Sieb Batterman

The accumulation and isolation of coniferin from a high-producing cell suspension of Linum flavum L.

Cell suspensions of Linum flavum L. contained large amounts (2 g·l−1) of the glucoside coniferin which was accumulated endogenously up to 12.4% on a dryweight basis. Callus material contained 5.6%, while in leaves of in-vitro-grown plantlets, the origin of the callus and suspension cultures, no coniferin could be detected. Leaf, callus and suspension material were compared for metabolite accumulation and associated enzyme activities. High coniferin contents corresponded with low 5-methoxypodophyllotoxin levels. A reciprocal relationship between β-glucosidase (E.C. 3.2.1.21) activity and coniferin accumulation was found. No relationship between peroxidase (E.C. 1.11.1.7) activity and coniferin accumulation or 5-methoxypodophyllotoxin could be demonstrated. Finally, a rapid and effective isolation procedure for coniferin was developed.

MUCUNA PRURIENS - IMPROVEMENT OF THE BIOTECHNOLOGICAL PRODUCTION OF THE ANTI-PARKINSON DRUG L-DOPA BY PLANT-CELL SELECTION

Routinely grown cell suspension cultures ofMucuna pruriens L. (Fabaceae) were able to endogenously accumulate the anti-Parkinson drug L-dihydroxyphenylalanine (L-dopa) in the range between 0.2 and 2% on a dry weight (DW) basis. The green colour that developed in light-exposed cultures, appeared to be a suitable marker to select cells with an increased L-dopa biosynthesis and/or phenoloxidase activity. For this purpose, saccharose concentrations from 0 to 4% (w/v), and light intensities of 1,000 and 2,000 lux, were involved in the selection procedure. After 6 months, photomixotrophic callus cultures with a rapid growth and a high L-dopa content of 0.9% (DW) were obtained on 2% saccharose and under 1,000 lux. The cell suspensions, derived from these calli, accumulated up to 6% (DW) L-dopa, which was the highest stable content ever measured in cultures ofM. pruriens. An L-dopa yield of approximately 1.2 g/l was calculated after 6 days of growth. In contrast, compared with the standard-grown parent cell line, the phenoloxidase activity, and consequently the bioconversion capacity as measured after entrapment in calcium alginate, of these high-producing cultures was approximately threefold lower.

High Peroxidase Activity In Cell Cultures of Artemisia Annua With Minute Artemisinin Contents

Abstract In callus and cell suspension cultures of Artemisia annua minute amounts of artemisinin were found. In these cultures, a high peroxidase activity was measured, intracellularly and, especially, in the medium. In vitro, artemisinin rapidly decomposed when incubated with a cell homogenate of A. annua or with spent culture medium, as well as in a solution of commercially obtained horseradish peroxidase (EC 1.11.1.7). In contrast, the peroxidase activity in the intact plant, containing considerable amounts of artemisinin, was low. We suggest that the high peroxidase activity in A. annua cell cultures contributes to their very low artemisinin contents.

Alkanes of the essential oil of Cannabis sativa

Abstract A waxy fraction obtained by column chromatography of the essential oil of Cannabis sativa consists of n-alkanes ranging from C9 to C39, 2-methyl and 3-methyl alkanes and some dimethyl alkanes. The qualitative and quantitative composition of this fraction has been compared with the alkane fraction obtained by extraction of the herb.

Continuous production of the pharmaceutical 7,8-dihydroxy N-di-n-propyl 2-aminotetralin using a phenoloxidase from cell cultures of Mucuna pruriens

Alginate-entrapped cells of Mucuna pruriens as well as the phenoloxidase isolated from the cell cultures, are able to ortho-hydroxylate several mono-, bi- and tri-cyclic monophenols. In this study, 7,8-dihydroxy N-di-n-propyl 2-aminotetralin, a catechol of pharmaceutical interest and difficult to prepare chemically, could be produced in considerable quantities by bioconversion of the precursor 7-hydroxy N-di-n-propyl 2-aminotetralin. A continuous flow system on a laboratory scale was used, which consisted of a phenoloxidase suspension in dialysis tubing as the biocatalysator in an airlift fermentor coupled with an aluminium oxide column for selective product isolation. Product formation continued for at least 50 h, resulting in ca. 130 mg product per liter, this being a bioconversion percentage of 25%. When the enzyme preparation was reused, 85% of the original activity was measured.