PRODUCTION AND PURIFICATION OF FIREFLY LUCIFERASE IN ESCHERICHIA-COLI

Authors
CHEN XJ REN SR JIN ZH ZHU SG
Citation
Xj. Chen et al., PRODUCTION AND PURIFICATION OF FIREFLY LUCIFERASE IN ESCHERICHIA-COLI, Biotechnology techniques, 10(2), 1996, pp. 89-92
Citations number
10
Categorie Soggetti
Biothechnology & Applied Migrobiology
Journal title
Biotechnology techniques → ACNP
ISSN journal
0951208X
Volume
10
Issue
2
Year of publication
1996
Pages
89 - 92
Database
ISI
SICI code
0951-208X(1996)10:2<89:PAPOFL>2.0.ZU;2-L
Abstract
A genetic recombinant stain of E.coli were induced to express and secr ete firefly luciferase. Cells, when broken by freeze/thawing, gave abo ut 2% of the total soluble protein as luciferase. The luciferase was p urified with ammonium sulphate fractionation and gel filtration chroma tography giving a luciferase product with high specific activity (10(6 ) light units/mg protein). SDS-PAGE of this product showed two active bands at 54 and 50 kDa, which corresponded to the-luciferases with and without a signal peptide on their N-terminals. The yield was more tha n one mg purified enzyme per 100 ml of fermentative liquid.