Abstract
Two‐phase bioreactor cultivation system was developed and applied for in sito recovery of extracellular galanthamine during the cultivation of Leucojum aestivum L. shoot culture in a modified column bioreactor system. The inclusion of an external circulation column with adsorbent resin Amberlite XAD‐4 as a second phase, on the 21st day of the beginning of cultivation resulted in 1.25 folds increase in biomass accumulation and maximal amounts of accumulated galanthamine of 6 mg/L (3.1 mg/L intracellular and 2.9 mg/L extracellular). It was demonstrated that the inclusion of a second phase at the cultivation of the L. aestivum shoot culture in a bubble column bioreactor with internal sections redirected the alkaloid metabolism to galanthamine synthesis and inhibits the synthesis of hemanthamine and lycorine type alkaloids. Our research demonstrated that the application of the two‐phase cultivation systems could be an important tool to increase the yields of valuable secondary metabolites in plant tissue culture‐based bioprocess.
Highlights
Amaryllidaceae alkaloid galanthamine is a long-acting, reversible, selective, and competitive acetylcholinesterase inhibitor that crosses the blood–brain barrier [1]
Many natural L. aestivum habitats have shrunk or fallen under threat due to the growing demand by the pharmaceutical companies and, the biosynthesis of galanthamine by plant in vitro systems has been considered as a prospective alternative approach for sustainable production of this valuable alkaloid
We noted that the two-phase process for in situ extraction of secreted galanthamine could be a prospective technological decision to increase the effectiveness of the modified column bioreactor
Summary
In sito galanthamine extraction during the cultivation of Leucojum aestivum L. shoot culture in two-phase bubble column cultivation system. Two-phase bioreactor cultivation system was developed and applied for in sito recovery of extracellular galanthamine during the cultivation of Leucojum aestivum L. shoot culture in a modified column bioreactor system. The inclusion of an external circulation column with adsorbent resin Amberlite XAD-4 as a second phase, on the 21st day of the beginning of cultivation resulted in 1.25 folds increase in biomass accumulation and maximal amounts of accumulated galanthamine of 6 mg/L (3.1 mg/L intracellular and 2.9 mg/L extracellular). It was demonstrated that the inclusion of a second phase at the cultivation of the L. aestivum shoot culture in a bubble column bioreactor with internal sections redirected the alkaloid metabolism to galanthamine synthesis and inhibits the synthesis of hemanthamine and lycorine type alkaloids. KEYWORDS absorption resin, bubble-column cultivation system, galanthamine, shoot cultures, summer snowflake
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