0.085MPa, time of drying 40h.Polyose from Polyporus frondosus fermenting liquor content 27.9% is obtained, contents of monosaccharides 2.21% under these processing condition.
Claims (7)
1. bi-membrane method extracts a method for polyose from Polyporus frondosus fermenting liquor, and it is characterized in that, the method is carried out according to following step:
(1) Grifola gigantea (Pers.) Piat. Fermented liquid is passed through microfiltration membrane, obtain micro-filtration dialyzate and mycelium, collect micro-filtration dialyzate; Wherein the aperture of microfiltration membrane is at 0.05-1 micron, and service temperature is 10-70 degree Celsius, operating pressure: entrance pressure is 2-8bar, goes out pressure for 0-5bar;
(2) the micro-filtration dialyzate in step (1) is passed through ultra-filtration membrane, obtain ultrafiltration dialysis liquid and ultrafiltration and concentration liquid, collect ultrafiltration and concentration liquid; Wherein, ultrafiltration selective retention molecular weight is the ultra-filtration membrane of 800-1000Da, and service temperature is 38-50 degree Celsius, operating pressure: entrance pressure is 1-8 bar, goes out pressure for 0.5-3bar;
(3) be that 30-70% obtains thickened pulp by ultrafiltration and concentration liquid in step (2) being concentrated to solid content under 40-80 degrees celsius;
(4) thickened pulp step (3) obtained, at 60-85 degree Celsius, is dried under the condition of more than vacuum tightness-0.08MPa, and drying to dry thing moisture content is mass percent 3-8%, to obtain final product.
2. bi-membrane method according to claim 1 extracts the method for polyose from Polyporus frondosus fermenting liquor, it is characterized in that the aperture of the microfiltration membrane adopted in described step (1) is at 0.05-0.5 micron.
3. bi-membrane method according to claim 1 extracts the method for polyose from Polyporus frondosus fermenting liquor, it is characterized in that the service temperature that in described step (1), micro-filtration adopts is 50-60 degree, operating pressure: entrance pressure is 2-4bar, goes out pressure for 1-2bar.
4. bi-membrane method according to claim 1 extracts the method for polyose from Polyporus frondosus fermenting liquor, it is characterized in that in described step (2), ultra-filtration membrane selective retention molecular weight is the ultra-filtration membrane of 900-1000Da.
5. bi-membrane method according to claim 1 extracts the method for polyose from Polyporus frondosus fermenting liquor, it is characterized in that the service temperature that described step (2) ultrafiltration membrance filter adopts is 40-45 degree, operating pressure: entrance pressure is 6-8bar, goes out pressure for 1-3bar.
6. bi-membrane method according to claim 1 extracts the method for polyose from Polyporus frondosus fermenting liquor, it is characterized in that in described step (1), micro-filtration film used is ceramic membrane or organic membrane.
7. bi-membrane method according to claim 1 extracts the method for polyose from Polyporus frondosus fermenting liquor, it is characterized in that in described step (2), the one in PAN, PVDF, PC, PES, PP or PE composite membrane selected by ultrafiltration mould material used.
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Cited By (6)
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| CN105669876A (en) * | 2016-03-02 | 2016-06-15 | 漳州片仔癀药业股份有限公司 | Extraction process and application of grifolan |
| CN105732836A (en) * | 2016-03-02 | 2016-07-06 | 漳州片仔癀药业股份有限公司 | Extraction process and application of polysaccharide of grifola frondosa to preparation of medicament for repairing gastric mucosa damage |
| CN105777931A (en) * | 2016-03-02 | 2016-07-20 | 漳州片仔癀药业股份有限公司 | Grifolan extracting method and pharmaceutical application thereof |
| CN109180832A (en) * | 2018-10-12 | 2019-01-11 | 绩溪县徽菜宝生物科技有限公司 | A kind of efficient Extraction Methods of Polysaccharides From Agrocybe aegerita |
| CN110559332A (en) * | 2019-10-11 | 2019-12-13 | 广东粤微生物科技有限公司 | Ganoderma lucidum and grifola frondosa hypoglycemic composition as well as preparation method and application thereof |
| CN112225827A (en) * | 2020-10-15 | 2021-01-15 | 广东粤微生物科技有限公司 | Extraction method of active polysaccharide of grifola frondosa, extracted active polysaccharide and application |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105669876A (en) * | 2016-03-02 | 2016-06-15 | 漳州片仔癀药业股份有限公司 | Extraction process and application of grifolan |
| CN105732836A (en) * | 2016-03-02 | 2016-07-06 | 漳州片仔癀药业股份有限公司 | Extraction process and application of polysaccharide of grifola frondosa to preparation of medicament for repairing gastric mucosa damage |
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| CN109180832A (en) * | 2018-10-12 | 2019-01-11 | 绩溪县徽菜宝生物科技有限公司 | A kind of efficient Extraction Methods of Polysaccharides From Agrocybe aegerita |
| CN110559332A (en) * | 2019-10-11 | 2019-12-13 | 广东粤微生物科技有限公司 | Ganoderma lucidum and grifola frondosa hypoglycemic composition as well as preparation method and application thereof |
| CN112225827A (en) * | 2020-10-15 | 2021-01-15 | 广东粤微生物科技有限公司 | Extraction method of active polysaccharide of grifola frondosa, extracted active polysaccharide and application |
| WO2022077853A1 (en) * | 2020-10-15 | 2022-04-21 | 广东粤微生物科技有限公司 | Method for extracting grifola frondosa active polysaccharide, and extracted active polysaccharide and use thereof |
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Application publication date: 20150107 |