Moss bioreactor

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A moss bioreactor with Physcomitrella patens Bioreaktor quer2.jpg
A moss bioreactor with Physcomitrella patens

A moss bioreactor is a photobioreactor used for the cultivation and propagation of mosses. It is usually used in molecular farming for the production of recombinant protein using transgenic moss. In environmental science moss bioreactors are used to multiply peat mosses e.g. by the Mossclone consortium to monitor air pollution.[ citation needed ]

Contents

Moss is a very frugal photoautotrophic organism that has been kept in vitro for research purposes since the beginning of the 20th century. [1]

The first moss bioreactors for the model organism Physcomitrella patens were developed in the 1990s to comply with the safety standards regarding the handling of genetically modified organisms and to gain sufficient biomass for experimental purposes. [2]

Functional principle

The moss bioreactor is used to cultivate moss in a suspension culture in agitated, and aerated liquid medium. The culture is kept under lighting with temperature and pH value held constant. The culture medium—often a minimal medium—contains all nutrients and minerals needed for growth of the moss. [3]

To ensure a maximum growth rate, the moss is kept at the protonema stage by continuous mechanical disruption, e.g. by using rotating blades. [4] Once the density of the culture has reached a certain threshold, the lack of nutrients and the increasing concentration of phytohormones in the medium triggers the differentiation of the protonema to the adult gametophyte. At this point the culture has to be diluted with fresh medium if it is intended for further use.

According to the intended yield, this basic principle can be adapted to various types and sizes of bioreactors. The cultivation chamber can, for example, consist of a column, a tube, or exchangeable plastic bags. [5]

In this moss bioreactor the peat moss Sphagnum palustre is cultivated Sphagnum palustre in Bioreactor.jpg
In this moss bioreactor the peat moss Sphagnum palustre is cultivated

Production of biopharmaceuticals

Various biopharmaceuticals have already been produced using moss bioreactors. [6] Ideally, the recombinant protein can be directly purified from the culture medium. [7] One example for this production method is factor H: this molecule is part of the human complement system. Defects in the corresponding gene are associated with human diseases such as severe kidney and retinal disorders. Biologically active recombinant factor H was produced in a moss bioreactor for the first time in 2011. [8] The enzyme alpha-galactosidase now is allowed to be produced in moss bioreactors by the German Federal Institute for Drugs and Medical Devices. [9] [10] It will be tested as enzyme replacement therapy in the treatment of Fabry's disease. The clinical trial phase 1 was completed in 2017. [11]

See also

Related Research Articles

<span class="mw-page-title-main">Genetically modified organism</span> Organisms whose genetic material has been altered using genetic engineering methods

A genetically modified organism (GMO) is any organism whose genetic material has been altered using genetic engineering techniques. The exact definition of a genetically modified organism and what constitutes genetic engineering varies, with the most common being an organism altered in a way that "does not occur naturally by mating and/or natural recombination". A wide variety of organisms have been genetically modified (GM), from animals to plants and microorganisms. Genes have been transferred within the same species, across species, and even across kingdoms. New genes can be introduced, or endogenous genes can be enhanced, altered, or knocked out.

<span class="mw-page-title-main">Protein production</span>

Protein production is the biotechnological process of generating a specific protein. It is typically achieved by the manipulation of gene expression in an organism such that it expresses large amounts of a recombinant gene. This includes the transcription of the recombinant DNA to messenger RNA (mRNA), the translation of mRNA into polypeptide chains, which are ultimately folded into functional proteins and may be targeted to specific subcellular or extracellular locations.

<span class="mw-page-title-main">Moss</span> Division of non-vascular land plants

Mosses are small, non-vascular flowerless plants in the taxonomic division Bryophytasensu stricto. Bryophyta may also refer to the parent group bryophytes, which comprise liverworts, mosses, and hornworts. Mosses typically form dense green clumps or mats, often in damp or shady locations. The individual plants are usually composed of simple leaves that are generally only one cell thick, attached to a stem that may be branched or unbranched and has only a limited role in conducting water and nutrients. Although some species have conducting tissues, these are generally poorly developed and structurally different from similar tissue found in vascular plants. Mosses do not have seeds and after fertilisation develop sporophytes with unbranched stalks topped with single capsules containing spores. They are typically 0.2–10 cm (0.1–3.9 in) tall, though some species are much larger. Dawsonia, the tallest moss in the world, can grow to 50 cm (20 in) in height. There are approximately 12,000 species.

<span class="mw-page-title-main">Agar plate</span> Petri dish with agar used to culture microbes

An agar plate is a Petri dish that contains a growth medium solidified with agar, used to culture microorganisms. Sometimes selective compounds are added to influence growth, such as antibiotics.

<span class="mw-page-title-main">Chinese hamster ovary cell</span> Cell line

Chinese hamster ovary (CHO) cells are an epithelial cell line derived from the ovary of the Chinese hamster, often used in biological and medical research and commercially in the production of recombinant therapeutic proteins. They have found wide use in studies of genetics, toxicity screening, nutrition and gene expression, particularly to express recombinant proteins. CHO cells are the most commonly used mammalian hosts for industrial production of recombinant protein therapeutics.

<span class="mw-page-title-main">Bioreactor</span> System that supports a biologically active environment

A bioreactor refers to any manufactured device or system that supports a biologically active environment. In one case, a bioreactor is a vessel in which a chemical process is carried out which involves organisms or biochemically active substances derived from such organisms. This process can either be aerobic or anaerobic. These bioreactors are commonly cylindrical, ranging in size from litres to cubic metres, and are often made of stainless steel. It may also refer to a device or system designed to grow cells or tissues in the context of cell culture. These devices are being developed for use in tissue engineering or biochemical/bioprocess engineering.

<span class="mw-page-title-main">Protoplast</span> Cell stripped of cell-wall

Protoplast, is a biological term coined by Hanstein in 1880 to refer to the entire cell, excluding the cell wall. Protoplasts can be generated by stripping the cell wall from plant, bacterial, or fungal cells by mechanical, chemical or enzymatic means.

<span class="mw-page-title-main">Growth medium</span> Solid, liquid or gel used to grow microorganisms or cells

A growth medium or culture medium is a solid, liquid, or semi-solid designed to support the growth of a population of microorganisms or cells via the process of cell proliferation or small plants like the moss Physcomitrella patens. Different types of media are used for growing different types of cells.

Pharming, a portmanteau of "farming" and "pharmaceutical", refers to the use of genetic engineering to insert genes that code for useful pharmaceuticals into host animals or plants that would otherwise not express those genes, thus creating a genetically modified organism (GMO). Pharming is also known as molecular farming, molecular pharming or biopharming.

A biopharmaceutical, also known as a biological medical product, or biologic, is any pharmaceutical drug product manufactured in, extracted from, or semisynthesized from biological sources. Different from totally synthesized pharmaceuticals, they include vaccines, whole blood, blood components, allergenics, somatic cells, gene therapies, tissues, recombinant therapeutic protein, and living medicines used in cell therapy. Biologics can be composed of sugars, proteins, nucleic acids, or complex combinations of these substances, or may be living cells or tissues. They are isolated from living sources—human, animal, plant, fungal, or microbial. They can be used in both human and animal medicine.

Industrial fermentation is the intentional use of fermentation in manufacturing processes. In addition to the mass production of fermented foods and drinks, industrial fermentation has widespread applications in chemical industry. Commodity chemicals, such as acetic acid, citric acid, and ethanol are made by fermentation. Moreover, nearly all commercially produced industrial enzymes, such as lipase, invertase and rennet, are made by fermentation with genetically modified microbes. In some cases, production of biomass itself is the objective, as is the case for single-cell proteins, baker's yeast, and starter cultures for lactic acid bacteria used in cheesemaking.

<span class="mw-page-title-main">Photobioreactor</span> Bioreactor with a light source to grow photosynthetic microorganisms

A photobioreactor (PBR) refers to any cultivation system designed for growing photoautotrophic organisms using artificial light sources or solar light to facilitate photosynthesis. PBRs are typically used to cultivate microalgae, cyanobacteria, and some mosses. PBRs can be open systems, such as raceway ponds, which rely upon natural sources of light and carbon dioxide. Closed PBRs are flexible systems that can be controlled to the physiological requirements of the cultured organism, resulting in optimal growth rates and purity levels. PBRs are typically used for the cultivation of bioactive compounds for biofuels, pharmaceuticals, and other industrial uses.

<i>Physcomitrella patens</i> Species of moss

Physcomitrium patens, the spreading earthmoss, is a moss (bryophyte) used as a model organism for studies on plant evolution, development, and physiology.

<span class="mw-page-title-main">Gene targeting</span> Genetic technique that uses homologous recombination to change an endogenous gene

Gene targeting is a biotechnological tool used to change the DNA sequence of an organism. It is based on the natural DNA-repair mechanism of Homology Directed Repair (HDR), including Homologous Recombination. Gene targeting can be used to make a range of sizes of DNA edits, from larger DNA edits such as inserting entire new genes into an organism, through to much smaller changes to the existing DNA such as a single base-pair change. Gene targeting relies on the presence of a repair template to introduce the user-defined edits to the DNA. The user will design the repair template to contain the desired edit, flanked by DNA sequence corresponding (homologous) to the region of DNA that the user wants to edit; hence the edit is targeted to a particular genomic region. In this way Gene Targeting is distinct from natural homology-directed repair, during which the ‘natural’ DNA repair template of the sister chromatid is used to repair broken DNA. The alteration of DNA sequence in an organism can be useful in both a research context – for example to understand the biological role of a gene – and in biotechnology, for example to alter the traits of an organism.

<span class="mw-page-title-main">Genetically modified plant</span> Plants with human-introduced genes from other organisms

Genetically modified plants have been engineered for scientific research, to create new colours in plants, deliver vaccines, and to create enhanced crops. Plant genomes can be engineered by physical methods or by use of Agrobacterium for the delivery of sequences hosted in T-DNA binary vectors. Many plant cells are pluripotent, meaning that a single cell from a mature plant can be harvested and then under the right conditions form a new plant. This ability is most often taken advantage by genetic engineers through selecting cells that can successfully be transformed into an adult plant which can then be grown into multiple new plants containing transgene in every cell through a process known as tissue culture.

<span class="mw-page-title-main">Ralf Reski</span> Plant biologist

Ralf Reski is a German professor of plant biotechnology and former dean of the Faculty of Biology of the University of Freiburg. He is also affiliated to the French École supérieure de biotechnologie Strasbourg (ESBS) and Senior Fellow at the Freiburg Institute for Advanced Studies.

<span class="mw-page-title-main">Somatic fusion</span> Genetic modification fusing plants into a hybrid

Somatic fusion, also called protoplast fusion, is a type of genetic modification in plants by which two distinct species of plants are fused together to form a new hybrid plant with the characteristics of both, a somatic hybrid. Hybrids have been produced either between different varieties of the same species or between two different species.

<span class="mw-page-title-main">Algae bioreactor</span> Device used for cultivating micro or macro algae

An algae bioreactor is used for cultivating micro or macroalgae. Algae may be cultivated for the purposes of biomass production (as in a seaweed cultivator), wastewater treatment, CO2 fixation, or aquarium/pond filtration in the form of an algae scrubber. Algae bioreactors vary widely in design, falling broadly into two categories: open reactors and enclosed reactors. Open reactors are exposed to the atmosphere while enclosed reactors, also commonly called photobioreactors, are isolated to varying extents from the atmosphere. Specifically, algae bioreactors can be used to produce fuels such as biodiesel and bioethanol, to generate animal feed, or to reduce pollutants such as NOx and CO2 in flue gases of power plants. Fundamentally, this kind of bioreactor is based on the photosynthetic reaction, which is performed by the chlorophyll-containing algae itself using dissolved carbon dioxide and sunlight. The carbon dioxide is dispersed into the reactor fluid to make it accessible to the algae. The bioreactor has to be made out of transparent material.

<span class="mw-page-title-main">Knockout moss</span> Genetically modified moss plant

A knockout moss is one kind of genetically modified moss, which are GM plants. One or more of the moss's specific genes are deleted or inactivated, for example by gene targeting or other methods. After the deletion of a gene, the knockout moss has lost the trait encoded by this gene. Thus, the function of this gene can be inferred. This scientific approach is called reverse genetics as the scientist wants to unravel the function of a specific gene. In classical genetics the scientist starts with a phenotype of interest and searches for the gene that causes this phenotype. Knockout mosses are relevant for basic research in biology as well as in biotechnology.

<span class="mw-page-title-main">International Moss Stock Center</span>

The International Moss Stock Center (IMSC) is a biorepository which is specialized in collecting, preserving and distributing moss plants of a high value of scientific research. The IMSC is located at the Faculty of Biology, Department of Plant Biotechnology, at the Albert-Ludwigs-University of Freiburg, Germany.

References

  1. Hohe, A.; Reski, R. (2005). "From axenic spore germination to molecular farming: one century of bryophyte in vitro culture". Plant Cell Reports. 23 (8): 513–521. doi:10.1007/s00299-004-0894-8. PMID   15558285.
  2. Reutter, K.; Reski, R. (September 1996). "Production of a heterologous protein in bioreactor cultures of fully differentiated moss plants" (PDF). Plant Tissue Culture and Biotechnology. 2 (3): 142–147. Archived from the original (PDF) on 2011-10-04.
  3. Hohe, A.; Reski, R. (June 2005). "Control of growth and differentiation of bioreactor cultures of Physcomitrella by environmental parameters". Plant Cell, Tissue and Organ Culture. 81 (3): 307–311. doi:10.1007/s11240-004-6656-z.
  4. Decker, E. L.; Reski, R. (April 2004). "The moss bioreactor". Current Opinion in Plant Biology. 7 (2): 166–170. doi:10.1016/j.pbi.2004.01.002.
  5. Homepage of greenovation GmbH, showing various types of moss bioreactors: Archived November 9, 2011, at the Wayback Machine
  6. Decker, Eva L.; Reski, Ralf (2008). "Current achievements in the production of complex biopharmaceuticals with moss bioreactors". Bioprocess and Biosystems Engineering. 31 (1): 3–9. doi:10.1007/s00449-007-0151-y. PMID   17701058.
  7. Baur, A.; Reski, R.; Gorr, G. (2005). "Enhanced recovery of a secreted recombinant human growth factor using stabilizing additives and by co-expression of human serum albumin in the moss Physcomitrella patens". Plant Biotechnology Journal. 3 (3): 331–340. doi: 10.1111/j.1467-7652.2005.00127.x . PMID   17129315.
  8. Büttner-Mainik, A.; Parsons, J.; Jérôme, H.; Hartmann, A.; Lamer, S.; Schaaf, A.; Schlosser, A.; Zipfel, P. F.; Reski, R.; Decker, E. L. (April 2011). "Production of biologically active recombinant human factor H in Physcomitrella". Plant Biotechnology Journal. 9 (3): 373–383. doi: 10.1111/j.1467-7652.2010.00552.x . PMID   20723134.
  9. "Homepage of Greenovation: Approval to begin a Phase I clinical trial in Europe for moss-aGal (agalsidase) produced in Mossbioreactors" . Retrieved 18 October 2015.
  10. Reski, Ralf; Parsons, Juliana; Decker, Eva L. (October 2015). "Moss-made pharmaceuticals: from bench to bedside". Plant Biotechnology Journal. 13 (8): 1191–1198. doi:10.1111/pbi.12401. PMC   4736463 . PMID   26011014.
  11. Greenovation gelingt der Durchbruch Portal goingpublic Archived 2018-01-11 at the Wayback Machine