title: Identification by high-performance liquid chromatography of tyrosine ammonia-lyase activity in purified fractions of Phaseolus vulgaris phenylalanine ammonia-lyase.
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.
authors.affiliation: Department of Clinical Biochemistry, Addenbrooke's Hospital, Cambridge, UK.
authors.forename: D A
authors.initials: DA
authors.lastname: Scott
authors.forename: P M
authors.initials: PM
authors.lastname: Hammond
authors.forename: G M
authors.initials: GM
authors.lastname: Brearley
authors.forename: C P
authors.initials: CP
authors.lastname: Price
mesh_terms: D000642:Ammonia-Lyases / Q000302:isolation & purification / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D002934:Cinnamates / Q000737:chemistry; D003373:Coumaric Acids / Q000737:chemistry; D007887:Fabaceae / Q000201:enzymology*; D010650:Phenylalanine Ammonia-Lyase / Q000302:isolation & purification / Q000378:metabolism*; D010946:Plants, Medicinal; D011422:Propionates; D013056:Spectrophotometry, Ultraviolet
chemical_list: D002934:Cinnamates; D003373:Coumaric Acids; D011422:Propionates; C029010:cinnamic acid; D000642:Ammonia-Lyases; C023208:L-tyrosine ammonia-lyase; D010650:Phenylalanine Ammonia-Lyase; C495469:p-coumaric acid