摘要
本论文包括四部分:
第一章固定化细胞技术及其应用研究进展
本章综述了固定化细胞的方法、载体材料及其应用,详细阐述了常用固定化细胞的载体材料,固定化细胞载体材料的改进,固定化细胞在食品与发酵工业、化学与医药工业、能源开发和环境工程上的应用,总结了固定化细胞的优异性能,并对未来的发展给予展望。
第二章酿酒酵母的分离鉴定
本章利用富集培养基、分离培养基、鉴定培养基等,通过稀释涂板法、平板划线分离法,从土壤中分离出1株酿酒酵母菌。对它的生理生化性质进行研究,并做了初步的鉴定,鉴定为内孢霉目,酵母属,酿酒酵母。
第三章固定化细胞材料的制备及其性能研究
本章包括两节:
第一节卡拉胶、瓜儿豆胶和膨润土固定化细胞材料的制备和性能研究
首次以卡拉胶、瓜儿豆胶和膨润土为载体固定化酿酒酵母,制备出具有高强度的卡拉胶/瓜尔豆胶/膨润土复合材料,利用正交设计优化了复合材料制备条件,并利用机械强度、扫描电镜、X射线衍射、红外、热分析对此复合材料进行了表征。
第二节卡拉胶、瓜儿豆胶和有机膨润土固定化细胞材料制备及其性能的研究
首次利用卡拉胶、瓜儿豆胶和有机膨润土为载体制备固定化酿酒酵母,制备了一种新型的卡拉胶/瓜尔豆胶/有机膨润土复合材料,利用正交设计优化了复合材料制备条件。并利用机械强度、扫描电镜、X射线衍射、红外、热分析对此复合材料进行了表征。
第四章固定化酿酒酵母细胞的制备及性能研究
制备了固定化酿酒酵母进行了酒精发酵试验,确定了其最佳发酵发酵条件。通过电镜可以看到复合材料的表面和内部均有大量的酿酒酵母细胞生长,用机械强度,SEM、IR等对固定化细胞进行了表征。通过试验室发酵试验得出酿酒酵母菌的最佳发酵条件:温度为30℃,pH值为5,时间为72h时固定化细胞发酵酒精的最大产率为15.1%
This article includes four parts:
Part I: A review on research and development of technology of immobilized cells and their applications.
In this part, the method of immobilized-cell preparation, the support of immobilized cells and their applications were looked back. Frequently used-support of immobilized cells and the application of immobilized cells in foodstuff and fermentation industry, chemical and medical industry, energy sources and environment engineering were discussed. Frequently used-support and the improvement on support of immobilized cells were emphasized. The excellent properties of immobilized cells and bright future were given.
Part II: Isolation and identification of Saccharomyces cerevisiae
In this part, Saccharomyces cerevisiae, which can produce alcohol, were isolated from the foot soil of maize. And also it was identified.
Part III: Studies on preparation and characterization of immobilized material
This part includes two subsection:
Section one: Studied on Preparation and characterization of montmorillonnite/ carrageen/ guar gum gel spherical beads
A new way to prepare composite gel spherical beads using montmorillonnite as a modifier was carried out. The beads were characterised by determining their mechanical strength, and by X-ray diffraction (XRD) and FT-IR spectroscopy. Their thermal stability and swelling kinetics were also determined.
Section two: Studied on Studied on Preparation and characterization of quaternary-montmorillonnite/ carrageen/ guar gum gel spherical beads.
A new way to prepare composite gel spherical beads using quaternary-MMT as a modifier was carried out. The beads were characterized with SEM, machine intensity, XRD and FTIR. The thermal stability of the composite gel spherical beads was also identified.
Part IV : Studies on preparation and characterization of living Saccharomyces cerevisiae cell immobilization
This study carried on the first step research to the possibility that turns living Saccharomyces cerevisiae cell immobilization. And the effect of storage time on the cell activity was also studied. And the optimum conditions of fermentation were determined. The temperature is 30℃, pH is 5, time is 72h.
引文
[1]Gerbsch N,Buchholz R.FEMS Microbiol Rev.1995,16:259-269
[2]Freeman A,Lilly M D.Enzyme Microb Technol 1998,23:335-45
[3]王家玲.环境微生物学.北京:高等教育出版社,1998.236-241
[4]罗贵民,曹淑桂,张今.酶工程.北京:化学工业出版社,2002,57
[5]王文仲.应用微生学.北京:中国医药科技出版社,1996,378-380
[6]Cassidy M B,Lee H,Trevors J T.J Ind Microbiol 1996,16:79-101
[7]Radovich J M.Enzyme Microb Technol 1985,7:2-10
[8]Conner M A and Allgeier R J.Adv Appl Microbial,1976,28:878-885
[9]Chibata Z and Tosa T.Appl Biochem.Bioeng,1983,4:1-9
[10]Scott C D.Enzyme Microb Technol,1987,9:66-73
[11]王建龙.细胞固定化技术与水污染控制.北京:科学出版社,2002,2-40
[12]杨文英,董学畅.细胞固定化及其在工业中的应用.云南民族学院学报(自然科学版),2001,10(3):406-410
[13]况金蓉,冯道伦,龚文琪.武汉理工大学学报,2001,23(11):88-91
[14]白凤武.生物工程进展.2000,20(2):32-36
[15]何忠效,静国忠,许左良等.生物技术概论.北京:北京师范大学出版社,2002,222-225
[16]梅庆和,姚善泾.现代化工,1998,1:19-22
[17]马子骏,陆志号.固定化技术及其应用.宁夏:宁夏人民出版社,1990,65-66
[18]尉迟力,夏仕文,李树本.甲烷生物催化氧化制甲醇--固定化细胞催化剂的制备及其催化性能的研究.催化学报,1997,18(6):503-507
[19]宋向阳,毛连山,杨富国,勇强,余世袁.林产化学与工业.2002,22(2):43-46
[20]田小光,彭万霖,于德水,张介弛,金永焕.微生物学通报,1995,22(5):282-284
[21]王洪祚,刘世勇.化学通报,1997,2:22-27
[22]段俊英.生物工程学报,1990,6(4):344
[23]李花子,王建龙,文湘华,施汉昌.环境科学研究,2002,15(5):26-27
[24]童群义,陈坚,堵国成,李华钟.工业微生物,2000,30(4):1-5
[25]王新,李培军,巩宗强,张海荣.农业环境保护,2001,20(2):120-122
[26]朱柱,李和平,郑泽根.重庆建筑大学学报,2000,22(5):95-101
[27]周帼萍,贾君,黄家怿.生物技术,2002,1:封2-封3
[28]薛胜伟,陈雪梅,康亮.江西化工,2002,(3):61-63
[29]李淑彬,钟英长.农业与环境生物学报,1999,5(4):422-426
[30]闵航,郑耀通,钱泽澍.环境科学,1995,15(5):10-14
[31]周华,杨正景,欧阳平凯.化学工程与工艺,1998,14(4):418-422
[32]潘光亮,黄慧,周华.工业微生物,1995,25(4):6-9
[33]江龙法,张所信,薛婉丽,刘红亚,孙进.淮海工学院学报,200l,10(2):50-53
[34]王延春,刘践,周瑞敏,马瑞德.激光生物学,1996,5(3):894-899
[35]吴斌,冯胜彦,陈蔚梅,郭明雄,艾建宇,熊晓然,吴显辉.环境科学学报,2003,23(1):74-78
[36]刘春芳.石化技术与应用,2003,21(1):67-72
[37]Yu～Kang Yuan,[P].US,6153416.2000-11-28
[38]唐学友,丁庆豹,邱蔚然.药物生物技术,2000,7(4):204-208
[39]李学梅,林建平,刘茉娥,岑沛霖.菌物系统,1998,17(4):318-326
[40]常秀莲.山东轻工业学院学报,1999,13(3):45-48
[41]R.Daraktchiev,V.Beschkov,N.Kolev,T.Aleksandrova,Bioprocess Engineering,1997,16:115-117
[42]J.C.Cotton,A.L.Pometto,III J.Gvozdenovic-Jeremic,Appl.Microbiol.Biotechnol.,2001,57:626-630
[43]王建龙,钱易.中国调味品,1995,2:2-5
[44]刘松青,武成荣.中国调味品,1998,3:10-12
[45]吴茂玉,许平,林春国.中国调味品,2001,8:15-18
[46]高放,秦克亮,赵玉秀,范秋昱,吴志龙,杨光礼.中国生化药物杂志,2000,21(6):286-288
[47]王克明,钟键江.菌物系统.,2003,22(1):123-127
[48]陈爱政,万宁.食品与发酵工业,28(8):42-44
[49]Roca E,Meinander N,Hahn,Hagerdal B,Biotech.and Bioeng.,1996,51:317-326
[50]何嘉波,吴宇澄.江苏食品与发酵,2000,103(4):4-7
[51]陈九武,赵学慧,吴思方.工业微生物,1997,27(3):27-31
[52]辛嘉英,崔俊儒,陈建波,李树本,夏春谷.生物工程学报,2002,17(1):89-93
[53]胥秀英,郑一敏,温寿祯,曾素芳.生物学杂志,2001,18(1):18-19
[54]何成,许建和,刘幽燕,庄英萍.化学研究与应用,2001,13(6):632-635
[55]娄文勇,宗敏华,范晓丹等.微生物学报,2002,42(4):485-489
[56]娄文勇,宗敏华,范晓丹等.生物化学与生物物理进展,2002,29(2):297-301
[57]吉爱国,高培基.药物生物技术,1999,6(1):14-19
[58]张建国,钱亚娟.生物工程学报,2000,16(2):188-192
[59]潘冰峰,戴学倩,冯青,李祖义.生物化学物理学报,1995,27(4):465-468
[60]Yamamoto K,Tosa T,Yamashita K,Chibata.L,Eur.J.Appl.Microbiol.,1976,3:169-175
[61]Takata I,Kayashima K,Tosa T,et al.,J.Ferment.Technol.,1982,60(5):431-437
[62]胡纯铿.华侨大学学报(自然科学版),1999,20(3):240-245
[63]王燕,张凤宝,宋正孝,张国亮,高志明,赵林,徐雅妮.化工学报,2002,53(7):747-751
[64]王燕,张凤宝,宋正孝,张国亮,王诤,赵林.过程工程学报,2002,2(2):132-136
[65]华蕾,汤一新,孙志浩,过鑫富,汪军.工业微生物,2001,31(4):5-8
[66]张朝晖,夏黎明,柯世省,岑沛霖.高校化学工程学报,1999,2(1):135-140
[67]Svetiana Pashova,Ludmila Slokoska,Ekaterina Krumova,Maria Angelova,Enzyme and Microbial Technology,1999,24:535-540
[68]S.Tao,L.Peng,L.Beihui,L.Deming,L.Zuohu,Bioprocess Enineering,1998,18:379-381
[69]王鲁燕,赵炎生,杨晓阳,武俊民.中国抗生素杂志,1998,23(6):464-466
[70]周华,姚忠,张玮,徐虹,韦萍.南京化工大学学报,2001,23(4):66-68
[71]陈斌,杨顺楷.应用与环境生物学报,1999,5(4):418-421
[72]姚汝华.微生物工程工艺原理,广州:华南理工大学出版社,1996,134-419
[73]Minoru S,Chiharu O,Mayuko T,Biotech.and Bioengine.,1997,53:214-219
[74]胡立勇,谢奕森,连颖开.中国医药工业杂志,2002,33(1):12-14
[75]牛红星,邱蔚然,丁庆豹.华东理工大学学报,2002,28(4):372-375
[76]余冬生.工业微生物,2002,32(2):34-36
[77]沈立新,魏东芝,张嗣良,王二力.华东理工大学学报,2002,28(1):24-27
[78]张宇昊,王颉,张伟,李长文等.酿酒科技,2004,125(05):72-74
[79]刘娜,石淑兰.现代化工,2005,25(02):19-23
[80]沈金龙,毛爱军,王远亮,江宁,董志扬.微生物学报,2004,44(04):507-510
[81]高德忠,王昌东,李丽华,张金生.辽宁石油化工大学学报,2004,24(02):23-26
[82]刘翔,何国庆.粮油加工与食品机械,2003,(08):67-69
[83]宋安东,张建威,吴云汉,马向东.农业工程学报,2003,19(04):278-281
[84]许凤,孙润仓,詹怀宇.纤维素科学与技术,2004,12(01):45-54
[85]马晓建,赵银峰,祝春进等.食品与发酵工业,2004,30(11):77-81
[86]李盛贤,贾树彪,顾立文.酿酒,2005,32(02):13-16
[87]张继泉,王瑞明,孙玉英.酿酒科技,2003,115(01):39-42
[88]O.C.Sitton,Gaddy J.L.,Biotechnology and Bioengineering,1980,ⅩⅫ:1735
[89]梅乐和,姚善泾,朱自强.化工学报,2002,53(5):493-497
[90]邓旭,岑沛霖.生物工程学报,1997,13(3):246-251
[91]T.Lebwau,T.Jouenne,G-A,Junter,Appl Microbial,1998,50:309-313
[92]Chatterjee S,Grethlein A J,Worden R M,Journal of Fermentation and Bioengineering,1996,81(2):158-162
[93]Narendra Kumar,Debabrata Das,Enzyme and Microbial Technology,2001,29:280-287
[94]田沈,杨静,张伟等.应用与环境生物学报.1999,(5):80-83
[95]J.Dela Nouee,D.Proulx.,Presented at 4.Internation Meeting of the Society for Applied Algology,Villeneved Ascp(France):15-17
[96]宋吟玲,田永静,樊颖果.苏州城建环保学院学报,1999,12(4):45-48
[97]Vanotti M.B.,Hunt P.G.,Tansactions of the Asae.,2000,43(2):405-414
[98]Nagadom H.,Takahasi T.,World Journal of Microbiology & Biotechnology,2000,16(1):57-62
[99]Nilssonl,etal.,Eur J Appl.Microbiol.Biotechnol.,1980,10:261
[100]Nilssonl,etal.,Eur J Appl.Microbiol.Biotechnol.,1982,14:86
[101]张彤,曹国民,赵庆祥.城市环境与城市生态,2000,13(2):17-20
[102]陈铭,周晓云.固定化细胞技术在有机废水处理中的应用与前景.水处理技术,1997,23(2):98-104
[103]S.Manohar,T.B.Karegouda.Appl.Microbiol.Biotechnol.,1998,49:785-792
[104]Lee S T,etal,Appl.Microbiol.Biotechno1.,1994,41:652-657
[105]黄霞,陈戈,邵林广等,中国环境科学,1995,15(1):1-4
[106]韩力平,王建龙,刘恒,施汉昌,钱易,环境科学,2001,22(1):78-80
[107]陈敏,罗启芳,环境科学,1993,15(3):11-14
[108]白珠,董晓燕,孙彦.微生物学通报,1996,23(3):140-143
[109]K.Czaczyk,K.Trojanowska,W.Grajek,Appl.Microbiol.Biotechno1.,1997,48:630-635
[110]Webb O F,Davison B H,Scott T C,Appl.Biochem.and Biotech.,1995,52:559-568
[111]谢晓航.酿酒科技,1999,3:44-46
[112]庞永新,魏芳萍,刁会敏,黄鹏,郭勇,贾德民.功能高分子学报,1995,8(2):213-219
[113]Yang P Y,Wang M L,Wat.Sci.Tech.,1990,22(3/4):343-350
[114]Ogbonna J C,Pham C B,Matsumu R A,et al.Biotechnol.Tech.,1989,3(6):421-424
[115]YANG P Y,Environ.Sci.Health.Part A,1991,26(8):1491-1512
[116]G.Annadurai,S.Mathalai Balan,Bioprocess Engineering,2000,22:101-102
[117]G.Annadurai,S.Rajesh Babu,K.P.O.Mahesh,T.Murugesan,Bioprocess Engineering,2000,22:493-501
[118]G.Annadurai,S.Mathalai Balan,T.Murugesan,Bioprocess Engineering,2000,22:101-107
[119]T.Branyik,G.Kuncova,J.Paca,Appl.Microbiol.Biotechnol.,2000,54:168-172
[120]刘丽霞,纪树兰,谭立扬.计算机与应用化学,2000,17(3):235-238
[121]田建民.太原理工大学学报,2000,31(1):74-78
[122]Angela Vecchio,Carlo Finoli,Damiano Di,Simine Vincenza,Andreoni Fresenius,J.Anal.Chem.,1998,361:338-342
[123]Y.Andre's,H.J.MacCordick,J.C.Hubert,Appl.Microbiol.Biotechnol.,1995,44:271-276
[124]C.Sreenivasulu,Y.Aparna,Bull.Environ.Contam.Toxicol.,2001,67:98-105
[125]李淑彬,钟英长.应用与环境生物学报,1999,5(4):422-426.
[126]张彤,曹国民,赵庆祥.城市环境与城市生态,2000,13(2):17-20
[127]田建民.生物吸附法在含重金属废水处理中的应用.太原理工大学学报,2000,31(1):74-78
[128]Calton G J,Wood L L,Updike M H,Lantz L,Hammn J P,Biotechnol.,1986,4:317-320
[129]Tanako T,Yamamoto K,Towprayoon S,Nakajima H,Sonomoto K,Yokozeki K,Kubota K,Tanaka A,Appl.Microbiol.Biotechnol.,1989,30(6):564-568
[130]刘和,王晓云.固定化微生物处理含酚废水研究.甘肃环境研究与监测,1990,13(2):76-79
[1]张宇吴,王颉,张伟,李长文等.半纤维素发酵生产燃料乙醇的研究进展.酿酒科技,2004,125(05):72-74
[2]刘娜,石淑兰.木质纤维素转化为燃料乙醇的研究进展.现代化工,2005,25(03):19-23
[3]许凤,孙润仓,詹怀宇.木质纤维原料生物转化燃料乙醇的研究进展.纤维素科学与技术,2004,12(01):45-54
[4]马晓建,赵银峰,祝养进等.以纤维素类物质为原料发酵生产燃料乙醇的研究进展,食品与发酵工业,2004,30(11):77-81
[5]李盛贤,贾树彪,顾立文.利用纤维素原料生产燃料酒精的研究进展.酿酒,2005,32(02):13-16
[6]张继泉,王瑞明,孙玉英.利用木质纤维素生产燃料酒精的研究进展.酿酒科技,2003,115(01):39-42
[7]沈金龙,毛爱军,王远亮,江宁,董志扬.纤维素酶在木质纤维素生物质转化中的应用研究.微生物学报,2004,44(04):507-510
[8]高德忠,王昌东,李丽华,张金生.燃料乙醇的性能及生产工艺.辽宁石油化工大学学报,2004,24(02):23-26
[9]刘翔,何国庆.利用木素纤维素生产燃料乙醇的微生物代谢工程.粮油加工与食品机械,2003,(08):67-69
[10]宋安东,张建威,吴云汉,马向东.利用酒糟生物质发酵生产燃料乙醇的试验研究.农业工程学报,2003,19(04):278-281
[11]刘向勇,张小华,鲍晓明等.酿酒酵母工业菌株胁迫条件耐受性分析.中国酿造,2006,(1):8-11
[12]周剑平,王治业,杨晖,魏甲乾.固定化酵母在清香型白酒发酵的应用研究.中国酿造,2006,(9):42-44
[13]郝士海.现代细菌学培养基和生化实验工册.北京:中国科学技术出版社,1992,346-368.
[14]陈天寿.培养基的制作与应用.北京:中国农业出版社,1995.509-514.
[15]张海彦.话说酵母菌.生物学教学.1994,(9):37
[16]沈萍,范秀容,李广武,微生物学实验.高等教育出版社.1981
[17]赵红梅,刘景武,张伟.耐高滲酵母的分离、筛选及鉴定.食品研究与开发,2006,(6):34-37
[18]杜连祥.工业微生物学实验技术.天津:天津科学技术出版社,1992
[19]吴帅,肖冬光,原通磊,赵旭.高耐性酿酒酵母菌种的筛选.酿酒科技,2006,(9):37-40
[20]陈士仪.酵母菌遗传学.北京:科学出版社,1989,27-38
[21]巴尼特 J A.酵母菌生理特性及鉴定手册.青岛:青岛海洋大学出版社,1990,80-118.
[22]中国科学院微生物研究所《常见与常用真菌》.北京:科学出版社,1978
1 Hsiue G H,Kuo W J,Huang Y P,et al.Microstructural and morphological charactericstics of PS-SiO_2nanocomposites.Polymer,2000,41:2813-2825
2 Kawasumi M,Hasgawa N,Kato M,et al.Preparation and mechanical propertiers of polypropylene-clay hybrid.Macromolecules,1997,30:6333-6338
3 Wu K H,Chang T C,Wang T Y,et al.Organic-inorganic hybrid material.Characterization and degadation of poly(imide-silica)hybrid.J.Polym.Sci.A:Polym.Chem,1999,37:2275-2284
4 Ciannelis E P.Polymer-layered silicate nanocomposites:synthesis,properties and applications.Appl.Organometal.Chem,1998,12:675-680
5 Liu L M,Oi Z N,Zhu X G.Studies on nylon 6/clay nanocomposites by melt-intercalation process.J.Appl.Polym.Sci.1999,71:1133-1138
6 王云普,王有朋,王利平,刘东杰,刘小军.微波合成有机坡缕石研究.非金属矿,2005,28(6):27-287
7 王璋,许时婴,汤坚.食品化学.北京:中国轻工业出版社,1999:77-80
8 P.A.Spagnuolo,D.G.Dalgleish,H.D.Goff,E.R.Morris.Kappa-carrageenan interaction in systems containing casein micelles and polysaccharide stabilizers.Food Hydrocolloids 19(2005):371-377
9 A.Berthold,K.Cremer,J.Kreuter.Preparation and characterization of chitosan microspheres as drug carrier for prednisolone sodium phosphate as model for anti-inflammatory drugs.J.Control.Release 39(1996):17-25.
10 L.P.Wang,Y.P.Wang,F.A.Zhang.Preparation and characterization of Ni~(2+)-montmorillonite/polyvinyl alcohol water-soluble nanocomposite film.Polymers and Polymer Composites 13(2005):839-846
11 P.Giunchedi,I.Genta,B.Conti,R.A.A.Muzzarelli,U.Conte.Preparation and characterization of ampicillin loaded methylpyrrolidinone chitosan and chitosan microspheres.Biomaterials 19(1998):157-161.
12 P.He,S.S.Davis,L.Illum.Chitosan microspheres prepared by spray drying.Int.J.Pharm.187(1999):53-65.
13 A.B.Madhan Kumar,K.Panduranga Rao.Preparation and characterization of pH-sensitive proteinoid microspheres for the oral delivery of methotrexate.Biomaterials.19(1998):725-732.
14 K.L.Shantha,D.R.K.Harding.Synthesis and characterization of chemically modified chitosan microspheres.Carbohydr.Polym.48(2002):247-253.
[1]章克昌.发展“燃料酒精”的建议.中国工程科学,2000,(06)
[2]薛正莲.玉米原料无蒸煮酒精发酵工艺的研究.工业微生物,1999,(04)
[3]谢林,郭少春,戴成民.高粱原料酒精发酵的应用实践与研究.酿酒科技,1999,(04)
[4]王瑞明,关凤梅,马霞等.固态发酵高产酒精酵母菌株的选育.酿酒科技,2002,(01)
[5]秦人伟,周悦,龙莹.耐高酒度酵母菌株的选育.酿酒科技,1994,(06)
[6]刘建军,姜鲁燕,赵祥颖等.高产酒精酵母菌种的选育.酿酒,2003,30(01)
[7]罗进贤,吴晓萍,张添元等.可降解淀粉和产生酒精的酵母工程菌的构建.工业微生物,2000,(04)
[8]方心芳.应用微生物学实验法.北京:中国财政经济出版社,1962
[9]大连轻工业学院等合编.食品分析.北京:中国轻工业出版社,1999
[10]熊子书.甘蔗糖蜜酒精酵母菌的筛选与应用.酿酒科技,1996,(01)
[11]马立安.耐高温酒精酵母菌驯化与筛选.湖北农学院学报,2000,(01)
[12]毛志群,张伟,檀建新等.高产酒精酵母的筛选及鉴定.食品与发酵工业.2003,29(02)
[13]曹俊峰,姚培鑫,马小魁.发酵甜高粱汁耐高浓度酒精酵母菌的选育.西北植物学报,2001,(05)
[14]杨勇.安琪牌耐高温干酵母在玉米酒精浓醪发酵生产中的应用.酿酒科技,1998,(02)
[15]寇运同,胡永松,王忠彦等.高温酵母发酵特性及功能的研究.酿酒科技,1995,(03)
[16]王梅,张彭湃,帅桂兰等.TTC在黄酒酵母选育中的应用.酿酒,2001,(05)
[17]秦成国.玉米粉高浓度酒精发酵的研究.酿酒科技,1997,82(04)
[18]赵华,赵树欣,才向东等.玉米原料酒精浓醪发酵技术的研究.酿酒科技,1998,(05)