沤麻废水处理池微生物多样性分析及其Fosmid文库酶活初步筛选  

Microbial diversity analysis of alkaline flax retting wastewater and enzyme activity screen of Fosmid library clones

作　　者：董亮[1] 俞勤丽 董志扬[2] 王丽[2] 王远亮[1] 

机构地区：[1]湖南农业大学食品科技学院,湖南长沙410128 [2]中国科学院微生物研究所,微生物资源前期开发国家重点实验室,北京100101

出　　处：《食品工业科技》2015年第21期167-172,177,共7页Science and Technology of Food Industry

基　　金：国家自然科学基金项目(青年基金31300007)

摘　　要：通过构建元基因组16S r RNA文库和Fosmid文库,对富含纤维素、半纤维素等大分子有机物的湖南沅江明星麻厂碱性沤麻废水处理池(p H9.35)微生物多样性进行了分析和新的工业用酶基因筛选,发现该碱性处理池原核微生物主要以细菌为主,包括厚壁菌门、变形菌门、拟杆菌门、绿菌门等四个细菌类群,其中变形菌门弯曲杆菌科硫磺单胞菌属细菌和腐败希瓦氏菌为优势菌群,其次为拟杆菌门和厚壁菌门。该环境古菌的生物量和丰度较少,以广古菌门马氏甲烷八叠球菌为优势类群。并且从该碱性池元基因组Fosmid文库中筛选出包括淀粉酶(36个)、脂肪酶(9个)、蛋白酶(4个)、纤维素酶(4个)在内的一系列食品和工业用酶活性克隆。为进一步开发和利用该碱性处理池微生物基因资源奠定了基础。The microbial samples from alkaline sludge of retting waste water（ p H9.35） which were riched in cellulose and hemicelluloses were collected. DNA was isolated and 16 S r RNA genes were amplified. The bacterias in the alkaline anaerobic sludge contained 13 OTUs belong to Proteobacteria,Firmicute and Bacteriodetes. The most abundant bacterias were Sulfuropirillum alkalitolerans and Shewanella hafniensis. The archaeas in the alkaline anaerobic sludge contained 3 OTUs belong to Euryarchaeota.The most abundant one was Methanosarcina mazei.Fosmid library was constructed and macromolecular organics hydrolysis activities include cellulose,hemicelluloses,starch,protein and lipid hydrolyze activities were screened. After two runs of screening,36 clones contain amylase,4 clones with cellulose hydrolysis activity,4 clones contain proteinase and 9 clones contain lipase were obtained.It is very helpful to further exploit and use these gene resources and enzymes in food industry from this alkaline envioronment.

关 键 词：沤麻废水处理 原核微生物多样性 16S r RNA文库 碱性酶 

分 类 号：TS201.3[轻工技术与工程—食品科学]