Thesis title: Production and Characterization of Keratinase Enzyme by Culturing Xanthomonas citri

Duration: 07/2019 - 02/2020

Thesis supervisor: Prof. Kiran Shinde (Retd.)

College (University): Vivekanand College, Kolhapur, India (Shivaji University, Kolhapur, India)

Description: In my bachelor's thesis, the process of isolating and characterizing keratinase enzyme from Xanthomonas citri involved several key steps. Initially, X. citri strains were isolated from citrus canker-infected samples using selective media and culture techniques. The isolated strains were then cultured in specific growth media optimized for keratinase production.

To induce keratinase production, various culture conditions, such as temperature, pH, and nutrient composition, were optimized through factorial experimental designs. Subsequently, the cultured supernatants were harvested and subjected to protein purification techniques, including ammonium sulfate precipitation and chromatography methods, to isolate the keratinase enzyme.

The purified enzyme was characterized using various biochemical assays to determine its molecular weight, substrate specificity, pH and temperature optima, and stability. Additionally, the enzyme's activity on poultry waste substrates was assessed through enzymatic assays and degradation studies.

The results of the study revealed the significant activity of the keratinase enzyme on poultry waste substrates, indicating its potential application in waste management practices. Further analysis involved identifying the specific amino acid residues and active sites involved in keratin degradation through protein sequencing and structural modeling techniques, providing insights into the enzyme's catalytic mechanism and potential for biotechnological applications.

Title: Understanding the Acetone-Butanol-Ethanol (ABE) Fermentation Metabolism of Clostridium Co-culture on Synthetic Lignocellulose

Duration: 12/2022 to 03/2023

Supervisor: Prof. V. S. Moholkar; Mentor: Dr. Karan Kumar

Department: School of Energy Science and Engineering

Institute: Indian Institute of Technology (IIT) Guwahati, Assam, India

Description: During this research internship at IIT Guwahati focused on elucidating mechanisms of Acetone-Butanol-Ethanol (ABE) fermentation from synthetic lignocellulosic biomass. Task formulation involved defining objectives centered on microbial fermentation and bioprocessing, with a specific emphasis on clostridial biobutanol fermentation. Techniques included utilizing Clostridium co-culture of C. acetobutylicum and C. pasteurianum to investigate enzymatic synthesis pathways for bio-alcohols from agro-biomass.

Analyses were comprehensive, evaluating lignocellulose bio-depolymerization efficiency through enzyme activity and substrate degradation assessments. Gas chromatography and mass spectrometry quantified acetone, butanol, and ethanol production during clostridial biobutanol fermentation. Enzymatic assays and biochemical analyses characterized the enzymatic synthesis of bio-alcohols, providing insights into enhancing ABE fermentation efficiency.

The research advanced the understanding of microbial fermentation and bioprocessing techniques, particularly in sustainable biofuel production contexts. Through meticulous experimentation and analysis, significant progress was made in unraveling the intricate mechanisms driving ABE fermentation from synthetic lignocellulosic biomass. My contributions to this research led to a publication on acetone-butanol-ethanol fermentation for enhanced biobutanol production in Elsevier’s Process Safety and Environmental Protection journal. 

Publication: https://www.sciencedirect.com/science/article/pii/S0957582024002441

Selected as a fully funded attendee for the Karyashala Workshop on Statistical Data Analysis and Modeling in Biophysics at the Indian Institute of Technology Hyderabad, I immersed myself in mastering essential statistical methodologies crucial for unraveling biophysical complexities. Through interactive sessions led by experts, I gained insights into mathematical modeling, statistical inference, and data-driven approaches. Emphasizing experimental design and real-life examples, the workshop equipped me with the practical skills necessary for addressing contemporary challenges in soft matter and biophysics research. This transformative experience not only deepened my understanding but also fueled my passion for scientific inquiry, marking a significant milestone in my academic journey.

As a fully funded attendee of the IBRO BMAI Workshop 2023 at the Indian Institute of Technology Delhi, I immersed myself in the convergence of neuroscience and artificial intelligence. Sponsored by International Brain Research Organization (IBRO) and DST – Kiran, supported by IAN, the workshop offered a vibrant platform for interdisciplinary learning. Through lectures, hands-on sessions, and networking opportunities, I delved into understanding the brain, translating neuroscience into AI, and exploring cutting-edge AI techniques. Emphasizing inclusivity, the workshop nurtured collaboration among diverse backgrounds. It ignited my passion for advancing brain research and harnessing AI's potential for healthcare. My experience was transformative, enriching, and profoundly inspiring. 

Selected as a fully funded participant for the DST SERB-sponsored National Level High-End Workshop/Karyashala on Field Techniques in Insect Observation and Collection at the Zoological Survey of India Western Regional Centre (ZSI WRC) in Pune, I immersed myself in immersive hands-on sessions led by Dr. Aparna Kalawate. From insect pinning to microscopic observation, I honed essential skills alongside esteemed naturalists and scientists. The field survey tour to Bhimashankar Wildlife Sanctuary provided invaluable practical exposure, where I learned ethical collection methods and preservation techniques, with a special emphasis on Lepidoptera. This transformative experience not only deepened my understanding of insect biodiversity but also instilled a profound connection with nature's wonders.