Yeast extract mediated autoinduction of lacUV5 promoter: an insight.
We report a simple and cost-effective autoinducible media component responsible for the autoinduction of proteins in Escherichia coli under lacUV5 promoter system. Yeast extract (YE) at high concentration was found to stimulate the expression of T7 RNA polymerase in BL21(DE3) cells while such an effect was not seen in BL21A1 cells. A systematic study on the effect of varying concentrations of YE indicated several folds higher expression of genes viz., human granulocyte colony stimulating factor (rhGCSF), human interferon alpha 2b (rhIFN-alpha2b) and Staphylokinase (rSAK) in BL21(DE3) cells in the absence of any specific inducer like IPTG or additional lactose. Additional investigations on the inducible component of the YE revealed the presence of significant amount of endogenous lactose as the contributory factor for the observed autoinduction phenomenon. This paper highlights the easy scalability of the use of the present media component for large-scale production in biotechnology industry.
Nair
Rahul
R
Lupin Limited, Biotechnology R & D, Gat #1156, Ghotawade Village, Mulshi Taluka, Pune 411042, India.
Salvi
Pankaj
P
Banerjee
Sampali
S
Raiker
Veena A
VA
Bandyopadhyay
Suman
S
Soorapaneni
Sudheerbabu
S
Kotwal
Prakash
P
Padmanabhan
Sriram
S
D003470:Culture Media / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005947:Glucose / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor / Q000378:metabolism; D006801:Humans; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000378:metabolism; D007785:Lactose / Q000378:metabolism; D008666:Metalloendopeptidases / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism*; D012995:Solubility / Q000187:drug effects
D003470:Culture Media; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D000242:Cyclic AMP; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus; D005947:Glucose; D007785:Lactose