重组大肠杆菌VG1(pTU14)产PHB的补料分批培养  

PRODUCTION OF POLY-β-HYDROXYBUTYRATE BY FED-BATCH CULTURE OF NOVEL RECOMBINANT Escherichia coli VG1(pTU14)

作　　者：于慧敏[1] 张延平 史悦[1] 杨胜利[2] 沈忠耀[1] 

机构地区：[1]清华大学化学工程系,北京100084 [2]中国科学院上海生物工程研究中心,上海200233

出　　处：《化工学报》2002年第7期742-746,共5页CIESC Journal

基　　金：国家"九五"攻关项目 (No .96-C0 3 -0 3 -0 2 );国家自然科学基金重点资助项目(No .2 983 410 3 );国家自然科学基金面上资助项目 (No .2 98760 2 1)~~

摘　　要：利用已构建的同时含有透明颤菌血红蛋白基因 (vgb)、λ噬菌体裂解基因 (S-RRz)和PHB合成基因(phbCAB)的产PHB基因工程大肠杆菌VG1(pTU14 ) ,在 5L发酵罐里进行补料分批培养 .研究发现 :碳氮比(C/N) ,DO(溶氧 )及碳源和氮源的浓度是影响菌体生长和PHB积累的重要因素 ;补料时间可以通过DO和 pH值两个参数的变化在线综合识别 ;流加策略应使发酵前期菌体大量生长 ,发酵后期PHB大量积累 .在优化条件下 ,对VG1(pTU14 )进行 6 0h的补料分批培养 ,菌体细胞浓度、PHB浓度和PHB占细胞干质的分数分别高达2 15 .9g·L-1、 193.7g·L-1和 89.7% .VG1(pTU14) is a novel recombinant Escherichia coli for Poly- β -hydroxybutyrate (PHB) production,which contains Vitreoscilla hemoglobin gene ( vgb ),lytic genes of phage λ( S -RRz ) and PHB biosynthetic genes ( phb CAB) in the cell.During the process of fed-batch culture of VG1 (pTU14),it was found that the level of dissolved oxygen (DO),the concentration ratio (C/N) and the range of carbon and nitrogen source were all important parameters affecting cell growth and PHB accumulation.DO and pH could be used to decide the time for the nutrition feeding.When C/N,carbon and nitrogen concentration were in the range of 10-20,10-40?g·L -1 ,1.0-2.0?g·L -1 respectively and DO was higher than 5%,the cell growth was enhanced.Otherwise,PHB accumulation would be enhanced too early to attain the high cell density culture.Finally,the optimal feeding strategy,by which the cell growth was emphasized in early process and the PHB accumulation was promoted in late process,was carried out.After 60?h fed-batch culture under the optimal conditions,cell concentration,PHB concentration and PHB content were reached up to 215.9?g·L -1 ,193.7?g·L -1 and 89.7%,respectively.

关 键 词：重组大肠杆菌VG1 PHB 透明颤菌血红蛋白基因 塑料 白色污染 微生物降解 补料 分批培养 λ噬菌体裂解基因 聚Β-羟基丁酸酯 

分 类 号：X705[环境科学与工程—环境工程] X172