Overview

Our final goal after the iGEM cycle will be to continue working on the idea and develop it to a stage where it can be taken up as a scalable solution. We have been told by multiple people with whom we discussed our project that our idea beholds a lot of potential as it is novel and sustainable. JetroEco fueling the aviation sector is our goal, and for that we have come up with different strategies for entering and capturing the market of aviation fuel.

A start-up cannot sustain itself on its own; at every stage, it needs input in a variety of forms- like financial, mentorship, infrastructure, legal etc. We have identified our stakeholders and tried to chalk out a plan to get our product- which is a novel Sustainable Aviation Fuel made from engineered yeast Yarrowia lipolytica.

This is the proposed timeline for our start-up:

Supporting Entrepreneurship

Structure of the Startup

Market Analysis

Sometimes timing is very crucial to capture a market. For us there is a lot of opportunity at this time as there are no commercial producers of SAF in India and the number is quite limited even across the world. With no existing competitors in the market, we could break in and present a lucrative solution. The two groups working on SAF in India at a noticeable scale are- CSIR-IIP, Dehradun and Praj Industries Limited, Pune. We have spoken to the chief scientists at both places and taken their inputs. Both were very keen on our idea and have shown interest in future collaborations and investments.

Currently there is a huge difference between the SAF prices in India and other countries and this allows us to work around the range.

SWOT Analysis

The SWOT stands for Strengths, Weaknesses, Opportunities and Threats. It is a project management tool to assess different options and determine the feasibility of each of our options. It allows the identification of internal and external factors to increase awareness of the factors that go into making a business decision or establishing a business strategy.

PESTEL

Once we have identified internal and external factors affecting our organisation, we now carefully differentiate into different categories and factors that may impact the business and what opportunities and risks are associated. It helps in understanding market growth or decline, business position, potential and direction for operations.

IP and Patenting

Any new development in research and technology that can potentially be scaled up for future manufacturing and end-user products can be protected under patent rights and Intellectual properties. However, it is a lengthy and tedious procedure to go through that involves multiple parties in between. We reached out to the Atal Incubation Centre (AIC) of IISER Pune to assist us in the process.

Business Model

Life Cycle Assessment

Life Cycle Analysis (LCA) is a comprehensive methodology used to assess the environmental impact of a product or process throughout its entire lifecycle, from raw material extraction to disposal or end use.

Purpose

LCA serves the crucial purpose of quantifying the environmental footprint associated with a specific product, in this case, Bio-Jet Fuel (BJF) derived from yeast through synthetic biology. By conducting an LCA, our goal will be to calculate and understand the carbon emissions and other environmental factors linked to BJF production and consumption, encompassing the entire journey from its creation to its utilization in aircraft (Well to Wake). Our chosen functional unit will be 1MJ of energy, allowing for a precise and standardized assessment. This data enables us to make informed comparisons between BJF and conventional jet fuel, supporting sustainable decision-making in aviation and reducing our carbon footprint.

The impact factor that we chose to study through this LCA was Global Warming Potential (GWP).

Methodology

Harnessing Yarrowia lipolytica's ability to thrive on agricultural residues as a growth medium offers a sustainable means to close the carbon cycle. By tapping into the carbon that's already in circulation through these residues, rather than relying on carbon derived from fossil fuels, we can effectively reduce the carbon footprint of Sustainable Aviation Fuel (SAF) production. This approach serves as a valuable method for carbon offsetting, helping us mitigate the environmental impact of SAF production and contribute to a more eco-friendly aviation industry.

Raw Materials

Raw materials for the Bio-Jet Fuel (BJF) LCA included Yarrowia lipolytica (yeast) and agricultural waste.

Processing

The processing steps involved in BJF production encompassed:

• Collecting agricultural residue and wastewater.
• Culturing Yarrowia lipolytica in agricultural wastewater mixture
• Illuminating the bioreactor with blue light to activate the photo-decarboxylase gene (cvFAP) (No CO2 emissions as it utilizes solar energy).
• Cell lysing through bead milling.
• Centrifugation for biomass separation.
• Hydroprocessing, which included isomerization, fragmentation, and aromatization.
• Purification of the mixture through fractional distillation to obtain the desired hydrocarbons.
• Transportation of BJF to airports using appropriate vehicles for jet fuel transportation (No direct CO2 emissions during transportation).

Usage and Emission

The final stage involved the usage of BJF as an energy source by airplanes, resulting in the direct release of carbon emissions (CO2) during combustion, while other stages of the process are considered indirect sources of CO2 emissions.

Our method of Bio-Jet Fuel (BJF) production using Yarrowia lipolytica and agricultural waste demonstrates a significant potential for reducing carbon emissions compared to traditional Sustainable Aviation Fuel (SAF) production. Here's how each step contributes to lower emissions:

• Raw Materials: Using agricultural waste as a feedstock instead of fossil fuels reduces the initial carbon footprint.
• Processing: Illuminating with blue light by utilizing solar energy for activation eliminates direct CO2 emissions.
• Usage and Emission: While combustion during usage leads to CO2 emissions, the indirect emissions from the production process are significantly reduced due to the use of renewable resources, efficient processing steps, and minimized transportation emissions.

Our method prioritizes carbon neutrality in production, making it a more environmentally friendly alternative to conventional SAF production, which typically relies on fossil fuels throughout its lifecycle. This leads to a lower net carbon footprint for the BJF we will be producing.

The net carbon emission of conventional jet fuel is 89 Kg CO2/MJ of energy whereas in the case of Jatropha-based HEFA fuel the net carbon emission is only about 35 Kg CO2/MJ of energy. Thus the Jatropha-based HEFA fuel can reduce the net carbon emissions by more than 60% compared to the conventional jet fuel.

Cost Analysis

Motivation

During the ideation phase of our project, experts underscored the potential benefits of our initiative in curbing the reliance on traditional aviation fuel. Furthermore, they emphasised the scalability of our project to advance the utilisation of sustainable biofuels. However, a critical challenge identified pertains to the economic competitiveness of our biofuel on a large scale. To attain viability, its cost must not significantly surpass that of conventional aviation fuel or Sustainable Aviation Fuel (SAF). This prompted a strategic examination of cost estimation methodologies within the confines of the iGEM cycle.

The current market price for conventional aviation fuel in India stands at approximately 1.07 lakh INR per kilolitre of fuel, equivalent to 107 INR per litre or 1.29 USD per litre of fuel [1]. Meanwhile, Sustainable Aviation Fuel (SAF) commands a price range that is 1.5 to 6 times higher than conventional aviation fuel [2].

Procedure

In order to compute the cost estimate, we took into account a lot of external factors, and had to make some assumptions. All our calculations were made based on a 100L capacity bioreactor. We incorporated media cost, electricity cost, labour cost, cost of purification by fractional distillation, and bioreactor cost. Through various aspects of our project, we were able to bring down the price significantly. We completed four cycles of cost calculations, keeping some assumptions and values the same, while changing others, based on other aspects of our project, with the aim of reducing the final production cost. We used the following equation for our calculations, which were made for a period of one year.

Final production cost of fatty acids per kg = Media cost + bioreactor cost + labour cost + electricity cost + cost of purification by fractional distillation

General Assumptions

• Bioreactor Cost: The airlift-type bioreactor we are considering falls within a cost range of 3,00,000 to 5,00,000 INR (3,606.30 to 6,010.50 USD), with an assumed average of 4,00,000 INR (4,808.40 USD) over a 10-year lifespan. For our calculations covering a one-year period, we normalised the cost to 40,000 INR (480.84 USD).
• Labour Cost: Utilising minimum wage rates in India, set at 16,000 INR (192.34 USD) per month for unskilled workers and 20,000 INR (240.42 USD) per month for skilled workers [4], we estimated the labour cost for operating a single 100L bioreactor over one year. Assuming employment of 2 unskilled workers and 8 skilled workers, the final labour cost for 10 labourers totals 1,92,000 INR (2,308.03 USD) annually.
• Electricity Cost: Electricity in India is priced at 6.19 INR (0.07 USD) per kWh [5]. Considering the power requirements for a single-use bioreactor (120V, 20A) over 21 days, the yearly electricity consumption is calculated to be 3,65,414.64 kWh, resulting in an annual cost of 22,61,916 INR (27,190.49 USD).
• Cost of purification: The purification method we intend to use is fractional distillation. To get an estimate for the equipment to perform this, we got inputs from some experts, and used an average value to be 40,000INR (480.84 USD).

• Culture cost: For our bioreactor cultures, glucose serves as a primary nutrient, obtained from Praj Industries at a cost of 30 INR (0.36 USD) per kg.

  In the case of the YPD Media composition, utilising 20g/L of glucose, each batch for a 100L bioreactor requires 2 kg of glucose, amounting to 60 INR (0.72 USD) per batch. With a batch duration of 4 days, we anticipate a total of 91 batches annually. Consequently, the yearly culture cost, excluding the minimal costs of peptones and dextrose, stands at 5460 INR (65.63 USD).

  Conversely, the optimised composition, determined through the results of the Design of Experiment, indicates an optimal glucose concentration of 70g/L. Employing the same pricing, the total culture cost for one year using this optimised composition is calculated to be 19,110 INR (229.72 USD).

  For a more in-depth understanding of the glucose concentration optimization process, please refer to the Design of Experiments section on the Measurements page.

• Yield: Two distinct assumptions govern our considerations regarding fatty acid yield, each tailored for specific calculations.

  In the first scenario, we presume complete stoichiometric conversion of glucose to C16 and C18 fatty acids, with a distribution ratio of 15% for C16 and 85% for C18. This assumes exclusive fatty acid production from the provided glucose. Consequently, 4.425 moles of glucose yield 1 mole of a general fatty acid, with a calculated mass of 278.1g per mole. This results in a yield of 350g of fatty acids per 1 kg of glucose.

  In the second assumption, we base our calculations on the yield of palmitate derived from a kinetic model we established. This assumption posits that the production of C18 fatty acids mirrors that of palmitate. According to this model, the yield is determined to be 994g of fatty acids for every 1 kg of glucose. Comprehensive details on the creation of the kinetic model are available in the Kinetic Modelling section on the Model page.

Iteration 1

Under conditions of YPD culture with 20g/L glucose and assuming complete stoichiometric conversion, the annual production cost for 63.7 kg of fatty acids is 25,39,376 INR (30,525.84 USD). The corresponding cost per kilogram is 39,865 INR (479.22 USD).

Iteration 2

Maintaining YPD culture conditions with 20g/L glucose, and now considering the yield from the kinetic model, the annual production cost for 164 kg of fatty acids remains at 25,39,376 INR (30,525.84 USD). The updated cost per kilogram is 15,484 INR (186.13 USD), representing a 2.6-fold reduction from the first iteration.

Iteration 3

Shifting to an optimised media with 70g/L glucose and assuming stoichiometric conversion, the annual yield of fatty acids increases to 223 kg. The total production cost is 25,53,026 INR (30,689.93 USD), resulting in a cost per kilogram of 11,449 INR (137.63 USD). This marks a 3.5-fold reduction from the first iteration and a 1.35-fold reduction from the second.

Iteration 4

Maintaining optimised media conditions with 70g/L glucose, and considering the yield from the kinetic model, the annual yield of fatty acids reaches 628 kg. The production cost remains at 25,53,026 INR (30,689.93 USD), yielding a cost per kilogram of 4065 INR (48.87 USD). This represents a significant reduction, approximately 10-fold from the first iteration, 4-fold from the second, and 3-fold from the third iteration.

Conclusion

Through cost analysis of our project, we tackled an essential aspect of the project, attempting to ensure that the implementation of our work can be competitive in the market. Through various aspects of the model such as Design of Experiment, Kinetic Modelling and its optimisation, we managed to bring the initial 200x SAF cost of our biofuel to 20x, a massive 10-fold slash. Through further improvements, in the future we hope to slash this price further and make it economically viable.