FED-BATCH CULTIVATION OF RECOMBINANT ESCHERICHIA-COLI JM103 AND PRODUCTION OF THE FUSION PROTEIN SPA=ECORI IN A 60-L WORKING VOLUME AIRLIFT TOWER LOOP REACTOR

被引:12
作者
BRANDES, L [1 ]
WU, XA [1 ]
BODE, J [1 ]
RHEE, JI [1 ]
SCHUGERL, K [1 ]
机构
[1] UNIV HANNOVER, INST TECH CHEM, W-3000 HANNOVER, GERMANY
关键词
ESCHERICHIA-COLI JM103; FUSION PROTEIN; TEMPERATURE INDUCTION; CHEMICAL INDUCTION; FED-BATCH; AIRLIFT TOWER LOOP REACTOR;
D O I
10.1002/bit.260420208
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
SPA=EcoRI fusion protein was produced by Escherichia coli JM103 carrying the multicopy expression plasmid pMTC48, the multicopy repressor plasmid pRK248, and the multicopy protection plasmid pEcoR4 in a 60-L working volume airlift tower loop reactor on M9 minimal medium with glucose. Cell mass concentration, total cell count, number of colony-forming units, specific growth rate, yield coefficient, and metabolite (acetate, pyruvate, succinate, lactate, ethanol) concentrations were monitored during the growth phase and gene expression. Gene expression was induced by temperature shift or chemically by isopropyl-thiogalactosidase in the airlift tower loop reactor (ALTR) at constant cultivation time and in a small stirred tank reactor at different cultivation times. During induction, the cultivation medium was supplemented with concentrated Luria-Bertani (LB) medium. The intracellular enzyme activity was evaluated as a function of the time after the start of the induction. It was found that the reduction of the glucose concentration and increase of the dissolved oxygen concentration reduced the acetate produced and increased the intracellular enzyme activity.
引用
收藏
页码:205 / 214
页数:10
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