Production of alkaloids

The reductive coupling of of dienes containing amine groups in the backbones allows for the production of alkaloid skeletons in relatively few steps [36,46,47]. Epilupinine 80 was formed in 51% yield after oxidation by treatment of the tertiary amine 81 with PhMeSiEh in the presence of catalytic 70 [46]. Notably, none of the trans isomer was observed in the product mixture (Eq. 11). The Cp fuMcTIIF was found to catalyze cyclization of unsubstituted allyl amine 82 to provide 83. This reaction proceeded in shorter time and with increased yield relative to the same reaction with 70 (Eq. 12) [47]. Substitution of either alkene prevented cyclization, possibly due to competitive intramolecular stabilization of the metal by nitrogen preventing coordination of the substituted olefin, and resulted in hydrosilylation of the less substituted olefin. 

Goodbody and co-workers (7/9) have examined the production of alkaloids in root and shoot cultures induced from seedlings of C. roseus. The pattern of alkaloids in the root cultures was similar to that of the roots from intact plants. Thus ajmalicine (39) and catharanthine (4) were produced, but no vindoline (3), a major leaf alkaloid, and no bisindole alkaloids. Similarly, the pattern of the alkaloid content of the shoot cultures was like that of the leaves of the intact plant, showing the presence of vindoline (3), catharanthine (4), and ajmalicine (39), with 3 predominating. A search for the bisindole alkaloids in the cultures indicated the presence of anhydrovinblastine (8) and leurosine (11) in the shoot cultures (2.6 and 0.3 xg/g fresh weight, respectively), but no vinblastine (1) or vincristine (2). 

In vitro production of alkaloids is possible also with the use of another in vitro technique organ culture. Root cultures are the most common concerning alkaloids, as this part of plants is of great importance for alkaloid synthesis. Root cultures that produce alkaloids have been studied as far back... 

Roberts, M. F. 1998. Production of alkaloids in plant cell culture. In Alkaloids. Biochemistry, Ecology, and Medicinal Applications (Roberts, M. F. and Wink, M., eds), pp. 159-197 and, Roberts, M. F. 1998. Enzymology of alkaloid biosynthesis. In Alkaloids. Biochemistry, Ecology, and Medicinal Applications (Roberts, M. F. and Wink, M., eds), pp. 109-146. New York - London Plenum Press. 

Verpoorte R, van der Heiden R and Memelink J (1998) Plant biotechnology and the production of alkaloids prospects of metabolic engineering. The Alkaloids, Chemistry and Pharmacology (ed Cordell GA) Vol 50. Academic, San Diego, pp 453-508. 

This section considers proven biochemical pathways within alkaloid-producing plants, as well as the stronger circumstantial evidence based on structural features (e.g., optical rotation, substitution patterns) and biosynthetic speculations not included in the material of Section II. The biochemical reaction products of alkaloids after administration to non-alkaloid-producing species are discussed in Section IV,B. Biological activity is discussed in Section VIII. 

O-methylflavinantine from Rhiziocarpa racemifera.147 The influence of day-length and of light intensity on the production of alkaloids in P. somniferum has been studied.148... 

The toxic effects observed with intact animals has its counterpart in the cytotoxic effect, which has been recorded for nearly 180 alkaloids (Table III). These data have been obtained by screening many natural products for anticancer activity. However, an alkaloid that can kill a cancer cell is usually also toxic for normal cells. Therefore, the data shown in Table III are another indication of the general toxicity of alkaloids toward animals. Because this toxicity applies also for herbivores, the production of alkaloids by plants can certainly be interpreted as a potent antiherbivore mechanism. 

The absorbance of this at the absorption maxima (405 to 410 nm) is measured. The alkaloid content is then calculated from a calibration graph. Hydrolysis products of alkaloids do not interfere unless present in threefold amounts. 

Better cell growth and a prevention in the rapid decrease in cell mass after substrate depletion in cell suspension cultures of T. rugosum was provided by light. In addition, continuous illumination enhanced the production of alkaloid in cell culture. Finally, light was found to significantly suppress the secretion of the major alkaloid berberine into the culture [23]. 

Many investigations have been carried out of gas chromatography of tobacco alkaloids in order to study the volatile compounds of the smoke, i.e. thermal decomposition products of alkaloids and other substances in tobacco. This chapter is mainly concerned with the gas chromatography of the alkaloids themselves and their metabolites in man, also to some extent with the decomposition products of the alkaloids. 

Production of Alkaloids When Inhibiting Respiration by the Creation of a Nutrient Deficiency... 

The process of the invention is therefore one for the production of alkaloid derivatives of lysergic acid which comprises fermenting under aerobic conditions an aqueous nutrient medium containing a source of carbon, nitrogen and mineral salt with a new strain of Claviceps paspali Stevens and Hall as herein before defined. 

As regards the production of alkaloid derivatives of lysergic acid, the present invention is not limited to the use of the described strains, but comprises also the mutants thereof, which may be obtained, e.g., by means of either a selection or a mutation by the action of U.V. rays or Roentgen rays or any other mutagenous substance or, particularly, by artificial infection of either embryos or grasses cultivated in vitro or plants of grasses cultivated both in vivo or in vitro and the said mutants are to be included in the definition of a new strain of Claviceps paspali Stevens and Hall. 

The behavior of the strains described in the present invention when grown in presence of Fe-i-+ and Zn++ in the medium, is different from that of the strains of Claviceps purpurea described by Stoll et al. (1957). These two elements may decrease the production of alkaloids markedly. 

In this medium, the average production of alkaloids reaches 1000 ug./mLi. after 7-9 days incubation. 

The fermentation is carried out according to the procedure in Example 1. After 7-9 days incubation, the production of alkaloids reaches the value of 1000 ug./mL. The same yield is obtained if tartaric acid, citric acid, malic acid, acetic acid, fumaric acid, succinic acid are used. 

The pH is adjusted to 5.2 with aqueous ammonia solution. The fermentation is carried out as described earlier. After 4-7 days culture the production of alkaloids reaches 1400-1600 ug./mL. 

The organisms grow best at temperatiu-es in the range of about 22° C. to about 28° C. Optimal production of alkaloids appears to occur at a temperature of about 23° C. to 24° C. 

For optimum production of alkaloids, it is important that the mycelium employed for the inoculation of a liquid culture medium be maintained on solid media exclusively prior to transfer to submerged culture. Thus, for example, yields are significantly depressed when the mycelium has been transferred from a submerged culture to surface culture on solid media prior to inoculation into submerged culture. The optimum route for preparation of inoculum therefore is from surface culture to submerged medium or from surface culture, through a series of transfers on surface culture, to submerged medium... 

It is of interest to note that addition of tryptophan to the culture medium does not appear to enhance the production of alkaloids by the new strain of Claviceps paspali employed herein. Also noteworthy is the fact that relatively high concentrations of iron appear to promote, or at least support, alkaloid synthesis by this strain. Thus, for example, concentrations of FeS04.7H20 in the range of 100 - 200 mg. per liter appear to be without detrimental effect. 

Rehacek Z, Kozova J. Production of alkaloids and differentiation in a submerged culture of Claviceps purpurea (Fr.) Tul. Folia Microbiol (Praha) 20(2) 112-117, 1975. 

Most field isolates of C. purpurea obtained in screening programs over the years yielded interesting strains producing different, specific alkaloids (or desired mixtures) at moderate rates in planta only, thus they were not suited for submersed production of alkaloids. Strain-improvement programs focused therefore on an increased capacity of strains to produce high amounts of a specific alkaloid in axenic culture. This was mainly achieved, as in other... 

Spalla C, Guicciardi A, Marnati MP, Oddo N. Protoplasts and production of alkaloids with a heterokaryotic strain of Claviceps purpurea. Abstracts. Fifth Int Ferment Symp. 

Sawada and coworkers 2 have reported preliminary studies on the induction of callus tissues from Aconitum ibukiense var. eizaenense Nakai. The production of alkaloids by these callus tissues was indicated by a positive Dragendorff test of the weak base fraction of an extract. These studies are significant in that such methods might be useful for the production of alkaloids from rare species of plants. 

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