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Purification of tyrosyl-tRNA synthetase by affinity chromatography.

1984; Society of Chemical Engineers, Japan; Volume: 17; Issue: 6 Linguagem: Inglês

10.1252/jcej.17.642

1881-1299

Eizô Sada, Shigeo Katoh, Tsuneo Inoue, Kazuo MATSUKURA, Masami Shiozawa, Akira Takeda,

Immune Response and Inflammation

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.