Project Inspiration
As you are reading this, you stand on your feet, and around you millions of others wade through their daily lives in this mile-a-minute world. Let us pause for a second and wonder how many of those million footfalls are ‘comfortable’ in the true sense.
A moisture-retaining, dark, warm setting, ideal for the micro-world is more than accounted for by your shoes. If worn for long periods regularly, these shoes cause bacteria and fungi to thrive giving to rise to deadly bacterial and fungal foot infections which are anything but skin-deep.
It is said that 20% of the entire population of India which accounts for about 200 million of us are likely to have Athlete’s foot, a serious foot fungal infection, at some point in life. And Athlete’s Foot is just the tip of the iceberg! There are many more such diseases to wreak havoc, giving rise to pain, inflammation and days gone by on medications.
Diabetic patients show higher tendency of suffering from stubborn, serious microbial infections of the lower limbs. A whopping 33 million are affected by DFU globally. Diabetic foot ulcers might lead to serious complications which might even lead to amputation of a limb, due to necrosis from infection.
EXISTING SOLUTIONS AND WHAT THEY LACK TO TREAT YOU
Imagine bad odour and a stubborn itch on the foot, being your top priority. Of course, we do have existing solutions such as foot creams, artificial fragrances, and other topical remedies for a foot sore. However, let us pause and think, what does that bottle of medicine actually do to you?
The most common drugs used to treat foot microbial infections such as terbinafine, clotrimazole, ketoconazole, among others, lead to various side-effects, gynaecomastia, hormonal imbalances such as adrenal insufficiency, and liver diseases among others.
WHAT IS IN YOUR BOTTLE OF PERFUME?
Bringing it down to the common-man's problems, foot creams and other topical remedies greatly increase consumer effort, besides having lower regions of applicability. A foot cream might not reach the crevices of your foot-skin, is quite washable and thus it reduces efficiency by a huge factor, not to mention the cost of it all and repeated expenditure on such drugs!
And on top of it all, all our topical, oral drugs and fragrances, that are ‘supposed to’ keep your infection and odour at bay, use harmful chemicals such as parabenes, toluene, triclosan, etc. which not only harm our skin, but also contribute to the carbon footprint.
AN INGENIOUS FOOT-CARE SOLUTION IS THE NEED OF THE HOUR
And on top of it all, all our topical, oral drugs and fragrances, that are 'supposed to' keep your infection and odour at bay, use harmful chemicals such as parabens, toluene, triclosan, etc. which not only harm our skin, but also contribute to the carbon footprint.
YOU CAN NEITHER SAVE YOUR FEET NOR THE EARTH! WHAT COMES OF IT ALL?
WHAT ARE WE BRINGING TO THE TABLE?
Team E.solei takes a step ahead of your everyday foot cream by designing such a foot insole, to be placed inside the shoe, which would house an ecosystem of bacteria. The bacteria are genetically modified to produce certain such volatile organic molecules which produce an overall protective, caring effect on the foot. The GM bacteria (our chassis) survive on a nutrient medium placed in the sole, producing organic volatiles which would escape through the porous medium into the foot environment.
We exploit the concept of Live Application wherein the GM bacteria survive on a medium, in proximity to the foot environment, while producing a fragrant molecule, which also has certain anti-microbial and anti-inflammatory properties. The molecule creates a healthy environment for the foot, while neutralising the bad odour.
Whenever Live Application comes into play, we have a fishbone in dealing with early depletion of media by the bacteria. Due to this, the bacteria cannot sustain itself on that media for a long time and linalool production stops after some time. This may lead to reduced turn-over of the product and repeated changes of insole with new media.
To tackle this, our product has an inbuilt regulating mechanism in the GM bacteria, which controls the population, having oscillations around a certain stable population, so that the production of linalool is maintained and usage of media is optimised. By doing this we prolong media depletion and we make the turn-over more efficient, reducing consumer effort.
The product also promises to have greater regions of applicability because the volatiles produced are small in size and readily escape to spread to the entire foot environment.
Finally through Live Application we ban use of chemicals in our product. We use a layer of self-sufficient live organism, whose maintenance doesn’t require introduction of harmful chemicals, nor increases carbon footprint!
WHAT WILL OUR PRODUCT LOOK LIKE?
E.solei comes in the form of a foot insole, split into several layers. Within the insole, a layer of biocompatible, nutrient-extractable polymer houses the GM bacteria. This layer is guarded by a non-porous layer at the bottom, which prevents the bacteria from leaking out. Another porous layer sandwiches the biopolymer layer from top, designed in such a way that the target volatile escapes but not the bacteria.
BUILDING UP
We chose Linalool as our target compound, which would be produced by the GM Bacteria. A mono-terpenoid, as well as a tertiary alcohol, Linalool is produced as a plant metabolite. It has a pleasant fragrance and is a volatile oil compound. Besides being involved industrially in the manufacturing of soaps, fragrances, food additives as flavours, insecticides, it also a part of complex pollination mechanism of various plant species to ensure reproduction and survival.
Linalool has antifungal and antibacterial, anti-inflammatory (reduces skin irritation) and wound-healing properties, as well as a characteristic pleasant fragrance of a lavender, making it an obvious choice in caring for the foot.
OUR CHASSIS: THE PROTAGONIST
Lactobacillus rhamnosus is a well-established pro-biotic, commonly found in the gut and known for its safe use in various health applications. It is known for producing antimicrobial compounds like 3-phenyllactic acid, which exhibits strong antifungal properties. This would complement the production of linalool, a compound with antifungal and anti-inflammatory properties, enhancing the foot-bed's efficacy in managing skin health and preventing fungal infections, such as Athlete's Foot.
MAPPING THE PATH
Our project can be traced by four milestones.
The course of E.solei can be divided into two primary modules:
PRODUCTION OF LINALOOL
SUSTAINED PRODUCTION OF LINALOOL BY REGULATING THE POPULATION LEVELS
TRACED IN FOUR MILESTONES
MODULE 1: PRODUCTION OF THE TARGET COMPOUND LINALOOL
Linalool is produced as a metabolite in plants through the Mevalonate Pathway (MEV Pathway). We aim to incorporate this pathway in our chassis and turn on the production of Linalool.
The MEV pathway intrinsically creates mono-terpenes with two isoprene units. We choose to redirect the pathway into producing our desired compound. For this we place genes of interest and transform them into our GM bacteria.
MODULE 2: SUSTENANCE OF LINALOOL PRODUCTION
In many of the treatments exploiting the concept of Live Application we face problems of small activation window of the product due to which the efficiency of the product goes down and we required repeated, frequent application.
Here we regulate the population level of the GM bacteria and hence by maintaining oscillations about a particular population level we optimize Linalool production and media usage. Hence we increase the depletion time for the media, sustaining linalool production and product usage for a longer period of time.
FUTURE PLANS
Creating a GMO with a mind of its own!
We plan to incorporate the above two modules mentioned in our chassis by co-transforming the plasmids housing the genes for the two modules. These co-transformed bacteria would behave as an automated machine where each would sense and halt its division in such a way so that the overall population is maintained. Our GMO is thus equipped to utilize the nutrient-media efficiently and produce Linalool for a longer time than other such products.
Ensuring Bio-Containment and Safety
The GMO is housed in biopolymer which simultaneously acts as a nutrient-extractable source for the GMO which lives on incorporated within that layer. We would alter pore size and porosity in such a way so as to ensure no leakage of bacteria outside the foot insole.
The bacteria being made dormant prior to embedding by standard Lyophilisation techniques are induced, signaling the start of usage of product and Linalool's emergence into the picture.
We design a kill-switch which would induce death in the bacteria whenever it detaches from its medium, ensuring no escape of GMO into the environment and thereby safety.
Polymers are cheap, and easily available and hence the perfect choice for providing a base camp for our GMO.
E.solei finally comes in as a foot insole, suitable for any type of shoe but more so as a small step we take for a whopping 9 million of us, out of which someone would be sitting right this second at the clinic, in pain with a foot fungal infection, wondering what an ingenious foot care solution would look like!
