Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
9604276 | Journal of Biotechnology | 2005 | 13 Pages |
Abstract
The development of a simplified process for the simultaneous disruption and direct selective purification of intracellular proteins from unclarified yeast disruptate has been investigated. The recovery of glyceraldehyde 3-phosphate dehydrogenase (G3PDH) from baker's yeast was selected as a potential demonstration of the generic applicability and practical feasibility of this integrated technique. The application of an adsorbent characterised by high density (UpFront steel-agarose; Ï = 2.65 g mlâ1) facilitated the combining of cell disruption operation (bead milling of 50% ww/v of yeast suspension at 7.2 l hâ1) with fluidised bed dye-ligand (Cibacron Blue 3GA) adsorption operated immediately downstream of the disrupter. The adoption of a polymer shielded, dye-ligand technique advanced recovery efficiency. It was demonstrated that G3PDH could be recovered with a yield of 67.5% bound activity and a specific activity of 40.2 IU mgâ1, after a single step elution with 0.15 M NaCl. The generic application of this approach has been evaluated.
Related Topics
Physical Sciences and Engineering
Chemical Engineering
Bioengineering
Authors
Tau Chuan Ling, Andrew Lyddiatt,