| Agaricus blazei Murill , a kind of important medical and edible mushroom, it is rich in proteins ,polysaccharides , polyunsaturated fatty acids and dietary fibers ,it also has the effect of inhibiting tumour,depressing the blood glucose , decreasing deposition of low density cholesterol and boosting organismimmunity and so on. For a long time, deep processing on Agaricus blazei Murrill was extraction ofpolysaccharides mainly, and proteins of Agaricus blazei, fibers of Agaricus blazei and the other by-productswere not utilized fully, the results caused large losses or wastes of quality protein and fiber. In addition, thestudy and development on functional oil in edible fungus was not reported even more.Availability of Agaricusblazei Murill is low, processing depth is insufficient, lacking high technology content, lacking high addedvalue and new product with great market potential. This research is on the base of the research to the domesticand foreign edible mushroom deep processing present situation and the development tendency, fullyconsidering the raw material comprehensive utilization and the influence of processing to environment, alsocritical technique of extracting Agaricus blazei Murill specificity ingredient and function factor, each kind offactor functional characteristic and the application has conducted the thorough research, implementing rawmaterial entire use, conforming to national initiative resource conservation and the environment friendlyproduction pattern, will provide the theory basis and the practical reference and the model for Agaricus blazeiMurill and even other edible mushroom comprehensive utilizations and the profound processing.The main contents and results as follow:1.ω-6 polyunsaturated fatty acids were extracted by supercritical fluid extraction technology from Agaricusblazei Murill , through the analysis of orthogonal test, the optimum condition was determined: extractingpressure was 25 MPa, extracting temperature was 50℃, extracting time was 60 min, CO2 volume was 25L·h-1.The yields of extraction were 4.03g·(100g)-1, lipid contents in Agaricus blazei Murill was 0.01% afterextraction. This research results provided good precondition for extracting and producing Agaricus blazeiMurill peptide and other functional factors.2. Compositional characteristic of Agaricus blazei Murill fatty acid after supercritical fluid extraction was typeofω-6 polyunsaturated fatty acid, and the content of linoleic acid was 52.4%, the content ofγ-linolenic acidwas 21.12%, the content of oleic acid was 12.89%, the content of palmitic acid was 7.98%, stearic acid wasnot detected out, the total cintents ofω-6 polyunsaturated fatty acids were 73.52%,it may be as the materialfor producing health food which was Agaricus blazei Murrillω-6 polyunsaturated fatty acid(ω-6APFA).3. Sixty rats wererandomly divided into control group, model group, natural soybean phospholipids capsule(NSPC)0.70g·kg-1 group,ω-6APFA 0.28, 0.14, 0.07 g·kg-1 groups(n=10). The other groups excepting control group were given high-fat diets for 14 d, on the fourteenth the rats began to be administered orally, the controlgroup and model group were administered distilled water 10ml·kg-1 at the same volume, after 14 days ofcontinuously administration ,rats were anesthetized, the blood extracted in abdominal artery , T-CHO, TG,HDL-C, LDL-C in sera were determined. At the same time, the activity of SOD in liver and the content ofMDA was determined, the fat accumulated coefficient was calculated. 72 male mice were randomly dividedinto control group, model group, NSPC 1.0g·kg-1 group,ω-6APFA 0.4, 0.2, 0.1 g·kg-1 groups (n=12). Micewere administered continuously,16h before the last administration ,except oontrol group, the other groups wasinjected 75% yolk physiological salt solution 0.5mL through the abdominal cavity, and began to starve ,1 hafter the last administration, blood was extracted in eyeball , sera total cholesterol and triglyceride weredetermine. Compare with model group, T-CHO and TG in rat of NSPC 0.70g·kg-1 andω-6APFA 0.28 g·kg-1 allreduced and HDL-C raised obviously(P＜0.001), activity liver SOD of NSPC 0.70g·kg-1 andω-6APFA0.28,0.14 g·kg-1 obvious raised (P＜0.05, P＜0.001 and P＜0.05), MDA reduced obviously(P＜0.01, P＜0.001, P＜0.01), fat accumulation coefficient of NSPC 0.70g·kg-1 andω-6APFA reduced obviously. Comparewith model mice, T-CHO and TG in acute hyperlipemia mice of NSPC 1.0g·kg-1 andω-6APFA0.4,0.2 g·kg-1reduced obviously(P＜0.05, P＜0.01 and P＜0.05).The results indicate thatω-6APFA has the function oflowering blood fat on hyperlipemia rats and mice.4. Sixty rats were randomly divided into control group, natural soybean phospholipids capsule(NSPC)0.70g·kg-1·d-1group ,ω-6APFA 0.07, 0.14,0.28g·kg-1·d-1groups (n=12).Rats in each group were exhibitedonce a day for 7 d,1 h after the last exhibition ,rats were injected chloral hydrate 300g·kg-1 in abdominal cavityto anesthetize, the back were fixed, the carotid of the right neck were separated, and the time of thrombosisformation were determined by experimental intracorporeal thrombosis surveyor. Another 60 mice wererandomly divided into control group, NSPC 1.0g·kg-1·d-1group,ω-6APFA0.1, 0.2,0.4 g·kg-1·d-1 groups(n=12).Mice were exhibited once a day for 7 d continuously, 1h after the last exhibition, blood was extracted in theorbit vein, the clotting time were determined through glass slides method. NSPC 0.7 g·kg-1·d-1 andω-6APFA0.14,0.28g·kg-1·d-1all had obvious inhibitory effect on thrombus formation in blood stasis mice,compare with model group, P＜0.05, andω-6APFA0.14,0.28g·kg-1·d-1 were superior to NSPC 0.7g·kg-1·d-1.NSPC 1.0 g·kg-1·d-1 andω-6APFA0.2,0.4g·kg-1·d-1 all could prolong the clotting time ,compare with modelgroup, NSPC 1.0 g·kg-1·d-1 was P＜0.05,ω-6APFA0.2, 0.4g·kg-1·d-1 were P＜0.01,the result showed thatω-6APFA0.2, 0.4g·kg-1·d-1 were superior to NSPC 1.0g·kg-1·d-1 group. The results indicate thatω-6APFAhad the function of inhibiting thrombus formation, promoting blood flow ability and lowering blood viscosity.5. Used defatted Agaricus biazei Murill through Supercritical fluid extraction as material, and used themicrowave extract technology to extract the Agaricus blazei Murill polysaccharides, the extract time wasshorten substantially, proportion of fluid to solid was reduced, and the following concentration energyconsumption was decreased. When Agaricus blazei Murill active polysaccharides were extracted through microwave technique, the influence of microwave power was biggest, next was the microwave time, and thenwas the material size, the influence of proportion of solid to fluid was the smallest, the optimum condition wasdetermined: microwave power was 600W, microwave time was 6 min, the proportion of solid to fluid was1 : 10, material size was 0.147 mm. According this technology to extract Agaricus blazei Murill activepolysaccharides, the extraction rate was 14.2%.The yield was raised 29.68% compared with reported before,and the volume of water was 1/3 or 1/2 of reported before, the extractive times were 1/30 of reported before,so the water ,times and energy were saved all.6. Removed proteins and purified Agaricus blazei Murill polysaccharides by D315 resin, and geted the idealresult, rate of removing proteins is 79.26%, rate of recovering in polysaccharides is 86.3%.7. PurifIed and separated Agaricus blazei Muril polysaccharides by Sephadex G75 gelatum, and removedproteins further, controled speed of flow, the flow was 0.3mL·min-1, obtainedβ-the glucosan which molecularweight was about 100000, the purity was 85%, the polysaccharides belong to this molecular weight scope hadhigher biological activity.8. Inspected the hydrolysis of the AL proteinase, the papain, the trypsin, the FW proteinase on Agaricus blazeiMuril protein, according to proteolysis ability and the molecular weight of enzymolysis product, the ALproteinase and the FW proteinase were determined to manufacture oligopeptide.9. Hydrolyzed the Agaricus blazei Muril protein with AL proteinase and the FW proteinase though two step.The best enzymolysis condition in AL proteinase was determined: pH was 8.5, the temperature was 55℃, theenzyme to the substrate was 2%, the substrate concentration was 5%, and hydrolisis time was 2h, the yield ofpeptide was 74.7%. Then used above hydrolysis product as material to hydrolyze through the FW proteinase,and pH was 7.0, the temperature was 50℃, the enzyme to substrate was 4%, the hydrolysis time was 1.0h,peptides yield were 80.6%.10. The examinating result indicated that the content of peptides was 94.2%, the total glucose was 0.1%, thefat has not pick out, moisture content was 5.2%, ash was 0.5% in Agaricus blazei Muril oligopeptides powder.Agaricus blazei Muril oligopeptides was rich in proline, the lysine, the phenylalanine, and the essential aminoacids content reached 50.91%.11. The finally enzymolysis products were purified by Sephadex G-25 gel filtration chromatography, theywere divided into three components, the molecular weight was lower than 5600Da.12. The functional test results showed T-CHO content and the TG content of blood serum were reducedobviously in high blood fat model animal due to Agaricus blazei Muril oligopeptides (ABO), and the decreaseof HDL-C content in the high blood fat model rats were inhibited obviously by ABO, the increase of bloodserum LDL-C content in the high blood fat model rats were reduced obviously. SOD level in the animal ABOwere raised obviously, and the liver MDA content were reduced. The situation of hyperlipaemia mousebecause of high fat diet and fat cumulation in liver due to DL-E were obvious improved by ABO. The coefficient of viscera in mouse spleen and the thymus were increased obviously by ABO, inchondriosis ofmononuclear phagocyte system in mice was enhanced, and mouse's survival times in water were lengthened.The results showed ABO had functions which of lowing fat, protecting the liver, enhancing the immunity, andlessening fatigue.13. The Agaricus blazei Muril by-product which after defatted, polysaccharides were extracted andoligopeptides were produced were used as material to manufacture high quality edible mushroom dietaryfibers, comprehensive utilization value of Agaricus blazei Muril was enhanced. Material physics andchemistry characteristic were improved by extrusive technology with high temperature and high pressure, theoptimum process condition was that moisture was 80%, extrusive temperature was 145℃, rate of feed was20Kg·h-1, speed of screw rotational was 220r·min-1. Content of soluble dietary fiber(SDF)in Agaricus blazeiMuril fibers without extrusion was 2.4%, the expansive power was 4.0 mL·g-1, holding water power was 3.1g·g-1 the bound water power was 2.7g·g-1; The SDF content after extrusion was 15.1%, the expansive powerwas 18.3mL·g-1, holding water power was 9.4 g·g-1, bounding water power was 6.4 g·g-1, and became highquality Agaricus blazei dietary fibers (HQADF).14. The normal mice and constipation model mice were selected and fed with HQADE The propellingfunction of HQADF to the gastrointestinal vermicular motion was monitored. The influence of the HQADF tothe defecation frequency and the time of the two group mice were detected. Results : the normal mice'sgastrointestinal vermicular motion was promoted obviously in dose of 3.50, 1.75g·kg-1, their defecationfrequency was improved and defecation time was shorted. The constipation mice's defecation frequency wasevidently improved and the most efficient dose was 3.5g·kg-1. The results showed that HQADF hadconspicuously anti-constipation function.15. The selected 50 mice were randomly divided into control group, alloxan model group, HQADF 3.50,1.75,0.88g·kg-1 dose group(n=10). Mice in each group were exhibited once a day for 15 days , on the sixthday ,mice in every group were injected alloxan physiological salt solution 80mg·kg-1 in caudal vena exceptcontrol group, mice in control group were injected physiological salt solution that is equal volume. 16 hbefore the last exhibition, mice in every group were starved.1h before the last exhibition, extracted blood inwhere eyeballs, separated sera and the contents of blood sugar were determined by auto biochemistry analyzer,mice were killed at once. The liver samples were obtained and the contents of liver glycogen were determined.Another 60 mice were divided into 5 groups with the method mentioned above. Each group was exhibitedonce day for 10 days continuously, 1h after the last exhibition, except control group, the others were injectedadrenalin hydrochloride 250μg·kg-1 through abdominal cavity, after 30 min, blood was extracted bydecapfitation, and the contents of blood sugar and the contents of liver glycogen were determined all by autobiochemistry analyzer. Results: the HQADF 3.50g·kg-1 had obvious inhibitory effect on the raise of bloodsugar in diabetes mice ( model group ) (P＜0.05 ) , it had obvious action on reducing the contents of liver glycogen in diabetes mice ( model group ) (P＜0.05 ) ,and 3.50g·kg-1 was superior to 1.75 g·kg-1 . TheHQADF with different dose had obvious action on inhibiting the raise of blood sugar in adrenine mice( model group ) (P＜0.001, P＜0.01, P＜0.05 ) ,and had obvious action on increasing the reduction of liverglycogen on adrenine mice ( model group ) ,and 3.50g·kg-1 was superior to 1.75g·kg-1. The results showedthat HQADF had the function of falling blood sugar in alloxan diabetes mice and hyperglycaemia miceinduced by adrenine.16. The selected 80 mice (half male, half female) were randomly divided into 5 groups: control group,adiposity model group, HQADF 3.50,1.75,0.88g·kg-1 dose group. Each group except the control group wasexhibited once a day for 35 days and was weighed once every 7 days. Another 80 mice (half male, half female)were grouped, fed and weighed in the same way as the above, 24 hours after the last exhibition, the mice werekilled, abdominal fat was weighed and the fat index number was calculated. Another 80 mice (half male, halffemale) were grouped, fed and weighed in the same way as the above, 24 hours after the last exhibition, theireyeballs were removed and the blood was extracted. Its biochemical indexes were determined byHITACHI7080 auto analyzer. The results show that: the HQADF can obviously reduce the adiposity mice'sweight, fat Index, the content of TC, TG, LDL-C and GLU, and improve their HDL-C in blood serum, soHQADF had great functions of losing weight and fat.17. The Agaricus blazei dietary fibers (ADF) through suitable processing can improve the bread quality. Theideal way was that extracted polysaccharide by microwave after supercritical fluid extraction or purify, thenoligopeptides were manufactured, the by-products were extruded. When the HQADF size was 0.175mm, theHQADF amount was 15%, the bread quality was the best, and the amount of bread additive was the least(1.0%). Because exrusion processing improved ADF quality, not only can add a great quantity dietarymushroom fibers in the dough, but also provided special health function for bread, in addition, the fiber sizewas not the smaller the better, so the energy consumption because mill fiber powder was reduced. |
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