CN103509130A - Novel green process for extracting ganoderma lucidum polysaccharides - Google Patents
Novel green process for extracting ganoderma lucidum polysaccharides Download PDFInfo
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- CN103509130A CN103509130A CN201310439842.2A CN201310439842A CN103509130A CN 103509130 A CN103509130 A CN 103509130A CN 201310439842 A CN201310439842 A CN 201310439842A CN 103509130 A CN103509130 A CN 103509130A
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Abstract
The invention discloses a novel green process for extracting ganoderma lucidum polysaccharides and in particular relates to a method for extracting ganoderma lucidum polysaccharides by a double membrane process. According to the method, ganoderma lucidum polysaccharides are obtained by separating a lucid ganoderma fermentation liquor using a micro-filtration membrane and an ultrafiltration membrane under special conditions. According to the invention, the conventional ganoderma lucidum polysaccharide extraction mode of water extraction and ethanol precipitation is changed, the purification effect of ganoderma lucidum polysaccharides is improved, and the consumption of ethanol during production is reduced, therefore the economic benefit is increased. Besides, with adoption of the method, the emission of filtrate wastewater is reduced and the social benefit is remarkably increased.
Description
Technical field
The invention belongs to microbial technology field, be specifically related to a kind of method of extracting fungi exocellular polysaccharide, more precisely a kind of novel method of extracting ganoderan based on membrane technique.
Background technology
The saccharan that ganoderan is comprised of other monose such as glucose, pectinose, wood sugar, Fucose, rhamnosyl, semi-lactosis, being to use modern biotechnology from glossy ganoderma fermentation filtrate, to extract the protein-polysaccharide material with physiologically active of refining, is one of effective constituent of glossy ganoderma.Experimental results show that, ganoderan has many-sided pharmacologically active: can improve immunity of organisms, accelerate blood microcirculation, improve blood oxygen ability, reduce the invalid oxygen-consumption under body stationary state, eliminate interior free yl, improve the sealing degree of body cell film, anti-radiation, improve liver, marrow, blood synthetic DNA, RNA, the ability of protein, prolongs life etc.The multiple pharmacologically active of glossy ganoderma is mostly relevant with ganoderan.
It is to carry out repeatedly alcohol alcohol precipitation after the glossy ganoderma fermentation filtrate after separation is concentrated to certain multiple that traditional ganoderan is produced, and removes monose wherein, improves polysaccharide content.But in alcohol precipitation process, because monose is often accompanied by polysaccharide simultaneously by under alcohol precipitation, monose clearance is lower, therefore needs repeatedly alcohol precipitation, expends alcohol and manually more, and production cost is higher; Meanwhile, the concentrated mode of traditional heat, consumes a large amount of steam, and production cost and quality product are had to considerable influence.
Summary of the invention
The object of the invention is in order to solve the deficiency of above-mentioned ganoderan separation purification method, thereby designed a kind of novel environment-friendly process that carrys out separation and purification ganoderan based on membrane technique, this technique had both improved the purification effect of ganoderan, had increased economic benefit; Reduce again the discharge of waste water, increased social benefit.
To achieve these goals, the present invention mainly carries out according to following step:
Pair embrane method is extracted a method for ganoderan, and the method comprises the steps:
1) Ganoderma fermentation liquid is passed through to microfiltration membrane, obtain micro-filtration dialyzate and mycelium, collect micro-filtration dialyzate; Wherein, the aperture of microfiltration membrane is at 0.05～1 μ m, and service temperature is 10～70 ℃, operating pressure: entrance pressure is 2～8bar, and going out to press is 0～5bar;
2) the micro-filtration dialyzate in step 1) is passed through to ultra-filtration membrane, obtain ultrafiltration dialysis liquid and concentrated solution, collect concentrated solution; Wherein, the ultra-filtration membrane that ultrafiltration selective retention molecular weight is 800～1000Da, service temperature is 38～50 ℃, operating pressure: entrance pressure is 1～8bar, going out to press is 0.5～3bar;
3) by step 2) middle ultrafiltration and concentration liquid concentrated thickened pulp that obtains under cold condition; Thickened pulp further cryogenic vacuum oven dry to dry thing moisture content is 3～8%, obtains.The dry thing of gained is ganoderan.
Preferably by step 2) in ultrafiltration and concentration liquid under 40～80 ℃ of conditions, to be concentrated to mycelium content be 30～70% to obtain thickened pulp; Again by the thickened pulp of gained at 60～75 ℃, under condition more than vacuum tightness-0.08Mpa, dry, dry to dry thing moisture content be 3～8%.
Preferably micro-filtration employing aperture is the microfiltration membrane of 0.05～0.5 μ m, and most preferably micro-filtration employing aperture is the microfiltration membrane of 0.05 μ m, and micro-filtration adopts the service temperature of 50～60 ℃, and entrance pressure is 2～4bar, and going out to press is 1～2bar.Preferably ultrafiltration adopts the ultra-filtration membrane that molecular weight cut-off is 900～1000Da, and ultrafiltration adopts the service temperature of 40～45 ℃, and entrance pressure is 6～8bar, and going out to press is 1～3bar.
Micro-filtration of the present invention film used is ceramic membrane or organic membrane.Ultrafiltration mould material used can be selected a kind of in PAN, PVDF, PC, PES, PP, PE composite membrane.
The ganoderan that Ganoderma fermentation liquid extracts is bio-active products because of it, comparatively responsive to temperature, if temperature surpasses 60 ℃, and 5 ℃ of every risings, ganoderan index declines 10%.Be no more than within the scope of 60 ℃, choose respectively 20,30,40,50,60 ℃ of 5 temperature and carried out the operation of membrane sepn, found that, at these 5 its physical and chemical indexs of the resulting ganoderan of temperature operation, be 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 it is larger to maintain the required water coolant of low temperature, temperature raises, and membrane flux increases, and operating process does not need water coolant or the few water coolant of needs.To sum up, 40～60 ℃ of preferred film filtration temperature scopes, wherein preferred microfiltration membrane is 50～60 ℃, 40～45 ℃ of ultra-filtration membranes.
The present invention adopts Ganoderma fermentation liquid to pass through microfiltration membrane, obtains micro-filtration dialyzate and the mycelium of fermented liquid, and mycelium directly cryodrying pulverizing is made fermentation glossy ganoderma bacterium powder; Micro-filtration dialyzate, by ultra-filtration membrane, obtains ultrafiltration dialysis liquid and concentrated solution, and ultrafiltration dialysis liquid can be used as fermentation ingredient water; After processing, two films can significantly reduce production costs.
Production method of the present invention provides a kind of new method for ganoderan separation and Extraction compared with the prior art, and the method has advantages of following outstanding:
1, micro-filtration improves mycelia yield except mycelium, and in operating process, machinery-free power is destroyed, and greatly degree has been preserved adenosine, N.F,USP MANNITOL composition in mycelium, and supporting cost is low, need not sheet frame, the device such as whizzer;
2, the filtering accuracy of ultra-filtration membrane is high, and monose clearance is high, improves ganoderan quality product; Meanwhile, in ultrafiltration dialysis liquid, contain monose, can be used as fermentation batching water, reduced sewage load;
3, with ultra-filtration membrane, carry out pre-concentration, realize cold concentration, reduce steam consumption, reduce production costs;
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, has greatly improved Working environment.
5, membrane technique is at home and abroad constantly promoted widely in recent years, and film cost is also in continuous decline, membrane separation and purification technology is applied in the leaching process of ganoderan 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, and protection of the environment, realizes automatization control and cleaner production, reduces production costs, and can also improve workman's Working environment, increases work efficiency.
6, adopt polysaccharide yield that the inventive method obtains up to 4.24g/L, polysaccharide content 28.41%, monose 2.14%.Polysaccharide determination method phenolsulfuric acid method, monose measuring method DNS method is measured.Compare with traditional water extraction, the extraction yield of ganoderan has improved at least 1.0 percentage points, and polysaccharide content has improved more than 10 percentage points, and contents of monosaccharides has reduced at least 3 percentage points, and extraction time shorten more than 3 times, cost significantly reduces; Extract whole process and avoided heat, thereby reduced the destruction of high temperature to polysaccharide; Instrument operation of equipment is convenient, and level of automation is higher, is convenient to scale operation.
Accompanying drawing explanation
Fig. 1 is the process flow sheet that two embrane methods are extracted ganoderan.
Embodiment
By specific embodiment, further illustrate the present invention below.But the detail of embodiment only, for explaining the present invention, should not be construed as limited overall technical solution.
Embodiment 1
Ganoderan extracting method of the present invention carries out in the steps below:
1, Ganoderma fermentation liquid source: administrative center provides and identifies by the preservation of Chinese agriculture microbial strains, Ganoderma lucidium, deposit number ACCC50059.Bacterial strain is transferred to 26～28 ℃ of cellar culture 5-6 days of slant medium, obtains the ganoderma strain capable of activation; The ganoderma strain capable having activated is inoculated in to seed culture medium, wherein seed culture medium consists of soybean cake powder 2.0%, sucrose 4.0%, yeast powder 2.0%, potassium primary phosphate 0.4%, sal epsom 0.09%, defoamer 0.1%, water preparation, pH value nature, 26～28 ℃ of shaking culture 2-3 days obtain glossy ganoderma primary seed solution; Primary seed solution is accessed to secondary seed medium, wherein secondary seed medium consists of soybean cake powder 3.0%, sucrose 3.0%, yeast powder 2.0%, potassium primary phosphate 0.4%, sal epsom 0.09%, defoamer 0.1%, water preparation, pH value nature, cultivate about 2 days for 26～28 ℃, inoculum size 1%, 1: 0.4～0.8vvm ventilates; Secondary seed medium is accessed to fermention medium, and wherein fermention medium consists of soybean cake powder 4.0%, sucrose 2.0%, yeast powder 2.0%, potassium primary phosphate 0.4%, sal epsom 0.09%, defoamer 0.1%, water preparation, pH value nature, cultivates 80-90h, inoculum size 3%-5% for 26～28 ℃, 1: the 0.3～0.5vvm that ventilates, fermentation ends is Ganoderma fermentation liquid.
2, micro-filtration filters: by above-mentioned Ganoderma fermentation liquid 7500Kg(fermented liquid mycelium content, be 3.7%) by micro-filtration, filter, obtain the micro-filtration dialyzate 6370Kg of glossy ganoderma and the wet mycelium of 1130Kg (water content is 76%), the transmittance of dialyzate is 83%, and wet mycelium directly cryodrying pulverizing makes fermentation glossy ganoderma bacterium powder.It is that aperture is the ceramic membrane of 50nm that micro-filtration filters mould material used, and concrete technology condition is: service temperature is 50～60 ℃, and entrance pressure is 3.0bar, goes out to press for 1.0bar, and pressure difference is 2.0bar.
3, ultra-filtration filters: the micro-filtration dialyzate 6370Kg of step 1 gained is obtained to ultrafiltration dialysis liquid 5640Kg and 730Kg ultrafiltration and concentration liquid by ultra-filtration filters, and the transmittance of dialyzate is that 95%, 5640Kg dialyzate can be used as fermentation batching water.Ultrafiltration mould material used is that molecular weight cut-off is the ultra-filtration membrane of the PVDF of 1000Da, and processing condition are: service temperature is 40 ℃, and entrance pressure is 7bar, goes out to press for 2bar, and pressure difference is 5bar.
4, cryoconcentration: be 0.09MPa by the ultrafiltration and concentration liquid 730Kg of step 2 ultrafiltration gained in vacuum tightness, thickening temperature is at 60 ℃ to continue lucidum concentrating liquid to be concentrated into the ganoderan thickened pulp 55.6Kg of solid content 59%.
5, low-temperature vacuum drying: the 55.6kg ganoderan thickened pulp that step 3 is obtained enters vacuum drying oven dries, and obtains 31.3kg ganoderan crude product (moisture content 4.2%).Concrete technology condition is: 75～85 ℃ of drying temperatures, vacuum tightness is-0.085Mpa, time of drying 40h.Under these processing condition, obtain ganoderma polyoses content 28.41%, contents of monosaccharides 2.14%.
Embodiment 2
Ganoderan extracting method of the present invention, carries out in the steps below:
1, micro-filtration filters: by the Ganoderma fermentation liquid 7500Kg(fermented liquid mycelium content in embodiment 1, be 3.7%) by micro-filtration, filter, obtain the micro-filtration dialyzate 6372Kg of glossy ganoderma and the wet mycelium of 1128Kg (water content is 76.1%), the transmittance of dialyzate is 84%, and wet mycelium directly cryodrying pulverizing makes fermentation glossy ganoderma bacterium powder.It is that aperture is the ceramic membrane of 100nm that micro-filtration filters mould material used, and concrete technology condition is: service temperature is 50～60 ℃, and entrance pressure 2.0bar goes out to press 1.0bar, and pressure difference is 1.0bar.
2, ultra-filtration filters: the micro-filtration dialyzate 6372Kg of step 1 gained is obtained to ultrafiltration dialysis liquid 5655Kg and 717Kg ultrafiltration and concentration liquid by ultra-filtration filters, and the transmittance of dialyzate is that 96%, 5655Kg dialyzate can be used as fermentation batching water.Ultrafiltration mould material used is that molecular weight cut-off is the ultra-filtration membrane of the PAN of 800Da, and processing condition are: service temperature is 42 ℃, and entrance pressure is 6.5bar, goes out to press for 2.0bar, and pressure difference is 4.5bar.
3, cryoconcentration: be 0.09MPa by the ultrafiltration and concentration liquid 717Kg of step 2 ultrafiltration gained in vacuum tightness, thickening temperature is at 60 ℃ to continue ganoderan to be concentrated into the ganoderan thickened pulp 53.8Kg of solid content 61%.
4, low-temperature vacuum drying: the 53.8kg ganoderan thickened pulp that step 3 is obtained enters vacuum drying oven dries, and obtains 31.8kg ganoderan crude product (moisture content 3%).Concrete technology condition is: 75～85 ℃ of drying temperatures, vacuum tightness is-0.085Mpa, time of drying 42h.Under these processing condition, obtain ganoderma polyoses content 28.35%, contents of monosaccharides 2.85%.
Embodiment 3
Ganoderan extracting method of the present invention, carries out in the steps below:
1, micro-filtration filters: the 7500Kg fermented liquid after glossy ganoderma fermentation (fermented liquid mycelium content is 3.7%) is filtered by micro-filtration, obtain the micro-filtration dialyzate 6374Kg of glossy ganoderma and the wet mycelium of 1126Kg (water content is 77%), the transmittance of dialyzate is 80%, and wet mycelium directly cryodrying pulverizing makes fermentation glossy ganoderma bacterium powder.It is that aperture is the ceramic membrane of 500nm that micro-filtration filters mould material used, and concrete technology condition is: service temperature is 50～60 ℃, and entrance pressure 2.3bar goes out to press 0.9bar, and pressure difference is 1.4bar.
2, ultra-filtration filters: the micro-filtration dialyzate 6374Kg of step 1 gained is obtained to ultrafiltration dialysis liquid 5651Kg and 723Kg ultrafiltration and concentration liquid by ultra-filtration filters, and the transmittance of dialyzate is that 94%, 5651Kg dialyzate can be used as fermentation batching water.Ultrafiltration mould material used is that molecular weight cut-off is the ultra-filtration membrane of the PP of 900Da, and processing condition are: service temperature is 45 ℃, and entrance pressure is 6.0bar, goes out to press for 1.8bar, and pressure difference is 4.2bar.
3, cryoconcentration: be 0.09MPa by the ultrafiltration and concentration liquid 723Kg of step 2 ultrafiltration gained in vacuum tightness, thickening temperature is at 60 ℃ to continue ganoderan to be concentrated into the ganoderan thickened pulp 58.5Kg of mycelium content 56%.
4, low-temperature vacuum drying: the 58.5kg ganoderan thickened pulp that step 3 is obtained enters vacuum drying oven dries, and obtains 30.9kg ganoderan crude product (moisture content 3.3%).Concrete technology condition is: 75～85 ℃ of drying temperatures, vacuum tightness is-0.085Mpa, time of drying 38h.Under these processing condition, obtain ganoderma polyoses content 28.05%, contents of monosaccharides 2.97%.
Claims (9)
1. two embrane methods are extracted a method for ganoderan, it is characterized in that the method comprises the steps:
1) Ganoderma fermentation liquid is passed through to microfiltration membrane, obtain micro-filtration dialyzate and mycelium, collect micro-filtration dialyzate; Wherein, the aperture of microfiltration membrane is at 0.05～1 μ m, and service temperature is 10～70 ℃, operating pressure: entrance pressure is 2～8bar, and going out to press is 0～5bar;
2) the micro-filtration dialyzate in step 1) is passed through to ultra-filtration membrane, obtain ultrafiltration dialysis liquid and concentrated solution, collect concentrated solution; Wherein, the ultra-filtration membrane that ultrafiltration selective retention molecular weight is 800～1000Da, service temperature is 38～50 ℃, operating pressure: entrance pressure is 1～8bar, going out to press is 0.5～3bar;
3) by step 2) middle ultrafiltration and concentration liquid concentrated thickened pulp that obtains under cold condition; Thickened pulp further cryogenic vacuum oven dry to dry thing moisture content is 3～8%, obtains.
2. the according to claim 1 pair of embrane method extracted the method for ganoderan, it is characterized in that the aperture of the microfiltration membrane that adopts in described step 1) is 0.05～0.5 μ m.
3. the method for according to claim 1 pair of embrane method extraction ganoderan, is characterized in that the service temperature that in described step 1), micro-filtration adopts is 50～60 ℃, operating pressure: entrance pressure is 2～4bar, and going out to press is 1～2bar.
4. the according to claim 1 pair of embrane method extracted the method for ganoderan, it is characterized in that described step 2) in the ultra-filtration membrane selective retention molecular weight ultra-filtration membrane that is 900～1000Da.
5. the according to claim 1 pair of embrane method extracted the method for ganoderan, it is characterized in that described step 2) in the service temperature that adopts of ultrafiltration membrance filter be 40～45 ℃, operating pressure: entrance pressure is 6～8bar, going out to press is 1～3bar.
6. the method for according to claim 1 pair of embrane method extraction ganoderan, is characterized in that in described step 1), micro-filtration film used is ceramic membrane or organic membrane.
7. the according to claim 1 pair of embrane method extracted the method for ganoderan, it is characterized in that described step 2) in ultrafiltration mould material used select a kind of in PAN, PVDF, PC, PES, PP, PE composite membrane.
8. the according to claim 1 pair of embrane method extracted the method for ganoderan, it is characterized in that in described step 3) step 2) in ultrafiltration and concentration liquid under 40～80 ℃ of conditions, to be concentrated to mycelium content be 30～70% to obtain thickened pulp.
9. the according to claim 1 pair of embrane method extracted the method for ganoderan, it is characterized in that the thickened pulp of gained in described step 3), at 60～85 ℃, dries under condition more than vacuum tightness-0.08Mpa, dry to dry thing moisture content be 3～8%, obtain.
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Cited By (7)
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| CN103897074A (en) * | 2014-03-12 | 2014-07-02 | 山东省食品发酵工业研究设计院 | Method for separating and purifying curdlan by use of membrane process |
| CN103951762A (en) * | 2014-05-23 | 2014-07-30 | 吉林农业大学 | Extraction and separation method of extracellular scleroglucan |
| CN104188889A (en) * | 2014-08-27 | 2014-12-10 | 仙芝科技(福建)股份有限公司 | Lucid ganoderma toothpaste |
| CN104262501A (en) * | 2014-09-30 | 2015-01-07 | 天津生机集团股份有限公司 | Method for extracting grifola frondosa fermentation broth polysaccharide by virtue of dual-membrane process |
| CN104788534A (en) * | 2015-05-08 | 2015-07-22 | 飞潮（无锡）过滤技术有限公司 | Ganoderan tripeptide filtering system process device |
| CN104877183A (en) * | 2015-05-29 | 2015-09-02 | 孙长顺 | Ganoderma-antradiacomphora polysaccharide and membrane permeation preparation process 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 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103897074A (en) * | 2014-03-12 | 2014-07-02 | 山东省食品发酵工业研究设计院 | Method for separating and purifying curdlan by use of membrane process |
| CN103897074B (en) * | 2014-03-12 | 2016-03-02 | 山东省食品发酵工业研究设计院 | A kind of method utilizing membrane process separation and purification curdlan |
| CN103951762A (en) * | 2014-05-23 | 2014-07-30 | 吉林农业大学 | Extraction and separation method of extracellular scleroglucan |
| CN104188889A (en) * | 2014-08-27 | 2014-12-10 | 仙芝科技(福建)股份有限公司 | Lucid ganoderma toothpaste |
| CN104188889B (en) * | 2014-08-27 | 2017-04-05 | 仙芝科技(福建)股份有限公司 | ganoderma toothpaste |
| CN104262501A (en) * | 2014-09-30 | 2015-01-07 | 天津生机集团股份有限公司 | Method for extracting grifola frondosa fermentation broth polysaccharide by virtue of dual-membrane process |
| CN104788534A (en) * | 2015-05-08 | 2015-07-22 | 飞潮（无锡）过滤技术有限公司 | Ganoderan tripeptide filtering system process device |
| CN104788534B (en) * | 2015-05-08 | 2018-06-12 | 飞潮（无锡）过滤技术有限公司 | A kind of ganoderma lucidum polysaccharide tripeptides point filter system process device |
| CN104877183A (en) * | 2015-05-29 | 2015-09-02 | 孙长顺 | Ganoderma-antradiacomphora polysaccharide and membrane permeation preparation process 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 |
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Application publication date: 20140115 |