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1、应用与环境生物学报2010,16(2):274~2782010-04-25ChinJApplEnvironBiol=ISSN1006-687XDOI:10.3724/SP.J.1145.2010.00274纤维素酶产生菌HS-F9的筛选鉴定和产酶条件优化*魏艳红1熊鹰1袁永泽1郑扬忠1胡柳如1张华1李风1杨江科2闫云君2刘德立1**1(华中师范大学生命科学学院遗传调控与整合生物学重点实验室武汉430079)2(华中科技大学生命科学与技术学院武汉430074)摘要为了选育高产纤维素酶菌株,通过刚果红鉴别培养基以及
2、滤纸条崩解实验测定,从牛粪堆肥中筛选到一株产纤维素酶的真菌HS-F9,根据菌株形态特性和18SrRNA基因序列分析,初步鉴定该菌为绿色木霉(Trichodermaviride).利用液体发酵培养产生纤维素酶,研究了碳源、氮源、培养时间、培养温度、培养基起始pH、接种量对菌株HS-F9产酶的影响.结果表明:产EG、CBH和FPA的最适碳源均为CMC-Na;EG和CBH在以蛋白胨为唯一氮源时酶活最高,FPa�e则在以黄豆粉为唯一氮源时酶活最高;产生EG、CBH和FPA的最适温度分别为30℃、30℃、33℃;最适起始
3、pH为3.0、3.0和4.0;EG和FPa�e的最适接种量为2%,CBH最适接种量达到了8%;培养时间均以5~6�为宜.在最适条件下培养,该菌株EG、CBH和FPa�e的酶活分别达到了5275.3U/��L、8502.1U/��L和3619.1U/��L,是未优化前的1.42、1.35和1.32倍.图6表2参23关键词纤维素酶;菌株筛选;产酶条件优化;酶活性CLCTQ925:Q936ScreeningandIdentificationofaCellulase-producingFungusandOptimiza
4、tionofItsFermentationConditions*WEIYanhong1,XIONGYing1,YUANYongze1,ZHENGYangzhong1,HULiuru1,ZHANGHua1,LIFeng1,YANGJangke2,YANYunjun2&LIUDeli1**(1LaboratoryofGeneticRegulationandIntegrativeBiology,CollegeofLifeSciences,HuazhongNormalUniversity,Wuhan430079,Chi
5、na)(2CollegeofLifeSciencesandTechnology,HuazhongUniversityofScienceandTechnology,Wuhan430049,China)AbstractInor�ertoerto�reeree�high-�high-�ieliel���traintrain���orcellula�e,ae,a�unguungu�(HS-F9)�(HS-F9)�ithhighcellula�eacti�itit���a�ia�i�olate�olate��ro��co
6、co��dungthroughcongoredidentificationmediumandthecollapseoffilter,andwasidentifiedasTrichodermaviride��anal�zingitsmorphologyand18SrRNAgenesequences.Differentconditionsincludingcarbonsource,nitrogensource,incubatingtime,growthtemperature,initialpHandinoculat
7、ionquantityfortheproductionofcellulasewerestudiedwithliquid-statefermentation.TheoptimumculturalconditionsofstrainHS-F9forthecellulaseproductionwereasfollows:TheoptimumcarbonsourceforproducingEG(endo-β-1,4-glucana�e�),CBH(exo-β-1,4-glucanasesorcellobiohy-dro
8、lase)andFPase(filterpaperenzyme)wasCMC-Na;theoptimumnitrogensourcewaspeptoneforEGandCBH,butsoyflourforFPase;theoptimumtemperaturesforEG,CBHan�FPa�e�ere30℃,30℃an�33℃,andtheoptimuminitialpHvaluesw