caa a22 4500 477101437 CHVBK 20180405111542.0 cr unu---uuuuu 170330e19960501xx s 000 0 eng 10.1007/BF00196893 doi (NATIONALLICENCE)springer-10.1007/BF00196893 Purification of hydroxycinnamoyl-CoA:tyramine hydroxycinnamoyltransferase from cell-suspension cultures of Solanum tuberosum L. cv. Datura [Elektronische Daten] [Hartwig Hohlfeld, Dierk Scheel, Dieter Strack] A pathogen-elicitor-inducible acyltransferase [tyramine hydroxycinnamoyltransferase (THT); EC 2.3.1], which catalyzes the transfer of hydroxycinnamic acids from hydroxycinnamoyl-CoA esters to tyramine in the formation of N-hydroxycinnamoyltyramine, was purified to apparent homogeneity from cell-suspension cultures of potato (Solanum tuberosum L. cv. Datura), with a 1400-fold enrichment, a 5% recovery and a final specific activity of 208 mkat·(kg protein)−1. Affinity chromatography on Reactive Yellow-3-Agarose using the acyl donor (feruloyl-CoA) as eluent was the decisive step in the purification sequence. The purified protein showed a native molecular mass of ca. 49 kDa. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the presence and in the absence of a reducing agent (2-mercaptoethanol) indicated that THT is a heterodimer in which the protein subunits (ca. 25 kDa) are non-covalently associated. The enzyme was stimulated fivefold by 10 mM Ca2+. The apparent K m value for tyramine was dependent on the nature of the hydroxycinnamoyl-CoA present. Thus, the K m value for tyramine was about tenfold greater (174 μM) in the presence of 4-coumaroyl-CoA than in the presence of feruloyl-CoA (20 μM). Springer-Verlag, 1996 Acyltransferase purification nationallicence Affinity chromatography (reactive dye) nationallicence Hydroxycinnamic acid amides nationallicence Phenylpropanoids nationallicence Solanum nationallicence PAL : phenylalanine ammonia-lyase nationallicence THT : hydroxycinnamoyl-CoA:tyramine hydroxycinnamoyltransferase nationallicence Hohlfeld Hartwig Institut für Pflanzenbiochemie, Weinberg 3, D-06120, Halle (Saale), Germany aut Scheel Dierk Institut für Pflanzenbiochemie, Weinberg 3, D-06120, Halle (Saale), Germany aut Strack Dieter Institut für Pflanzenbiochemie, Weinberg 3, D-06120, Halle (Saale), Germany aut Planta Springer-Verlag 199/1(1996-05-01), 166-168 0032-0935 199:1<166 1996 199 425 https://doi.org/10.1007/BF00196893 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00196893 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hohlfeld Hartwig Institut für Pflanzenbiochemie, Weinberg 3, D-06120, Halle (Saale), Germany aut NATIONALLICENCE 700 1- Scheel Dierk Institut für Pflanzenbiochemie, Weinberg 3, D-06120, Halle (Saale), Germany aut NATIONALLICENCE 700 1- Strack Dieter Institut für Pflanzenbiochemie, Weinberg 3, D-06120, Halle (Saale), Germany aut NATIONALLICENCE 773 0- Planta Springer-Verlag 199/1(1996-05-01), 166-168 0032-0935 199:1<166 1996 199 425 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer