Contents 1 Executive Summary 2 The Opportunity 3 Our Solution 4 The Competition and Our Advantage 5 Marketing Strategies 6 Milestones 7 Company Structure 8 Advisory Board

Executive Summary

New developments in synthetic biology have allowed scientists to explore innovative ways of producing important, high-value chemicals from what was once seen as industrial waste. Paper mills and recycling plants, as a byproduct of their operation, produce a waste sludge composed of paper fibres too short for further processing. As this waste is seen as having negative value (requiring money to be disposed of), this is an attractive source of potentially exploitable cellulose. On the other end, aromatics represent a potentially lucrative chemical endpoint, having high price per unit mass and a wide variety of applications as feedstocks in a number of industries. Where others see waste, we at Upcycled Aromatics see opportunity. Our mission is to create an innovative, green, and inexpensive source of vital, high-demand aromatic chemicals by using the principles of synthetic biology.

Our proposed process has two parts: in the first, cellulose from waste sludge from recycling plants is converted into glucose; in the second, glucose from the first part is used as a feedstock for the biosynthesis of aromatic chemicals from genetically engineered bacteria.

In the glucose-aromatic conversion, we plan to use a single metabolic pathway in the biosynthesis of our product, with “on and off switches” at each “step”. This gives us the freedom to produce any intermediary compounds in addition to the natural end product. The current proposed pathway will allow us to produce either shikimate or cinnamic acid derivatives using the method described above, and should the market favour it, give us the flexibility to produce any other substance along the pathway.

Both shikimic acid and cinnamic acid (and its derivatives) are high-value, high-demand chemicals with steady markets and high potential for growth. Current commercial production of shikimic acid by extraction from Chinese star anise is effectively monopolized, and as seen from recent shortages in 2005, highly susceptible to the effects of a bad harvest. Meanwhile, cinnamic acid, an important precursor in the production of sweeteners and many pharmaceuticals, is produced mainly from synthesis of potentially harmful and polluting organic solvents. Upcycled Aromatics, with its innovative syn-bio approach, can provide a reliable and green source of both these vital compounds, produced from a feedstock for which there is minimal competition.

Once our project has been given the go-ahead for commercial scale-up, we plan to install our first plant directly downstream of either a paper mill or recycling facility. This will minimize transportation costs of the sludge, allowing us to spend money saved on enzymes needed for the cellulose-glucose conversion. Eventually, we hope to move towards a model with centralized production and an extensive distribution network to move goods to and from our suppliers and our plants.

The management of the company will consist of the current members of the University of Alberta iGEM Entrepreneurial team. With our extensive backgrounds in biochemistry, chemical engineering and genetics, we as a team have the knowledge and the skillset to make Upcycled Aromatics a success. Furthermore, we have also assembled a board of advisors that will provide us with the scientific and business expertise as we move forward. They include Dr. Chris Dambrowitz, who has had managerial experience with MDS Sciex (now AB Sciex), Blue Heron Biotechnology (Bothell WA), Visible Genetics (Toronto ON) and Atomic Energy of Canada Ltd (Chalk River ON), Dr. Dominic Sauvageau, who is doing research in bioprocessing strategies in microbes and has been involved with the start-up company Laborium Biopharma, Dr. Anthony Briggs of the University of Alberta Faculty of Business, who holds a M. S. from MIT Sloan and is an expert in the intellectual property of biotechnology, and Glen Penny, who is involved with a venture capital firm currently operating in the USA.

The Opportunity

Due to environmental concern, government regulations and economic considerations, there has been a consistent increased effort in recycling endeavours over the past years. Several hundred million metric tons of municipal solid waste is disposed of in Canada and the United States annually. Paper products alone account for approximately 45% of municipal solid waste by weight before recycling (N. Lark et al.). As a result, recycling paper products not only affects the upstream processes in paper production (where raw materials are acquired), but also has consequences on the downstream portion of paper use (waste-disposal).

Recycled paper processing plants use paper as their feedstock and recover fibre that can be used to produce new paper products. Paper fibre cannot, however, be recycled endlessly. It is generally accepted that a fibre can be used four to six times before it becomes too short to be utilized in new paper products. This unusable fibre accounts for 15-20% of the recycled fibres and is typically disposed of in a waste stream to landfills (N. Lark et al.).

The major constituent of these paper fibres is cellulose, a homopolysaccharide made up of β-D-glucose. Ergo, the unusable, short fibre in the waste stream of paper recycling plants is a significant source of potentially exploitable cellulose. Aromatics represent a potentially lucrative chemical endpoint for this cellulose, having high price per unit mass and a sustainable market in both the pharmaceuticals and cosmetics industries. The conversion of this waste into valuable industrial chemicals is a relatively unexplored business opportunity and is desirable from the standpoint of green and clean processing.

Our Solution

As technologies continue to improve, there has been increasing interest in exploring the use of biomass as a renewable resource. Biomass is typically a by-product of an industrial process and considered waste. In particular, the paper sludge produced by paper recycling plants is presently disposed of via landfill or burnt. This is a significant source of potentially exploitable cellulose that can not only be used to produce high-value chemicals but will also have a positive impact on waste management. Additionally, paper sludge is an attractive feedstock compared to other lignocellulosic biomass because is processed prior to its utilization and requires no pre-treatment as a result.

Many studies have concentrated on the conversion of the cellulose in the paper waste to different types of value-added chemicals. Primarily, it has been considered for conversion to ethanol (Yamshita et al. 2006; Vamvuka et al. 2009; Kang et al. 2010, 2011) and lactic acid (Marques et al. 2008; Mukhopadhyay 2009). Our business plan focuses on the production of aromatics from this cellulose. Aromatics represent a potentially lucrative chemical endpoint, having high price per unit mass and a sustainable market in both the pharmaceuticals and cosmetics industries.

Our company’s proposed process has two parts: in the first, cellulose from waste sludge from recycling plants is converted into glucose; in the second, glucose from the first part is used as a feedstock for the production of aromatic chemicals. In the glucose-aromatic conversion, we plan to use only a single metabolic pathway with “on and off switches” at each “step”. This gives us the freedom to produce any intermediary compounds in addition to the natural end product. The current proposed pathway will allow us to produce either shikimate or cinnamic acid derivatives using the method described above. Possible switch activators include temperature pH, or the addition of an inhibiting chemical such as phosphate.

The Competition and Our Advantage

Currently, pharmaceuticals giant Roche produces the majority of the world's supply of shikimic acid, holding an effective monopoly on the market. Their method of extraction involves isolating the compound from Chinese star anise, at a yield of approximately 30g of seed to very 1g of shikimate[cite]. Furthermore, as demonstrated by the Tamiflu shortages announced by Roche in 2005, a bad harvest will lead inevitably lead to mass shortages in the drug supply.

With this in mind, academics have disputed the infeasibility of producing the chemical by other means. Alternative routes to Tamiflu or sources shikimic acid may prove to be possible sources of competition for Upcycled Aromatics' major market. They include the usage of aminoshikimic acid, biosynthesized by genetically modified baceria, over shikimic acid as a chemical starting point [1], as well as extraction of shikimic acid from pine needles [2]. However, commercial viability on scale-up has yet to be proven for most proposed processes.

Cinnamic acid, converted to its ester form, is an organic chemical with a wide variety of applications ranging from sweeteners to pharmaceuticals. Although its value may be lower on a per gram basis when compared to shikimic acid, a wider variety of applications may mean increased profits depending on current demand. The scale of annual global production is in the thousands on tons as of the year 2000, and is likely to grow as demand for artificial sweeteners increases [3].

Major producers include Bayer (Germany), DSM (Netherlands), and Kay Fries (USA) [ibid].

Typically, the chemical is produced by a condensation of benzaldehyde and acetic anhydride in the presence of a sodium acetate catalyst, and can yield over 80% cinnamic acid based on consumed benzaldehyde [4]. Our process avoids the use of potentially polluting organic solvents in favour of converting something with negative value (paper sludge) into a desirable high-value platform chemical.

Our proposed feedstock, paper and recycling mill sludge, has little to no market competition due to its widespread perception as waste with negative value. The conversion of sludge to glucose shows promise (Banerjee 2011). Other possible uses include feedstock for the production of ethanol (Furstein and Sacakerelles, 2003; Yamshita et al. 2006; Vamvuka et al. 2009; Kang et al. 2010, 2011) and lactic acid (Marques et al. 2008; Mukhopadhyay 2009), but other possible applications remain undeveloped.

Upcycled Aromatics can potentially provide a practical, reliable source of aromatic chemicals. Other biosynthetic approaches lack key accompanying technologies with which to scale up into industrial production, but our tight integration between our genetics and our process gives us a significant leg up on the competition. Further advantages lie in our flexible process, our choice of feedstock, and our tight integration between process and genetics.

As mentioned previously, it is impossible for a recycling plant to process the same paper more than a few times before the fibres are deemed too short and disposed of. Our revolutionary strategy is to utilize these once useless fibres as the feedstock for our process, ultimately turning what was once waste into valuable industrial chemicals important in both the pharmaceutical and cosmetics industries.

Our strategy ultimately allows us to save significantly on operating costs, and since our initial prototype facility will be located at a recycling plant onsite, costs associated with the feedstock will be negligible if not zero. Many people are currently using cellulose as a feedstock, for example, as a precursor to bioethanol, but with Upcycled Aromatics' novel process, we are able to virtually eliminate any and all associated transportation costs, which constitute a huge energy sink in most other cellulosic approaches. Our revolutionary idea, while increasing our margins, also gives us the opportunity to funnel the money saved into other endeavours, such as expansion or R&D.

The flexibility our process offers us the option to respond to shifting markets. Although the value of shikimic acid is many times higher than that of cinnamic acid and its derivatives, any lull in demand can be compensated for by simply flipping our genetic switch and changing our product. On the other hand, any future spikes in the price of shikimic acid (perhaps due to another pandemic scare) will allow us to take advantage of such opportunities. Furthermore, the switchability of Upcycled Aromatics will insulate us from price crashes in any one of our possible products. Furthermore, the semi-permanent, modular nature of the process allows us to avoid costs in the transportation of glucose, and allows us to be consistent in the design and quality of our "tack-on" plants.

Process and genetics in Upcycled Aromatics have been developed in tandem. Such a relationship results in a great deal of synchronization between these two very different areas, ensuring that we will never experience any disconnect between biology and engineering. As these feed back onto one another in an iterative process, improvements in one will always be a fruit of and perhaps a seed for improvements in the other.

Marketing Strategies

Marketing Analysis and Exploration

• Target on a marketing consultant to get early stage market research information.

Potential consultant companies: Matrix Fine Chemicals, StrategyMark, MarketsandMarkets, etc.

Estimated Cost: $4000

• Explore the world market by giving competitive prices and services.

Company Construction

• Package the company: set up professional website, company logo, and branding.

Estimated Cost: $1000

• Easy reachable sales and technical persons (online, on call, “24 hours a day”) for products and services.

• Keep persons getting feedback from the recycle plants and customers.

Media and Advertisement

• Write articles for trade magazines among the recycling industry and fine chemicals industry to get credibility and exposure.

Recycling industry magazines: Solid Waste and Recycling, Recycling Today, Recycling International, etc.

Fine chemicals industry magazines: Speciality Chemicals, Chemical Week, Canadian Chemical News, etc.

Estimated Cost: Free

• Send out technical or product updates and newsletters through emails to clients

Estimated Cost: Free

• Social Media: Facebook, Twitter, YouTube, etc.

Estimated Cost: Free

Media and Advertisement

• Other than collaborate with recycle plants, we also need to form strong networks with our buyers down the manufacturing chain. These buyers including pharmaceutical companies, generic drug companies, food producing companies and fragrances producers, etc. We can attract these buyers by giving discount or promotion packages on long term contracts.

• Participate in trade associations and conferences such as Canadian Generic Pharmaceutical association, Annual World Drug Manufacturing Summits, Paper Recycling Conference & Trade Show, and Canadian Waste and Recycling Expo. During the conferences, we can provide promotional materials, like paper and pens with our company name, logo, and contact information on them, to make impression on our potential business partners.

Estimated Cost: $16000/year

Community Involvement and Reputation Building

• Enter biotech or business award competition to build reputations. As now the 2012 International Genetically Engineered Machine (iGEM) Entrepreneurial competition is our initial opportunity to present our company to the public.

• Sponsor local sports teams. Like buying them team shirts. Thus, we can advertise our business as well as showing strong interaction with the community.

Estimated Cost: $5000/year

• Charity donation.

• As our product is eventually paper consuming, we might want to set up events like annual tree planting to show our concern about the environment.

Estimated Cost: $1000/year

Milestones

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Company Structure

We, Upcycled Aromatics, are composed of unique, brilliant, and well-experienced engineers,scientists,business professionals and our advisers. Each team member are well-trained in their specialized fields. Not only are they well experienced, but also, each of them hold the Upcycled Aromatics vision: *insert here*. Even though each division has its own separate duties, but each of us are achieving the same goal:____.

• Business Division

• Responsible for the business portion of the start-up company

• Some of the responsibilities include: accounting, marketing, market analysis, finance, and risk managements

• The skills that our business division have are maintain a healthy relationship with our customers and suppliers, enhance our services, assess our financial needs, and promote our products

• Research and Development

• Responsible for the science and research portion of the company

• As our process is improving each day, our scientists are responsible for finding the new ways to minimize our waste output, optimizing out products and process.

• Some of the skills we have are project management, critical thinking, creativity, and problem solving

• Company members: Jermey Morries, Kelly Holdstock

• Engineering

• Responsible for the overall process from recycled waste paper to our products
• The optimize our process, the engineers are involved in design, implementation, equipment selection, and maintenance
• The skills that our engineers bring to our company are creativity, critical thinking, project management, integrity, and passion

• Advisory Board

• Responsible for providing us with constructive feedbacks
• Providing support for all divisions
• Overlook our company and recommend any improvements

Advisory Board

Director

• Responsible for overlooking the company.
• Solve or advisor any new ways to run the company

Science Director

• Responsible for advising and making important decisions regarding to the science and research portion of Upcycled Aromatics
• Conducts reviews on the genetics and biology process in the conversion system
• Maintain a healthy and safe working conditions for our team

Engineering Director

• Approving our process, equipments, and designs
• Ensuring our team is following the regulations
• Advises the engineers on improving the process and designs

Business Director

• Responsible for overlooking and reviewing the business plans, marketing strategies, risk management and the market size
• Maintaining a identify any possible future markets for our products
• Advising our company on how to expand nationally