用不同诱导剂和生物合成前体处理的 Lepechinia meyenii 细胞悬液中鼠尾草酸和鼠尾草酚的控制生产,Chemistry & Biodiversity

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用不同诱导剂和生物合成前体处理的 Lepechinia meyenii 细胞悬液中鼠尾草酸和鼠尾草酚的控制生产
Chemistry & Biodiversity ( IF 2.3 ) Pub Date : 2022-12-23 , DOI: 10.1002/cbdv.202200733
Nemesio Villa-Ruano ₁ , Néstor Hernández-Silva ₂ , Jenaro Leocadio Varela-Caselis ₃ , Sergio Alberto-Ramirez-Garcia ₄ , Clemente Mosso-González ₅

Affiliation

麻雀是一种药用植物，专门用于生物合成不同类型的抗氧化剂，包括二萜鼠尾草酸 (CA) 酸和鼠尾草酚 (CS)。在此，我们介绍了在这种药用植物中进行的植物组织培养方法的结果，特别强调了用于 CA 和 CS 生产的细胞悬浮系统的生成和评估。研究了蔗糖浓度、温度、pH 和紫外线照射的影响。此外，还对不同浓度的微生物诱导剂（水杨酸、绿脓素、Glucanex 和几丁质）、非生物诱导剂模拟物（聚乙二醇和 NaCl）和生物合成前体（甲瓦龙酸内酯、香叶基香叶醇和 miltiradiene/abietatriene）进行了分批培养评估20天。酿酒酵母。我们的结果表明，CA 和 CS 的最大积累 (Acc _max ) 主要是由于与氧化应激相关的刺激，例如紫外线照射 (Acc _max , 6.2 mg L ⁻¹ ) 聚乙二醇 (Acc _max , 6.5 mg L ^{− 1} ) 分别模拟干旱和盐胁迫的NaCl (Acc _max , 5.9 mg L ^-1 )。然而，细菌诱导剂绿脓素也可有效增加两种二萜的产量（Acc _max，6.4 mg L ^-1). 值得注意的是，上游生物合成前体（如香叶基香叶醇和 miltiradiene/abietatriene）的掺入产生了最佳结果，Acc _max分别为 8.6 和 16.7 mg L ^-1。^{含有 100 mg L −1}香叶基香叶醇、50 mg L ⁻¹米替拉迪烯/松香三烯 (95 : 5 %) 和 5 g L ⁻¹聚乙二醇的优化批次培养物，在 30 天内用 6 分钟紫外线脉冲处理，导致 Acc_最大值为 26.7 mg在第 18-24 天，CA 的L ^-1和CS 的17.3 mg L ^{-1 。}与没有诱导的分批培养相比，该策略允许将 CA 和 CS 的数量增加七倍（Acc _max, 4.3 毫克·升^-1 )。

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Controlled Production of Carnosic Acid and Carnosol in Cell Suspensions of Lepechinia meyenii Treated with Different Elicitors and Biosynthetic Precursors

Lepechinia meyenii is a medicinal plant specialized in the biosynthesis of different types of antioxidants including the diterpenes carnosic (CA) acid and carnosol (CS). Herein we present the results of plant tissue culture approaches performed in this medicinal plant with particular emphasis on the generation and evaluation of a cell suspension system for CA and CS production. The effect of sucrose concentration, temperature, pH, and UV-light exposure was explored. In addition, diverse concentrations of microbial elicitors (salicylic acid, pyocyanin, Glucanex, and chitin), simulators of abiotic elicitors (polyethylene glycol and NaCl), and biosynthetic precursors (mevalonolactone, geranylgeraniol, and miltiradiene/abietatriene) were evaluated on batch cultures for 20 days. Miltiradiene/abietatriene obtainment was achieved through a metabolic engineering approach using a recombinant strain of Saccharomyces cerevisiae. Our results suggested that the maximum accumulation (Acc_max) of CA and CS was mainly conferred to stimuli associated with oxidative stress such as UV-light exposure (Acc_max, 6.2 mg L⁻¹) polyethylene glycol (Acc_max, 6.5 mg L⁻¹) NaCl (Acc_max, 5.9 mg L⁻¹) which simulated drought and saline stress, respectively. Nevertheless the bacterial elicitor pyocyanin was also effective to increase the production of both diterpenes (Acc_max, 6.4 mg L⁻¹). Outstandingly, the incorporation of upstream biosynthetic precursors such as geranylgeraniol and miltiradiene/abietatriene, generated the best results with Acc_max of 8.6 and 16.7 mg L⁻¹, respectively. Optimized batch cultures containing 100 mg L⁻¹ geranylgeraniol, 50 mg L⁻¹ miltiradiene/abietatriene (95 : 5 %) and 5 g L⁻¹ polyethylene glycol treated with 6 min UV light pulse during 30 days resulted in Acc_max of 26.7 mg L⁻¹ for CA and 17.3 mg L⁻¹ for CS on days 18–24. This strategy allowed to increase seven folds the amounts of CA and CS in comparison with batch cultures without elicitation (Acc_max, 4.3 mg L⁻¹).

更新日期：2022-12-23

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