| Polysaccharides are the key medicinal components from Ganoderma lingzhi,and?-glucan is the most active ingredients of G.lingzhi polysaccharide,which have various pharmacological effects.At present,?-glucan from G.lingzhi was seldom reported.In this paper,?-glucan from G.lingzhi as the research objiect by making a comparative study on extraction technology of ?-glucans from G.lingzhi fruiting bodies and mycelia,and the preparation technology of ?-glucan from G.lingzhi was studied.This provides the technical foundation for further study on its physiological activity.The main results are as follows:1.?-glucans from G.lingzhi fruiting bodies were extracted using ethanol-enzyme pretreatment combined with the typical water extraction method.Based on the ethanol-enzyme pretreatment system was optimized,results showed that the key parameters of ethanol-enzyme pretreatment system were 60%of ethanol(v/v),1.5%of enzyme(m/v,g/mL),45? of enzymolysis temperature and 8.0 of enzymolysis pH.2.Based on the ethanol-enzyme pretreatment method,testing the process of ultrasonic assisted extraction of ?-glucans from G.lingzhi fruiting bodies.Determination of ultrasound power(A),ratio of water to material(B)and ultrasound time(C)as the key factors of ultrasonic extraction technology of ?-glucans from G.lingzhi fruiting bodies by single factor test.And ultrasonic extraction technology of ?-glucans from G.lingzhi fruiting bodies were further optimized by Box-Behnken test design,and the regression model which can predict yield of P-glucan from G.lingzhi fruiting bodies was obtained:Y2=+0.50+0.009A+0.016B+0.023C-0.00175AB-0.014AC-0.045BC-0.058A2-11B2-0.078C2.The optimal levels of the three factors were ultrasound power 360W,ratio of water to material 25:1(v/m,mL/g)and ultrasound time 51 min.Under these conditions,the yield of?-glucan from G.lingzhi fruiting bodies was up to 0.500(mg/g).The validation tests showed that the result predicted by the model was similar to the actual extraction results,so the model can be used in practical application.Based on the above experiment,comparing the ethanol-enzyme pretreatment combined with typical water extraction method,ultrasonic extraction method and microwave assisted extraction method.The results showed that the yield of ?-glucan from G.lingzhi fruiting bodies by ultrasonic extraction method was the highest.3.The traditional ethanol reflux pretreatment method is suitable of ultrasonic extraction technology of ?-glucan from G.lingzhi mycelia.Determination of ultrasound power(A),ratio of water to material(B)and ultrasound time(C)as the key factors of ultrasonic extraction technology of(3-glucans from G.lingzhi mycelia by single factor test.And ultrasonic extraction technology of(3-glucans from G.lingzhi mycelia were further optimized by Box-Behnken test design,and the regression model which can predict yield of ?-glucan from G.lingzhi mycelia was obtained:Y3= +3.77+0.17A+0.18B+0.015C-0.020 AB+0.070AC-0.020BC-0.51A2-0.30B2-0.20C2.The optimal levels of the three factors were ultrasound power 248W,ratio of water to material 26:1(v/m,mL/g)and ultrasound time 60min.Under these conditions,the yield of ?-glucan from G.lingzhi mycelia was up to 3.77(mg/g).The validation tests showed that the result predicted by the model was similar to the actual extraction results,so the model can be used in practical application.In addition,based on the above experiment,comparing the traditional ethanol reflux pretreatment combined with typical water extraction method,ultrasonic extraction method and microwave assisted extraction method.The results showed that the yield of ?-glucan from G.lingzhi mycelia by ultrasonic extraction method was the highest.4.Based on the amyloglucosidase treatment was optimized,results showed that the key parameters of amyloglucosidase treatment were 1.5%of enzyme amount(m/v,g/mL)(amyloglucosidase),50 ? of enzymolysis temperature,4.0 of enzymolysis pH and 50min of enzymolysis time.Trypsin-sevag method was used as the best method to de-protein of?-glucan from G.lingzhi.Deproteinization rate was 84.35%,and retention rate of ?-glucan was 81.53%.The D301G macroporous adsorption resin was the best decoloring agent.And the key parameters of decolorization of ?-glucan from G.lingzhi were further optimized by single factor test and orthogonal test.The optimum decoloring conditions were:resin amount 3.0g,temperature 50?,speed 160r/min,time 3.5h,pH 5.0.The decolorization rate obtained in the experiment was 68.35%,and retention rate of ?-glucan was 84.67%.5.The crude ?-glucan from G.lingzhi was prepared by amyloglucosidase treatment,deproteinization,decoloration and dialysis desaiting,then the products of ?-glucan from G.lingzhi is a pale yellow powder,dissolve in water and dissolve easily in hot water.The yield of products to the crude ?-glucan from G.lingzhi was 31.07%,and the yield of products to the original G.lingzhi mycelia was 5.00%.And the yield of ?-glucan of the products of ?-glucan from G.lingzhi was 4.94%,compared with the yield of ?-glucan from the original G.lingzhi mycelia(0.376%,3.76mg/g),?-glucan was effective concentration. |
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