Beta-peltatin 6-O-methyltransferase from suspension cultures of Linum nodiflorum.
S-Adenosyl-L-methionine:beta-peltatin 6-O-methyltransferase was isolated and characterized from cell suspension cultures of Linum nodiflorum L. (Linaceae), a Linum species accumulating aryltetralin lignans such as 6-methoxypodophyllotoxin. The enzyme transfers a methyl group from S-adenosyl-L-methionine to the only free OH-group of beta-peltatin in position 6 thus forming beta-peltatin-A methylether. This reaction is a putative biosynthetic step in the biosynthesis of 6-methoxypodophyllotoxin from deoxypodophyllotoxin. The enzyme has a pH-optimum at pH 7.7 and a temperature optimum at 40 degrees C. The enzyme activity is strongly inhibited by MnSO(4), FeCl(3), FeSO(4) and ZnSO(4) as well as S-adenosyl-homocysteine. Mg(2+) and EDTA did not influence the methylation of beta-peltatin. Substrate saturation curves were obtained for S-adenosyl-methionine and beta-peltatin and apparent K(m)-values of 15 microM and 40 microM, respectively, were determined for these substrates. Substrate inhibition was observed for beta-peltatin. No other lignan substrate tested nor caffeic acid were accepted. The suspension cell line of Linum nodiflorum was characterized with respect to growth, medium alterations and lignan production as well as activity of SAM:beta-peltatin 6-O-methyltransferase. Highest specific activities of beta-peltatin 6-O-methyltransferase were determined on day 7 of the culture period corresponding to the highest levels of 6-methoxypodophyllotoxin on days 7 to 12.
Kranz
Kerstin
K
Institut für Pharmazeutische Biologie, Philipps-Universität Marburg, Deutschhausstrasse 17 A, D-35037 Marburg, Germany.
Petersen
Maike
M
D004365:Drugs, Chinese Herbal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019597:Flax / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development; D006863:Hydrogen-Ion Concentration; D017705:Lignans / Q000032:analysis / Q000096:biosynthesis; D008780:Methyltransferases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism*; D011034:Podophyllotoxin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D012436:S-Adenosylmethionine / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature; D013997:Time Factors
D004365:Drugs, Chinese Herbal; D004791:Enzyme Inhibitors; D017705:Lignans; C014451:deoxypodophyllotoxin; D012436:S-Adenosylmethionine; D008780:Methyltransferases; D011034:Podophyllotoxin