Qualification:

 Research:

Major area of work: Gene cloning and recombinant protein expression, Biophysical chemistry, Structural Biology, Proteomics. Proteins are produced by DNA recombinant technology by optimizing their production in biologically active form or by refolding them from the inclusion bodies. Their structure and function are studis by using various biochemical and biophysical methods including spectroscopic (Circular dichroism, Fluorescence spectroscopy) and thermodynamic techniques (DSC) for understanding the kinetics and thermodynamics associated with molecular processes. Also, we produce high throughput transcriptomics and proteomics data for understanding the molecular and cellular processes and the underlysing mechanisms.

Antimicrobial proteins: We are working on animal originated antimicrobial proteins like lysozyme, lactoferrin, lactadherin, xanthine oxidoreductase and genes/proteins involved in molybdenum cofactor (Moco) synthesizing metabolic pathway. We are also targeting the antimicrobial resistance via engineering the bacteriophage originated endolysins to improve their activity against the gram negative pathogens, since these organisms are impervious owing to their outer membrane. We are also interested in proteins of probiotic origin like surface proteins to serve as antimicrobial agent against drug-resistant microbes.

Figure shows the differences in the FAD and NAD binding sites between cattle (A) and buffalo (B) xanthine oxidoreductase (Kaustubh et al., 2012, PLOS ONE).

Proteolytic enzymes: Another area is production of recombinant aminopeptidases (PepL, PepI, PepN) and other proteolytic enzymes (cell lenvelope protease PepO2) of lactic acid bacteria.  Recombinant leucyl aminopeptidase (PepL) has been found very active but possesses lower thermal stability. Currently, we are trying to engineer PepL to enhance its thermal stability. These enzymes could find an application in dairy, food and pharmaceutical industry for the production of protein hydrolysate or production of bioactive peptides.
Figure: Active site of PepL of Lactobacillus casei. The balls in magenta color indicate cobalt ion.

Protein associated with animal reproduction and health: We also have interest in understanding the important genes/proteins involved in animal reproduction, e.g., those involved in sperm-oocyte binding like sperm lysozyme-like proteins (SLLP) and their receptors, and fertility augmenting factors like growth and differentiation factor (GDF9). We try to understand the role of proteins in binding of sperm with oocytes by using biophysical and structural modeling work. The work should provide a better understanding of sperm-oocyte binding and possible reason and treatment of infertility arising because of those reasons in farm animals.

Figure: The sperm protein (shown in yellow color) binds in the active site of the receptor localized on the oocyte. The receptor has been shown in grey color with zinc (shown in magenta color) sitting in its active site channel.

Animal Proteomics: Transcriptomics, proteomic and other studies are important to understand the dynamics and adatation at system level in response to external/internal stimuli or genetic variations. We are analysing the proteome and transcriptome of farm animals to understand production and reproduction processes. Our interest is to understand spermatogenesis process and expression of X and Y chromosome located genes; factors responsible for semen freezing potential in cross-bred bulls, and pashmina fibre growth and development to name a few by analysing the differential transcriptome and proteome. The study is likely to provide important biomarkers, biosynthetic pathways and biological processes involved in gene expression regulation at transcrition and protein level.

Teaching:

Publications:

1. Bathla S, Sindhu A, Kumar S, Dubey SK, Pattnaik S, Rawat P, Chopra A, Dang A, Kaushik JK, Mohanty AK.(2020). Tandem Mass Tag (TMT)-based quantitative proteomics reveals potential targets associated with onset of Sub-clinical Mastitis in cows. Sci Rep. 10: 9321. doi: 10.1038/s41598-020-66211-6
2. Jaswal S, Anand V, Kumar S, Bathla S, Dang AK, Kaushik JK, Mohanty AK. (2020). In-depth proteome analysis of more than 12,500 proteins in buffalo mammary epithelial cell line identifies protein signatures for active proliferation and lactation. Sci Rep. 10: 4834. doi: 10.1038/s41598-020-61521-1.
3. Choudhary, S., Janjanam, S., Kumar, S., Kaushik, J.K., Mohanty, A.K. (2019). Structural and functional characterization of buffalo oviduct-specific glycoprotein (OVGP1) expressed during estrous cycle. Bioscience reports, 39 (12).
4. Singh, S., Kalra, S., Bubber, P., Datta, T.K., Mohanty, A.K., Kaushik, J.K. (2019). Functional analysis of recombinant buffalo lactoferrin and monoferric lobes and their cytotoxic effect on buffalo mammary epithelial cells. BioMetals 32 (5): 771-783.
5. Singh, S., Choudhary, S., Anand, V., Jaswal, S., Verma, A.K., Kumar, S., Kaushik, J.K., Mohanty, A.K. (2019). New insights into the catalytic inactivity of mammary gland protein-40, a chitinase-like protein expressed during mammary gland involution. Molecular Biology Reports 46 (2): 2243-2257.
6. Lotfan, M. Choudhary, S., Yadav, M.L., Tripathi, I., Bhan, S.S, Bathla, S., Rawat, P., Kumar, S., Mohanty, T.K., Kaushik, J.K., Mohanty, A.K. (2019). Characterization of buffalo native pregnancy-associated glycoprotein: mass spectrometry-based glycan composition analysis, sugar-binding characteristics and proteolytic. Journal of Proteins and Proteomics 10 (1): 23-32.
7. Bhat, B., Singh, A., Iqbal, Z., Kaushik, J.K., Rao, A.R., Ahmad, S.M., Bhat, H.,  Ayaz, A., Sheikh, F.D., Kalra, S., Shanaz, S., Mir, M.S., Agarwal, P.K., Mohapatra, T., Ganai, N.A. (2019). Comparative transcriptome analysis reveals the genetic basis of coat color variation in Pashmina goat. Scientific Reports 9 (1): 1-9.
8. Singh, K.S., Kumar, S., Mohanty, A.K., Grover, S., Kaushik, J.K. (2018). Mechanistic insights into the host-microbe interaction and pathogen exclusion mediated by the Mucus-binding protein of Lactobacillus plantarum. Scientific Reports 8 (1): 1-10
9. Ali, S., Malakar, D., Kaushik, J.K., Mohanty, A.K., Kumar, S. (2018). Recombinant purified buffalo leukemia inhibitory factor plays an inhibitory role in cell growth. PloS ONE 13 (6), e0198523.
10. Bisht, S., Singh, K.S., Choudhary, R., Grover, S., Kumar, S., Mohanty, A.K., Kaushik, J.K. (2018). Expression of fibronectin-binding protein of L. acidophilus NCFM and in vitro refolding to adhesion capable native-like protein from inclusion bodies Protein Expression & Purification, 145: 7-13. doi: 10.1016/j.pep.2017.11.007. PMID: 29229289.
11. Ali, S.A., Kaur, G., Kaushik, J.K., Malakar, D., Mohanty, A.K., Kumar, S. (2017). Examination of pathways involved in leukemia inhibitory factor (LIF)-induced cell growth arrest using label-free proteomics approach. Journal of Proteomics, 168: 37-52. doi: 10.1016/j.jprot.2017.07.014. PMID: 28755912
12. Choudhary, S., Kumaresan, A., Kumar, M., Chhillar, S., Malik, H., Kumar, S., Kaushik, J.K., Datta, T.K., Mohanty, A.K. (2017). Effect of recombinant and native buffalo OVGP1 on sperm functions and in vitro embryo development: a comparative study. Journal of Animal Science & Biotechnology, 8: 69-80. DOI 10.1186/s40104-017-0201-5. PMID: 28883914
13. Singh, K.S., Choudhary, R., Bisht, S., Grover, S., Kumar, S., Mohanty, A.K. and Kaushik, J.K. (2017). Expression of recombinant truncated domains of mucus-binding (Mub) protein of Lactobacillus plantarum in soluble and biologically active form. Protein Expression & Purification, 135: 54-60. https://doi.org/10.1016/j.pep.2017.04.015. PMID: 28499579
14. Chaudhari, D.D., Singh, R., Mallappa, R.H., Rokana, N., Kaushik, J.K., Bajaj, R.,  Batish, V.K., Grover, S. (2017). Evaluation of casein & whey protein hydrolysates as well as milk fermentates from Lactobacillus helveticus for expression of gut hormones. The Indian Journal of Medical Research. 146 (3): 409.
15. Kaur, G., Ali, S.A., Pachauri, S., Malakar D., Kaushik, J.K., Mohanty A.K., Kumar S. (2017). Buffalo Leukemia Inhibitory Factor Induces Differentiation and Dome-Like Secondary Structures in COS-1 Cells. Cytogenetics & Genome Research. 151: 119-130. https://doi.org/10.1159/000465507. PMID: 28441662
16. Tyagi, A., Kumar, A., Mohanty, A.K., Kaushik, J.K., Grover, S., Batish, V.K. (2017). Expression of buffalo chymosin in Pichia pastoris for application in mozzarella cheese. LWT-Food Science and Technology.

Book Chapters:

1. R. Kumar & J. K. Kaushik. (2015). Molecular Biology of adaptation of Starter lactic acid Bacteria to dairy System. In Fermented Milk and Dairy Products Eds. A. K. Puniya, CRC Press, Pp: 133-166.
2. J. K. Kaushik (2009). Structure and Stability of Proteins from Hyperthermophiles. In Advances in Microbiology, Chemistry & Technology of Dairy Functional Foods & Nutraceuticals. Editors: R. Singh, S. K. Tomar, G. Goel (Eds). ISBN 81-8321-009-0. Agrotech Publishing Academy, Udaipur, India. Pp. 100-107.
3. J. K. Kaushik (2011). Adaptation at Molecular Level in Hyperthermophiles. In Functional Dairy Foods: Concepts and Applications. S. K. Tomar, R. Singh, A. K. Singh, S. Arora, R. R. B. Singh (Eds). ISBN: 81-89304-90-9. Satish Serial Publishing House, New Delhi, India. Pp 183-189. 

Technologies Developed/ Patents Granted (if any):
Patent: Method for regulating fertility (Submitted)