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M ETABOLISM O F PLANT C E L L S IN CULTURE
INVESTIGATIONOF SUPEROXIDEDISMUTASE INCALLUS CULTUREOF
PANAXGINSENG
N.V.KIRILLOVA,YU.V.KOMOV
St.PetersburgStale ChemicalPharmaceuticalAcademy 197376St.Petersburg
Prof.Popov Str., 14 E -m ail:
Protection of aerobic cells from toxic oxygen radicals achieved, mainly,
by enzymes o f antioxidant system, key component o f which is superoxide
dismutase (SOD).We studied SOD in process o f growth of cell culture of
Rauvolfia serpentina before. In this work activity and some chemical and
physical properties o f SOD in some stages of development of cell culture of
ginseng were examined. Dynamics of level o f activity o f SOD was studied
for 30 days of cell growth in every 4-5 days. It was established that there are
2 peaks o f enzyme activity on 20th and 35th days o f growth, these data
correlates with mitotic activity o f cells of ginseng. It was obtained 2 forms o f
enzyme from cells o f different age. On early stage of cell development and
after 30th day of cell growth two isoforms had same electrophoretic mobility
(Rf1=0.28; Rf2=0.72). However, on 20th days there was a lock o f isoform 2
and appearance of isoform 3 of this enzyme (Rf3=0.94).
For isolation and purification of SOD gel-filtration on G-75 and ion-
exchange chromatography on aminodiacetate cellulose were used.
As a result high-purified enzyme was obtained. It was homogeneous
according to the electrophoresis data, and some physical and chemical
properties were investigated (optimal pH was 7.8; temperature optimal was
20-24 C).
Molecular weight of native SOD was 27.5-30 kDa. According to the
electrophoresis data, enzyme was dimer. Its protomers had molecular weights
16-17 kDa.
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