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KMID : 1094720140190050892
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Biotechnology and Bioprocess Engineering
2014 Volume.19 No. 5 p.892 ~ p.899
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Lipase purification by affinity precipitation with a thermo-responsive polymer immobilized Cibacron Blue F3GA ligand
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Ding Zhaoyang
Zheng Kai
Cao Xuejun
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Abstract
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A thermo-responsive polymer (PNNB) was synthesized with lower critical solution temperature 27.5°C and over 95% recovery. The adsorption of porcine pancreatic lipase on Cibacron Blue F3GA-conjugated PNNB (PNNB-CB) closely followed the bi-Langmuir adsorption isotherm. The maximum adsorption capacity was found at pH 5.0, with a ligand density of 18.4 μmol/g polymers. The optimized eluent was a 0.01 M phosphate buffer solution at pH 8.0 containing 20% ethylene glycol. Six adsorptiondesorption recycles indicated excellent reusability of the affinity adsorbent. PNNB-CB was applied to separate porcine pancreatic lipase from its crude material giving a lipase activity recovery of 81.6% with a 16-fold purification factor. Lipase could be purified to single-band purity, according to gel electrophoresis. The purification strategy is therefore feasible and efficient for purifying proteins of interest.
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KEYWORD
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affinity precipitation, thermo-responsive polymer, lipase, cibacron blue F3GA
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