桑黄生物学特性及主要活性成分研究
摘要
本文以开发利用桑黄这一新兴的药用菌物资源，探明其菌丝体生长条件、活性成分为目的，对桑黄的主要生物学特性进行测定，对其主要活性成分多糖进行分析，以正交试验确定桑黄菌丝体多糖的提取工艺，为桑黄的活性成分研究、开发新药及人工栽培桑黄子实体提供了科学依据。
药用真菌桑黄(Phellinus igniarius)的最新药理活性研究发现其在抗肿瘤方面具有巨大的潜力，其菌丝体多糖含量为3.75mg/g干菌丝体。但由于对其菌丝体的生长条件还没有完全掌握，子实体的栽培技术还未成功，致使对桑黄各方面的研究进展很慢，阻碍了这一新兴中药资源的开发进度。
为了最大限度地保存菌种的活力，以提高菌丝体的质量及菌丝体内活性成分的累积，本文通过对比研究，进一步对其生长基质进行筛选，明确了两种适于桑黄菌丝生长的固体培养基：以马铃薯为氮源、蔗糖为碳源的培养基较适用于菌丝收集，以蛋白胨为氮源、可溶性淀粉为碳源的培养基较适用于菌种的保藏。同时通过对不同生长时期菌丝体形态的观察描绘，为桑黄的品种鉴定提供依据，可避免因品种混杂而影响桑黄的生物活性。
为了掌握液体培养桑黄菌丝体对环境因子的生物利用度和对活性酶的生产能力，本文首次对桑黄液体培养菌丝体的八种胞外酶的活性进行了测定，培养天数为20d，隔天取样，一次性测定各种胞外酶的活性。测定结果表明桑黄具有完整的胞外酶系，具备人工栽培桑黄子实体所需的条件。淀粉酶、CMC酶、纤维素酶、漆酶、愈创木酚氧化酶、多酚氧化酶的活性高峰分别出现在第10d、第12d、第12d、第14d、第16d、第16d，说明桑黄对淀粉类物质、纤维素类、木质素类物质均具有降解能力。培养液中的蛋白质及还原糖的高峰期分别出现在第14d和第16d，表明蛋白质的收集宜在第14d进行，还原糖的收集宜在第16d进行。
本文对液体培养桑黄菌丝体的主要活性成分多糖进行了研究，首次利用lg9(3~4)正交试验法对桑黄菌丝体多糖的提取工艺进行研究，从级差的大小可以看出影响桑黄菌丝体粗多糖提取率的因素主要是浸提次数，其次分别是浸提时间、
吉林农业大学硕士学位论文
桑黄主要生物学特性及多糖的研究
浸提温度、浸提比。确定最佳工艺组合为浸提比为1:50，90℃提取180而n，浸
提3次。本实验采用蛋白酶法、三氛乙酸法及Sevag法结合的方法脱蛋白，最后
粗多糖中蛋白质含量低于4%，表明该方法高效可行且节约资源。
首次对菌丝体的石油醚提取物进行定性分析，确定该菌丝体中含有二丁基夯
基甲苯、棕桐酸、9，12一十八碳二烯酸甲醋，二十碳烷及二十六碳烷，这五种物质
均为在桑黄菌丝体中首次发现。
本论文以通过固体培养及液体发酵培养获得的桑黄菌丝体为主要研究对象，
对其主要生物学特性，即菌丝体的生长及胞外酶活力进行了研究，为菌丝体的液
体发酵生产及子实体栽培莫定了基拙;同时对菌丝体的多糖及醚溶性成分进行了
研究，为开发利用桑黄这一药用菌物资源及开发新药提供了科学依据，使其能尽
早应用于医疗及保健品事业中。
In order to devolping and utilizing Phellinus igniarius resources, the main biologic characteristics and polysaccharide of Phellinus igniarius were studied. Meanwhile the optimum method suitable for mass production of polysacchride was established. The basis data on studying active compositions, developing new medicine and culturing fruit bodies from mycelia of Phellinus igniarius were provided.
Lately research indicated that Phellinus igniarius had the activity of antitumor and the polysaccharide content of mycelia was 37.5%. But because of having not mastered the growth conditions of mycelia and not succeeded in culturing fruit bodies, it was retarded the pace of developing and utilizing the new medical source.
The biological characteristics of mycelia from Phellinus igniarius and culture media were studied. Two kinds of culture media were suitable for the growth of mycelia. The result indicated that the culture medium with potato as nitrogen source and saccharose as carbon source was suitable for collecting mycelia, and the culture medium with peptone as nitrogen source and solvable amylum as carbon source was suitable for conservation. Meanwhile the depiction of mycelia could contribute the identification among species.
Eight extracellular anzymes of mycelia from Phellinus igniarius cultured by shaking were studied for the first time. The extracellular anzymes were measured within 20 days. The result showed that the fruit bodies of Phellinus igniarius could be cultured because of having the whole enzymic system. The peak times of amylase, CMC enzyme,
cellulose, laccase, guaiacol oxidase and polyphenol oxidase were the 10th day, the 12th day, the 12th day, the 14th day, the 16th day and the 16th day respectively. It indicated that Phellinus igniarius has the capability of discomposing amylum, cellulose and lignose. The peak time of protein and reductic saccharide were the 14th and the 16th. It showed that optimum time of collecting protein and reductic saccharide were the 14th day and the 16th day respectively.
Polysaccharide of mycelia from Phellinus igniarius cultured by shaking were studied. Lg9(34) orthogonal experiment were carried out to find the optimum method of extracting polysaccharide from mycelia. The magnitude of range indicated that the factors which effected extracting ratio were times, time, temperature and proportion. The optimum condition was: proportion 1:50, temperature 90, time 180min and two times. A mixed method composed of proteinase, trichloroactic acid and Sevag was established which had high efficiency and spared sources. It could make the content of protein in polysaccharide under 4%.
A substance extracted from mycelia with benzinum was studied for the first time. Five compositions were found: butylated hydroxytoluene, n-hexadcanoic acid, 9,12-octadecadienoic acid methyl ester , eicosane and hexacosane.
The paper provided scientific basis data for researching mainly biologic characteristics and active compositions of mycelia from Phellinus igniarius cultured with solid and liquid culture media. It provided information in cultivation of its mycelia and fruit body. Also it had practical significance for accelerating the speed of research, creating economic and social benefits..
药用真菌桑黄(Phellinus igniarius)的最新药理活性研究发现其在抗肿瘤方面具有巨大的潜力，其菌丝体多糖含量为3.75mg/g干菌丝体。但由于对其菌丝体的生长条件还没有完全掌握，子实体的栽培技术还未成功，致使对桑黄各方面的研究进展很慢，阻碍了这一新兴中药资源的开发进度。
为了最大限度地保存菌种的活力，以提高菌丝体的质量及菌丝体内活性成分的累积，本文通过对比研究，进一步对其生长基质进行筛选，明确了两种适于桑黄菌丝生长的固体培养基：以马铃薯为氮源、蔗糖为碳源的培养基较适用于菌丝收集，以蛋白胨为氮源、可溶性淀粉为碳源的培养基较适用于菌种的保藏。同时通过对不同生长时期菌丝体形态的观察描绘，为桑黄的品种鉴定提供依据，可避免因品种混杂而影响桑黄的生物活性。
为了掌握液体培养桑黄菌丝体对环境因子的生物利用度和对活性酶的生产能力，本文首次对桑黄液体培养菌丝体的八种胞外酶的活性进行了测定，培养天数为20d，隔天取样，一次性测定各种胞外酶的活性。测定结果表明桑黄具有完整的胞外酶系，具备人工栽培桑黄子实体所需的条件。淀粉酶、CMC酶、纤维素酶、漆酶、愈创木酚氧化酶、多酚氧化酶的活性高峰分别出现在第10d、第12d、第12d、第14d、第16d、第16d，说明桑黄对淀粉类物质、纤维素类、木质素类物质均具有降解能力。培养液中的蛋白质及还原糖的高峰期分别出现在第14d和第16d，表明蛋白质的收集宜在第14d进行，还原糖的收集宜在第16d进行。
本文对液体培养桑黄菌丝体的主要活性成分多糖进行了研究，首次利用lg9(3~4)正交试验法对桑黄菌丝体多糖的提取工艺进行研究，从级差的大小可以看出影响桑黄菌丝体粗多糖提取率的因素主要是浸提次数，其次分别是浸提时间、
吉林农业大学硕士学位论文
桑黄主要生物学特性及多糖的研究
浸提温度、浸提比。确定最佳工艺组合为浸提比为1:50，90℃提取180而n，浸
提3次。本实验采用蛋白酶法、三氛乙酸法及Sevag法结合的方法脱蛋白，最后
粗多糖中蛋白质含量低于4%，表明该方法高效可行且节约资源。
首次对菌丝体的石油醚提取物进行定性分析，确定该菌丝体中含有二丁基夯
基甲苯、棕桐酸、9，12一十八碳二烯酸甲醋，二十碳烷及二十六碳烷，这五种物质
均为在桑黄菌丝体中首次发现。
本论文以通过固体培养及液体发酵培养获得的桑黄菌丝体为主要研究对象，
对其主要生物学特性，即菌丝体的生长及胞外酶活力进行了研究，为菌丝体的液
体发酵生产及子实体栽培莫定了基拙;同时对菌丝体的多糖及醚溶性成分进行了
研究，为开发利用桑黄这一药用菌物资源及开发新药提供了科学依据，使其能尽
早应用于医疗及保健品事业中。
In order to devolping and utilizing Phellinus igniarius resources, the main biologic characteristics and polysaccharide of Phellinus igniarius were studied. Meanwhile the optimum method suitable for mass production of polysacchride was established. The basis data on studying active compositions, developing new medicine and culturing fruit bodies from mycelia of Phellinus igniarius were provided.
Lately research indicated that Phellinus igniarius had the activity of antitumor and the polysaccharide content of mycelia was 37.5%. But because of having not mastered the growth conditions of mycelia and not succeeded in culturing fruit bodies, it was retarded the pace of developing and utilizing the new medical source.
The biological characteristics of mycelia from Phellinus igniarius and culture media were studied. Two kinds of culture media were suitable for the growth of mycelia. The result indicated that the culture medium with potato as nitrogen source and saccharose as carbon source was suitable for collecting mycelia, and the culture medium with peptone as nitrogen source and solvable amylum as carbon source was suitable for conservation. Meanwhile the depiction of mycelia could contribute the identification among species.
Eight extracellular anzymes of mycelia from Phellinus igniarius cultured by shaking were studied for the first time. The extracellular anzymes were measured within 20 days. The result showed that the fruit bodies of Phellinus igniarius could be cultured because of having the whole enzymic system. The peak times of amylase, CMC enzyme,
cellulose, laccase, guaiacol oxidase and polyphenol oxidase were the 10th day, the 12th day, the 12th day, the 14th day, the 16th day and the 16th day respectively. It indicated that Phellinus igniarius has the capability of discomposing amylum, cellulose and lignose. The peak time of protein and reductic saccharide were the 14th and the 16th. It showed that optimum time of collecting protein and reductic saccharide were the 14th day and the 16th day respectively.
Polysaccharide of mycelia from Phellinus igniarius cultured by shaking were studied. Lg9(34) orthogonal experiment were carried out to find the optimum method of extracting polysaccharide from mycelia. The magnitude of range indicated that the factors which effected extracting ratio were times, time, temperature and proportion. The optimum condition was: proportion 1:50, temperature 90, time 180min and two times. A mixed method composed of proteinase, trichloroactic acid and Sevag was established which had high efficiency and spared sources. It could make the content of protein in polysaccharide under 4%.
A substance extracted from mycelia with benzinum was studied for the first time. Five compositions were found: butylated hydroxytoluene, n-hexadcanoic acid, 9,12-octadecadienoic acid methyl ester , eicosane and hexacosane.
The paper provided scientific basis data for researching mainly biologic characteristics and active compositions of mycelia from Phellinus igniarius cultured with solid and liquid culture media. It provided information in cultivation of its mycelia and fruit body. Also it had practical significance for accelerating the speed of research, creating economic and social benefits..
引文
[1] 黄年来．中国食用菌百科．农业出版社[M]，1993：53
[2] Lee,S.Y. Characterization and production of antitumor polysaccharide from basidiomycetes[J]. Biotech. News,1996,3:399～403
[3] Mizinu,T., Wsaa, T. Antituuor active polysaccharide isolated from the fruiting body of Hercium erinaceum, an edible and medicinal mushroom called yamabushitake or houtou[J]. Biosci.Biotech. Biochem. 1992,56:347～348
[4] Shon,Y.H., Nam, K.S. Antimutagencity and induction of anti-carcinogenic phase·enzymes by basidiomycetes[J]. J. Ethonpharm.2001,77:103～109
[5] 杨新美．中国食用菌栽培学．北京：农业出版社[M]，1989：92～113
[6] 张树庭，Miles P.G.著．杨国良，张金霞，秦永生等译．食用菌及其栽培[M]．保定：河北大学出版社，1992：54～60
[7] 张雪岳编著．食用菌学[M]．重庆大学出版社，1988：72
[8] 杨新美主编．食用菌研究法[M]．北京：中国农业出版社，1998：237～240
[9] 陈立国，杨新美．黑木耳Auricularia auricular(L.Ex Hook)Underw纤维素酶和酚氧化酶的初步研究．武汉：华中农业大学硕士学位论文，1981
[10] 陈方顺，于青，林立仙等．木屑栽培香菇胞外纤维素酶和磷酸二酯酶的活性变化[J]．食用菌学报，1999，6(3)：8～12
[11] 尉晓宇．食用菌酶学的应用研究[J]．中国食用菌，1990，9(5)：15～16
[12] 村尾泽夫，王家玉译．应用蛋白酶抑制剂提高食用菌产量[J]．食用菌，1986，2：34～35
[13] John S.Holcenberg．用作药物的酶[J]．生物化学译丛(第2辑)，1980：11～14
[14] 黄年来主编．自修食用菌学[J]．南京大学出版社，1987：124
[15] W费迪南德著，王志美，何忠效，孟震译．酶分子[M]．科学出版社，1980：12～14
[16] 张丽萍，苗春艳，许李艳等．金顶侧耳多糖Pc-4P的结构确定与抗肿瘤活性的研究[J]．真菌学报，1995，14(1)：69～74
[17] 庄名扬．拱状灵芝多糖的分离与鉴定．天然产物研究与开发[J]，1993，5(2)：23～38
[18] 靳菊情，丁东宁，王晓美等．黑石耳多糖的初步化学研究[J]．中国药学杂志，1996，31(9)：520～522
[19] 陈西广等．安络小皮伞水溶性多糖的研究．Ⅰ．甘露聚糖FP1的纯化与结构研究[J]．真菌
学报，1990，9(2)：155～160
[20]沈萍萍，付庭治，曹幼琴．红栓菌固培物多糖的分离纯化与分析[J]．中国药科大学学报，1996，27(7)：441～444
[21]朱必凤，马海燕，赵发清．鸡纵菌的液体培养及其多糖物质的研究．真菌学报[J]，1996，15(1)：42～47
[22]林玉满．短裙竹荪多糖Dd-S3P的分离纯化及其性质研究[J]．生物化学杂志，1997，13(1)：99～102
[23]许燕萍，笪远峰，杨伟珠等．凤尾侧耳多糖的分离纯化及其抗氧化作用[J]．中国生化药物杂志，1997，18(2)：59～62
[24]肖盛元，郭顺星．国内真菌多糖结构研究现状[J]．天然产物研究与开发，2000(12)：81～85
[25]盖新杰，张翼伸．蛹虫草胞外多糖的研究．Ⅰ．半乳甘露聚糖(CM—1)的纯化和结构研究[J]．真菌学报，1992，11(4)：300～307
[26]曹锦花，李华．多糖免疫调节作用的研究进展[J]．中国生化药物杂志，1999，20(2)：104～105
[27]董群，方积年．多糖在医药领域中的应用[J]．中国药学杂志，2001，36(10)：649～651
[28]Wang X.B, Ng TB, Ooi VEC, etc. A polysaccharide-peptide complex from cultured mycelia of the mushroom Tricholoma mongolicum with immuno-enhancing and antitumor activities[J]. Biochem Cell Biol.1996,74(1):95～97
[29]Lee KH, Lee JW, Han MD, etc. Correlation between anticomplementary and antitumor activity of the crude polysaccharide from Granoderma Luciclum[J]. Snaop Misaengmul Hakhoechi, 1994, 22(1):45
[30]黄芳，蒙义文．活性多糖的研究进展[J]．天然产物研究与开发．1999，11(5)：90～98
[31]卯晓岚．中国经济真菌[M]．1998年版：535
[32]Dombrovska O.M.;Kostyshyn S.S..Biotransformatin of lignocellulse by fungi Pleurotus floridae(Fries)Kummer and Phellinu signiarius (Linnearus:Fries) Quelet-the pathogens of white rot in trees[J].Ukr BiokhimZh, 1998,70(1):68.
[33]张万国，胡晋红，蔡溱．桑黄增强人外周血单个核细胞产生(?)—干扰素的研究[J]．基层中药杂志，2002，16(3)：5～6
[34]A Manual of Pharmacology(Sollmann,T.) [M].1957, 8Ed.:394.
[35]U.S.Dispensatory[M].1955,25Ed.:1534.
[36]张万国，胡晋红，蔡溱．桑黄增强人外周血单个核细胞产生—干扰素的研究[J]．基层中药杂志，2002，(16)3：5～6
[37]张万国，胡晋红，蔡溱等．桑黄抗大鼠肝纤维化与抗脂质过氧化[J]．中成药，2002，24(4)：281～283
[38]温克，陈劲，李红．桑黄等四种抗癌药物抗癌活性比较[J]．吉林大学学报，2002，28(3)：247～249
[39]Ki-Bum Park, Hyo-Cheol Ha, So-yeun Kim etc. Induction of Anticarcinogenic Enzymes of Waxy Brown Rice Cultured with Phellinus igniarius 26005[J].Mycobiology,2002,30(4):213～218
[40]杨全，王琦．桑黄的液体发酵及其粗多糖抗肿瘤作用的研究．长春：吉林农业大学硕士学位论文，2002
[41]卯晓岚，蒋长坪．西藏大型经济真菌93年版[M]：470．
[42]刘金荣，江发涛，李艳．药用真菌桑黄甾类成分的提取与鉴定[J]．农垦医学，1998，20(3)：141
[43]Gazaryan,I.G.;Reshetnikova, I.A.;Vererkin,A.N.etc. Peroxidase from the fungi PheUinus igniarius[J]. Dokl. Akad. Nauk. 1993,329(5):663～5 Biochem J(Russ)
[44]Dombrovska, O. M.; Kostyshyn, S. S.. Biotuansformation of lignocellulose by whiterot fungi Pleurotus florida (Fries) Kummer and Phellinus igniarius (Linearus:Fries) Quelet Ukr[J].Biokhim. Zh. 1998,10(1):68～73
[45]莫顺燕，杨永春，石建功．桑黄化学成分研究[J]．中国中药杂志，2003，28(4)：339～341
[46]李国俊，吴国用，崔基成等．裂蹄木层孔菌菌丝培养及其应用研究[J]．中国食用菌，1998，(17)5：11～15
[47]吉林农业大学药用植物栽培教研室编．药用真菌学实验：14～18
[48]徐锦堂主编．中国药用真菌学[M]．北京医科大学，中国协和医科大学联合出版社出版，1997：194
[49]池玉杰，潘学仁、7种木层孔菌属真菌的培养特性[J]．菌物系统，2001，20(3)：378～380
[50]盛骤．线性代数与数里统计[M]．1988：293～299
[51]张惟杰．复合多糖的生化研究技术[M]．上海科技出版社，1987，12：211～215
[52]Staub AN. Removal of proteins from polysaccharides[J]. Methods in Carbohudr.Chem, 1956:5～6
[53]汪家政，范明主编．蛋白质技术手册[M]．科学出版社，2000：60
[54]鲁子贤主编．蛋白质和酶学研究方法[M]．科学出版社，1989：1
[2] Lee,S.Y. Characterization and production of antitumor polysaccharide from basidiomycetes[J]. Biotech. News,1996,3:399～403
[3] Mizinu,T., Wsaa, T. Antituuor active polysaccharide isolated from the fruiting body of Hercium erinaceum, an edible and medicinal mushroom called yamabushitake or houtou[J]. Biosci.Biotech. Biochem. 1992,56:347～348
[4] Shon,Y.H., Nam, K.S. Antimutagencity and induction of anti-carcinogenic phase·enzymes by basidiomycetes[J]. J. Ethonpharm.2001,77:103～109
[5] 杨新美．中国食用菌栽培学．北京：农业出版社[M]，1989：92～113
[6] 张树庭，Miles P.G.著．杨国良，张金霞，秦永生等译．食用菌及其栽培[M]．保定：河北大学出版社，1992：54～60
[7] 张雪岳编著．食用菌学[M]．重庆大学出版社，1988：72
[8] 杨新美主编．食用菌研究法[M]．北京：中国农业出版社，1998：237～240
[9] 陈立国，杨新美．黑木耳Auricularia auricular(L.Ex Hook)Underw纤维素酶和酚氧化酶的初步研究．武汉：华中农业大学硕士学位论文，1981
[10] 陈方顺，于青，林立仙等．木屑栽培香菇胞外纤维素酶和磷酸二酯酶的活性变化[J]．食用菌学报，1999，6(3)：8～12
[11] 尉晓宇．食用菌酶学的应用研究[J]．中国食用菌，1990，9(5)：15～16
[12] 村尾泽夫，王家玉译．应用蛋白酶抑制剂提高食用菌产量[J]．食用菌，1986，2：34～35
[13] John S.Holcenberg．用作药物的酶[J]．生物化学译丛(第2辑)，1980：11～14
[14] 黄年来主编．自修食用菌学[J]．南京大学出版社，1987：124
[15] W费迪南德著，王志美，何忠效，孟震译．酶分子[M]．科学出版社，1980：12～14
[16] 张丽萍，苗春艳，许李艳等．金顶侧耳多糖Pc-4P的结构确定与抗肿瘤活性的研究[J]．真菌学报，1995，14(1)：69～74
[17] 庄名扬．拱状灵芝多糖的分离与鉴定．天然产物研究与开发[J]，1993，5(2)：23～38
[18] 靳菊情，丁东宁，王晓美等．黑石耳多糖的初步化学研究[J]．中国药学杂志，1996，31(9)：520～522
[19] 陈西广等．安络小皮伞水溶性多糖的研究．Ⅰ．甘露聚糖FP1的纯化与结构研究[J]．真菌
学报，1990，9(2)：155～160
[20]沈萍萍，付庭治，曹幼琴．红栓菌固培物多糖的分离纯化与分析[J]．中国药科大学学报，1996，27(7)：441～444
[21]朱必凤，马海燕，赵发清．鸡纵菌的液体培养及其多糖物质的研究．真菌学报[J]，1996，15(1)：42～47
[22]林玉满．短裙竹荪多糖Dd-S3P的分离纯化及其性质研究[J]．生物化学杂志，1997，13(1)：99～102
[23]许燕萍，笪远峰，杨伟珠等．凤尾侧耳多糖的分离纯化及其抗氧化作用[J]．中国生化药物杂志，1997，18(2)：59～62
[24]肖盛元，郭顺星．国内真菌多糖结构研究现状[J]．天然产物研究与开发，2000(12)：81～85
[25]盖新杰，张翼伸．蛹虫草胞外多糖的研究．Ⅰ．半乳甘露聚糖(CM—1)的纯化和结构研究[J]．真菌学报，1992，11(4)：300～307
[26]曹锦花，李华．多糖免疫调节作用的研究进展[J]．中国生化药物杂志，1999，20(2)：104～105
[27]董群，方积年．多糖在医药领域中的应用[J]．中国药学杂志，2001，36(10)：649～651
[28]Wang X.B, Ng TB, Ooi VEC, etc. A polysaccharide-peptide complex from cultured mycelia of the mushroom Tricholoma mongolicum with immuno-enhancing and antitumor activities[J]. Biochem Cell Biol.1996,74(1):95～97
[29]Lee KH, Lee JW, Han MD, etc. Correlation between anticomplementary and antitumor activity of the crude polysaccharide from Granoderma Luciclum[J]. Snaop Misaengmul Hakhoechi, 1994, 22(1):45
[30]黄芳，蒙义文．活性多糖的研究进展[J]．天然产物研究与开发．1999，11(5)：90～98
[31]卯晓岚．中国经济真菌[M]．1998年版：535
[32]Dombrovska O.M.;Kostyshyn S.S..Biotransformatin of lignocellulse by fungi Pleurotus floridae(Fries)Kummer and Phellinu signiarius (Linnearus:Fries) Quelet-the pathogens of white rot in trees[J].Ukr BiokhimZh, 1998,70(1):68.
[33]张万国，胡晋红，蔡溱．桑黄增强人外周血单个核细胞产生(?)—干扰素的研究[J]．基层中药杂志，2002，16(3)：5～6
[34]A Manual of Pharmacology(Sollmann,T.) [M].1957, 8Ed.:394.
[35]U.S.Dispensatory[M].1955,25Ed.:1534.
[36]张万国，胡晋红，蔡溱．桑黄增强人外周血单个核细胞产生—干扰素的研究[J]．基层中药杂志，2002，(16)3：5～6
[37]张万国，胡晋红，蔡溱等．桑黄抗大鼠肝纤维化与抗脂质过氧化[J]．中成药，2002，24(4)：281～283
[38]温克，陈劲，李红．桑黄等四种抗癌药物抗癌活性比较[J]．吉林大学学报，2002，28(3)：247～249
[39]Ki-Bum Park, Hyo-Cheol Ha, So-yeun Kim etc. Induction of Anticarcinogenic Enzymes of Waxy Brown Rice Cultured with Phellinus igniarius 26005[J].Mycobiology,2002,30(4):213～218
[40]杨全，王琦．桑黄的液体发酵及其粗多糖抗肿瘤作用的研究．长春：吉林农业大学硕士学位论文，2002
[41]卯晓岚，蒋长坪．西藏大型经济真菌93年版[M]：470．
[42]刘金荣，江发涛，李艳．药用真菌桑黄甾类成分的提取与鉴定[J]．农垦医学，1998，20(3)：141
[43]Gazaryan,I.G.;Reshetnikova, I.A.;Vererkin,A.N.etc. Peroxidase from the fungi PheUinus igniarius[J]. Dokl. Akad. Nauk. 1993,329(5):663～5 Biochem J(Russ)
[44]Dombrovska, O. M.; Kostyshyn, S. S.. Biotuansformation of lignocellulose by whiterot fungi Pleurotus florida (Fries) Kummer and Phellinus igniarius (Linearus:Fries) Quelet Ukr[J].Biokhim. Zh. 1998,10(1):68～73
[45]莫顺燕，杨永春，石建功．桑黄化学成分研究[J]．中国中药杂志，2003，28(4)：339～341
[46]李国俊，吴国用，崔基成等．裂蹄木层孔菌菌丝培养及其应用研究[J]．中国食用菌，1998，(17)5：11～15
[47]吉林农业大学药用植物栽培教研室编．药用真菌学实验：14～18
[48]徐锦堂主编．中国药用真菌学[M]．北京医科大学，中国协和医科大学联合出版社出版，1997：194
[49]池玉杰，潘学仁、7种木层孔菌属真菌的培养特性[J]．菌物系统，2001，20(3)：378～380
[50]盛骤．线性代数与数里统计[M]．1988：293～299
[51]张惟杰．复合多糖的生化研究技术[M]．上海科技出版社，1987，12：211～215
[52]Staub AN. Removal of proteins from polysaccharides[J]. Methods in Carbohudr.Chem, 1956:5～6
[53]汪家政，范明主编．蛋白质技术手册[M]．科学出版社，2000：60
[54]鲁子贤主编．蛋白质和酶学研究方法[M]．科学出版社，1989：1