In sito galanthamine extraction during the cultivation of Leucojum aestivum L. shoot culture in two‐phase bubble column cultivation system

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.

Recent Progress in Amaryllidaceae Biotechnology

“…As stated in the Introduction, the optimization of the production process of Amaryllidaceae alkaloids is well documented, especially in the case of galanthamine [ 24 , 27 , 29 , 30 , 31 , 32 , 33 ]. It is clear that the main stages of such optimization are: optimization of the nutrient medium for maximal yields of the target alkaloids; selecting the appropriate cultivation system and improving its design; optimization of the environmental conditions of cultivation and process control, management and modeling.…”

“…Results received from this experimental work revealed that more than 40% of alkaloids were secreted into the culture liquid [ 24 ] and this required further improvement of the design of the cultivation system to a two-phase bioreactor cultivation system. An essential part of this cultivation system was an external circulation column filled with adsorbent resin [ 33 ]. Using this system for the cultivation of L. aestivum shoots, metabolism of Amaryllidaceae alkaloids was redirected preliminary to galanthamine synthesis.…”

“…Using this system for the cultivation of L. aestivum shoots, metabolism of Amaryllidaceae alkaloids was redirected preliminary to galanthamine synthesis. The results obtained demonstrated that the two-phase bioreactor cultivation system is an important tool for improving the yields of secondary metabolites in plant in vitro system-based bioprocess and could be the basis for the development of large-scale Amaryllidaceae alkaloid production processes [ 24 , 33 ].…”

“…A significant success has been demonstrated in the development of in vitro systems for production of Amaryllidaceae alkaloids [ 23 , 24 , 25 , 26 , 27 , 28 ]. Optimization of the production process is also well documented, starting with nutrient media optimization [ 29 ], development of appropriate bioreactor design [ 24 , 30 , 31 ], and optimization of the cultivation conditions [ 27 , 32 , 33 ]. In 2018, Laurain-Mattar and Ptak published a book chapter [ 28 ] which is very informative and helpful to scientists working in this area.…”

In vitro regeneration of bulblet using two and four bulb-scales explants of summer snowflake (Leucojum aestivum L.)

“…Leucojum aestivum L., popularly known as summer snowflake, is an example of valuable plants which, despite its extremely high industrial potential, has received little attention regarding its protection and propagation (Kohut et al, 2007). During the past three decades, many natural L. aestivum habitats have fallen under threat due to the increasing demand by the pharmaceutical companies (Ivanov et al, 2019), while Ekıcı (2017) claimed that this plant species has the potential for an alternative tourism type, i.e. flora tourism, due to its visual impacts on the landscape.…”