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A simple method for the purification of over-expressed extracellular sucrase of Zymomonas mobilis from a recombinant Escherichia coli |
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| Authors: | Gurunathan Sangiliyandi Paramasamy Gunasekaran |
| Affiliation: | (1) Department of Microbial Technology, School of Biological Sciences, Madurai Kamaraj University, Madurai, 625 021, India;(2) Department of Molecular Genetics, Weizmann Institute of Science, Rehovot , 76100, Israel |
| Abstract: | The over-expressed extracellular sucrase (SacC) of Zymomonas mobilisfrom 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. coli proteins. The enzyme was further purified 14 fold with a 55% yield from the cellular extract of E. coli by hydroxyapatite chromatography. The purified 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 °C. |
| Keywords: | over-expression protein purification sucrase Zymomonas mobilis |
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