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1、安徽农业大学学报,2017,44(3):482-486JournalofAnhuiAgriculturalUniversity[DOI]10.13610/j.cnki.1672-352x.20170524.014网络出版时间:2017/5/2411:23:52[URL]http://kns.cnki.net/kcms/detail/34.1162.S.20170524.1123.028.html热泉土壤来源的纤维素降解菌的分离与初步鉴定*王旭,王志,蔡梦宇,周育(安徽农业大学茶与食品科技学院,茶树生物学与资源利用国家重点实验室,合肥230036)摘要:通过富集培
2、养方法,从5份热泉土壤中筛选得到6株具有羧甲基纤维素(CMC)降解活性的菌株。采用原核生物保守16SrRNA核糖体基因序列分析,发现它们分别隶属于假单胞菌属、芽孢杆菌属和类芽孢杆菌属。在此基础上,对6株纤维素降解菌的生长温度、pH值、抗生素敏感性、盐离子耐受性以及羧甲基纤维素酶(CMCase)-1活力做了初步研究。结果表明,芽孢杆菌属的菌株WP-6和WP-3的CMCase活力最强,胞内酶活力最高达72.0U·mL,-1假单胞菌株WP-2的CMCase活力最弱,胞内外酶活力均小于5.0U·mL。生理学结果显示,6个降解菌株都具有嗜热特点及不同程度的耐盐特性,给后续
3、高温及盐离子耐受性纤维素酶开发提供了较好的材料。关键词:降解菌;羧甲基纤维素酶;富集;分离;酶活力中图分类号:Q939.9文献标识码:A文章编号:1672352X(2017)03048205Isolationandidentificationofcellulose-degradingbacteriafromthehotspringsoilWANGXu,WANGZhi,CAIMengyu,ZHOUYu(StateKeyLaboratoryofTeaPlantBiologyandUtilization,SchoolofTea&FoodScienceandTec
4、hnology,AnhuiAgriculturalUniversity,Hefei230036)Abstract:Sixcellulose-degradingbacterialstrainswereisolatedandidentifiedfromfivehotspringsoilsbytheenrichmentandculture-dependentmethod.Sixisolateswereidentifiedbyanalyzingthe16SrRNAgenese-quence,andtheisolateswereallocatedtothreegenera
5、ofPseudomonassp.,Bacillussp.andPaenibacillussp.Moreover,thegrowthtemperature,pHvalue,NaCltolerance,antibioticsusceptibilityandCMCaseactivityofthesixisolatesweredeterminedandcompared.TheresultsshowedthatBacillusspp.WP-3andWP-6displayedthehighestCMCase,andPseudomonassp.WP-2displayedthe
6、lowestCMCasetocarboxymethylcellulose(CMC)-1amongthetestedstrains.ThehighestCMCaseactivitywas72.0U·mL,whichwasobtainedfromintracellularof-1strainWP-6,andthelowestCMCaseactivitywasabout5.0U·mL,whichwasobservedfromstrainWP-2.Physiologicalinvestigationindicatedthatthesixstrainswerethemes
7、ophilicandhalotolerant,andthecharacteris-ticswereinterestingtofurtherapplicationstudies.Keywords:degradingbacteria;CMCase;enrichment;isolation;identification维素酶是一类能够降解木质纤维素或半纤维素生产过程中产生的副产物,中国每年秸秆产量约为88[3]酶系的总称,包括多组分的复合生物催化剂,主要6×10~7×10t之多。传统农业对于秸秆的处理由β-1,4-葡聚糖外切酶[EC.3.2.1.91]、β-1,4-葡
8、聚糖方式主要是喂养牲畜、