Substrate gradient formation in the large-scale bioreactor lowers cell yield and increases by-product formation

被引:162
作者
Bylund, F [1 ]
Collet, E [1 ]
Enfors, SO [1 ]
Larsson, G [1 ]
机构
[1] Royal Inst Technol, Dept Biochem & Biotechnol, S-10044 Stockholm, Sweden
关键词
D O I
10.1007/s004490050427
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
A heterogeneous micro-environment Mi as identified in a 12 m(3) bioreactor with a height-to-diameter ratio of 2.5. The reactor was aerated by a ring sparger and stirred by three Rushton turbines. E. coli cells were cultivated in minimal medium to a cell density in the order of 30 g/l. Samples of glucose, the growth limiting component fed to the process, were taken at three levels in the bioreactor (top/middle/bottom). These showed that glucose concentration declined away from the feedpoint. The gradients depended on the mixing characteristics of the feedpoint, and concentrations of up to 400 times the mean value were found when feed was added to a relatively stagnant mixing zone. This resulted in up to 20% lower biomass yield compared to the bench scale. Gradients also affected the by-product formation, resulting in acetate formation in the large-scale bioreactor. IPTG induction of a recombinant protein was shown to influence important cell parameters and considerably increased the yield of carbon dioxide per glucose added, indicating an increased maintenance. The product formation rate was, however, not notably affected by the scale-up.
引用
收藏
页码:171 / 180
页数:10
相关论文
共 21 条
[1]   Impact of plasmid presence and induction on cellular responses in fed batch cultures of Escherichia coli [J].
Andersson, L ;
Yang, SJ ;
Neubauer, P ;
Enfors, SO .
JOURNAL OF BIOTECHNOLOGY, 1996, 46 (03) :255-263
[2]   RAPID ENUMERATION OF VIABLE MICROORGANISMS BY STAINING AND DIRECT MICROSCOPY [J].
BETTS, RP ;
BANKES, P ;
BANKS, JG .
LETTERS IN APPLIED MICROBIOLOGY, 1989, 9 (05) :199-202
[3]  
CARILLI A, 1961, Sci Rep Ist Super Sanita, V1, P177
[4]   EFFECT OF GROWTH-RATE ON STABILITY AND GENE-EXPRESSION OF RECOMBINANT PLASMIDS DURING CONTINUOUS AND HIGH CELL-DENSITY CULTIVATION OF ESCHERICHIA-COLI TG1 [J].
HELLMUTH, K ;
KORZ, DJ ;
SANDERS, EA ;
DECKWER, WD .
JOURNAL OF BIOTECHNOLOGY, 1994, 32 (03) :289-298
[5]   PRODUCTION OF RECOMBINANT HUMAN GROWTH-HORMONE IN ESCHERICHIA-COLI - EXPRESSION OF DIFFERENT PRECURSORS AND PHYSIOLOGICAL-EFFECTS OF GLUCOSE, ACETATE, AND SALTS [J].
JENSEN, EB ;
CARLSEN, S .
BIOTECHNOLOGY AND BIOENGINEERING, 1990, 36 (01) :1-11
[6]   Rapid sampling, cell inactivation and evaluation of low extracellular glucose concentrations during fed-batch cultivation [J].
Larsson, G ;
Tornkvist, M .
JOURNAL OF BIOTECHNOLOGY, 1996, 49 (1-3) :69-82
[7]  
Larsson G, 1996, BIOPROCESS ENG, V14, P281
[8]   High cell-density culture of Escherichia coli [J].
Lee, SY .
TRENDS IN BIOTECHNOLOGY, 1996, 14 (03) :98-105
[9]   MIXING AND OXYGEN-TRANSFER IN CONVENTIONAL STIRRED FERMENTERS [J].
MANFREDINI, R ;
CAVALLERA, V ;
MARINI, L ;
DONATI, G .
BIOTECHNOLOGY AND BIOENGINEERING, 1983, 25 (12) :3115-3131
[10]   Response of guanosine tetraphosphate to glucose fluctuations in fed-batch cultivations of Escherichia coli [J].
Neubauer, P ;
Ahman, M ;
Tornkvist, M ;
Larsson, G ;
Enfors, SO .
JOURNAL OF BIOTECHNOLOGY, 1995, 43 (03) :195-204