高产降脂活性物质的红曲霉选育及应用
|
摘要
对收集的红曲霉菌株进行筛选，得到一株Monacolin K产量较高的红曲霉菌株M_9，其产量为15.82 μg／ml。以此菌株为原始出发菌株，孢子通过紫外、紫外—盐酸羟胺、紫外—LiCl、HNO_2、HNO_2—盐酸羟胺和HNO_2—LiCl复合诱变，得到突变株M_(9x)，其Monacolin K产量为25.32 μg／ml，较出发菌株提高了60％。
对突变株M_(9x)的原生质体制取及其原生质体再生进行了探索。其原生质形成和再生最佳条件为：以豆芽汁培养液为菌丝培养基，采用60h菌龄的菌丝体，以0.6MMgSO_4作为渗稳剂，用1.5％混合酶(蜗牛酶：纤维素酶=7：3)在30℃处理2h，原生质体形成数可以达到1.61×10~7个／ml，再生率为8.49％。
采用He-Ne激光诱变突变株M_(9x)原生质体，获得一株Monacolin K产量为103.73 μg／ml的突变株M_(9y)，其产量约是原始出发菌株M_9的6.6倍，经酯酶同工酶分析，其酶带条纹数、迁移率等均发生了明显变化。
以突变株M_(9y)、米曲霉A_1为菌种，用大米、豆粕、麸皮等为原料，分别进行单菌种及混合菌种制曲，制曲成熟后，并对其进行发酵。结果表明：可用大米、豆粕为原料培养突变株M_(9y)。米豆比例1.5：1最佳。在制曲过程中对两种工艺进行了酶学研究，混合制曲能显著提高中性蛋白酶的活力。酸性蛋白酶及糖化酶活力也有不同程度的提高。同时对两种发酵产品进行了比较：混合菌种制曲发酵工艺酿制的新型调味品，在氨基酸态氮生成率、感官及理化指标等方面均优于单菌种制曲混合发酵工艺所酿制的调味品。最后测定了混合菌种制曲发酵工艺酿制的调味品中的Monacolin K含量，其含量为21.56 μg／ml。
The collected Monascus strains were screened, a strain named Monascus sp. with high productivity of Monacolin K was obtained, whose yield reached 15.82μ/ml. Monascus sp. M9 was used as the original strain, whose spores were treated with ultraviolet ultraviolet-NH2HO ·HC1. ultraviolet-LiC1. HNO2. HNO2-NH2HO ·HC1 and HNC2-LiCl. A mutant strain named Monascus sp. M9x was gained, whose yield of Monacolin K increased 60% in comparison with the original strain, reaching 25.32μ /ml.
Next, the paper focused on the formation and regeneration of the protoplast from Monascus sp. M9x. The results showed that the soybean sprout juice was used to culture mycelium and the mycelium age of 60h was used as operated material were easy to form protoplast. Under the action of 1.5% mixed enzyme (Snail enzyme: Cellulose enzyme =7:3) with 0.6M MgSo4 sucrose used as osmotic stabilizer, the protoplasts reached 1.61 ×10~7/ml, and the regeneration rate was 8.49%.
A mutant strain called Monascus sp. M9y was achieved using irradiating protoplasts by He-Ne laser (632.8nm). Its yield of Monacolin K increased as much as 6.6 times than that of the original strain, which was 103.73μ/ml. The esterase (EST) isoenzyme electropheresis bands of the mutant strain Monascus sp. M9y compared with that of the original strain changed dramatically.
Monascus sp. M9y and Aspergillus Oryzae A1 were adopted as culture strains. Rice. soybean chips and wheat bran were used as raw materials. Single stain and mixed strains were cultured respectively on them. Immediately, they were fermented after maturing. Results showed that Monasucus sp. M9y can be cultured employing riee and soybean chips. The optimum proportion was 1.5: 1. By study on the enzymes of the two culture technologys in the course of culture, results showed that the'mixed strains culture technology can increase the activity of neutral protease remarkably, at the same time the activities of acid protease and saccharifying enzyme were raised in different degree, comparing with the single strain culture technology.
The new condiment was produced by fermenting the mature koji produced with mixed strains culture technology. The other was produced by fermenting the mature koji produced with single strain culture. By comparing the two condiments, the former was better than the latter in amino acid nitrogen producing rate. frems of sense. and physical & chemical index. And the contend of Monacolin K in the former was21.52/ml.
对突变株M_(9x)的原生质体制取及其原生质体再生进行了探索。其原生质形成和再生最佳条件为：以豆芽汁培养液为菌丝培养基，采用60h菌龄的菌丝体，以0.6MMgSO_4作为渗稳剂，用1.5％混合酶(蜗牛酶：纤维素酶=7：3)在30℃处理2h，原生质体形成数可以达到1.61×10~7个／ml，再生率为8.49％。
采用He-Ne激光诱变突变株M_(9x)原生质体，获得一株Monacolin K产量为103.73 μg／ml的突变株M_(9y)，其产量约是原始出发菌株M_9的6.6倍，经酯酶同工酶分析，其酶带条纹数、迁移率等均发生了明显变化。
以突变株M_(9y)、米曲霉A_1为菌种，用大米、豆粕、麸皮等为原料，分别进行单菌种及混合菌种制曲，制曲成熟后，并对其进行发酵。结果表明：可用大米、豆粕为原料培养突变株M_(9y)。米豆比例1.5：1最佳。在制曲过程中对两种工艺进行了酶学研究，混合制曲能显著提高中性蛋白酶的活力。酸性蛋白酶及糖化酶活力也有不同程度的提高。同时对两种发酵产品进行了比较：混合菌种制曲发酵工艺酿制的新型调味品，在氨基酸态氮生成率、感官及理化指标等方面均优于单菌种制曲混合发酵工艺所酿制的调味品。最后测定了混合菌种制曲发酵工艺酿制的调味品中的Monacolin K含量，其含量为21.56 μg／ml。
The collected Monascus strains were screened, a strain named Monascus sp. with high productivity of Monacolin K was obtained, whose yield reached 15.82μ/ml. Monascus sp. M9 was used as the original strain, whose spores were treated with ultraviolet ultraviolet-NH2HO ·HC1. ultraviolet-LiC1. HNO2. HNO2-NH2HO ·HC1 and HNC2-LiCl. A mutant strain named Monascus sp. M9x was gained, whose yield of Monacolin K increased 60% in comparison with the original strain, reaching 25.32μ /ml.
Next, the paper focused on the formation and regeneration of the protoplast from Monascus sp. M9x. The results showed that the soybean sprout juice was used to culture mycelium and the mycelium age of 60h was used as operated material were easy to form protoplast. Under the action of 1.5% mixed enzyme (Snail enzyme: Cellulose enzyme =7:3) with 0.6M MgSo4 sucrose used as osmotic stabilizer, the protoplasts reached 1.61 ×10~7/ml, and the regeneration rate was 8.49%.
A mutant strain called Monascus sp. M9y was achieved using irradiating protoplasts by He-Ne laser (632.8nm). Its yield of Monacolin K increased as much as 6.6 times than that of the original strain, which was 103.73μ/ml. The esterase (EST) isoenzyme electropheresis bands of the mutant strain Monascus sp. M9y compared with that of the original strain changed dramatically.
Monascus sp. M9y and Aspergillus Oryzae A1 were adopted as culture strains. Rice. soybean chips and wheat bran were used as raw materials. Single stain and mixed strains were cultured respectively on them. Immediately, they were fermented after maturing. Results showed that Monasucus sp. M9y can be cultured employing riee and soybean chips. The optimum proportion was 1.5: 1. By study on the enzymes of the two culture technologys in the course of culture, results showed that the'mixed strains culture technology can increase the activity of neutral protease remarkably, at the same time the activities of acid protease and saccharifying enzyme were raised in different degree, comparing with the single strain culture technology.
The new condiment was produced by fermenting the mature koji produced with mixed strains culture technology. The other was produced by fermenting the mature koji produced with single strain culture. By comparing the two condiments, the former was better than the latter in amino acid nitrogen producing rate. frems of sense. and physical & chemical index. And the contend of Monacolin K in the former was21.52/ml.
引文
[1]傅金泉．中国红曲及其实用技术．北京．中国轻工业出版社．1997．4：1～3
[2]傅金泉．中国古代红曲文化及现化新技术．酿酒科技．2001(1)：79～80
[3]方元超，杨柳．红曲霉研究进展．江苏食品与发酵．1999(1)：19～21
[4]中国科学院微生物研究所．常见与常用真菌．科学出版社．北京．1978．2：95～97
[5]刘波．中国药用真菌．山西人民出版社．太原．1978．4：6～20
[6]戴芳澜．真菌的形态与分类．科学技术出版社．北京．1987．5：78～89
[7]张纪忠．微生物分类学．复旦大学出版社．上海．1990：189～244
[8]俞大绂，李季伦．微生物学．科学出版社．北京．1988．5：293～297
[9]Yasuhiro,shinobu G, shigeo H.Nonel and Carboxypeptidase manufacture with Monascus. Jpn. Kokai Tokkyo Ko-ho 62,158,452,482 1989.6
[10]贾波，周立平．红曲霉研究之现状及发展前景．中国食品添加剂2002(6)：32～33
[11]吴衍庸．酯化酶技术与白酒增香．酿酒科技．1999(5)：24～26
[12]Nishikawa, Jiro.cellular fatty acids composition of the genus MOnascus .J.Basic Microbiol, 1989,29(6),369～374
[13]Kranz,Christiane.Biotrans formation of fatty acids in mixtures to methyl ketones by monascus purpurues,Appl, Mircrobiol Bictechnol, 1992,36(4),436～439
[14]Lumyong, Saisanom, Organicacis production by Monascus araneosus. Ahu9087.Microb util.Renewabl Resour. 1990.7 166～174
[15]陈陶声．近代工业微生物学(下册)[M]．上海科学出版社．上海．1982．5：553～562
[16]郭东川．红曲色素的两种新结构[J]．真菌学报．1993(1)：65～70
[17]傅亮．红曲色素在红曲发酵代谢中生理功能的探讨．食品科学．1998．19(10)：10
[18]杨能．生物物理与生物化学进展．1992．19(2)：81～85
[19]Endo A, Monacolin K. A new hypoeholesterolemic agent produced by a Monascus species[J]. Antibiotics, 1979,(8):852～854
[20]End A, Dihydromonacolin L and Monacolin X. New metabolites those inhabit cholesterd biosynthesis[J]. Antibiotics. 1985,(3); 321～331
[21]Endo A. Monacolin S,J,L, New inhibitors of cholesterol biosynthesis produced by Monaseus rubber[J]. Antibiotics, 1985.420～422
[22]Alberts A W.Mevinolin, A highly potent competitive inhibitor of byalroxynethyl glutaryl- coenzyme A reductuse and cholesterol-lowering agent[j]proc Natl Acad Sci USA. 1980, 3957～3961
[23]Endo A. Monacolin M. A new inhibitor of oholesterol biosynthesis[J]Antibiotics 1986,1670～1673
[24]马美荣．红曲有效生理活性物质的研究现状与进展．酿酒科技．1999．5：26～28
[25]Nobufusa Serizawa, Keiko Najagawa, Yoshio Tsujita, et al, 3α-hydroxy-ML-236B(3α- hydroxycompactin), microbial transformation ofML-236B(Compactin), J.Antibiotics, 1983,36,608～610
[26]Endo A, Yoshinori Negishi, Takasshi Iwashita, et al, Biosynthesis of ML-236B(Compactin)and Monacolin K, J.Antibiotics, 1985,38 444～448
[27]L.R.Treiber,R.A.Reamer, C,S.Rooney, et al, Origin of Monacolin I from Aspergillus terresus cultures,J.Antibiotics, 1989,42,30～36
[28]Daisuke Komagata, Hideaki Shimada, Shigco Murakawa, et al, Biosynthesis of Monacolins:conversion of Monacolin Lto Monacolin Jby a monozygenase of Monascus rubber.J.Antibiotics, 1989,42,407～412
[29]Inaminc,Edward S,Hensens; et al,U.S.Patent 04940727,Jul, 1990
[30]Smith;Robert L,Halczenko,et al,U.S.Patemt 05116870,May 1992
[31]文镜，常平．红曲及洛伐他汀的生理活性和测定方法研究进展．中国食品添加剂．2001(3)：7～14
[32]陈松生，毛宁．红曲霉的麦角固醇研究[J]．食品与发酵工业1995．(6)：18～23
[33]孟昭赫，张国柱．真菌毒素图解．人民卫生出版社．北京．1979．4
[34]Kuju. Food coloring product. Pct int.Appl.wo9514786
[35]许赣荣，卢晨，穆晓青等．部分红曲霉菌株产桔霉素的研究．无锡轻工大学学报．2001(1)：58～61
[36]宫慧梅，赴树欣，邹海晏等．用双向薄层层析检测红曲中的桔霉素．酿酒科技2002(2)：83～85
[37]迁启介，周恒刚．红曲对血压的调节作用[J]．酿酒科技．1997．(3)：75
[38]Anya Y,Nakanish H,Ishikawa T, et al. Monascus for Manufacture of antioxidant. Jpn Kokai Tokkyo Koho,Jp08 85,789[95,85,789]
[39]Hiromitsu O. et al Jpn kokai 76,128,415(1976)
[40]黄谚谚，毛宁，陈松生．红曲霉发酵产物抗疲劳作用的研究．食品科学．1998(9)：9～11
[41]路秀玲，赵树欣，宫慧梅．红曲霉的应用研究现状与展望．食品科技．2001(1)：43～45
[42]宋洪涛，郭涛等．中药红曲对高血脂症鹌鹑模型的降血脂作用．中草药．1998．29(5)：317～319
[43]曹军卫，马辉文．微生物工程．，科学出版社．北京．2002．8：24～26
[44]沈萍．微生物学．高教出版杜．北京．2000．7：231～232
[45]向洋．激光生物学．湖南科学技术出版社．长沙．1995：73～226
[46]陈五岭等．He-Ne激光在香菇生产菌株选育中的应用—一菌株的诱变及选育．光子学报．1997(11)：972～973．
[47]郭爱莲，徐金贵，杨琳．He-Ne激光选育高木质素降解率的白腐真菌LX．光子学报．2001(6)：684～685
[48]洪智勇，毛宁．激光对红曲霉的诱变作用．激光杂志．2002(5)：53～54
[49]Salton,M.E,Nature,1952,170:746
[50]谌斌．紫红曲的原生质体紫外线诱变育种．广西科学院学报．1999(2)：86～89
[51]郭爱莲，郭廷巍，朱宏莉等．He-Ne激光诱变黑曲霉Sx的原生质体．光子学报．1999(7)：603～605
[52]王卫卫，任鹏康等．He-Ne激光对亚麻酸产生菌少根根霉的诱变作用．光子学报．2002(2)：158～160
[53]施巧琴，吴松刚．工业微生物育种．福建科学技术出版社．福建．1991：231～235
[54]胡能书．同工酶技术及其应用．湖南科学技术出版社．长沙．1985．6：54～126
[55]王镜岩．生物化学(上册)．高教出版社．北京．2002
[56]陈五怜，杨晓燕．He-Ne激光辐射红霉素链霉菌诱变育种的研究．光子学报．1998(6)：539～541
[57]文铁桥，赵学慧．酵母菌属间原生质体融合构建高温酵母菌株．微生物学报．1999．(2)：141～144
[58]胡卫红，陈有为等．CO_2激光辐射对酿酒酵母菌的诱变作用．微生物学通报．2000．27(1)：36～38
[59]冯树，周樱桥，张忠泽．微生物混合培养及其应用．微生物学通报．2001(3)：92-93
[60]山东大学微生物系．微生物工程．山东大学出版社．济南．199l：315～325
[61]Shimizu H, Mizuguchi T, TanakaE,etal .Appl Environ Microbiol, 1999,65(7):3134～3141
[62]Beaubien A, KeitaL, Jolicoeur C, Appl Microbiol, 1987,53(10):2567～2578
[63]GlancerM. Process Biochemistry, 1989,24(3):109～113
[64]中科院微生物所编著．常见与常用真菌．科学出版社．北京．
[65]高嘉安，郭东，王德辉等．产Monaeolin K红曲霉菌的筛选及发酵条件的研究．吉林农业大学学报．1997(3)：85～90
[66]马美荣，樊游，方京．降胆固醇红曲霉菌种的筛选．无锡轻工大学学报．1999(3)：66～68
[67]胡晓清，陈福生，邢淑靖等．红曲中桔霉素的薄层层析分析．食品科学．2003(5)：126～129
[68]王玮萍，吴思方，曾莹．红曲霉红色素高产菌株的诱变选育．中国酿造．2003(3)：28～30
[69]孙剑波，周东坡．微生物原生质体技术．微生物学通报．2002(7)：2～5
[70]彭常安，李焕勇．红曲色素液体深层发酵工艺研究．芜湖职业技术学院学报．2001(1)：30～33
[71]Nakasa T, Suzuki M Taxonomic studies on Debaromyces hansenii (ZDRF) Lodder et Kreger van Rij and related species I. Chemotaxonomic investigations. J Gen Appl Microbiol 1985 31: 49～69
[72]邢旺兴，宓鹤鸣，陈士景等．中药红曲基原真菌的酯酶同工酶分析．微生物通报．2000(6)：437～439
[73]李多伟，尉亚辉，王义潮等．生物技术实验与仪器操作指南．西北大学出版社．西安．1996：12～14
[73]中华人民共和国专业标准．ZBX 6625—87
[74]中华人民共和国专业标准．ZB X 6630—87
[75]郭勇．酶工程．中国轻工业出版社．北京．1994．5：311～315
[76]中山大学生物系生化微生物学教研室编．生物化学技术导论．人民教育出版社．北京1979：61～63
[77]上海市酿造科学研究所．发酵调味品生产技术．中国轻工业出版社．北京1998：72～78
[78]中华人民共和国专业标准．ZB X66005-87
[79]中华人民共和国国家标准．GB/T 5009.9—1985
[80]中华人民共和国行业标准．SB 101-80
[81]杨建雄．生物化学与分子生物学实验技术教程．科学出版社．北京．2002．5：96～97
[82]周东坡．微生物原生质体融合．黑龙江科技出版社．哈尔滨．1990：79～80
[83]赵谋明．调味品．化学工业出版社．北京．2001．1：7～9
[2]傅金泉．中国古代红曲文化及现化新技术．酿酒科技．2001(1)：79～80
[3]方元超，杨柳．红曲霉研究进展．江苏食品与发酵．1999(1)：19～21
[4]中国科学院微生物研究所．常见与常用真菌．科学出版社．北京．1978．2：95～97
[5]刘波．中国药用真菌．山西人民出版社．太原．1978．4：6～20
[6]戴芳澜．真菌的形态与分类．科学技术出版社．北京．1987．5：78～89
[7]张纪忠．微生物分类学．复旦大学出版社．上海．1990：189～244
[8]俞大绂，李季伦．微生物学．科学出版社．北京．1988．5：293～297
[9]Yasuhiro,shinobu G, shigeo H.Nonel and Carboxypeptidase manufacture with Monascus. Jpn. Kokai Tokkyo Ko-ho 62,158,452,482 1989.6
[10]贾波，周立平．红曲霉研究之现状及发展前景．中国食品添加剂2002(6)：32～33
[11]吴衍庸．酯化酶技术与白酒增香．酿酒科技．1999(5)：24～26
[12]Nishikawa, Jiro.cellular fatty acids composition of the genus MOnascus .J.Basic Microbiol, 1989,29(6),369～374
[13]Kranz,Christiane.Biotrans formation of fatty acids in mixtures to methyl ketones by monascus purpurues,Appl, Mircrobiol Bictechnol, 1992,36(4),436～439
[14]Lumyong, Saisanom, Organicacis production by Monascus araneosus. Ahu9087.Microb util.Renewabl Resour. 1990.7 166～174
[15]陈陶声．近代工业微生物学(下册)[M]．上海科学出版社．上海．1982．5：553～562
[16]郭东川．红曲色素的两种新结构[J]．真菌学报．1993(1)：65～70
[17]傅亮．红曲色素在红曲发酵代谢中生理功能的探讨．食品科学．1998．19(10)：10
[18]杨能．生物物理与生物化学进展．1992．19(2)：81～85
[19]Endo A, Monacolin K. A new hypoeholesterolemic agent produced by a Monascus species[J]. Antibiotics, 1979,(8):852～854
[20]End A, Dihydromonacolin L and Monacolin X. New metabolites those inhabit cholesterd biosynthesis[J]. Antibiotics. 1985,(3); 321～331
[21]Endo A. Monacolin S,J,L, New inhibitors of cholesterol biosynthesis produced by Monaseus rubber[J]. Antibiotics, 1985.420～422
[22]Alberts A W.Mevinolin, A highly potent competitive inhibitor of byalroxynethyl glutaryl- coenzyme A reductuse and cholesterol-lowering agent[j]proc Natl Acad Sci USA. 1980, 3957～3961
[23]Endo A. Monacolin M. A new inhibitor of oholesterol biosynthesis[J]Antibiotics 1986,1670～1673
[24]马美荣．红曲有效生理活性物质的研究现状与进展．酿酒科技．1999．5：26～28
[25]Nobufusa Serizawa, Keiko Najagawa, Yoshio Tsujita, et al, 3α-hydroxy-ML-236B(3α- hydroxycompactin), microbial transformation ofML-236B(Compactin), J.Antibiotics, 1983,36,608～610
[26]Endo A, Yoshinori Negishi, Takasshi Iwashita, et al, Biosynthesis of ML-236B(Compactin)and Monacolin K, J.Antibiotics, 1985,38 444～448
[27]L.R.Treiber,R.A.Reamer, C,S.Rooney, et al, Origin of Monacolin I from Aspergillus terresus cultures,J.Antibiotics, 1989,42,30～36
[28]Daisuke Komagata, Hideaki Shimada, Shigco Murakawa, et al, Biosynthesis of Monacolins:conversion of Monacolin Lto Monacolin Jby a monozygenase of Monascus rubber.J.Antibiotics, 1989,42,407～412
[29]Inaminc,Edward S,Hensens; et al,U.S.Patent 04940727,Jul, 1990
[30]Smith;Robert L,Halczenko,et al,U.S.Patemt 05116870,May 1992
[31]文镜，常平．红曲及洛伐他汀的生理活性和测定方法研究进展．中国食品添加剂．2001(3)：7～14
[32]陈松生，毛宁．红曲霉的麦角固醇研究[J]．食品与发酵工业1995．(6)：18～23
[33]孟昭赫，张国柱．真菌毒素图解．人民卫生出版社．北京．1979．4
[34]Kuju. Food coloring product. Pct int.Appl.wo9514786
[35]许赣荣，卢晨，穆晓青等．部分红曲霉菌株产桔霉素的研究．无锡轻工大学学报．2001(1)：58～61
[36]宫慧梅，赴树欣，邹海晏等．用双向薄层层析检测红曲中的桔霉素．酿酒科技2002(2)：83～85
[37]迁启介，周恒刚．红曲对血压的调节作用[J]．酿酒科技．1997．(3)：75
[38]Anya Y,Nakanish H,Ishikawa T, et al. Monascus for Manufacture of antioxidant. Jpn Kokai Tokkyo Koho,Jp08 85,789[95,85,789]
[39]Hiromitsu O. et al Jpn kokai 76,128,415(1976)
[40]黄谚谚，毛宁，陈松生．红曲霉发酵产物抗疲劳作用的研究．食品科学．1998(9)：9～11
[41]路秀玲，赵树欣，宫慧梅．红曲霉的应用研究现状与展望．食品科技．2001(1)：43～45
[42]宋洪涛，郭涛等．中药红曲对高血脂症鹌鹑模型的降血脂作用．中草药．1998．29(5)：317～319
[43]曹军卫，马辉文．微生物工程．，科学出版社．北京．2002．8：24～26
[44]沈萍．微生物学．高教出版杜．北京．2000．7：231～232
[45]向洋．激光生物学．湖南科学技术出版社．长沙．1995：73～226
[46]陈五岭等．He-Ne激光在香菇生产菌株选育中的应用—一菌株的诱变及选育．光子学报．1997(11)：972～973．
[47]郭爱莲，徐金贵，杨琳．He-Ne激光选育高木质素降解率的白腐真菌LX．光子学报．2001(6)：684～685
[48]洪智勇，毛宁．激光对红曲霉的诱变作用．激光杂志．2002(5)：53～54
[49]Salton,M.E,Nature,1952,170:746
[50]谌斌．紫红曲的原生质体紫外线诱变育种．广西科学院学报．1999(2)：86～89
[51]郭爱莲，郭廷巍，朱宏莉等．He-Ne激光诱变黑曲霉Sx的原生质体．光子学报．1999(7)：603～605
[52]王卫卫，任鹏康等．He-Ne激光对亚麻酸产生菌少根根霉的诱变作用．光子学报．2002(2)：158～160
[53]施巧琴，吴松刚．工业微生物育种．福建科学技术出版社．福建．1991：231～235
[54]胡能书．同工酶技术及其应用．湖南科学技术出版社．长沙．1985．6：54～126
[55]王镜岩．生物化学(上册)．高教出版社．北京．2002
[56]陈五怜，杨晓燕．He-Ne激光辐射红霉素链霉菌诱变育种的研究．光子学报．1998(6)：539～541
[57]文铁桥，赵学慧．酵母菌属间原生质体融合构建高温酵母菌株．微生物学报．1999．(2)：141～144
[58]胡卫红，陈有为等．CO_2激光辐射对酿酒酵母菌的诱变作用．微生物学通报．2000．27(1)：36～38
[59]冯树，周樱桥，张忠泽．微生物混合培养及其应用．微生物学通报．2001(3)：92-93
[60]山东大学微生物系．微生物工程．山东大学出版社．济南．199l：315～325
[61]Shimizu H, Mizuguchi T, TanakaE,etal .Appl Environ Microbiol, 1999,65(7):3134～3141
[62]Beaubien A, KeitaL, Jolicoeur C, Appl Microbiol, 1987,53(10):2567～2578
[63]GlancerM. Process Biochemistry, 1989,24(3):109～113
[64]中科院微生物所编著．常见与常用真菌．科学出版社．北京．
[65]高嘉安，郭东，王德辉等．产Monaeolin K红曲霉菌的筛选及发酵条件的研究．吉林农业大学学报．1997(3)：85～90
[66]马美荣，樊游，方京．降胆固醇红曲霉菌种的筛选．无锡轻工大学学报．1999(3)：66～68
[67]胡晓清，陈福生，邢淑靖等．红曲中桔霉素的薄层层析分析．食品科学．2003(5)：126～129
[68]王玮萍，吴思方，曾莹．红曲霉红色素高产菌株的诱变选育．中国酿造．2003(3)：28～30
[69]孙剑波，周东坡．微生物原生质体技术．微生物学通报．2002(7)：2～5
[70]彭常安，李焕勇．红曲色素液体深层发酵工艺研究．芜湖职业技术学院学报．2001(1)：30～33
[71]Nakasa T, Suzuki M Taxonomic studies on Debaromyces hansenii (ZDRF) Lodder et Kreger van Rij and related species I. Chemotaxonomic investigations. J Gen Appl Microbiol 1985 31: 49～69
[72]邢旺兴，宓鹤鸣，陈士景等．中药红曲基原真菌的酯酶同工酶分析．微生物通报．2000(6)：437～439
[73]李多伟，尉亚辉，王义潮等．生物技术实验与仪器操作指南．西北大学出版社．西安．1996：12～14
[73]中华人民共和国专业标准．ZBX 6625—87
[74]中华人民共和国专业标准．ZB X 6630—87
[75]郭勇．酶工程．中国轻工业出版社．北京．1994．5：311～315
[76]中山大学生物系生化微生物学教研室编．生物化学技术导论．人民教育出版社．北京1979：61～63
[77]上海市酿造科学研究所．发酵调味品生产技术．中国轻工业出版社．北京1998：72～78
[78]中华人民共和国专业标准．ZB X66005-87
[79]中华人民共和国国家标准．GB/T 5009.9—1985
[80]中华人民共和国行业标准．SB 101-80
[81]杨建雄．生物化学与分子生物学实验技术教程．科学出版社．北京．2002．5：96～97
[82]周东坡．微生物原生质体融合．黑龙江科技出版社．哈尔滨．1990：79～80
[83]赵谋明．调味品．化学工业出版社．北京．2001．1：7～9