CN102120779B - Efficient extraction technology of ganodorma lucidum fermention mycelium homogeneous polysaccharides - Google Patents
Efficient extraction technology of ganodorma lucidum fermention mycelium homogeneous polysaccharides Download PDFInfo
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Abstract
The invention provides an efficient extraction technology of ganodorma lucidum fermention mycelium homogeneous polysaccharides. The method comprises the following steps: stirring ganodorma lucidum fermentation liquor to boil and extract without separating mycelium; filtering the obtained fermentation liquor to separate out mycelium, performing ultrafiltration separation to the separated filtrate to obtain ganodorma lucidum polysaccharides, adding water in the separated mycelium to pulp, adding water in the obtained slurry to stirr, boil and extract, centrifuging the extracting solution to obtain supernatant, performing ultrafiltration separation to the supernatant to separate out ganodorma lucidum polysaccharides; and combining the ganodorma lucidum polysaccharides obtained from the filtrate and the ganodorma lucidum polysaccharides obtained from the mycelium, and concentrating and drying the polysaccharides to obtain the product. By adopting the method provided by the invention to extract ganodorma lucidum polysaccharides, the dry matter extraction rate is more than 40%, the polysaccharide product extraction rate is 89% and the polysaccharide content is more than 30%; and if primary alcohol precipitation is performed, the polysaccharide content is up to more than 50%.
Description
Technical field
The present invention relates to the extraction process of polysaccharide, be specifically related to fermented hyphostroma of Ganoderma homogeneous polysaccharide high efficiency extraction technology.
Background technology
Glossy ganoderma is the general name of red sesame G.lucidumkarst of Basidiomycetes polyporaceae (Polyporaceae) ganoderma (Ganodelma) fungi and purple sesame G.japonicrn L loyd; Claiming Ganoderma lucidum seu Japonicum, refreshing sesame, sesame grass, celestial grass, auspicious grass again, is the complete stool of red sesame of polyporaceae plant or purple sesame.Glossy ganoderma originates in east Asia, and China distributes the widest in Jiangxi, and glossy ganoderma is treasured medicinal material as the Chinese tradition that has thousands of years medicinal histories; Possess very high pharmaceutical use, through the modern pharmacology research confirmation of scientific research institution's many decades, glossy ganoderma is for enhancing human immune; Blood sugar regulation, controlling blood pressure, auxiliary tumor chemoradiotherapy; Liver protecting, aspects such as promote sleep all have significant curative effect.
The chemical ingredients of glossy ganoderma is comparatively complicated, and because of used bacterial classification, the bacterial classification place of production, cultivating method, extraction process, formulation method different different.Contain polyose, ucleosides, furan derivative, steroid ferment class, alkaloids, protein, polypeptide, amino acids, triterpenes, sesquiterpene, organic germanium, inorganic salt etc. in the sporophore of glossy ganoderma, mycelium and the spore.
Ganoderan is one of main effective constituent of glossy ganoderma, has antitumor, immunomodulatory, hypoglycemic, anti-oxidant, reducing blood-fat and anti-aging effects.
Ganoderan is made up of three strands of monose chains, and its configuration and DNA, RNA are similar, are a kind of spirrillum steric configuration things, mainly fix with hydrogen bond between the spiral layers.There is significant difference in ganoderan aspect some physico-chemical property such as monose composition, glycosidic link configuration, molecular weight, specific rotation, solubleness.Have in more than the 200 kind of ganoderan what extracted, great majority are different polysaccharide, promptly except that containing glucose, also contain other monose such as semi-lactosi, seminose, pectinose, wood sugar, Fucose, rhamnosyl; Because the source is different, molecular weight can not wait from tens thousand of to hundreds thousand of, and most with branch, the part polysaccharide also contains peptide chain.Scarcely be dissolved in high concentration ethanol, and be dissolved in hot water.Temperature is too high, can cause the degraded of ganoderan, glycosidic link easy fracture in the polysaccharide body and form a plurality of sugar monomers, and sugar monomer be do not have active.
Traditionally, glossy ganoderma is to obtain through field acquisition or artificial culture, but glossy ganoderma wild resource shortage, the tame cycle is long, and floor space is big, receives season limit again, thereby has influenced making full use of of glossy ganoderma.Adopt the ganoderma lucidum liquid fermentation culture method to have short, advantage such as cost is low, output is big of cycle.But extract polysaccharide all is to adopt Ganoderma fermentation liquid, filters out Ganoderma mycelium, after the drying, is carrying out hot water extraction, the fermentation filtered liq passes into disuse, and does not perhaps separate polysaccharide, and extraction efficiency is low, wastes time and energy.In addition, adopt water boiling and extraction ethanol sedimentation method, consuming time longer, can't isolate the ganoderan that molecular weight is concentrated, extract is main with Crude polysaccharides.Influence finished product polysaccharide content and drug effect.Except containing needed macromolecular polysaccharide, also contain small molecules dextrin in a series of micromolecular polysaccharides, the substratum etc. in Ganoderma fermentation liquid and the mycelium, the existence of these materials has certain influence to product quality.
Summary of the invention
The objective of the invention is defective, a kind of fermented hyphostroma of Ganoderma polysaccharide extracting process is provided to prior art.
The present invention is primarily aimed at fermented hyphostroma of Ganoderma extracting effective components ganoderan, in fermentation, just carries out the first time and extracts, and separates with ultrafiltration process then.Save the energy, shorten man-hour, and content improves, and obtains the polysaccharide that molecular weight is concentrated, and curative effect is high.The whole process of ultrafiltration membrance filter method carries out in closed system, need not heating, avoids and alleviated the influence to the ganoderan nutritive ingredient of heat and oxygen.
Fermented hyphostroma of Ganoderma polysaccharide extracting process provided by the invention comprises the following steps:
1) Ganoderma fermentation liquid is not being separated stirring decoction extraction under the mycelial situation.
Preferable, stirring decocts the method for extracting and does, is heated to boiling, is incubated 1.4-1.6 hour.Stirring velocity preferred 58-62 time/minute.
Further, heating and extracting is directly carried out in fermentor tank.
2) with the filtering fermentation liquor separation of mycelial after the heating and extracting; Filtrating ultra-filtration and separation after the separation goes out ganoderan; After mycelium after the separation added the water mill slurry, slurries added water and stir the decoction extraction, extracting solution centrifuging and taking supernatant, and the supernatant ultra-filtration and separation goes out ganoderan.
Preferable, slurries stir and decoct the method for extracting is that the water that adding mycelium dry weight 5.8-6.2 doubly measures is heated to boiling, is incubated 1.4-1.6 hour.Stirring velocity preferred 58-62 time/minute.
Preferable, the used ultra-filtration membrane of ultrafiltration is selected the ultra-filtration membrane of ability molecular weight cut-off greater than 6000D, is 0.05 micron-0.1 micron ultra-filtration membrane like the aperture.
Repeatedly loop ultrafiltration is essentially 0 until the dialyzate soluble solids.
Ganoderan that 3) will from filtrating, obtain and the ganoderan that from mycelium, obtains merge, and concentrated after drying obtains product.
Concentrate and to adopt vacuum concentration, vacuum tightness 0.07-0.09MPA, temperature 60-85 degree centigrade.Preferably, control vacuum tightness is between the 0.08-0.09MPa, between the temperature 65-85 ℃.
Drying means can be spraying drying, and inlet temperature 172-180 ℃, temperature out 67-75 ℃.
Further, in step 3), after the enrichment step, can increase the alcohol precipitation treatment step, can further improve the polysaccharide content of finished product.Alcohol precipitation can adopt routine, is the ethanol of 90--95% (concentration of volume percent) as adding concentration, triplication, and the post precipitation throw out adds the liquid concentrator that hot water is adjusted to proportion 1.15, spraying drying.
Method of the present invention is a combining with fermentation technology, at fermentation termination, because nitrogenous source in the substratum and carbon source organism; Digested by Ganoderma, be converted into the small molecules metabolite, consider that fermented liquid still has the ability of dissolving ganoderan; After killing Ganderma lucidum, directly heat up and extract the polysaccharide in the mycelium, the polysaccharide in the Ganoderma lucidum mycelium somatocyte is discharged in the fermented liquid; Refilter fermented liquid, fermented liquid separates through membrane filtration again, removes small-molecule substance residual in the fermented liquid respectively.The polysaccharide of molecular weight cut-off more than 6000.The mycelium of spinning extracts lucidum mycelium polysaccharide adopting hot water to decoct process for extracting, and extracting solution is being isolated intracellular polyse through membrane filtration.Two portions polysaccharide merges, and gets the lucidum mycelium polysaccharide of content 30% after the drying.Improve yield and ganoderma polyoses content, reduced production cost, improved the effect of ganoderan.
Adopt method of the present invention to extract ganoderan, the soluble solids extraction yield is more than 40%, and detecting polysaccharide product extraction yield is 89%, and polysaccharide content is handled if carry out an alcohol precipitation process more than 30%, and polysaccharide content reaches more than 50%.
Description of drawings
Fig. 1, the lucidum mycelium polysaccharide technological process of production
Fig. 2, fermented liquid and extracting solution ultrafiltration schematic flow sheet
Embodiment
Below enumerate embodiment with further elaboration the present invention, should be understood that instance is not to be used to limit protection scope of the present invention.
Embodiment 1
The acquisition of Ganoderma fermentation liquid: reference " the 3rd academic nd Annual Meeting selected works of CCOA (volume two) " 2004, the method for the research of ganoderma lucidum mycelium liquid layer fermentation technique (Zu Liyawusong mound, Cheng Shu peak Tang Fang) record obtains.
The ganoderan process for extracting:
1. fermentation post: fermentation termination heats up, and reaches boiling, stirs (60 times/minute) insulation after 1.5 hours, reach extract purpose after, Plate Filtration, filtered liq changes the ultra-filtration membrane post with the smart filter of tubular-bowl centrifuge, the mycelium filter residue is gone round and round a millstone and is starched the post.
2.1 adopt the ultra-filtration membrane equipment of molecular weight cut-off greater than 6000D, start ultra-filtration membrane equipment, be controlled at into film pressure 0.15-0.20MPA; Temperature be controlled at the 25-45 degree/℃; Liquid flow rate 70-90L/ minute, detecting the dialyzate solid substance almost nil was terminal point, and the time is approximately 4-6 hour.The polysaccharide dope changes the vacuum concentration post.
2.2 the mycelium filter residue adds suitable quantity of water (add entry 30-50%, press mycelium dry weight and calculate), stainless steel paste mill grinding; Reach the 100-200 order, put into extractor and add six times of water gagings, be heated to more than 95 degree; Stir (60 times/minute) and decoct extraction 1.5 hours, whizzer separates, and parting liquid send the ultra-filtration membrane post; By behind the 2.1 method ultrafiltration membrance filters, the polysaccharide dope changes the vacuum concentration post.
3 ultrafiltration membrance filter liquid concentration: open vacuum pump and recirculated water cooling pump; When treating that vacuum tightness rises to 0.04MPa, in vaporizer, add extracting solution to liquid level, continue to vacuumize at following visor middle part; When treating vacuum tightness to 0.08MPa; Open steam cock, control vacuum tightness keeps liquid level in the visor scope between the 0.08-0.09MPa, between the temperature 65-85 ℃.When liquid concentration to proportion is 1.05-1.10 (60 ℃), filter fast in (aperture 80 orders), filtrating is put in the spraying drying basin, uses in order to spraying drying.
4 spraying dryings
EAT is controlled at 175 ± 3 ℃, and air outlet temperature is controlled at 70 ± 3 ℃.Collect ganoderan extract and pack in the PE bag, weigh, every barrel is no more than the 25kg packing.
Polysaccharide detection method: with reference to " the ganoderan detection method under 2010 editions glossy ganoderma items of Chinese pharmacopoeia
Calculate the polysaccharide content of efficiency of pcr product and unit extract by following formula:
Detecting polysaccharide product extraction yield is 89%, and the product polysaccharide content is 30%.Handle if before dry, increase an alcohol precipitation process, the product polysaccharide content reaches more than 50%.
Claims (9)
1. a fermented hyphostroma of Ganoderma extraction method of polysaccharides comprises the following steps:
1) Ganoderma fermentation liquid is not being separated stirring decoction extraction under the mycelial situation, when stirring the decoction extraction, the method for decoction is: be heated to boiling, be incubated 1.4-1.6 hour;
2) will stir the filtering fermentation liquor separation of mycelial that decocts after extracting; Filtrating ultra-filtration and separation after the separation goes out ganoderan; After mycelium after the separation added the water mill slurry, slurries added water and stir the decoction extraction, extracting solution centrifuging and taking supernatant, and the supernatant ultra-filtration and separation goes out ganoderan;
Ganoderan that 3) will from filtrating, obtain and the ganoderan that from mycelium, obtains merge, and concentrated after drying obtains product.
2. fermented hyphostroma of Ganoderma extraction method of polysaccharides according to claim 1 is characterized in that, in the step 1), stirs and decocts when extracting, and stirring velocity is 58-62 time/minute.
3. fermented hyphostroma of Ganoderma extraction method of polysaccharides according to claim 1 is characterized in that step 2) in, slurries stir and decoct when extracting, and the method for decoction is: add the water that mycelium dry weight 5.8-6.2 doubly measures, be heated to boiling, be incubated 1.4-1.6 hour.
4. fermented hyphostroma of Ganoderma extraction method of polysaccharides according to claim 1 is characterized in that step 2) in, slurries stir and decoct when extracting, and stirring velocity is 58-62 time/minute.
5. fermented hyphostroma of Ganoderma extraction method of polysaccharides according to claim 1 is characterized in that step 2) in, the used ultra-filtration membrane of ultrafiltration select can molecular weight cut-off greater than the ultra-filtration membrane of the material of 6000D.
6. like the said fermented hyphostroma of Ganoderma extraction method of polysaccharides of claim 5, it is characterized in that used ultra-filtration membrane aperture is 0.05 micron-0.1 micron.
7. fermented hyphostroma of Ganoderma extraction method of polysaccharides according to claim 1 is characterized in that step 2) in, repeatedly loop ultrafiltration does not contain soluble solids basically until dialyzate.
8. fermented hyphostroma of Ganoderma extraction method of polysaccharides according to claim 1 is characterized in that, after step 3) concentrates, increases the alcohol precipitation treatment step.
9. fermented hyphostroma of Ganoderma extraction method of polysaccharides according to claim 1 is characterized in that, in the step 3), and simmer down to vacuum concentration, vacuum tightness 0.07-0.09MPa, temperature 60-85 degree centigrade; Drying means is a spraying drying, inlet temperature 172-180 ℃, and temperature out 67-75 ℃.
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| CN102827304A (en) * | 2012-09-17 | 2012-12-19 | 江苏神华药业有限公司 | Method for extracting trametes versicolor exopolysaccharide through two-membrane process |
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| CN102676609B (en) * | 2011-12-27 | 2014-04-09 | 西北农林科技大学 | Method for efficiently preparing inhomogenous Marasmius androsaceus polysaccharide through membrane separation technology |
| TWI543765B (en) * | 2015-05-25 | 2016-08-01 | 長庚生物科技股份有限公司 | Method to prepare ganoderma lucidum polysaccharides possessing anti-obesity properties |
| CN104945528A (en) * | 2015-07-08 | 2015-09-30 | 广州白云山汉方现代药业有限公司 | Ganoderma lucidum spores polysaccharides preparing method |
| CN110922495A (en) * | 2019-12-14 | 2020-03-27 | 安徽金寨乔康药业有限公司 | Method for preparing antioxidant active polysaccharide by using oak culture medium |
| CN111995693A (en) * | 2020-07-28 | 2020-11-27 | 安徽利民生物科技股份有限公司 | Extraction method of solid-state fermentation ganoderma lucidum polysaccharide |
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