World Journal of Microbiology & Biotechnology 15: 465469, 1999.

1999 Kluwer Academic Publishers. Printed in the Netherlands.

465

Biotransformation of codeine to morphine in freely suspended cells

and immobilized cultures of Spirulina platensis
S. Ramachandra Rao, Usha Tripathi and G.A. Ravishankar*
Plant Cell Biotechnology Department, Central Food Technological Research Institute, Mysore 570 013, India
*Author for correspondence: Tel.: 091-821-516 501, Fax: 091-821-517 233, E-mail: pcbt@cscftri.ren.nic.in
Received in revised form 22 January 1999; accepted 15 April 1999

Keywords: Biotransformation, codeine, immobilization, morphine, Spirulina platensis

Summary
Both freely suspended cells and immobilized cultures of Spirulina platensis, a blue-green alga, biotransformed
exogenously fed codeine, an opium alkaloid, to morphine. The external addition of codeine to the culture medium
did not aect the growth of S. platensis. Immobilization of Spirulina in a calcium alginate gel matrix was optimized
by using 2% (w/v) sodium alginate and reducing the concentration of nutrients of Zarrouk's medium, which caused
destabilization of the calcium alginate gel. The accumulation of morphine increased gradually and reached maxima
of 330 lg 100 ml)1 culture at 105 h in freely suspended and 351 lg 100 ml)1 at 96 h in immobilized Spirulina
cultures. Accumulation of morphine was detected only in the medium, whereas cells did not show accumulation.
The immobilized Spirulina cultures showed marginally higher conversion of codeine to morphine over freely suspended cultures.
Introduction
Biotransformations may be exploited in the synthesis of
products which are normally extracted from the plant,
or in the formation of novel compounds (Suga & Hirata
1990; Pras 1992; Hamada & Furuya 1996; Kutney
1997). The immobilized plant and microbial systems
have been applied to the industrial production of useful
compounds such as food additives and drugs (Brodelius
1988; Scragg 1991). Morphine is the major alkaloid
component of opium, which is the dried latex material
from cut seed capsules of opium poppy, Papaver somniferum, used as most potent analgesic in therapeutics
(Bruce et al. 1995). Numerous reports of the production of morphinans, thebaine, codeine and morphine
from cell cultures of P. somniferum and Papaver bracteatum occur in the literature, although yields are low
compared with the high yields of plants. The structure of
codeine and morphine is shown in Figure 1. The conversion of thebaine into neopine and codeinone to codeine was reported in cell suspension cultures of
P. somniferum (Tam et al. 1982). The bioconversion of
codeine to morphine was demonstrated in isolated
capsules of P. somniferum (Hsu et al. 1985). The enzymatic reduction of codeinone to codeine was reported in
in vitro cell-free systems of P. somniferum and P. bracteatum (Hodges & Rapoport 1980). The conversion of
codeinone to codeine has been shown in calcium alginate immobilized cell cultures (Furuya et al. 1984;
Furusaki et al. 1988) and in polyurethane foam immo-

bilized cell cultures of P. somniferum (Corchete & Yeoman 1989). Similarly, many workers reported on
microbial transformation of Papaver alkaloids, in the
fungus Cunninghamella spp. (Gibson et al. 1984) and in
Streptomyces griseus (Kunz et al. 1985).
O-Demethylation is an important reaction step in the
preparation of drugs, which are dicult or hazardous to
achieve by chemical means utilizing toxic reagents with
low yields (Gibson et al. 1984; Hsu et al. 1985). The use
of microalgae for biotransformation has been occasionally investigated. This approach may be potentially safer
and more ecient for production of drugs. Codeine is
not a constituent of algal cells. Hence, its biotransformation is a novel method wherein a low value compound
could be biotransformed into high value morphine.
So far, there is no report on biotransformation of
codeine to morphine in microorganisms. The advantage
of the present investigation is production of morphine
using photosynthetic organism as a biocatalyst.
The objective of the present investigation was to study
biotransformation of externally fed codeine in freely
suspended as well as immobilized cultures of S. platensis.
Materials and Methods
Microorganism
The organism used in this study was an axenic culture of
Spirulina platensis, CFTRI strain.