Protoberberines biosynthesis

RUEFFER, M AMANN, M, ZENK, M.H., S-Adenosyl-L-methionine columbamine-O-methyl transferase, a compartmentalized enzyme in protoberberine biosynthesis. Plant Cell Rep., 1986,3,182-185. 

Genes of protoberberine biosynthesis are abundantly expressed in rhizomes of Thalictrum flavum, but were also active in roots and other organs (Samanani et al. 2005). In roots, transcripts were localized in the immature endodermis and root pericycle. In rhizomes transcripts were found in the protoderm of leaf primordial. As known from other plants, these data show that the sites of synthesis are not identical with the sites of accumulation. In many instances, a long-distance transport must occur. If this is the case, alkaloids have to pass several biomembranes. ABC-transporters and H+-alkaloid antiporters can be involved (see Chapter 1). 

The enzymes involved in the late steps of protoberberine biosynthesis, beginning with the berberine bridge enzyme all have a high pH-optimum of 8.9. This fact led to an investigation of the subcellular localization of these enzymes. After... 

M. Amann, N. Nagakura and M.H. Zenk, (S)-Tetrahydroprotoberberine oxidase, the final enzyme in protoberberine biosynthesis. Tetrahedron Lett. 25 (1984), 953-954. 

Amann M, Nagakura N, Zenk MH (1984) (S)-Tetrahydroprotoberberineoxidase, the final enzyme in protoberberine biosynthesis. Tetrahedron Lett 25 953-954 Antoun MD, Roberts MF (1975) Enzymic studies with Papaver somniferum. Planta Med 28 6-11 Barton DHR, Hesse HR, Kirby GW (1963) The origin of the berberine bridge carbon. Proc Chem Soc 267-268... 

Steffens P, Nagakura N, Zenk MH (1984) The berberine bridge forming enzyme in tetrahydro-protoberberine biosynthesis. Tetrahedron Lett 25 951-952 Sumimoto Chemicals Co Ltd (1982) Berberine alkaloid production by tissue culture. Jpn Kokai Tokkyo Koho 57,144,992 (Cl C 12pl7/18)... 

The protoberberine alkaloids (5-75) play important roles as precursors in the biosynthesis of a variety of related isoquinoline alkaloids such as protopine, phthalideisoquinoline, spirobenzylisoquinoline, rhoeadine, inde-nobenzazepine, secoberbine, and benzo[c]phenanthridine alkaloids. Chemical transformations of protoberberines to these alkaloids are particularly interesting and exciting from the biogenetic viewpoint and further from ready availability of starting protoberberines in nature or synthesis. 

Biosynthesis of the spirobenzylisoquinoline alkaloid ochotensimine (282) via the quinomethide intermediate (Scheme 49) was proposed by Shamma and Jones (7J0). On the basis of this hypothesis, several biomimetic transformations of phenolic protoberberines to spirobenzylisoquinolines have been realized by the base-induced rearrangement via the quinomethide. 

Lee MK, Kim HS. Inhibitory effects of protoberberine alkaloids from the roots of Coptis japonica on catecholamine biosynthesis in PC12 cells. Planta Med 1996 62 31-34. 

This observation was explained by the assumption that a portion of the protoberberine was formed via norreticuline (96) present in the same incubation mixture and derived from enzymic demethylation of reticuline. Reaction of 96 with an unlabeled one-carbon fragment and subsequent ring closure would then lead to C-8 unlabeled protoberberines. The authors suggest that this one-carbon fragment may be derived from S-adenosyl-methionine, and that the product of its combination with 96 may be converted directly to 91 or 94 without the intermediacy of free reticuline (99). If their assumption is correct, the conversion of norreticuline to the protoberberine alkaloids may not involve the formation of reticuline itself, a suggestion that is at variance with the known intermediacy of reticuline in the biosynthesis of alkaloids of this group. 

Protoberberine Alkaloids.—In the course of the bioconversion of the proto-berberine scoulerine (65) into chelidonine (62) and phthalide-isoquinolines, e.g. narcotine (63), C-13 becomes oxidized.61 Ophiocarpine (68), with a hydroxy-group at C-13, represents an intermediate stage in the modification of the protoberberine skeleton, and results62 of tracer experiments have shown that scoulerine (65) is also to be included in the biosynthesis of this alkaloid. Tetrahydro-protoberberine (67) is also a precursor, its incorporation indicating that C-13 hydroxylation is a terminal step. As for other protoberberine derivatives,63 nandinine (64) was not assimilated,62 and it follows then that (65) is probably converted into (67) by way of isocorypalmine (66). 

Using chirally tritiated samples of the protoberberine (67) it has been established that hydroxylation of (67) to give (68) occurs with loss of the 13-pro-R hydrogen atom, i.e. normal retention of configuration, and does not involve an enamine intermediate since tritium is not lost from C-14 during the course of this biotransformation.62 It is to be noted that similar results, associated with C-13, have been observed61 for narcotine (63) and chelidonine (62) biosynthesis, except that here the 13-pro-S proton is removed. 

The probability63 that (69) lies along the pathway to protoberberine and derived alkaloids, between scoulerine (65) and stylopine (70), has been supported by the observation that tritiated (69) is a precursor for protopine (73), and also for corynoline (78).64 Evidence previously obtained for the intermediacy of the metho-salt of stylopine [as (71)] in the biosynthesis of chelidonine (62)63 and protopine (73)63,65 has been affirmed, and it is apparently the a-form (71) and not the /3-form that is involved. [The authors are mistaken in assuming that ( —)-stylopine has the R-configuration at C-14 cf. ref. 63]. 

S)-Reticuline is a branch-point intermediate in the biosynthesis of most BAs. Most work has focused on branch pathways leading to the benzophenanthridine (e.g., sanguinarine), protoberberine (e.g., berberine), and morphinan (e.g., morphine and codeine) alkaloids.19 Most enzymes involved have been isolated, many have been purified, and four corresponding cDNAs have been cloned.19 The first committed step in benzophenanthridine and protoberberine alkaloid biosynthesis involves the conversion of (S)-reticuline to (5)-scoulerine by the berberine bridge enzyme (BBE) (Fig.7.2). BBE was purified from Berberis beaniana,20 corresponding cDNAs were cloned from E. californica and B. stolonifera,21 22 and BBE genes have been isolated from P. somniferum and E. californica.23,24... 

S)-Reticuline is also the precursor for the biosynthesis of benzophenanthri-dine (e.g. sanguinarine, marcarpine), protoberberine, berberine, palmatine) and morphinan alkaloids (morphine, codeine) (see next few paragraphs). 

Samanani, N., Park, S.-U., Facchini, P.J. (2005) Cell type-specific localisation of transcripts encoding nine consecutive enzymes involved in protoberberine alkaloid biosynthesis. Plant Cell, 17, 915-26. 

Berberine bridge enzyme catalyses are specialized step in protoberberine alkaloid biosynthesis similar genes and proteins are, however, widely present in higher plants, indicating common ancestry (Fig. 7.17c). BBE or similar proteins, which share a number of common conserved sites (Table 7.8), could also be found in fungi and bacteria. A similar pattern can be seen in the distribution of CR (Fig. 7.17d, Table 7.9). 

Speculations on the biosynthesis of berberine date back to the beginning of the century (121). Most of the early proposals recognized the structural relationship of the protoberberine alkaloids with the simpler benzylisoquinoline bases, from which it was supposed that they are derived. The additional carbon atom necessary for the formal conversion... 

Berberine chloride was evaluated for antimalarial activity against Plasmodium falciparum in vitro (two clones of human malaria Plasmodium falciparum D-6 [Sierra Leone clone] and W-2 (Indochina clone) and Plasmodium berghei in vivo (mice). The alkaloid exhibited an antimalarial potency equivalent to that of quinine in vitro, but was inactive in vivo. The results were consistent with those of others who have found berberine to be a potent inhibitor in vitro of both nucleic acid and protein biosynthesis in P. falciparum, and have demonstrated a strong interaction of berberine with DNA. In addition, the lack of in vivo antimalarial activity in mice observed with berberine and other protoberberine alkaloids agrees with clinical reports that have claimed berberine to be inactive as an antimalarial drug [228]. 

---