Szopa Agnieszka, Kokotkiewicz Adam, Luczkiewicz Maria, Ekiert Halina
Chair and Department of Pharmaceutical Botany, Jagiellonian University, Collegium Medicum, ul. Medyczna 9, 30-688 Kraków, Poland.
Chair and Department of Pharmacognosy, Faculty of Pharmacy, Medical University of Gdansk, al. gen. J. Hallera 107, 80-416 Gdańsk, Poland.
J Biotechnol. 2017 Apr 10;247:11-17. doi: 10.1016/j.jbiotec.2017.02.007. Epub 2017 Feb 14.
Schisandra chinensis (Chinese magnolia vine) is a rich source of therapeutically relevant dibenzocyclooctadiene lignans with anticancer, immunostimulant and hepatoprotective activities. In this work, shoot cultures of S. chinensis were grown in different types of bioreactors with the aim to select a system suitable for the large scale in vitro production of schisandra lignans. The cultures were maintained in Murashige-Skoog (MS) medium supplemented with 3mg/l 6-benzylaminopurine (BA) and 1mg/l 1-naphthaleneacetic acid (NAA). Five bioreactors differing with respect to cultivation mode were tested: two liquid-phase systems (baloon-type bioreactor and bubble-column bioreactor with biomass immobilization), the gas-phase spray bioreactor and two commercially available temporary immersion systems: RITA and Plantform. The experiments were run for 30 and 60 days in batch mode. The harvested shoots were evaluated for growth and lignan content determined by LC-DAD and LC-DAD-ESI-MS. Of the tested bioreactors, temporary immersion systems provided the best results with respect to biomass production and lignan accumulation: RITA bioreactor yielded 17.86g/l (dry weight) during 60 day growth period whereas shoots grown for 30 days in Plantform bioreactor contained the highest amount of lignans (546.98mg/100g dry weight), with schisandrin, deoxyschisandrin and gomisin A as the major constituents (118.59, 77.66 and 67.86mg/100g dry weight, respectively).
五味子是具有抗癌、免疫刺激和保肝活性的治疗相关二苯并环辛二烯木脂素的丰富来源。在本研究中,五味子的茎尖培养物在不同类型的生物反应器中生长,目的是选择一个适合大规模体外生产五味子木脂素的系统。培养物在添加了3mg/l 6-苄基腺嘌呤(BA)和1mg/l 1-萘乙酸(NAA)的Murashige-Skoog(MS)培养基中维持。测试了五种在培养模式上不同的生物反应器:两种液相系统(气球型生物反应器和带有生物质固定化的鼓泡柱生物反应器)、气相喷雾生物反应器和两种市售的临时浸没系统:RITA和Plantform。实验以分批模式运行30天和60天。对收获的茎尖进行生长评估,并通过LC-DAD和LC-DAD-ESI-MS测定木脂素含量。在所测试的生物反应器中,临时浸没系统在生物质生产和木脂素积累方面提供了最佳结果:RITA生物反应器在60天的生长期间产生了17.86g/l(干重),而在Plantform生物反应器中生长30天的茎尖含有最高量的木脂素(546.98mg/100g干重),其中五味子醇甲、脱氧五味子醇甲和戈米辛A为主要成分(分别为118.59、77.66和67.86mg/100g干重)。