| Manchester - iGEM 2022

Implementation

Implementation of our project in the real world is vital for us to be able to make an impact. Here, we considered our proposed end users of BloomAid, the future goals for the project and the next steps for successful implementation. For further information regarding the entrepreneurial aspects of our implementation, please click below:

For a detailed description of our project, please click below:

Implementation summary

The proposed end users of BloomAid bacteria would mainly be the companies that utilize microalgae to produce various valuable compounds (such as omega-3 or biofuel) from lipid precursors. Our bacteria would be incorporated and co-cultivated with microalgae in the bioreactors that are already being used by these companies. To implement our project in the real world, we would first make an application to relevant regulatory authorities, providing them with a thorough risk assessment as well as evidences that the implementation does not violate any health and safety legislations. On top of that, we must ensure that appropriate containment measures are in place, such as the security of the bioreactors. Other safety measures must also be conducted when storing, transporting, and disposing the bacteria. Finally, we must consider the effect our bacteria would bring about to the natural biosphere in the case of accidental release.

Legal considerations and approval

The first step to implement our genetically modified organisms (GMOs) in the market is to ensure that it abades by the the Genetically Modified Organisms (Contained Use) Regulations of 2014 from the Health and Safety Executive (HSE). This legal framework regulates the use and commercialization of contained GMOs in England, Scotland, and Wales (with a 2015 amended version applying to Northern Ireland). If BloomAid seeks to reach the UK markets, it must provide satisfactory evidence to the relevant legal bodies which includes the HSE and the Department for Environment, Food and Rural Affairs (DEFRA) in England and Wales. For Scotland, this includes the HSE and the Scottish Ministers and the HSENI and the Department of the Environment in Northern Ireland) of its viability.

The first step to obtaining the rights of commercialisation is to make an application to the relevant regulatory authorities (mentioned above depending on the region of the UK). In order to make this application, we must provide:

A risk assessment highlighting the associated risks to humans and the environment, of the contained use of BloomAid to: Our team must provide an analysis of the effects that the accidental release of our GMO would have on ecosystems and human health.

Declare where our GM organism sits within the classification system (Genetically Modified Organisms - Contained Use Regulations, 2022)

Provide evidence that the implementation of the GMO does not violate any other piece of legislation such as the Health and Safety at Work Act of 1974, the Management of Health and Safety at Work Regulations of 1999, the Carriage of Dangerous Goods legislation, and the Control of Substances Hazardous to Health Regulations of 2002.

The first goal of BloomAid is to provide evidence to the GM inspectorate that our GM organism possesses the relevant characteristics to ensure a safe and effective operation of the trial. In brief, evidence of the GMO matching the description set in the application for containment, a stable and appropriate administrative structure, safe mechanisms of storage, transport, and disposal of the organism.

Proposed end users of our product

Our final product will be our modified bacteria, which, when co-cultivated with microalgae, has the ability to increase microalgal growth and the rate of lipid accumulation within microalgae. These lipids are the precursors of biofuel, therefore once these lipids are extracted they can undergo chemical conversion to biofuels.

Therefore, our proposed end users include:

Microalgae cultivation-based companies: As fuel companies around the world start to look towards investing into greener and more renewable fuels, these microalgae cultivation companies would benefit from our dual system which both assists in the increase of microalgae biomass and biofuel production. An example would be AlgaEnergy AlgaEnergy based in Madrid. AlgaEnergy utilises microalgae in many applications - ranging from an energy source to omega-3 supplements.

Microalgal biofuel production companies. Our bacteria has the benefit of changing mode (from microalgal growth to lipid accumulation mode) using light, where the wavelength of light used is in the blue region, where this colour of light is already used in microalgal biofuel plants, making it easily integrable into pre-existing systems. An example of such a company is Solix Biofuels based in Colorado, United States. Solix Biofuels work on growth and harvesting of algae along with extraction of the oil which is later converted into biofuel. Solix would benefit from our engineered organism as it can help increase the amount of algae harvested and increase the yield of subsequent extracted oil.

Future goals for the project

Moving on from what we have achieved towards this project during the course of iGEM, we have a few goals for the future to move this project forwards:`

All our experiments during the course of iGEM have been performed in E. coli as this is a widely used model organism. However, E. coli may not be the most suitable organism for our purpose. We believe our proof of concept organism is azotobacter, because this bacteria has nitrogen fixing abilities, making them ideal candidates to grow on wastewater as wastewater is naturally abundant in nitrogen and dissolved oxygen. This class of bacteria is also known to have a symbiotic relationship with microalgae (Villa, Ray and Barney, 2014). Azotobacter species would be better suited candidates to survive in wastewater compared to E. coli as azotobacter have a long history in wastewater (Patil et al., 2010). An example of a species that could be used is Azotobacter Vinelandii because it has previously been genetically studied (Dos Santos, 2018).

During the course of the project, we designed individual parts encoded on separate plasmids so that we can test our system in a modular manner. In our real world application, all the genes would be integrated into the bacterial genome.

General safety and hazard considerations

The topic of bioethics raises several main points that need to be considered. Anyone who is planning on introducing GM organisms into the real world would need to consider how they would regulate the possible accidental release of the GM bacteria into open waterways.

Additionally, with regards to the potential environmental hazards, the released GM bacteria could horizontally transfer the induced genes to the endogenous bacteria that are present in the waterway, inducing a potential disease or pest burden through playing more pronounced ecological roles than the wild types. The auxin phytohormone that we are producing could leak uncontrolled into the open waterway, disturbing the normal biosphere. The bacteria could also contribute to the production of virus-tolerant bacteria which would have potential drastic consequences upon the human health and the environmental integrity and induce unknown harms.

Bioethics

Another issue that needs to be taken into consideration is concerning the general ethical aspects surrounding a GM organism. We need to take into account the aforementioned environmental and human health, as well as the fact that the GM strains of the bacteria may be established as persistent populations, which would have susquent subtle and long-term effects on biological communities and natural ecosystems (Weale, 2010). Therefore, we need a risk assessment in order to take into consideration different factors such as genetic contamination, the competition with natural species, ecosystem impacts, increased selection pressure on target and nontarget organisms (Prakash, Verma, Bhatia and Tiwary, 2011).

Another aspect that needs to be considered with microalgal biofuels in particular, is the topic of the ethics of producing a potential unemployment issue in fossil fuel and crop biofuel industries, as well as inducing associated costs to the companies that our project will be implemented towards. Third world countries may not be able to implement our solution due to more stringent policies surrounding GMOs due to less accessibility to knowledge regarding GMOs that first world countries have. Third world countries are also less able to pour in investments in building bioreactors and other facilities, where our project would be implemented. This raises an ethical dilemma regarding how only first world countries would be able to implement our solution and possibly be able to implement biofuels into society faster. Additionally, biofuels usually cannot be used in standard vehicles as these vehicles are built to support petrochemical fuels, such as octane, rather than biofuels such as biodiesel (long chain fatty acid esters). Developing countries would find it difficult to implement the use of vehicles that support this type of fuel (Bailey, 2013).

However, our project relies on increasing the efficiency of microalgal biofuels, which does not require large amounts of land. One bottleneck in implementation of biofuels, particularly in developing countries, is that they cannot afford to give up large portions of land that is used to grow agricultural crops, to produce biofuels (Biofuels: benefits and risks for developing countries, 2022). This could possibly lead to a problem in food scarcity. However, microalgae can grow in bioreactors fitted with LEDs, therefore does not need to take up land that is used for agricultural purposes.

Refernece list

Hse.gov.uk. 2022. Genetically Modified Organisms - Contained Use Regulations. [online] Available at: <https://www.hse.gov.uk/biosafety/gmo/gmo-regulations-email.htm> [Accessed 27 September 2022].

Dos Santos, P., 2018. Genomic Manipulations of the Diazotroph Azotobacter vinelandii. Methods in Molecular Biology, pp.91-109.

Villa, J., Ray, E. and Barney, B., 2014. Azotobacter vinelandii siderophore can provide nitrogen to support the culture of the green algae Neochloris oleoabundans and Scenedesmus sp. BA032. FEMS Microbiology Letters, 351(1), pp.70-77.

Patil, S., Patil, C., Salunke, B., Salunkhe, R., Bathe, G. and Patil, D., 2010. Studies on Characterization of Bioflocculant Exopolysaccharide of Azotobacter indicus and Its Potential for Wastewater Treatment. Applied Biochemistry and Biotechnology, 163(4), pp.463-472.

Penningtonslaw.com. 2022. Regulation of GMOs: time to modify the law?. [online] Available at: <https://www.penningtonslaw.com/news-publications/latest-news/2021/regulation-of-gmos-time-to-modify-the-law> [Accessed 27 September 2022].

SciDev.Net. 2022. Biofuels: benefits and risks for developing countries. [online] Available at: <https://www.scidev.net/global/opinions/biofuels-benefits-and-risks-for-developing-countr/> [Accessed 27 September 2022].

Bailey, R., 2013. The Trouble with Biofuels: Costs and Consequences of Expanding Biofuel Use in the United Kingdom. [ebook] Available at: <https://www.chathamhouse.org/> [Accessed 27 September 2022].

Weale, A., 2010. Ethical arguments relevant to the use of GM crops. New Biotechnology, 27(5), pp.582-587.

Prakash, D., Verma, S., Bhatia, R. and Tiwary, B., 2011. Risks and Precautions of Genetically Modified Organisms. ISRN Ecology, 2011, pp.1-13.