Elsevier

Analytical Biochemistry

Volume 124, Issue 1, 15 July 1982, Pages 117-124
Analytical Biochemistry

Affinity partitioning with polymer-bound Cibacron blue F3G-A for rapid, large-scale purification of phosphofructokinase from baker's yeast

This paper is dedicated to Professor A. E. Braunstein on occasion of his 80th birthday.
https://doi.org/10.1016/0003-2697(82)90228-7Get rights and content

Abstract

Phosphofructokinase has been isolated in homogenous form from baker's yeast. The first two steps, fractional precipitation with polyethylene glycol and affinity partitioning in aqueous biphasic systems containing Cibacron blue F3G-A-polyethylene glycol, gave a 58-fold purification within 3 h. In these steps the amount of contaminating proteases was reduced by 2 orders of magnitude. After concentration using DEAE-cellulose followed by gel chromatography, homogeneous enzyme was obtained. The advantages of affinity partitioning for large-scale preparations are discussed.

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    Permanent address: Institute of Physiological Chemistry, Karl Marx University, 701 Leipzig, Liebigstrasse 16, German Democratic Republic.

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