Dr. Arati works at the National Centre for Biological Sciences in TIFR, India. Her primary work is in the class of non-coding RNAs called riboswitches, which directly bind cellular metabolites to control the expression of downstream genes.

Dr. Arati gave extremely insightful inputs with regards to safety, structural modelling and implementation of the project. In terms of structural modelling she advised that we needed to be extremely careful in interpreting our results when performing metal-protein docking, primarily because a metal ion by itself is an extremely small entity and current modelling softwares may not be that sophisticated to be able to accurately dock it with the protein and measure the binding affinity. Her rationale made us realise an improved strategy for molecular docking by measuring the best binding in terms of conformational stability of the complex after binding instead of trying to assess affinity. This helped us improve our understanding of current limitations and methods in structural modelling techniques.

In terms of the implementation, we also discussed progressing towards cell-free systems for the biosensor instead of a whole-cell system. She pointed out the crucial limitations of a cell-free system: even for the proper functioning of the machinery without the cell, it would require optimum pH, temperature, and salt conditions, among others. In the absence of a cell, these aren’t automatically regulated and could hamper the system’s functioning in the presence of environmental samples. On the other hand, whole-cell systems can regulate these conditions better, even in ecological samples, making them more reliable and stable. However, biocontainment is guaranteed in cell-free systems, unlike whole-cell ones. This tradeoff would have to be resolved by properly designing a contained environment like the microfluidic array for the biosensor.

She also posed interesting evolutionary questions regarding what kind of selection pressures this would put on our engineered E. coli , whether exposure to high concentrations of lead for recovery and detection could cause it to adapt to develop resistance to the heavy metal over time and what kind of implications this could have.

Dr. Krishna Ravi Srinivas is the Managing Editor of the journal ‘Asian Biotechnology and Development Review’ and is affiliated with Research and Information System for Developing Countries. He has been a visiting scholar at the University of Pennsylvania, a visiting scholar at Indiana University, Bloomington, anda Post-Doc Research Fellow at South Center, Geneva. He has published extensively on intellectual property rights, climate change and technology transfer, open source innovation, and traditional knowledge.

He was extremely kind and motivating for our team, especially since he was the first stakeholder that we interacted with. He helped us understand the current state of synthetic biology in India and abroad and drew parallels and differences between the two since he had varied experiences. We discussed how synthetic biology is progressing in India and what we can do as students to promote and accelerate its growth. We identified that poor science communication is among the primary reasons for people, especially those who are not well-versed with life sciences, to be averse to the idea of synthetic biology. Terms like “unnatural,” often used in media to describe synthetic biology innovations, makes it seem like a scary unregulated affair. We discussed how it was essential to improve methods of communication of engineering biology along with stressing on discussions on bioethics and safety to help this field progress further. He reflected on our education and communication initiatives and appreciated the amount of feedback we looked for from our stakeholders as well as the students and teachers receiving our developed SynBio content.

Prof. Ravikrishnan Elangovan is an Associate Professor in the Department of Biochemical Engineering and Biotechnology at the Indian Institute of Technology Delhi. He works on single-molecule biophysics, fluorescence spectroscops, molecular motors, and skeletal muscle mechanics. He also has great experience in microfluidics, especially in the domain of biology.

He made us realize that the microfluidic device we wished to design for the biosensor would require a lot of precision as well as optimization and might not turn out the way we want it to in merely the first trial. Also, due to the unavailability of resources and limited time, it would be difficult for us to fabricate such a device. As an alternative, he introduced to us a very logical approach that might even turn out to be better than making a microfluidic device, i.e., immobilization of cells using calcium alginate / low melting agarose. He pointed out that all we need are cells trapped in a porous medium enough for the exchange of nutrients and gases from the surroundings, which would allow it to perfectly fit into our idea.

Prof. Sanjay Ghosh is a Senior Assistant Professor of Synthetic Biology at IBAB(Institute of Bioinformatics and Applied Biotechnology). His primary research is in developing translatable technologies and products using synthetic biology approaches. He works on various genome editing technologies and DNA assembly methods to engineer microorganisms, mosquitoes, and human cells.

Prof. Sanjay gave us extremely innovative suggestions, like using quorum sensing in our recovery system, apart from the biosensor. Forming a biofilm of lead adsorbing bacteria could be an exciting application of our system once we characterize its ability to interact with lead. We could use this to develop more efficient lead adsorbing strategies. He also suggested using a consortium of our lead adsorbing bacteria expressing different lead-binding proteins to test if a combination could outperform individual types of the engineered bacteria. These were among the most out-of-the-box problems we had thought about in our project as possible implementations of it in the future.

Dr. Shyam Pingle is a senior Occupational Health Physician in India who is passionate about Occupational Health, Workplace Wellness, and Community Health. Dr. Pingle has worked as Chief of Medical Services in various industries. He is an active participant in promoting and advocating occupational health in India and other developing countries.

He provided insights on health issues faced by people working in heavy-metal-based industries. He brought to our notice the shortfalls in current regulations. Currently, checkups for blood lead levels occur regularly, but they lack quality control and execution. Following this, we were suggested to work on a solution post lead detection, i.e., towards recovery systems for such industries. Dr. Pingle liked our project and invited us to present our work at OCCUCON (Occupational Health Conference), held in January 2023 in Delhi. The Indian Association of Occupational Health (IAOH) organized the conference.

Prof. Srikanth Mutnuri is a professor of the Applied and Environmental Biotechnology Laboratory at the Dept. of Biological Sciences in BITS Pilani. His main research is on Sustainable Development Goals with a major emphasis on Clean water Sanitation, Zero hunger, Affordable & Clean Energy, Climate Action, and Responsible consumption and production. He had researched BLLs in extensive mining districts of India.

During our interaction with him, we explained our project, and he was deeply impressed by it seeing us leaping toward solving lead poisoning in India. He informed us of ignorance towards this sensitive topic and listed its reasons. One of the reasons he mentioned that centralized lead testing only in high-end labs is one of the key contributors to this ignorance. He told us that our Biosensor could go a long way to bridging this gap as it provides the decentralized basis for the quantification of lead which authorities can integrate with local wastewater treatment plants for water treatment for lead and make quick decisions. He also made us aware of the potential exposure in mechanical shops in India where lead acid batteries used in cars are handled improperly. He suggested using our lead recovery system in series with industrial treatments currently being used for lead treatment. He told us about the use of MBBR (Moving Bed Biofilm Reactor), which are being used in bioaugmentation.

Dr. Swathi Alagesan is a Department of Science and Technology Fellow Faculty at IBAB (Institute of Bioinformatics and Applied Biotechnology). Her current research is focused on generating various synthetic biology tools and using the insights obtained from flux analysis to engineer microorganisms for optimising behaviour.

She has experience in developing a biosensor to detect Arsenic poisoning in blood. She says that metal recovery is the next big thing to work on, as a lot of work has already been done on detection. She believes that in the future, we could integrate a thresholding metal detection circuit with lead-recovery cells. This won’t increase the complexity like that of an oscillating sensor while also helping us achieve two objectives. Additionally, a circuit to express the short domain of PbrR as the surface protein for lead detection could also help improve efficiency by reducing the bacterial load. Initially, we planned to use heat shock to weaken the lead-protein binding and recover lead from the bacteria. However, Dr. Swathi recommended using agents like HNO3 to precipitate lead, as heat might denature the protein and make the system unsustainable. We followed her advice in our wet lab and received positive results.