A simple method for the purification of over-expressed extracellular sucrase of Zymomonas mobilis from a recombinant Escherichia coli

Authors
Sangiliyandi, G Gunasekaran, P
Citation
G. Sangiliyandi et P. Gunasekaran, A simple method for the purification of over-expressed extracellular sucrase of Zymomonas mobilis from a recombinant Escherichia coli, BIOTECH LET, 22(13), 2000, pp. 1059-1062
Citations number
13
Categorie Soggetti
Biotecnology & Applied Microbiology",Microbiology
Journal title
BIOTECHNOLOGY LETTERS
ISSN journal
01415492 → ACNP
Volume
22
Issue
13
Year of publication
2000
Pages
1059 - 1062
Database
ISI
SICI code
0141-5492(200007)22:13<1059:ASMFTP>2.0.ZU;2-5
Abstract
The over-expressed extracellular sucrase (SacC) of Zymomonas mobilis from a recombinant Escherichia coli (pZSP62) carrying the sacC gene was purified partially by repeated cycles of freezing and thawing. This method separated the highly expressed recombinant protein from the bulk of endogenous E. co li proteins. The enzyme was further purified 14 fold with a 55% yield from the cellular extract of E. coli by hydroxyapatite chromatography. The purif ied enzyme had a Mr of 46 kDa by SDS-PAGE. Its km value for sucrose was 86 mM and was optimal at pH 5.0 and at 36 degrees C.