通过高效液相色谱法鉴定菜豆苯丙氨酸解氨酶纯化组分中的酪氨酸解氨酶活性。

Identification by high-performance liquid chromatography of tyrosine ammonia-lyase activity in purified fractions of Phaseolus vulgaris phenylalanine ammonia-lyase.

作者信息

Scott D A, Hammond P M, Brearley G M, Price C P

机构信息

Department of Clinical Biochemistry, Addenbrooke's Hospital, Cambridge, UK.

出版信息

J Chromatogr. 1992 Jan 17;573(2):309-12. doi: 10.1016/0378-4347(92)80134-c.

DOI:10.1016/0378-4347(92)80134-c

PMID:1601965

Abstract

Activities of phenylalanine ammonia-lyase (PAL) and tyrosine ammonia-lyase (TAL) were assessed at each stage of a three-step purification of PAL. Assays were performed by high-performance liquid chromatographic (HPLC) separation and ultraviolet detection of reaction products. Use of HPLC permitted assay of low activities of PAL and TAL for periods up to approximately four and two days, respectively. HPLC also facilitated the accurate quantitation of the product of the TAL reaction, trans-p-coumaric acid, which was observed to isomerize readily under experimental conditions. PAL and TAL were associated throughout the purification procedure, with TAL activity at 0.6-1.3% of PAL activity. It was concluded that, contrary to previous reports, TAL and PAL activities are mediated by the same enzyme, or else by chromatographically very similar enzymes.

摘要

在苯丙氨酸解氨酶（PAL）三步纯化的每个阶段，都对苯丙氨酸解氨酶（PAL）和酪氨酸解氨酶（TAL）的活性进行了评估。通过高效液相色谱（HPLC）分离和反应产物的紫外检测进行测定。使用HPLC分别允许在长达约四天和两天的时间内测定PAL和TAL的低活性。HPLC还便于对TAL反应产物反式对香豆酸进行准确定量，观察到该产物在实验条件下很容易异构化。在整个纯化过程中，PAL和TAL都相关联，TAL活性为PAL活性的0.6 - 1.3%。得出的结论是，与先前的报道相反，TAL和PAL活性由同一种酶介导，或者由色谱上非常相似的酶介导。