Stability and Partition of Alcohol Dehydrogenase in Polyethylene Glycol/Phosphate Affinity Aqueous Two-Phase Systems Using a Triazine Dye

JOURNAL OF CHEMICAL ENGINEERING OF JAPAN

Wei-Hong Wang¹  Isao Komasawa¹  Ryoichi Kuboi¹ 

[1] Department of Chemical Engineering, Osaka University

关键词: Extraction;    Affinity Ligand;    Biopolypner. Alcohol Dehydrogenase;    Denaturation;    Hydrophobicity;    Triazine Dye;    Aqueous Two-Phase;    Conformational Change;   

DOI  :  10.1252/jcej.25.134

来源: Maruzen Company Ltd

【 摘 要 】

References(15)Cited-By(1)The conformational changes in baker’s yeast alcohol dehydrogenase (ADH) caused by the interaction of the affinity ligand Procion Blue MX-R, is investigated at various pH values in polyethylene glycol/potassium phosphate aqueous two-phase systems (ATPS). The affinity partitioning of ADH is effectively carried out at neutral pH without any damage to the structural conservation. However, the protein becomes unstable at alkaline PH and is further denatured in the presence of Procion Blue MX-R. The oligomeric ADH is completely dissociated to the monomer, and the tertiary structure of the subunits is affected by the interaction of the Blue ligand. Hydrophobic residues such as tryptophan initially buried inside may become accessible to a solvent and make denatured ADH more hydrophobic than in the native state. This affinity ligand promotes partition of the denatured fraction to the top phase, and of the active fraction to the bottom phase, at alkaline pH. The analysis of partitioning behavior in ATPS could be utilized for sensitive detection of the conformational changes in protein structure.

【 授权许可】

Unknown