Purification of glucose-6-phosphate dehydrogenase from baker’s yeast in aqueous two-phase systems with free triazine dyes as affinity ligands |
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| Authors: | Yan Xu Michele Vitolo Cristina Northfleet de Albuquerque Adalberto Pessoa Jr |
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| Institution: | 1. Department of Chemical Engineering, Hebei Institute of Technology, 063009, Tangshan, Hebei, P.R. China
2. Biochemical and Pharmaceutical Technology Department, FCF/USP, PO Box 66083, CEP 05315-970, S?o Paulo, SP, Brazil
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| Abstract: | To improve the selectivity of glucose-6-phosphate dehydrogenase (G6PDH) extraction by an aqueous two-phase system, a simple
and inexpensive affinity aqueous two-phase system using unbound reactive triazine dyes as ligands was introduced. In a polyethylene
glycol (PEG)/hydroxypropyl starch (PES) system, the unbound free triazine dyes, Cibacron Blue F3GA and Procion Red HE3B, partitioned
unevenly in the top PEG-rich phase and thus showed an affinity effect on G6PDH, but no influence on hexokinase. The various
parameters investigated were pH of the system, buffers, molecular weight of PEG, and ligand type and concentration. A two-step
affinity extraction process was established for the purification of G6PDH from baker’s yeast. The total yield of G6PDH was
66.9% and purification factor was 2.35. |
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| Keywords: | Aqueous two-phase systems affinity partitioning triazine dyes glucose-6-phosphate dehydrogenase baker’ s yeast |
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