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Lipase purification by affinity precipitation with a thermo-responsive polymer immobilized Cibacron Blue F3GA ligand

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Abstract

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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Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, Department of Bioengineering, East China University of Science and Technology, Shanghai, 200-237, China

    Zhaoyang Ding, Kai Zheng & Xuejun Cao

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  1. Zhaoyang Ding
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  2. Kai Zheng
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  3. Xuejun Cao
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Correspondence to Xuejun Cao.

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Ding, Z., Zheng, K. & Cao, X. Lipase purification by affinity precipitation with a thermo-responsive polymer immobilized Cibacron Blue F3GA ligand. Biotechnol Bioproc E 19, 892–899 (2014). https://doi.org/10.1007/s12257-014-0015-7

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  • DOI: https://doi.org/10.1007/s12257-014-0015-7

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