Recombinant gene expression in Escherichia coli cultivation using lactose as inducer.
The use of lactose as inducer of foreign gene expression under control of the lac UV5 promoter was investigated in recombinant Escherichia coli. Chicken muscle troponin C (TnC) gene was transcripted by T7 RNA polymerase and expressed in bioreactor cultivations after a feed-forward controlled fed-batch growth phase. Cell concentrations of 22 g l-1 dry cell weight (DCW)--before induction started--were used to achieve a TnC content of 19.5% of total cell protein through an induction strategy that combined the addition of a specific lactose amount of 4.7 g g-1 DCW divided into three pulses and the addition of yeast extract (1 g l-1) together with the second and the third lactose pulses. The results presented suggest that the residual lactose concentration plays an important role on the production of the heterologous protein.
Gombert
A K
AK
Departamento de Engenharia Química, Escola Politécnica da Universidade de Sao Paulo, Brazil.
Kilikian
B V
BV
D000818:Animals; D002645:Chickens; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D005285:Fermentation; D005690:Galactose / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects*; D007785:Lactose / Q000494:pharmacology*; D009132:Muscles / Q000737:chemistry; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins; D014336:Troponin / Q000235:genetics; D020107:Troponin T; D014764:Viral Proteins
D011994:Recombinant Proteins; D014336:Troponin; D020107:Troponin T; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D007785:Lactose; D005690:Galactose