Yeast extract mediated autoinduction of lacUV5 promoter: an insight

被引:18
作者
Nair, Rahul [1 ]
Salvi, Pankaj [1 ]
Banerjee, Sampali [1 ]
Raiker, Veena A. [1 ]
Bandyopadhyay, Suman [1 ]
Soorapaneni, Sudheerbabu [1 ]
Kotwal, Prakash [1 ]
Padmanabhan, Sriram [1 ]
机构
[1] Lupin Ltd, Biotechnol R&D, Pune 411042, Maharashtra, India
关键词
ESCHERICHIA-COLI; CYCLIC-AMP; GENE-EXPRESSION; RNA-POLYMERASE; FERMENTATIONS;
D O I
10.1016/j.nbt.2009.08.002
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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-alpha 2b) 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.
引用
收藏
页码:282 / 288
页数:7
相关论文
共 18 条
[1]   Yeast extract stimulates glucose metabolism and inhibits lipolysis in rat adipocytes in vitro [J].
Edens, NK ;
Reaves, LA ;
Bergana, MS ;
Reyzer, IL ;
O'Mara, P ;
Baxter, JH ;
Snowden, MK .
JOURNAL OF NUTRITION, 2002, 132 (06) :1141-1148
[2]   Effect of yeast extract on the expression of thioredoxin-human parathyroid hormone from recombinant Escherichia coli [J].
Fu, Xiang-Yang ;
Wei, Dong-Zhi ;
Tong, Wang-Yu .
JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY, 2006, 81 (12) :1866-1871
[3]   MINOR ELEMENT COMPOSITION OF YEAST EXTRACT [J].
GRANT, CL ;
PRAMER, D .
JOURNAL OF BACTERIOLOGY, 1962, 84 (04) :869-+
[4]   Spontaneous cAMP-dependent derepression of gene expression in stationary phase plays a role in recombinant expression instability [J].
Grossman, TH ;
Kawasaki, ES ;
Punreddy, SR ;
Osburne, MS .
GENE, 1998, 209 (1-2) :95-103
[5]   Growth rate regulation of lac operon expression in Escherichia coli is cyclic AMP dependent [J].
Kuo, JT ;
Chang, YJ ;
Tseng, CP .
FEBS LETTERS, 2003, 553 (03) :397-402
[6]  
Larry E., 1971, Proceedings of the National Academy of Science, V68, P1828
[7]   High yielding recombinant Staphylokinase in bacterial expression system-cloning, expression, purification and activity studies [J].
Mandi, Nagnath ;
Soorapaneni, Sudheerbabu ;
Rewanwar, Sachin ;
Kotwal, Prakash ;
Prasad, Bhaskarjyoti ;
Mandal, Goutam ;
Padmanabhan, Sriram .
PROTEIN EXPRESSION AND PURIFICATION, 2009, 64 (01) :69-75
[8]  
Narang A, 2009, NAT REV MICROBIOL, V7, DOI [10.1038/nrmicro1932-c3, 10.1038/nrmicro1932-c1]
[9]   The relationship between external glucose concentration and cAMP levels inside Escherichia coli: Implications for models of phosphotransferase-mediated regulation of adenylate cyclase [J].
NotleyMcRobb, L ;
Death, A ;
Ferenci, T .
MICROBIOLOGY-SGM, 1997, 143 :1909-1918
[10]  
Schanus E.G., 1994, US Patent, Patent No. 5362869