title: Separation of transglutaminase by thermo-responsive affinity precipitation using l-thyroxin as ligand.
abstract: Transglutaminase (TGase) is widely used in the food industry. In this study, TGase was purified by affinity precipitation using l-thyroxin, coupled to a thermo-responsive polymer (PNBN), as an affinity ligand. The lower critical solution temperature and recovery of the affinity polymer were 31.0 °C and 99.6 %, respectively. The optimal adsorption condition was 0.02 mol/L phosphate buffer (pH 5.0). The recoveries 99.01 % (protein) and 98.85 % (activity) were obtained by 0.2 mol/L Gly-NaOH buffer (pH 10.0) as the elution agent. Circular dichroism spectroscopy and FortéBio Octet system were used to explore the interactions between l-thyroxin and TGase. The results show that l-thyroxin is suitable for affinity precipitation of TGase. The purity of the final product was verified using sodium dodecyl sulfate polyacrylamide gel electrophoresis.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, Department of Bioengineering, East China University of Science and Technology, Shanghai, 200237 China.
authors.forename: Sipeng
authors.initials: S
authors.lastname: Li
authors.affiliation: State Key Laboratory of Bioreactor Engineering, Department of Bioengineering, East China University of Science and Technology, Shanghai, 200237 China.
authors.forename: Zhaoyang
authors.initials: Z
authors.lastname: Ding
authors.affiliation: State Key Laboratory of Bioreactor Engineering, Department of Bioengineering, East China University of Science and Technology, Shanghai, 200237 China.
authors.forename: Xuejun
authors.initials: X
authors.lastname: Cao
keywords: Affinity precipitation; Thermo-responsive polymer; Transglutaminase; l-Thyroxin
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