Time-resolved NMR metabolomics of plant cells based on a microfluidic chip

Maisch, Jan; Kreppenhofer, Kristina; S, Buchler; Merle, Christian; Sobich, Shukhrat; Görling, Benjamin; Luy, Burkhard; Ahrens, Ralf; Guber, A.E.; Nick, Peter

doi:10.1016/j.jplph.2016.06.004

Cited by 13 publications

(14 citation statements)

References 22 publications

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“…One study focused on biotechnological production of secondary metabolites in plants and presented a biomimetic approach in which a real tissue is simulated by a microfluidic chamber system. 155 This study provided proof-of-principle results that a microfluidic system of cultivated plant cells can mimic the metabolic flows in a real plant tissue. Another study investigated the metabolic evolution and cultivar-dependent metabolite variation in soybean leaves using NMR-based metabolomics.…”

Section: Applicationsmentioning

confidence: 76%

Recent Advances in NMR-Based Metabolomics

Gowda¹,

Raftery

2016

Anal. Chem.

136

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Section: Applicationsmentioning

confidence: 76%

Recent Advances in NMR-Based Metabolomics

Gowda¹,

Raftery

2016

Anal. Chem.

136

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“…Moreover, the strategy might be further improved by integrating temporal dynamics to address cell proliferation and metabolic activity differently. For this purpose, microfluidic strategies [52] will be used as platform to integrate different plant-cell based metabolic modules.…”

Section: Discussionmentioning

confidence: 99%

Combination of Plant Metabolic Modules Yields Synthetic Synergies

et al. 2017

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The great potential of pharmacologically active secondary plant metabolites is often limited by low yield and availability of the producing plant. Chemical synthesis of these complex compounds is often too expensive. Plant cell fermentation offers an alternative strategy to overcome these limitations. However, production in batch cell cultures remains often inefficient. One reason might be the fact that different cell types have to interact for metabolite maturation, which is poorly mimicked in suspension cell lines. Using alkaloid metabolism of tobacco, we explore an alternative strategy, where the metabolic interactions of different cell types in a plant tissue are technically mimicked based on different plant-cell based metabolic modules. In this study, we simulate the interaction found between the nicotine secreting cells of the root and the nicotine-converting cells of the senescent leaf, generating the target compound nornicotine in the model cell line tobacco BY-2. When the nicotine demethylase NtomCYP82E4 was overexpressed in tobacco BY-2 cells, nornicotine synthesis was triggered, but only to a minor extent. However, we show here that we can improve the production of nornicotine in this cell line by feeding the precursor, nicotine. Engineering of another cell line overexpressing the key enzyme NtabMPO1 allows to stimulate accumulation and secretion of this precursor. We show that the nornicotine production of NtomCYP82E4 cells can be significantly stimulated by feeding conditioned medium from NtabMPO1 overexpressors without any negative effect on the physiology of the cells. Co-cultivation of NtomCYP82E4 with NtabMPO1 stimulated nornicotine accumulation even further, demonstrating that the physical presence of cells was superior to just feeding the conditioned medium collected from the same cells. These results provide a proof of concept that combination of different metabolic modules can improve the productivity for target compounds in plant cell fermentation.

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“…The authors have developed a microfluidic chip for measuring different metabolites (sucrose, fructose, glucose and myo-inositol) in the flow-through that correspond to the growth state of the tobacco BY-2 cell suspension and the metabolic activity of the cell suspension. For example, the levels of glucose and myo-inositol were very low during the first four days of the cultivation, which corresponded to intensive proliferation, while after day five till the end of the process the levels of glucose and myo-inositol increased due to the decreased catabolic activity and proliferation phase [80].…”

Section: Metabolomics For High-resolution Monitoring Of Secondary Metmentioning

confidence: 99%

“…Focusing on the PPP it was established that the oxidative steps from this pathway were duplicated in the cytosol and plastids, with flux through these reactions occurring largely in the cytosol [79]. An example of NMR spectroscopy used as a real-time monitoring tool of physiological parameters, including cell viability and growth during the cultivation of plant cell suspensions, is described by Maish et al [80]. The authors have developed a microfluidic chip for measuring different metabolites (sucrose, fructose, glucose and myo-inositol) in the flow-through that correspond to the growth state of the tobacco BY-2 cell suspension and the metabolic activity of the cell suspension.…”

Section: Metabolomics For High-resolution Monitoring Of Secondary Metmentioning

confidence: 99%

Green (cell) factories for advanced production of plant secondary metabolites

Marchev

Yordanova

Georgiev

2020

Critical Reviews in Biotechnology

113

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For centuries plants have been intensively utilized as reliable sources of food, flavoring, agrochemical and pharmaceutical ingredients. However, plant natural habitats are being rapidly lost due to climate change and agriculture. Plant biotechnology offers a sustainable method for the bioproduction of plant secondary metabolites using plant in vitro systems. The unique structural features of plant-derived secondary metabolites, such as their safety profile, multi-target spectrum and "metabolite likeness," have led to the establishment of many plant-derived drugs, comprising approximately a quarter of all drugs approved by the Food and Drug Administration and/or European Medicinal Agency. However, there are still many challenges to overcome to enhance the production of these metabolites from plant in vitro systems and establish a sustainable large-scale biotechnological process. These challenges are due to the peculiarities of plant cell metabolism, the complexity of plant secondary metabolite pathways, and the correct selection of bioreactor systems and bioprocess optimization. In this review, we present an integrated overview of the possible avenues for enhancing the biosynthesis of high-value marketable molecules produced by plant in vitro systems. These include metabolic engineering and CRISPR/Cas9 technology for the regulation of plant metabolism through overexpression/repression of single or multiple structural genes or transcriptional factors. The use of NMR-based metabolomics for monitoring metabolite concentrations and additionally as a tool to study the dynamics of plant cell metabolism and nutritional management is discussed here. Different types of bioreactor systems, their modification and optimal process parameters for the labor industrial-scale production of plant secondary metabolites are specified.

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Time-resolved NMR metabolomics of plant cells based on a microfluidic chip

Cited by 13 publications

References 22 publications

Recent Advances in NMR-Based Metabolomics

Recent Advances in NMR-Based Metabolomics

Combination of Plant Metabolic Modules Yields Synthetic Synergies

Green (cell) factories for advanced production of plant secondary metabolites

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