PURIFICATION OF TYROSYL-tRNA SYNTHETASE BY AFFINITY CHROMATOGRAPHY

JOURNAL OF CHEMICAL ENGINEERING OF JAPAN

TSUNEO INOUE²  EIZO SADA²  MASAMI SHIOZAWA¹  SHIGEO KATOH²  AKIRA TAKEDA¹  KAZUO MATSUKURA¹ 

[1] Research and Development Center, Unitika Ltd.;Department of Chemical Engineering, Kyoto University

关键词: Adsorption;    Affinity Chromatography;    Biochemical Engineering;    Chromatography;    Elution Curve;    Enzyme;    Ionic Strength;    Mass Transfer;    Purification;    Tyrosyl-tRNA Synthetase;   

DOI  :  10.1252/jcej.17.642

来源: Maruzen Company Ltd

【 摘 要 】

References(9)Cited-By(6)Stable tyrosyl-tRNA synthetase (EC 6.1.1.1) was isolated from thermophilic bacterium Bacillus stearothermophilus and purified by use of two consecutive affinity chromatographic procedures on a large scale. The specific activity increased 25, 900-fold after the purification. With a Matrex Blue A column, used in the finalstep affinity Chromatography, the effects of factors influencing the performance of Chromatography were studied. The column diameter was scaled up from 1.65cm to 18.4cm without appreciable change in the elution profiles. The degree of purification decreased gradually and the elution profile became sharper with the increase in concentration of potassium chloride from 0.1 to 0.6 mol ⋅ 1-1. Variation of the elution profile with liquid velocity was also studied. The experimental elution curves showed general agreement with the calculated values based on the mass transfer theory.

【 授权许可】

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