Team:NIT Warangal/Implementation

iGEM NIT_Warangal

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Problem

Animal Feed

Gossypol is a phenolic toxin produced by pigment glands in cotton stems, leaves, seeds, and flower buds. Cottonseed meal is a by-product of cotton that is used as animal feed because it is rich in proteins and fat. However, gossypol toxicity limits cottonseed use in animal feed. High concentrations of free gossypol may be responsible for acute clinical signs of gossypol poisoning which include respiratory distress, impaired body weight gain, anorexia, weakness, apathy, and death after several days. However, the most common toxic effect is the impairment of male and female reproduction. Another important toxic effect of gossypol is its interference with immune function, reducing an animal's resistance to infections and impairing the efficiency of vaccines.

The preventive procedures used currently involve the treatment of cottonseed products to decrease the concentrations of free gossypol through the use of heat, irradiation and pressure in the processing of these products[1]. Agronomic selection has produced cotton varieties devoid of glands producing gossypol[2], but these varieties are less grown because they are not as productive and are more vulnerable to attacks by insects.

Malnutrition

An attribute of cotton not widely recognized is that for every 1 kg of fibre, the plant produces 1.65 kg of seed. This makes cotton the third-largest field crop in terms of edible oilseed tonnage in the world. Cottonseed contains 17% fat, 21% crude fibre, and 24% crude protein on a dry matter basis[3]. 44 million metric tons (MT) of cottonseed (9.4 million MT of available protein) produced each year could provide the total protein requirements of half a billion people for 1 year (50 g/day rate) if the seed were safe for human consumption. In humans, gossypol lowers blood potassium to dangerous levels, resulting in fatigue and even paralysis[4].

India has zero tolerance for (genetically modified) GM products and has extensive regulations on GM crops. Indian authorities had issued 7,660 non-GM certificates to 98 countries in 2019 on products ranging from sweet corn, soya, and basmati rice[5].

Our solution

With our project, we aim to reduce the free gossypol concentrations in cottonseed meal (CSM) enabling it to be effectively used as animal feed or consumed by humans in the form of cottonseed or cottonseed-derived products. This is done by enzymatic treatment of cotton seeds pre CSM production. Therefore, outstepping the problems associated with GMOs in the country.

Proposed users

We plan on providing an enzyme, by the name BBa_K4077000 that can be used in bioprocess systems that refine cotton seeds for oil and CSM for animals as well as humans. The end-users for our project would be cotton farmers, cotton industries, biodiesel industries and food and nutrition industries which would later utilize those enzymes for the breakdown of gossypol. We expect utilization of this enzyme in all cottonseed derived products making them non-toxic and nutritional.

Our project reduced gossypol levels for 10 grams of CSM using crude laccase enzyme for biodegradation of gossypol. Crude glycerol (also known as glycerin) is a major byproduct in the biodiesel manufacturing process. In general, for every 100 pounds of biodiesel produced, approximately 10 pounds of crude glycerol are created. As the biodiesel industry is rapidly expanding, a glut of crude glycerol is being created[6]. Our project can be implemented on a large scale to utilize crude glycerol from biodiesel industries as a medium for growth of recombinant plasmid which would reduce the environmental impact of crude glycerol.

Inspiration

Inspired by Prof. Keerthi Rathore who spelt out the nutrition problems on a global scale and especially in India. He told us that cottonseed is abundant in protein but its gossypol toxicity makes it unsuitable for consumption. Ms. Surbhi Guha threw light on the scaling-up strategies of our product. She recommended steps to advance into a start-up and provided us with strategies to follow through.

Implementation Steps

Having talked to several farmers in Warangal, we realised they are unaware of the toxicity and nutritional value of cottonseed; they sell the seeds produced to local shop keepers who package and sell them to the locals.

From this interaction, we decided the first step in implementing our project would be to educate farmers on the issues and the possible applications of treated CSM with reference to farming. This can be done through government initiatives to teach farmers such as Kisan Suvidha, an app targeted to farmers, providing relevant information to them quickly in multiple Indian languages.

Secondly, we would like to market our product as an eco-friendly and sustainable source of nutrition to the general public. To secure funding, we would contact Government bodies and NGOs. And finally, we would scale up the project by collaborating with the aforementioned industries, resulting in the large scale production of the enzyme which we believe would be sold to farmers, cotton industries, and so on.

Challenges and future solutions

One of the possible challenges we are likely to face is the stabilization of the enzyme in a raw form. This could be done by extending the project towards making the enzyme more stable and usable in a wide variety of conditions. Further, we are planning to visualize the stability of the enzyme via molecular modelling and simulations. We also plan on performing point mutation studies that help us better understand and modify the functional enzyme, such that the product is as efficient as possible.

References:

Sponsors

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iGEM NIT Warangal
National Institute of Technology, Warangal
Warangal, Telangana, INDIA - 506004

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