Albrecht J, Jansen I, Kula M R
Institut für Enzymtechnologie der Heinrich-Heine-Universität Düsseldorf, Federal Republic of Germany.
Biotechnol Appl Biochem. 1993 Apr;17(2):191-203.
The purification of (R)-oxynitrilase (EC 4.1.2.10) from Linum usitatissimum (flax) has been improved considerably. The enzyme is obtained from seedlings in 60% yield by fractional salt precipitation followed by ion-exchange and hydrophobic-interaction chromatography. Final gel-permeation chromatography yields a protein with a specific activity of 53 units/mg at pH 4.1. The N-terminal sequence is reported and microheterogeneity demonstrated. The substrate range was investigated using (R)-oxynitrilase immobilized on Eupergit and t-butyl methyl ether as solvent. The addition of HCN to various aliphatic ketones and aldehydes is catalysed by the enzyme, while aromatic ketones are not converted. (R)-Butan-2-one cyanohydrin was synthesized on a preparative scale and the product characterized.
亚麻（Linum usitatissimum）中（R）-氧腈酶（EC 4.1.2.10）的纯化方法有了显著改进。通过分级盐析，然后进行离子交换和疏水相互作用色谱法，从幼苗中获得该酶，产率为60%。最终的凝胶渗透色谱法得到一种蛋白质，在pH 4.1时比活性为53单位/毫克。报告了其N端序列并证明了微不均一性。使用固定在Eupergit上的（R）-氧腈酶和叔丁基甲基醚作为溶剂研究了底物范围。该酶催化向各种脂肪族酮和醛中添加HCN，而芳香族酮则不被转化。以制备规模合成了（R）-丁-2-酮氰醇并对产物进行了表征。
