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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Improved sanguinarine production via biotic and abiotic elicitations and precursor feeding in cell suspensions of latex-less variety of Papaver somniferum with their gene expression studies and upscaling in bioreactor
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Improved sanguinarine production via biotic and abiotic elicitations and precursor feeding in cell suspensions of latex-less variety of Papaver somniferum with their gene expression studies and upscaling in bioreactor

机译：通过基因表达研究和生物反应器放大，通过生物和非生物诱因以及无乳胶罂粟品种细胞悬液中前体补料改善了血红素碱的生产

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Elicitors play an important role in challenging the plant defense system through plant-environment interaction and thus altering the secondarymetabolite production. Culture filtrates of four endophytic fungi, namely, Chaetomium globosum, Aspergillus niveoglaucus, Paecilomyces lilacinus, and Trichoderma harzianum were tested on embryogenic cell suspensions of latex-less Papaver somniferum in dosedependent kinetics. Besides this, abiotic elicitors salicylic acid, hydrogen peroxide, and carbon dioxide were also applied for improved sanguinarine production. Maximum biomass accumulation (growth index (GI)=293.50±14.82) and sanguinarine production (0.090±0.008 % dry wt.) were registered by addition of 3.3 %v/v T. harzanium culture filtrate. Interestingly, it was further enhanced (GI=323.40±25.30; 0.105±0.008 % dry wt.) when T. harzanium culture filtrate was employed along with 50 μM shikimate. This was also supported by real-time (RT) (qPCR), where 8–9-fold increase in cheilanthifoline synthase (CFS), stylopine synthase (STS), tetrahydroprotoberberine cis-N-methyltransferase (TNMT), and protopine 6-hydroxylase (P6H) transcripts was observed. Among abiotic elicitors, while hydrogen peroxide and carbon dioxide registered low level of sanguinarine accumulation, maximum sanguinarine content was detected by 250 μM salicylic acid (0.058±0.003 % dry wt.; GI=172.75±13.40). RT (qPCR) also confirms the downregulation of sanguinarine pathway on CO_2 supplementation. Various parameters ranging from agitation speed (70 rpm), impeller type (marine), media volume (2 l), inoculum weight (100 g), and culture duration (9 days) were optimized during upscaling in 5-l stirred tank bioreactor to obtain maximum sanguinarine production (GI=434.00; 0.119±0.070 % dry wt.). Addition of 3.3 %v/v T. harzanium culture filtrate and 50-μM shikimate was done on the 6th day of bioreactor run.
机译：引发剂在通过植物与环境的相互作用挑战植物防御系统中发挥着重要作用，从而改变了次级代谢产物的产生。在无胶乳罂粟的胚发生细胞悬液中，对四种内生真菌，即球壳拟南杆菌，普通曲霉，淡淡拟青霉和哈茨木霉的培养滤液进行了剂量依赖性动力学测试。除此之外，非生物引发剂水杨酸，过氧化氢和二氧化碳也被用于改善血红素碱的生产。通过加入3.3％v / v的T.harzanium培养滤液，记录最大的生物量积累（生长指数（GI）= 293.50±14.82）和血红碱碱产量（0.090±0.008％干重）。有趣的是，当与50μMiki草酸酯一起使用哈氏锥虫培养滤液时，其进一步增强（GI = 323.40±25.30;干重0.15％±0.008％）。实时（RT）（qPCR）也支持了这一点，其中乙酰胆碱合酶（CFS），stylopine合酶（STS），四氢原小ine碱顺式-N-甲基转移酶（TNMT）和protopine 6-羟化酶增加了8-9倍（P6H）成绩单被观察到。在非生物引发剂中，虽然过氧化氢和二氧化碳的血红素碱含量较低，但最大的血红素碱含量却是通过250μM水杨酸（0.058±0.003％干重; GI = 172.75±13.40）检测到的。 RT（qPCR）还证实了补充二氧化碳时Sanguinarine途径的下调。在5-l搅拌釜生物反应器升级至生产过程中，优化了搅拌速度（70 rpm），叶轮类型（海洋），培养基体积（2 l），接种量（100 g）和培养时间（9天）等各种参数。获得最大的血红素碱产量（GI ＝ 434.00; 0.119±0.070％干重）。在生物反应器运行的第6天，添加了3.3％v / v的T.harzanium培养滤液和50-μMiki草酸酯。

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