产纤维素酶菌株的分离_筛选及酶活性测定[1].pdf

《产纤维素酶菌株的分离_筛选及酶活性测定[1].pdf》由会员上传分享，免费在线阅读，更多相关内容在行业资料-天天文库。

1、安徽农业科学,JournalofAnhuiAgri.Sci.2009,37(17):7855-7857,7859责任编辑罗芸责任校对张士敏产纤维素酶菌株的分离·筛选及酶活性测定3吴琳,景晓辉,黄俊生(中国热带农业科学院环境与植物保护研究所,海南儋州571737)摘要[目的]寻找更多高效降解纤维素的新菌种及其科学利用方法。[方法]利用“采样—培养—分离单菌落—初筛—复筛—测OD值”的方法筛选出分解纤维素能力较强的菌株。[结果]经反复培养和划线分离从80份样品中初选出35株具有分解纤维素能力的菌株。其中10株由白转绿,长势较好;6株深绿色,长势一般;4株绿色,长势较好。这20株都产孢子

2、,被初步鉴定为绿色木霉。用刚果红培养基进行复筛选,得到9株透明圈较大的菌株。将这9株菌株再进行发酵培养,用DNS法进行酶活测定得到2株酶活较强的菌株。这2株菌株被鉴定为棘孢木霉。通过滤纸崩解测试筛选出20株降解纤维素能力较强的菌种。DNS法酶活测定结果表明采自甘蔗堆积处和甘蔗地的2个菌株的产酶活性最高。[结论]采自甘蔗堆积处和甘蔗地的2个菌株可以作为降解纤维素的新菌种。关键词纤维素降解菌;分离;筛选;纤维素中图分类号S188文献标识码A文章编号0517-6611(2009)17-07855-03IsolationandScreeningofStrainsProducingCellu

3、laseandTheirEnzymeActivityDeterminationWULinetal(InstituteofEnvironmentandPlantProtection,ChineseAcademyofTropicalAgriculturalScience,Danzhou,Hainan571737)Abstract[Objective]Thepurposewastofindmorenewhigh2efficiencycellulose2decomposingstrainsandtheirutilizationmethods.[Method]Thestrainswithst

4、rongercellulose2decomposingabilitywerescreenedoutbythemethod“sampling—culture—isolationofsinglecolony—primaryscreening—secondaryscreening—determinationofODvalue”.[Result]Thirty2fivestrainswithcellulose2decomposingabilitywereisolatedfrom80samplesthroughrepeatedcultureandstreakinginprimaryscreen

5、ing.Amongthem,10strainsturnedfromwhitetobegreenandtheirgrowthpotentialswerebetter;6strainsweredarkgreenandtheirgrowthpotentialswerecommon;4strainsweregreenandtheirgrowthpotentialswerebetter;allthese20strainspro2ducedsporesandwereidentifiedtobeTrichodermaviridepreliminarily.Thecongo2redmediawer

6、eusedand9strainswithbiggertransparentcirclesweregotinsecondaryscreening.These9strainswereculturedbyfermentationandtheirenzymeactivitiesweredeterminedbyDNSmethod,then2strainswithstrongerenzymeactivityweregot.These2strainswereidentifiedtobeTrichodermaasperellum.Twentystrainswithstrongercellulose

7、2decomposingabilitywerescreenedoutthroughfilter2paperdisintegrationtest.TheresultsofenzymeactivitydeterminationbyDNSmethodshowedthattheinulinase2producingactivitiesofthe2strainscollectedfromthestackingplaceofsugarcaneandsugarcanefieldwe