According to the National Centre for Biotechnology Information, xylose makes up 18-30% of the composition of lignocellulose, and the rest is mostly glucose. This 18-30% xylose is often wasted but can instead be used as a carbohydrate feedstock source for applications such as energy production or manufacturing of new products. This can happen if a method to better harness xylose can be applied to various sources of biomass waste. If achieved, businesses and individuals would not only find a solution to reduce their waste but may also sustainably produce new products. Our improved xylose utilising E. coli strain was created for this reason.
Our primary end users would be various commercial Synthetic Biology Laboratories, who by using our xylose utilising strain, can then use it in their own systems to increase the efficiency of production of their target products. This strain could also act as a building block for future scientific research. Eventually, in the future, we hope our project will also extend to secondary end users who use new products (as developed by our primary end users) to solve real-world applications to solve big problems, make new products and reduce waste!
For example, food catering services are potential secondary end users for our technology. They may incorporate our xylose engineering strains into applications where food waste is a problem such as in school canteens and large food chain productions. That is, they often produce large amounts of bio waste, and while some can have a waste management system, others may not. To gain further insight into how catering services would benefit from our project, we interviewed Chartwells Catering about bio-waste from food production as found here
Farmers are another possible secondary end user for our strain. In the process of production, farmers often produce excessive amounts of biomass waste as a byproduct of farming. This can be waste such as straw stubble which is often burnt or requires additional resources to manage. Through the use of our xylose utilising strain we can help farmers access the xylose sugar component in the waste to make new products. This provides an inexpensive, sustainable and ultimately beneficial alternative to current agricultural waste management systems where biomass is then ‘upcycled’ into new products. In order to learn more about the issue, we interviewed Mr Heilmann about current challenges faced by the farming industry in bio-waste management, found here .
Food processing facilities are another potential secondary end user for our project. The food manufacturing process is often specific and required to operate to certain benchmarks that result in large amounts of food that may be edible, still going to waste. Given the wide range of biowaste available through food processing facilities, they serve as a prominent option alongside farmers and catering services capable of producing bioenergy if they can harness all the carbohydrates, including xylose, from the biomass sources.
Upon comparing all three options, our team decided that it was clear that Farmers would be the designated secondary end users of our strain. This is because there is abundant biomass waste available across the farming sector, including straw, that has limited market value today. The farmers could benefit the most from our strain by collaborating with scientists who can use our xylose utilising strain to make new products. The farmers would provide the feedstock needed to make products in a sustainable way and increase the value of their crop, bringing in more income as well as reducing waste.
In order to test our thinking about waste management and the potential of using waste in a sustainable way to add value, we conducted a market segmentation analysis on catering services, farmers, and food processing facilities, which can be downloaded here
Market segmentation has various benefits that allow our team to best manage our implementation within the real world. In addition to supporting our own research on this project, market segmentation analysis provides valuable insight into our potential future customers, improves our business focus, and ideally allows us to then target the customer that would benefit the most from our technology.
From our market analysis, we found farmers are the most suitable secondary end user or customer, followed closely by catering services, and food processing facilities. In addition to the insight our ranking provided, it highlighted catering services as the potential ‘next step’ of our project as we extend our benefit to the wider world.
The vision of user interaction starts with our secondary end user, a typical farmer. In agriculture, factors such as pests, disease, and weather affect whether the crops meet contracted product specifications and cause the wastage of crops, generating large amounts of bio-waste. According to the World Wide Fund for Nature (WWF), This bio-waste is often burnt, producing damaging greenhouse gases such as carbon dioxide, methane and nitrous oxide, contributing to 18% of all greenhouse production worldwide and accelerating the rate of climate change around the globe. Our business model will focus on providing a cheap alternative to managing the amount of bio-waste that is produced through agricultural practices.
Over time, farmers internationally produce increasing amounts of bio-waste, and seek more environmentally and economically sustainable, efficient and reliable alternatives to waste disposal. Thus there is a considerable demand for solutions and possibilities, which is why this project has been initiated by our team. This trending demand for sustainable production and maximisation of profit from organic waste matter in the farming industry has been validated through our Human Practices interview with Mr Heilman found here
Our next element of user interaction, synthetic biology researchers and scientists, come into play next as another key stakeholder. Due to the evolving nature of scientific discovery and advancement in constantly being able to add on and change existing parts, we plan for our xylose utilising strain of E. coli to be a chassis organism for future implementation into the various industries of agriculture. Thus, we are bridging the ideal future of synthetic biology applications that entails effective waste management and energy utilisation with the current reality, wherein our individual strain can create a complete product, previously discarded, of benefit to farmers and other agricultural stakeholders.
As a part of our real world implementation, we plan to licence our xylose-utilising E. coli strain to external researchers who can then add their parts onto our strain. Although there is a general usage for these new strains, we particularly envision its prominence in clean energy production.
An example of this is the production of bioethanol from lignocellulosic biomass, which has already proved its viability in the biofuel industry. We envision the workflow as follows: a farmer can send their bio-waste to a bio-waste processing facility where after undergoing a pretreatment process, the lab’s new engineered strain is applied. The biomass then undergoes a series of processes that result in the production of bioenergy, e.g. bioethanol (Fig. 1). This bioethanol is then transported back to the farmer who can now repeat the process; any bioenergy produced through other sources of bio-waste will also be compensated accordingly.
Obviously there are inherent challenges associated with this process, such as transportation costs and additional management workload for farmers. However, we have validated farmers’ trend towards maximising profit from every aspect of their agricultural process through our Integrated Human Practices efforts, and thus the repurposing of what would otherwise be organic waste matter into sustainable and profitable products empowers them to achieve this goal.
The bio-energy applications of our new E. coli strain extend beyond bioethanol, as sustainable hydrogen production is another example. The 2017 Macquarie_Australia iGEM team has developed a hydrogen gas producing gene cluster (BBa_K2300001)that utilises the pentose phosphate pathway to ultimately produce clean hydrogen from carbohydrate sources. The project primarily focused on utilising glucose as a carbon source, but xylose can also be catabolised, using our engineered construct, into xylulose-5-phosphate, then lead into the pentose phosphate pathway. This presents another useful opportunity for our new xylose engineered strain which could serve as a chassis organism and would diversify the range of viable sugar sources to include xylose and xylitol. This means that when lignocellulosic bio-waste is supplied as the source for hydrogen production, the bacteria are able to capitalise on both the glucose AND now xylose content for a higher yield and growth rate. This process will follow a similar pipeline as the bioethanol workflow, involving pretreatment, sugar extraction etc.
Overall, our new E. coli strain leads to multitudinous real-world implementations especially in the field of bio-energy by improving the efficiency of xylose utilisation and hence enhancing the industrial viability of lignocellulosic bio-waste as sources.
Our project will assume a for-profit business model as investment is critical in early stages of business development. We will also seek to attain funding through government grants, donations and crowdfunding. Eventually, we aim to return profit through charging licensing fees for the genetic sequence of our bacterial chassis.
In terms of grants, the Australian government offers Business Research and Innovation Initiative (BRII) and the Emerging Industry Infrastructure Fund (EIIF), which provide financial support for startup companies looking to resolve pressing challenges in recent times. They provide funding for feasibility studies and proof of concept, up to around one million Australian dollars for each company. Also, Commonwealth Scientific and Industrial Research Organisation (CSIRO), a government agency for scientific research and innovation, provides the BioFoundry program which allows collaborating researchers to utilise their state-of-the-art laboratory space. Furthermore, the Australian government has identified synthetic biology as a promising field that will have transformative impacts on the various industry sectors. Overall, there are a diverse range of funding opportunities our business can apply for.
The ultimate vision would be to develop a bio-energy gene cluster in-house, combine it with the improved chassis, and apply appropriate hardware for a comprehensive bio-energy generating device. This will be the direction the business will be heading towards, as the commercialisation of a complete package that can be directly sold to end users will immensely elevate the levels of profit and open the door to a broad range of consumers.
As researchers looking to commercialise and distribute our engineered strain, licensing is a required element.
Dealing with genetic modified organisms in an unauthorised manner is considered to be an offence under the Gene Technology Act 2000, Part 4, Division 2 (Dealings with GMOs must be licensed), a licence must be obtained for either the intentional or unintentional release of a GMO into the environment.
With a licence we will be permitted to conduct experiments, propagate, grow, raise, culture and transport the xylose strain that has been developed by the team, particularly in the bio-waste facility, where the xylose and glucose components of the biowaste will be broken down into usable bioenergy.
The implementation of the xylose strain will be completely ethical, as there are legal repercussions in the incorrect release or disposal of the strain. This is outlined, under Gene Technology Act 2000, Part 5 (Licensing system), division 4 (Initial consideration of licences for dealings involving intentional release of a GMO into the environment), section 50A. The strain will be restricted from passing on its genetic material into the environment, through correct disposal methods.
Prior to the implementation of the strain, there will be documented data on the genetic material of the strain and the geographical area in which the strains are implemented, including information on the people who are involved and the duration of time they are involved in the project for. This will allow the reliable marketing of the product strain in the future economy as the strain is developed and implemented under a licensed applicant where ethical, social and economical considerations have been critically analysed and evaluated.
If the project is carried out further the application must assess the severity of the threats and provide additional documentation such as public consultation, engagement with Traditional Owners, or specific subject matter agencies.This is outlined under Gene Technology Act 2000, where the state and local government, Gene Technology Technical Advisory Committee, Commonwealth authority or agency and the Environment Minister (James Griffin), must be taken into consideration.
The Application for a licence (DIR licence) is for dealing with a non plant GMO involving intentional release of the GMO into the environment. This is applied for the risk assessment and management (RARMP) in relation to Section 50A, of the Gene Technology Act 2000, and a decision can be concluded within 255 working days. Under this permit, the applicant (the users of the xylose strain), need to report the details of the unintended effects during the release of the strain, propose a contingency plan that needs to be upheld if the GMO is found in areas that leak out of the bio waste facility and annually report the activities that are being conducted to make sure that they are legally permissible.
In terms of social licensing, we would like to advance our human-centred approach by incorporating more public feedback on our solution. During the latter stages of our project, we plan to hold forums to allow the public to voice their opinions and concerns about each phase in transforming our strain, thus retaining a degree of public input to our project that is imperative to the evolving nature of synthetic biology to better shape our plan of action. Additionally, this would allow for a greater focus on the scientific communication aspect of our project, wherein educating the larger population about the potential of synthetic biology can raise public awareness of the issue, and thus halt the growing issue of bio-waste in agricultural industries.
Social licensing extends to indigenous communities. Given that they are the traditional custodians of the land and have a deep spiritual connection to their country, their acceptance of our project and actions is paramount. Due to this it is vital that we as a team approach their communities and inform them of our project in a timely manner, as we must not continue without their permission. Through communicating with indigenous communities and fusing their ideas with our own, we may also further enrich our project, allowing for public awareness of increasing bio-waste spread in both their communities and the wider community.
As our project involves the release of a bacterial strain into the environment, it is necessary to account for the impacts of an unintended transfer of antibiotic resistance through forms of horizontal gene transmission. This can result in a shift in ecosystem dynamics and loss in biodiversity, where other, potentially pathogenic bacteria can gain antibiotic resistance from the plasmids of our xylose utilising strain. Additionally, if the plasmids remain intact, despite the host bacteria being dead, they still have the potential to transfer antibiotic resistance to other cells, meaning that effective controls are crucial.
Although our team is using an attenuated E.Coli strain that struggles to survive outside of ideal lab conditions, the issue brought by the potential transfer of antibiotic resistance must still be discussed in accordance with modern biocontainment practices. Biocontainment is the process of confining a potentially dangerous organism to limit the likelihood of its accidental release in the environment, thereby protecting ecosystem dynamics and biodiversity alike.
In addition to processing bio-waste and using the strain in a separate facility to reduce risk and likelihoods of transmission, Sweet Genes will also operate under best practice guidelines that allow for a safer, more controlled environment. Some more notable practices that our team will follow include:
In preparation for the real world implementation, our team researched many biocontainment methods that would be suitable for our bio-waste processing facility. In doing so, we aim to abide to best practice biocontainment standards that ensure a safe and equally sustainable operation .
Some challenges that may need to be considered would be transportation (primarily of the biomass and end product), as well as the fact that our attenuated E. coli strain may find it difficult to survive under long periods of time outside of lab conditions. Although we aim to address this challenge through bio-waste collection facilities, certain factors will need to be taken into consideration to ensure the survival of the strain.
As we expanded our knowledge into the intricacies of our project, we began to uncover the various ethical considerations of using an artificial strain in the natural world. As synthetic biology is a relatively new field of study, ethical discussions regarding the issue have had a limited time to formulate and present themselves.
GMOs such as our strain often raise ethical concerns regarding the ‘unnaturalness’ of the technology. This concern amongst many others was raised by Weale 2010, where although his arguments tend more towards crops instead of bacteria in our case, the concerns are still equally applicable.
Although arguments can be made that the artificial nature of the strain makes it unethical to be used in the natural world, concerns regarding the ‘unnaturalness’ of GMOs stem from ideas that it is immoral to tamper with the ‘essence’ of a species within the natural order. However, as described by Weale, this immorality only rose from extreme cases outside of crop and bacteria modification such as the origins of BSE. This is as it is often discussed that if the first cattle was not unnaturally fed bone-meal containing BSE, the disease would not have spread. Stemming from misconceptions within the public, although this is not an example of synthetic biology it describes why the impact of GMOs can be exaggerated to the extent that they are considered unethical as a consequence of artificial intervention. However in reality, small instances such as genetically modified bacteria in our case will not cause an impact significant enough to disrupt ecosystem dynamics at a large scale.
Additionally, the use of genetically modified organisms also brings about ethical concerns regarding the safety of their use. Generally, this is seen in the deliberate misuse of the organism in the intent of causing harm or damage in any shape or form. As Weale describes, in such cases GMOs can present themselves as more ‘problematic than advantageous’.
As our team researched a variety of ethical issues throughout the project, we learned the significance of understanding how ethical values play into the real world, as well as the importance of their communication to the general public in a clear manner. This will avoid exaggerated misconceptions regarding the unethical ‘unnaturalness’ of GMOs as seen in the example of BSE, as well as aid the public in establishing a strong ethical understanding of the world around them.
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