Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/71614
Type: Artigo de periódico
Title: S-ADENOSYL-L-METHIONINE-THEOBROMINE 1-N-METHYLTRANSFERASE, AN ENZYME CATALYZING THE SYNTHESIS OF CAFFEINE IN COFFEE
Author: MAZZAFERA, P
WINGSLE, G
OLSSON, O
SANDBERG, G
Abstract: This paper presents data on the purification and N-terminal sequence of S-adenosyl-L-methionine: theobromine 1-N-methyltransferase (STM), the enzyme responsible for the methylation of theobromine leading to caffeine formation in coffee. STM was purified from developing endosperms of immature fruits by DEAF-cellulose, hydrophobic interaction and affinity chromatography, using S-adenosyl-L-homocysteine as a ligand. The enzyme showed an apparent M(r) of ca 54 000 and ca 60 000 determined by gel filtration and SDS-PAGE, respectively. A pI of 5.1 was obtained by liquid chromatofocusing and 4.8 by isoelectrofocusing in polyacrylamide gel electrophoresis. Using theobromine as substrate, the K-m value for S-adenosyl-L-methionine was 10 mu M, being competitively inhibited by S-adenosyl-L-homocysteine (K-i = 4.6 mu M). STM is a bifunctional enzyme since it also methylated 7-methylxanthine, the immediate precursor of theobromine in the caffeine biosynthesis pathway. The specific activity of STM with 7-methylxanthine was ca 55% of that determined with theobromine. The K-m values obtained for theobromine and 7-methylxanthine were 0.196 and 0.496 mM, respectively. STM was also purified from leaves using the same procedures used for endosperms, plus an additional chromatography on a Mono Q column; theobromine was used as substrate. Finally, the N-terminal sequence for the first 20 amino acids was obtained for STM purified from endosperms. No similarities were found with other methyltransferase sequences or other known proteins.
Subject: COFFEA ARABICA
COFFEE BEANS
COFFEE LEAVES, CAFFEINE
N-METHYLTRANSFERASES
Country: Inglaterra
Editor: Pergamon-elsevier Science Ltd
Rights: fechado
Identifier DOI: 10.1016/S0031-9422(00)89570-9
Date Issue: 1994
Appears in Collections:Unicamp - Artigos e Outros Documentos

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