“ EnergHyve “
A composite ecosystem that can economically produce Hydrogen to fuel the next generation of sustainable energy
My project study led to the creation of this writing, which focuses on developing hydrogen harnessing systems that are sustainable using synthetic biology
The project that came in second place was a finalist, feel free to critique and reach out for feedback
Abstract
We aim to create an innovative continuous composite system that is compatible with other systems utilizing hydrogen to generate energy. We must also consider the cycle of by-products back into the fuel, the genetic mechanism that operates within it, anaerobic reactor optimisation, and the economically viable production of Biofuel using genetically engineered species. This technique incorporates a blend of various types of bacteria E-coli Clostridium Butylicum Clostridium pasteurianum And Rhodobacter sphaeroides Within a controlled environment. Our system is reliant on fermentation, catabolic breakdown, and photosynthetic pathways. Through precise metabolomic and reactor optimization methods, we aim to create a biological system that can convert organic substrates into hydrogen efficiently, with the goal of minimizing environmental impacts. Hydrogen is a viable option for decarbonizing sectors that are resistant to direct electrification, such as heavy industry and long-haul transport, while also improving energy security and resilience. We aim to capture the essence of a circular economy that is environmentally friendly while keeping the cycling of by-products in our consortium clear
Inspiration
Despite Formula-E being an attempt to tackle climate change, can It be considered a viable sustainable solution for F1 racing? This question was asked because we are interested in hydrogen powered vehicles that use cleaner substrates as fuel. As a result, it was natural to wonder about the future of shaping how we manage the world, particularly with regards to fuel and economic viability for cycling. These two main ideas were the focus of the team’s brainstorming sessions
The profound recognition of the transformative potential of energy vectors has led to the inspiration for economic hydrogen production. By utilizing synthetic biology, it is feasible to construct a comprehensive system that can produce hydrogen with greater efficiency, sustainability, and cost-effectiveness, which will aid in the generation of energy from the current industrial sector and contribute to ensuring societal progress and improved resilience. HICEVs are the primary focus of our conversation, as they offer unique advantages to industries due to their inherent characteristics, operational necessities, decarbonization potential, and incentive to embrace innovation. Our focus is on stakeholders such as long-hauling trans-European/continental trucks, maritime cargo ships, trains and aviation, refining, chemical production, metallurgy industries, and the grounded industries. The relevance of this statement is due to Hydrogen energy’s compatibility with various SDGs, such as Goal 7 and 13, which prioritize affordability and cleanness. [3] [4]
Status-Quo
The hydrogen-economy has been increasingly popular in recent years, partly due to a need to address existing renewable methods. The solution is a promising one, but it comes with its own set of challenges in terms of economic and infrastructure viability, fuel cell converters that exploit electrolysis, material inefficiency, algae culture systems, and traditional steam methane reforming
Hydrogen, a widely advertised clean energy source, has been produced using traditional methods that have significant adverse health and environmental effects, leading to an increased net carbon footprint. The use of SMR and other conventional hydrogen production methods has significant environmental and health risks, which renders them ineffective for their intended purpose
Our research is in its early stages, but we are actively working on enhancing production speeds, efficiency, and flexibility. HICE-powered cars have already been demonstrated in proofs of concept, and several companies are working on this technology independently. HICE can function as a transitional technology for hydrogen fuel cells, leading to an environmentally friendly future
Our Proposal
Dark fermentation and photosynthesis are utilized by two interrelated and mutually beneficial parts of the Hydrogen production system
Recombinant E. C oli K12 serves as a platform for gene introduction, plasmid development, screening, selection, and gene expression. The presence of Formate Dehydrogenase (FDH) is natural in certain strains of E. C oli. The gene that encodes (fdhF) will be determined by E coli. Itself. The lac-Z type genotype of it means that its efficiency and functionality can be tested with little effort. FDH requires the presence of selenocysteine and molybdenum. FDH and FHL, which are referred to as Hyc3/E in the literature, function simultaneously and appear to be membrane bound. Hydrogen is made by utilizing an electron obtained from the breakdown of formate
2H+ + 2e‐ ↔ H2.
[22]
E. T he production of Coli FDH-H can go up to 2.3 mol H2/mol formate. E. C oli is a more mature and well-understood expression system, which makes it easier to create variants. FDH-H’s pH is approximately 7 when pH balanced. 5–8. A temperature of 5 and a maximum temperature range of 30-40°C is present. The activity coefficient of FDH-H is high, reaching up to 50 U/mg. The absence of FDH-O and FDP-N will theoretically result in a threefold increase in our yield, as we have an additional substrate to act on that we can eliminate through selectional gene knockout. [23] [24] [25] [26]
E n the same E. C oli K12 contains a Recombinant pyruvate, PFL, which can be utilized to economically produce H2 by coupling its activity with hydrase enzyme. The gene for PFL can be separated from anaerobic C cells. PflA and pflanzenb.) Pasteurium has a higher rate of H2 production than E.). Coli. Nonetheless, EKGs can be improved through genetic engineering and the optimization of growth conditions. Coli’s H2 production facility is available. The organism can undergo screening at a later stage. CP6013_3048, AP6012_2343, and CFK120 are the genes that regulate pyruvate formate lyase (PFL) in Clostridium pasteurium. A protein that can serve as a formate transporter is encoded in the PFL operon
Figure out the process of calculating levels in mathematics E. C oli Is regulated during glucose fermentation. The lines that represent the cytoplasmic membrane are parallel to the ones below. The transformations of metabolites are indicated by black straight or curved full lines, while the dashed black line represents gaseous diffusion across the membrane, and the light green lines represent regulatory protein interactions
The PFL substance is formed by converting pyruvate into acetyl-Co A. F ormate undergoes further processing by a FHL complex that includes formate dehydrogenase and hydrogenases, ultimately leading to H2 and CO2 production. The mechanism is based on radical chemistry. A glycyl radical is utilized as a critical catalytic ingredient in PF L. T he radical that is formed by removing a hydrogen atom from pyruvate also produces an acetyl intermediate, which is then converted into ACEtYl-Co A. D ue to the presence of photosynthetic bacteria, a certain amount of CO2 will be diverted to part two of the HPS system. These bacteria can utilize this by-product to produce their own glucose
[27]
The HPS second phase consists of a Hybrid system of non-photosynthetic and photosynchastic bacteria. It has the potential to raise the amount of hydrogen that can be generated. A range of carbohydrate types can be metabolized by C. B utyricum. By harnessing light energy, the photosynthetic counterpart facilitates energetically harmful reactions such as the transformation of anaerobic bacteria’s organic acids into hydrogen. By harnessing light, they can surpass the positive free energy barrier and utilize organic acids as substrates for hydrogen production
A hybrid biological hydrogen generation system utilizes photosynthetic and anaerobic bacteria to produce hydrogen by undergoing the degradation of carbohydrates without using light. The utilization of organic acids (Butyrate, Acetate, and Formate) by photosynthetic bacteria could result in the production of hydrogen. The energy profile of hydrogen production by anaerobic and photosynthetic bacteria is depicted in the figure below. The digestion of carbohydrates by anaerobic bacteria results in the acquisition of energy and electrons. Anaerobic digestion’s ability to decompose organic acids into hydrogen was limited by the absence of reactions with negative free energy. Anaerobic digestion does not completely break down glucose into hydrogen and carbon dioxide. By using light to overcome the positive free energy reaction, photosynthetic bacteria can utilize organic acids as a source of hydrogen. Both types of bacteria combined not only reduce the light energy requirement of photosynthetic bacteria but also increase hydrogen production [14] [15]. The excess formate from this site could be converted into HPS1 for H2 production, or the process could occur in a self-contained anaerobic chamber. [1]
Take note of both the basic substrate and the workflow> Both systems possess an uniform product mechanism
Energetics of Photosynthetic Bacteria
Bacteria that break down carbohydrates during anaerobic digestion produce organic acids like butyrate, acetate, and formate. Only reactions with a negative free energy change can occur under anaerobic conditions, so the breakdown of glucose to produce hydrogen and carbon dioxide cannot proceed to completion through anagaaganism. The energy needed for such a process is greater than that consumed by anaerobic bacteria
However, photosynthesis enables photosynthetic bacteria to break the positive free energy barrier and convert organic acids into hydrogen gas. Light energy is utilized by the photosynthetic bacteria to carry out reactions that are not suitable for anaerobic bacteria due to their energetic disadvantages
Experimental Modelling
The objective is to find inspiration for our experimental setup through this
Feedstock Wet cocopeat can be used to suspend molases, which has a high surface area for hosting
Preliminary push and optimization of the reactor:: Through genetic and metabolic engineering, recombinant strains can be produced, leading to increased hydrogen production. Many organisms with uptake hydrogenases have a poor hydrogen yield, as some of the hydrogen they produce is used up. Eliminating genes that regulate uptake hydrogenases is a method to increase hydrogen production. The overexpression of hydrogenevolving hydrogenases, shutting down metabolic pathways that compete for hydrogen, and overexpressing cellulains, hemicellulas, or lignades to maximize glucose availability are other ways to increase hydrogen production. Nonmetabolic engineering techniques that increase fermentative hydrogen yields include heat treatment, sparging, and operational controls E. C oli [20]. Furthermore, adequate mixing is advantageous for the transfer of hydrogen from fermentation to headspace. Bacteria produce hydrogen using compounds other than formate and glucose. The use of thiosulfate resulted in a two-fold increase in hydrogen production when E molecule is immobilized, as demonstrated. By limiting The consumption of glucose by other metabolic pathways, coli cells were able to produce hydrogen; an increase in fructose levels and The addition of succinate resulted in similar effects. Additionally, low levels of nitrogen (1 mM) are crucial for increasing hydrogen yields (2 mol H2 per mole of glucose), and high amounts of ammonium resulted in a sharp decrease in hydrogen production. [21]
Function
E. C oli (K12 Strain)
It is a strain that has been tested and manipulated before, and it can be cloned and tested without difficulty. The ligation method we have previously employed will be utilized by us 2018: U CB erkeley — E. B iosynthesis utilizing Coli K12 is conducted at the University of Washington in 2017. Col I K12 for biofuel production [13]
Plate for enumerating and transmitting the gene in the HPS
To increase H2 production, we are introducing two genes into the same organism through gene expression to create net compounds
Clostridium pasteurianum CP6013_3048, AP6012_2343, and CFK120 are the genes that regulate pyruvate formate lyase (PFL) in Clostridium pasteurium. A protein that can serve as a formate transporter is encoded in the PFL operon
The gene for PFL can be separated from anaerobic C cells. PflA and pflanzenb.) Pasteurium has a higher rate of H2 production than E.). Coli Rhodobacter sphaeroides Use the organic acids derived from anaerobic bacteria as substrates for hydrogen generation. By using light, these bacteria break the energy barrier and facilitate the conversion of organic acids into hydrogen. The positive free energy barrier that exists for converting organic acids like butyrate, acetate, and formate H2 is broken by light conversion Clostridium Butylicum The process of fermentation involves breaking down substrate in the absence of oxygen. The formation of organic acids, such as butyrate, acetate, and formate, occurs through this process
Sidenote The realization that more output can be attained through the use of a more sophisticated system has led us to compromise on performance in certain areas for the sake of simplicity. Acetobutylicum and T. T o ensure compatibility with Lazarus ‘guidelines, we use kodakarensis in HPS1, but exclude less complex systems
Estimated time
Ordering plasmids, creating recombinants to be implanted in the chassis, pre-qualification for human activities
Cloning experiments, Biochemical Testing, and In-vitro analysis
Production of all essential GMOs. Lab-based experiments are followed by repeated analysis and optimization of the output. Continuous inputs from professors
The optimization and co-working of the two systems, along with scaling and upstream processing. Work on Scientific communication
Human Practices
Team lead:
Yatin Prakash
The topics covered include literature review and compilation, Experimental Design, workflow construction, Statistical Programming (R), Scientific Writing, PyMol visualisation, Basic Wet lab Techniques, Team Leader, Experimental Design, Wet laboratory methodology, and more
Bibliography
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