Factors Influencing Biotransformations

By: Pharma Tips | Views: 4969 | Date: 01-May-2011

Many substrates are harmful to cultured cells. So it is necessary to decrease the toxicity in order to increase the yield of the products.

1. Improvements cell viability

            Many substrates are harmful to cultured cells. So it is necessary to decrease the toxicity in order to increase the yield of the products.

            Yokoyama et al. (1990) proposed that sugar could increase cell viability during glycosylation of phenolic compounds. They reported that sucrose (6%w/v) improved the cell viability to a large extent when hydroquinone was biotransformed to arbutin by Catharanthus roseus. Arbutin production was enhanced by 2-3 folds, with the increase being directly dependent on the concentration of sucrose. The exogenously added sucrose was not metabolized and the concentration remained unchanged. The reason for the effect is probably related to the effect of these sugars to act as the scavenger of hydroxyl radicals presumed to be generated on the surface or within the cell. A sugar is known to be specific scavengers of hydroxyl radical and not reacts with superoxide. These assumption leads to the conclusion that antioxidants can improve cell viability and increase the product formation in the biotransformation of phenolics. Yokoyama (1991) reported that antioxidants such as Gallic acid, Ascorbic acid, Cystein and Tannins at 200mg/l could increase the production of arbutin when hydroquinone was added to the cell cultures of Chatharanthus roseus.

2. Selection of Plant Species
           The capacity for biotransformation is diverse among plant species. Tabatas et al. (1988) reported that among seven species of plant cell cultures, the datura had only capacity to biotransform coumarins, flavonoids, anthraquinones and phenolic acids. Datura had the greatest capability to glycosylate coumarins, where as Mallotas japonica did not biotransform coumarins. Different biotransformation can be found among strains of the same species in culture. Courtois et al (1988) showed 3.5  fold difference in the biotransformation rate of tryptamine to serotonin by Peganum harmala cell cultures.
            There are several reports on effect of culture age on glucosylation biotransformation. The culture age yielding glucosylation is variously reported. The lag phase or exponential phase,  when pentachlorophenol was glucosylated using soybean cell cultures or late exponential phase when umbelliferone was glucosylated by Datura innoxia or exponential phase when salicyl alcohol was glucosylated by gardenia jasminoide. However these results do not directly reflect the biotransformation ability of the cells.(15)

3. Immobilized Plant Cells
            Immobilization of plant cells has some distinct advantages e.g. reuse of the expensive biocatalyst, continuous process and process control is simplified. polyurethane foam immobilized cells of Pyrethrum somniferum biotransformed codeinone to codeine with increased efficiency (57-79%) as compared to suspension cultures. Using alginate immobilized cells of the same species, the biotransformation efficiency was 70.4% compared with cell cultures of 60.8%. It was interesting to note that 88% of the converted codeine was excreted into the medium in both experiments. It was observed that the immobilization does not effect rather decreases biotransformation ability.

4. Root Culture
            Cell suspension cultures are generally believed to have excellent biotransformation capacity, especially for glucosylation. However, furuya et al (1989) has found that the root cultures showed higher glycosylation activity than cell culture. Panax ginseng root cultures are able to covert (R,S) -2 phenyl propionic acid into four kinds of glycosyl conjugates where as the cell cultures biotransformed 2-phenyl propionic acid into only (R,S) -2-phenyl propionyl-β-d-glucopyranoside.(16)

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