3D-printed autoclavable plant holders to facilitate large-scale protein production in plants

IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Engineering in Life Sciences Pub Date : 2022-08-15 DOI:10.1002/elsc.202200001
Ling Chuang, Anton Enders, Sascha Offermann, Janina Bahnemann, Jakob Franke
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Abstract

The Australian tobacco plant Nicotiana benthamiana is becoming increasingly popular as a platform for protein production and metabolic engineering. In this system, gene expression is achieved transiently by infiltrating N. benthamiana plants with suspensions of Agrobacterium tumefaciens carrying vectors with the target genes. To infiltrate larger numbers of plants, vacuum infiltration is the most efficient approach known, which is already used on industrial scale. Current laboratory-scale solutions for vacuum infiltration, however, either require expensive custom-tailored equipment or produce large amounts of biologically contaminated waste. To overcome these problems and lower the burden to establish vacuum infiltration in new laboratories, we present here 3D-printed plant holders for vacuum infiltration. We demonstrate that our plant holders are simple to use and enable a throughput of around 40 plants per hour. In addition, our 3D-printed plant holders are made from autoclavable material, which tolerate at least 12 autoclave cycles, helping to limit the production of contaminated waste and thus contributing to increased sustainability in research. In conclusion, our plant holders provide a simple, robust, safe and transparent platform for laboratory-scale vacuum infiltration that can be readily adopted by new laboratories interested in protein and metabolite production in Nicotiana benthamiana.

Practical application

Transient expression in Nicotiana benthamiana provides a popular and rapid system for producing proteins in a plant host. To infiltrate larger numbers of plants (typically >20), vacuum infiltration is the method of choice. However, no system has been described so far which is robust to use and can be used without expensive and complex equipment. Our autoclavable 3D-printed plant holders presented here will greatly reduce the efforts required to adopt the vacuum infiltration technique in new laboratories. They are easy to use and can be autoclaved at least 12 times, which contributes to waste reduction and sustainability in research laboratories. We anticipate that the 3D printing design provided here will drastically lower the bar for new groups to employ vacuum infiltration for producing proteins and metabolites in Nicotiana benthamiana.

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3d打印的可高压灭菌植物支架,以促进植物中的大规模蛋白质生产
作为蛋白质生产和代谢工程的平台,澳大利亚烟草植物benthamiana正变得越来越受欢迎。在该系统中,通过将农杆菌悬浮液携带携带目标基因的载体浸润本菌属植物,实现基因的短暂表达。为了渗透更多的植物,真空渗透是已知的最有效的方法,已经在工业规模上使用。然而,目前实验室规模的真空渗透解决方案要么需要昂贵的定制设备,要么会产生大量受生物污染的废物。为了克服这些问题,降低在新实验室建立真空渗透的负担,我们在这里提出了3d打印的真空渗透装置支架。我们证明我们的植物支架使用简单,每小时可实现约40个植物的吞吐量。此外,我们的3d打印植物支架由可高压灭菌的材料制成,可承受至少12个高压灭菌循环,有助于限制污染废物的产生,从而有助于提高研究的可持续性。总之,我们的植物支架为实验室规模的真空渗透提供了一个简单、可靠、安全和透明的平台,可以很容易地被对benthamiana蛋白质和代谢物生产感兴趣的新实验室采用。本拟烟的瞬时表达为植物寄主中蛋白质的快速生成提供了一种流行的系统。为了渗透更多的植物(通常是20),真空渗透是首选的方法。然而,到目前为止,还没有一种系统可以可靠地使用,并且可以在没有昂贵和复杂设备的情况下使用。我们在这里展示的可高压灭菌的3d打印植物支架将大大减少在新实验室采用真空渗透技术所需的努力。它们易于使用,可以至少蒸12次,这有助于减少浪费和研究实验室的可持续性。我们预计,这里提供的3D打印设计将大大降低新群体使用真空渗透来生产本烟中蛋白质和代谢物的门槛。
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来源期刊
Engineering in Life Sciences
Engineering in Life Sciences 工程技术-生物工程与应用微生物
CiteScore
6.40
自引率
3.70%
发文量
81
审稿时长
3 months
期刊介绍: Engineering in Life Sciences (ELS) focuses on engineering principles and innovations in life sciences and biotechnology. Life sciences and biotechnology covered in ELS encompass the use of biomolecules (e.g. proteins/enzymes), cells (microbial, plant and mammalian origins) and biomaterials for biosynthesis, biotransformation, cell-based treatment and bio-based solutions in industrial and pharmaceutical biotechnologies as well as in biomedicine. ELS especially aims to promote interdisciplinary collaborations among biologists, biotechnologists and engineers for quantitative understanding and holistic engineering (design-built-test) of biological parts and processes in the different application areas.
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