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P R O D U C T IO N O F B IO A C T IV E S U B S T A N C E S B Y U N C O N V E N T IO N A L
T E C H N O L O G IE S ________________________ '
EXTRACELLULARSYNTHESISOFAJMALINE
M.G.Smirnova,N.V.Kirillova, L.A.Troitskaya, V.P.Komov
St.Petersburg State Chemical PharmaceuticalAcademy, 197376St.Petersburg
Prof.PopovStr. 14, E-mail:
Ajmaline is a monoterpenoid indole alkaloid that is widely used in
cardiology as a high effective antiarrhythmic the same time
comp lex chem ical structure o f this synthesis.Today some enzymes
catalyzing the formation of Rauwolfia alkaloids are intensively studied,e.g.
17-0-acetylajmalin:acetylesterase (AAE).This enzyme catalyzes the reaction
o f deacetylation of acetylajmaline leading to the formation of ajmaline and
acetic observed that maxima o f AAE activity were reached by the
15th,45th and 70th day o f tissue growth.The scheme o f AAE isolation and
purification included supernatant production.precipitation with ammonium
sulfate,gelfiltration on G-150,ion-exchange chromatography on DEAE-
sephacel and CM-sephadex.Using this scheme o f AAE isolation allowed us
to reach 400-fold purification with 30% yield of the enzyme.The purified AAE
was homogeneous according to the electrophoresis data, its spesific activity
was 87-187 nkat/mg.For extracellular synthesis of ajmaline in vitro highly
purified enzyme was used. Its optimal activity was at pH=6,0; 40°C,duration
o f incubation was 10 min.Qualitative and quantitative methods of analysis
(thin layer chromatography and HPLC) were used fo r the determination of
17-0-acetylajmaline and ajmaline in cell culture a result,we
demonstrated the possibility to use preparation ofAAE for ajmaline synthesis
in v itro : using our experiment scheme ajmaline increase in Rauwolfia cell
extract was 10-11 %.
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