Purification and Properties of Putrescine Hydroxycinnamoyl Transferase from Tobacco (Nicotiana tabacum) Cell Suspensions.

摘要: The enzyme putrescine hydroxycinnamoyl transferase (PHT) was purified 400-fold in 7.1% yield from tobacco (Nicotiana tabacum L. cv Xanthi) cell suspensions to a final specific activity of 45 nanokatal per milligram protein. purification procedure involved conventional chromatography techniques (anion exchange chromatography, gel permeation, and hydroxylapatite chromatography) followed by on caffeoyl-cysteamine-Sepharose. This led considerable enrichment 50 kilodalton protein that could be further near homogeneity chromatofocalization (apparent isoelectric point = 8). PHT repeatedly found associated with this protein, although approximately 66% the enzymic lost during chromatofocalization. Purified exhibited same properties as unpurified extract. It not for used other aliphatic diamines (mainly diaminopropane cadaverine) substrates. most efficient phenolic substrate caffeoyl-CoA, but cinnamoyl-, feruloyl-, sinapoyl-, p-coumaroyl-CoA were also conjugated putrescine, decreasing order activity. use artificial p-fluorocinnamoyl-CoA.