Biosynthesis of podophyllotoxin in Linum album cell cultures

Abstract

Cell cultures of Linum album Kotschy ex Boiss. (Linaceae) showing high accumulation of the lignan podophyllotoxin (PTOX) were established. Enzymological studies revealed highest activities of phenylalanine ammonia-lyase, cinnamyl alcohol dehydrogenase, 4-hydroxycinnamate:CoA ligase and cinnamoyl-CoA:NADP oxidoreductase immediately prior to PTOX accumulation. To investigate PTOX biosynthesis, feeding experiments were performed with [2-(13)C]3',4'-dimethoxycinnamic acid, [2-(13)C]3',4'-methylenedioxycinnamic acid (MDCA), [2-(13)C]3',4',5'-trimethoxycinnamic acid, [2-(13)C]sinapic acid, [2-(13)C]- and [2,3-(13)C(2)]ferulic acid. Analysis of the metabolites by HPLC coupled to tandem mass spectrometry revealed incorporation of label from ferulic acid into PTOX and deoxypodophyllotoxin (DOP). In addition, MDCA was also unambiguously incorporated intact into PTOX. These observations suggest that in L. album both ferulic acid and methylenedioxy-substituted cinnamic acid can be incorporated into lignans. Furthermore, it appears that, in this species, the hydroxylation of DOP is a rate-limiting point in the pathway leading to PTOX. Electronic supplementary material to this paper can be obtained by using the Springer LINK server located at http://dx.doi.org/wo.1007/s00425-002-0834-1.

Induction of lignans and phenolic compounds in cell culture of Linum album by culture filtrate of Fusarium graminearum

“…The conversion of matairesinol into deoxypodophyllotoxin (DOP), the first aryltetralin lignan, has yet to be fully characterized, although the pathway for matairesinol to yatein, an intermediate of the matairesinol to PTOX biosynthesis, was reported. Furthermore, DOP-7-hydroxylase is thought to be responsible for the hydroxylation at positions 7 of DOP to give PTOX (Seidel et al 2002;Federolf et al 2004) (Figure 1).…”

Next Generation Sequencing in Predicting Gene Function in Podophyllotoxin Biosynthesis

“…Several of these known steps have been subsequently confirmed using Podophyllum and Linum species (20,22). On the other hand, putative downstream steps converting (Ϫ)-matairesinol (9b) into (Ϫ)-podophyllotoxin (1b) have only been reported using crude enzymatic assays; no genes have yet been identified or the enzymes purified to homogeneity (23)(24)(25)(26)(27).…”

Optimization of podophyllotoxin extraction method fromLinum albumcell cultures

“…Additionally, the PTOX production achieved in this work and recovered by the optimized extraction method (<0.37 mg/g dw) suggests that L. album cell cultures constitute a promising biotechnological system for producing this anti-cancer agent. When comparing our PTOX production with others previously reported (Smonlly et al, 1998;Seidel et al, 2002), we can infer that our system should be optimized to be competitive at an industrial level. …”