{"title":"合理优化批量生物反应器培养紫罗兰植物细胞,实现其主要生物活性成分的可持续生产","authors":"R. Babu, Smita Srivastava","doi":"10.1007/s11240-024-02829-y","DOIUrl":null,"url":null,"abstract":"<p><i>Viola odorata</i> plant extracts are extensively used in indigenous medicine to treat respiratory diseases. <i>V. odorata’s</i> limited availability in nature, due to geographical conditions and adulteration, makes continuous supply of high-quality plant raw material challenging. This results in low-quality plant extracts that can have inconsistent and sometimes adverse effects when used in medicine. As an effective solution, in vitro plant biomass cultivation conserves plant diversity, ensuring high-quality raw materials with uniform quality and quantity. Toward this, the current study focuses on establishing bioprocess strategy to achieve maximum. <i>V. odorata</i> biomass productivity up to bioreactor level. In this context, in vitro culture conditions were rationally optimised, focusing upon both statistical optimization and one-factor-at-a-time (OFAT) approach, for improved biomass productivity. Inoculum density, macronutrient concentration, shaking speed, temperature, pH and light intensity were systematically optimized to achieve the maximum biomass. This led to increased biomass production from 10.2 ± 0.8 g DW L<sup>− 1</sup> to 21.7 ± 0.8 g DW L<sup>− 1</sup> within 12 days of the batch cultivation period. Optimized culture conditions were implemented in three different types of bioreactors and cultivation in a stirred tank reactor produced ∼ 19.7 g DW L<sup>− 1</sup> of maximum biomass within a 12 day batch cultivation period. Further, the metabolite profile of the bioreactor cultivated biomass (alcoholic extract) was compared with that of the natural plant biomass using gas chromatography-mass spectrometry-based metabolomics. The antibacterial activity of the plant extract was tested against three bacteria responsible for respiratory tract infections, and their antioxidant activity was compared with that of Vitamin E, wherein the results demonstrated comparable activity. The novelty of this study is in the fact that the <i>V. odorata</i> plant cells were cultivated in bioreactors for the first time for potential medicinal applications. 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In this context, in vitro culture conditions were rationally optimised, focusing upon both statistical optimization and one-factor-at-a-time (OFAT) approach, for improved biomass productivity. Inoculum density, macronutrient concentration, shaking speed, temperature, pH and light intensity were systematically optimized to achieve the maximum biomass. This led to increased biomass production from 10.2 ± 0.8 g DW L<sup>− 1</sup> to 21.7 ± 0.8 g DW L<sup>− 1</sup> within 12 days of the batch cultivation period. Optimized culture conditions were implemented in three different types of bioreactors and cultivation in a stirred tank reactor produced ∼ 19.7 g DW L<sup>− 1</sup> of maximum biomass within a 12 day batch cultivation period. Further, the metabolite profile of the bioreactor cultivated biomass (alcoholic extract) was compared with that of the natural plant biomass using gas chromatography-mass spectrometry-based metabolomics. The antibacterial activity of the plant extract was tested against three bacteria responsible for respiratory tract infections, and their antioxidant activity was compared with that of Vitamin E, wherein the results demonstrated comparable activity. The novelty of this study is in the fact that the <i>V. odorata</i> plant cells were cultivated in bioreactors for the first time for potential medicinal applications. 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引用次数: 0
摘要
臭紫罗兰(Viola odorata)植物提取物被广泛应用于治疗呼吸系统疾病的本土药物中。由于地理条件和掺假等原因,臭紫罗兰在自然界中的供应有限,因此持续供应高质量的植物原料具有挑战性。这就导致低质量的植物提取物在用于医药时可能会产生不一致的效果,有时甚至会产生不良影响。作为一种有效的解决方案,体外植物生物质栽培可以保护植物多样性,确保高质量的原材料具有统一的质量和数量。为此,目前的研究重点是建立生物过程策略,以实现最大的。臭腥草的生物量生产率达到生物反应器水平。在此背景下,对体外培养条件进行了合理优化,重点是统计优化和一次一因素(OFAT)方法,以提高生物量生产率。对接种物密度、常量营养素浓度、振荡速度、温度、pH 值和光照强度进行了系统优化,以获得最大生物量。这使得生物量产量在批量培养期的 12 天内从 10.2 ± 0.8 g DW L- 1 提高到 21.7 ± 0.8 g DW L- 1。在三种不同类型的生物反应器中实施了优化的培养条件,在搅拌罐反应器中的培养在 12 天的批量培养期内产生了 19.7 g DW L- 1 的最大生物量。此外,利用基于气相色谱-质谱联用技术的代谢组学方法,将生物反应器培养的生物质(酒精提取物)的代谢物谱与天然植物生物质的代谢物谱进行了比较。此外,还测试了植物提取物对三种呼吸道感染细菌的抗菌活性,并将其抗氧化活性与维生素 E 的抗氧化活性进行了比较,结果表明两者的活性相当。这项研究的新颖之处在于首次在生物反应器中培养了臭腥草植物细胞,并将其用于潜在的医药应用。它为植物生物质的可持续生产提供了一种替代方法,从而保护了天然植物资源。
A rationally optimised batch bioreactor cultivation of Viola odorata plant cells for sustainable production of its key bioactive principles
Viola odorata plant extracts are extensively used in indigenous medicine to treat respiratory diseases. V. odorata’s limited availability in nature, due to geographical conditions and adulteration, makes continuous supply of high-quality plant raw material challenging. This results in low-quality plant extracts that can have inconsistent and sometimes adverse effects when used in medicine. As an effective solution, in vitro plant biomass cultivation conserves plant diversity, ensuring high-quality raw materials with uniform quality and quantity. Toward this, the current study focuses on establishing bioprocess strategy to achieve maximum. V. odorata biomass productivity up to bioreactor level. In this context, in vitro culture conditions were rationally optimised, focusing upon both statistical optimization and one-factor-at-a-time (OFAT) approach, for improved biomass productivity. Inoculum density, macronutrient concentration, shaking speed, temperature, pH and light intensity were systematically optimized to achieve the maximum biomass. This led to increased biomass production from 10.2 ± 0.8 g DW L− 1 to 21.7 ± 0.8 g DW L− 1 within 12 days of the batch cultivation period. Optimized culture conditions were implemented in three different types of bioreactors and cultivation in a stirred tank reactor produced ∼ 19.7 g DW L− 1 of maximum biomass within a 12 day batch cultivation period. Further, the metabolite profile of the bioreactor cultivated biomass (alcoholic extract) was compared with that of the natural plant biomass using gas chromatography-mass spectrometry-based metabolomics. The antibacterial activity of the plant extract was tested against three bacteria responsible for respiratory tract infections, and their antioxidant activity was compared with that of Vitamin E, wherein the results demonstrated comparable activity. The novelty of this study is in the fact that the V. odorata plant cells were cultivated in bioreactors for the first time for potential medicinal applications. It provides an alternative method for sustainable production of plant biomass, thereby conserving the natural plant sources.
期刊介绍:
This journal highlights the myriad breakthrough technologies and discoveries in plant biology and biotechnology. Plant Cell, Tissue and Organ Culture (PCTOC: Journal of Plant Biotechnology) details high-throughput analysis of gene function and expression, gene silencing and overexpression analyses, RNAi, siRNA, and miRNA studies, and much more. It examines the transcriptional and/or translational events involved in gene regulation as well as those molecular controls involved in morphogenesis of plant cells and tissues.
The journal also covers practical and applied plant biotechnology, including regeneration, organogenesis and somatic embryogenesis, gene transfer, gene flow, secondary metabolites, metabolic engineering, and impact of transgene(s) dissemination into managed and unmanaged plant systems.