Bioprospecting and Characterization of Cellulases and PET-Degrading Enzymes

Our lab focuses on the bioprospecting and characterization of enzymes involved in cellulose degradation and polyethylene terephthalate (PET) hydrolysis, aiming to drive innovations in sustainable biotechnology. By integrating computational modeling with experimental validation, we investigate cellulases and PET-degrading enzymes to elucidate their structural features, catalytic mechanisms, and key determinants of activity, stability, and substrate specificity.

• Konar, A. et al. (2022) 'A processive GH9 family endoglucanase of Bacillus licheniformis and the role of its carbohydrate-binding domain,' Applied Microbiology and Biotechnology, 106(18), pp. 6059–6075. https://doi.org/10.1007/s00253-022-12117-4.

• Sinha, S.K., Datta, M. and Datta, S. (2021) 'A glucose tolerant β-glucosidase from Thermomicrobium roseum that can hydrolyze biomass in seawater,' Green Chemistry, 23(18), pp. 7299–7311. https://doi.org/10.1039/d1gc01357b.

• Aich, S. et al. (2017) 'Genome-wide characterization of cellulases from the hemi-biotrophic plant pathogen, Bipolaris sorokiniana, reveals the presence of a highly stable GH7 endoglucanase,' Biotechnology for Biofuels, 10(1). https://doi.org/10.1186/s13068-017-0822-0.

• Sinha, S.K. and Datta, S. (2016) 'β-Glucosidase from the hyperthermophilic archaeon Thermococcus sp. is a salt-tolerant enzyme that is stabilized by its reaction product glucose,' Applied Microbiology and Biotechnology, 100(19), pp. 8399–8409. https://doi.org/10.1007/s00253-016-7601-x.

Understanding the mechanism and engineering of cellulolytic enzymes

In our lab, we focus on understanding the mechanisms of cellulolytic enzymes and engineering them for enhanced industrial applications. Combining computational approaches, such as molecular dynamics and docking studies, with experimental wet-lab techniques, we aim to design more efficient cellulases. Our research involves introducing targeted mutations to improve enzyme efficiency, glucose tolerance, and thermal stability. By analyzing enzyme-substrate interactions and identifying structural weak points, we engineer cellulases that perform better under industrial conditions. These improved enzymes have potential applications in biofuel production, bioremediation, and other biotechnological processes, contributing to more sustainable and cost-effective solutions.

• Sengupta, S. et al. (2024) 'Rational Engineering of a β-Glucosidase (H0HC94) from Glycosyl Hydrolase Family I (GH1) to Improve Catalytic Performance on Cellobiose,' The Journal of Physical Chemistry B, 128(36), pp. 8628–8640. https://doi.org/10.1021/acs.jpcb.4c03464.

• Aich, S. and Datta, S. (2020) 'Engineering of a highly thermostable endoglucanase from the GH7 family of Bipolaris sorokiniana for higher catalytic efficiency,' Applied Microbiology and Biotechnology, 104(9), pp. 3935–3945. https://doi.org/10.1007/s00253-020-10515-0.

• Sahu, S. et al. (2023) 'Thermal Sensitivity of the Enzymatic Activity of β-Glucosidase: Simulations Lend Mechanistic Insights into Experimental Observations,' Biochemistry, 62(23), pp. 3440–3452. https://doi.org/10.1021/acs.biochem.3c00387.

• Konar, S. et al. (2020) 'The effect of ionic liquid on the structure of active site pocket and catalytic activity of a β-glucosidase from Halothermothrix orenii,' Journal of Molecular Liquids, 306, p. 112879. https://doi.org/10.1016/j.molliq.2020.112879.

Engineering Fatty Acid Metabolism of E. coli for Biofuel Production

Our lab is focused on engineering the fatty acid metabolic pathways of Escherichia coli to enhance the microbial production of biofuels. By leveraging synthetic biology and metabolic engineering techniques, we modify key enzymes and regulatory networks to redirect fatty acid flux toward the synthesis of energy-dense compounds such as fatty acid esters, alkanes, and alcohols. These efforts aim to optimize yield, reduce production costs, and enable the development of sustainable, renewable biofuels as an alternative to fossil fuels.

• Ghosh, S., Pooja, N. and Datta, S. (2023) 'Strain Improvement Strategies of Industrially Important Microorganisms,' in Industrial Microbiology and Biotechnology, pp. 499–518. https://doi.org/10.1007/978-981-99-2816-3_17.

• Konar, A. and Datta, S. (2022) 'Strain Improvement of Microbes' in Industrial Microbiology, and Biotechnology,' in Springer eBooks, pp. 169–193. https://doi.org/10.1007/978-981-16-5214-1_6.

Immobilization of Enzymes for Enhanced Reusability and Stability

Our research focuses on the immobilization of enzymes, particularly cellulases, onto solid supports such as nanoparticles, covalent organic frameworks (COFs), polymers, and peptide-based materials. This approach enhances the reusability and stability of enzymes, making them suitable for various industrial and biomedical applications. By immobilizing enzymes, we enable efficient one-pot catalysis, improve their performance under diverse reaction conditions, and extend their functional lifespan. Additionally, we explore the use of immobilized enzymes for targeted drug delivery, leveraging the unique properties of the solid supports to achieve precision and efficacy. This innovative strategy combines the advantages of biocatalysis with material science to address challenges in sustainability and healthcare.

• Mukherjee, I. et al. (2018), 'Recyclable Thermoresponsive Polymer−β-Glucosidase Conjugate with Intact Hydrolysis Activity,' Biomacromolecules, 19(6), pp. 2286-2293. https://doi.org/10.1021/acs.biomac.8b00258.

• Paul, S; Gupta, M. et al. (2023), 'Covalent Organic Frameworks for the Purification of Recombinant Enzymes and Heterogeneous Biocatalysis,' Journal of the American Chemical Society, 146(1), pp. 858–867. https://doi.org/10.1021/jacs.3c11169.

• Paul, S., Gupta, M., Dey, K., et al. (2023), 'Hierarchical covalent organic framework-foam for multi-enzyme tandem catalysis,' Chemical Science, 14(24), pp. 6643–6653. https://doi.org/10.1039/d3sc01367g.

• Gupta, M. et al., (2025), Heterogeneous biocatalysis by magnetic nanoparticle immobilized biomass-degrading enzymes derived from microbial cultures.' JMC B  https://doi.org/10.1039/D4TB02011A

• Padhy, A; Gupta, M, et al. 'Lysosome Specific Delivery of β-glucosidase Enzyme using Protein-glycopolypeptide Conjugate via Protein Engineering and Bioconjugation.'  Bioconjugate Chemistry https://doi.org/10.1021/acs.bioconjchem.4c00430

• Paul, S; Gupta M. et al. 'Coprecipitation of Enzyme-Encapsulated Covalent Organic Framework for Biocatalysis.' Journal of the American Chemical Society JACS (Just Accepted)

Design of Genetic Networks Using Synthetic Biology to Understand Enzyme Dynamics:

Our lab research explores the design of genetic networks using synthetic biology to gain deeper insights into enzyme dynamics, specifically focusing on cellulases. By integrating quorum sensing mechanisms, we aim to regulate the expression of cellulase enzymes in response to environmental cues, optimizing their production and activity. This approach enables us to better understand how cellulases interact within microbial communities and their efficiency in biomass degradation. Through the systematic manipulation of genetic circuits and signaling pathways, we seek to enhance cellulase performance for industrial applications, such as biofuel production and waste recycling.

Optimization of Translational and Secretory Pathways for Enhanced Protein Expression  

Our research focuses on optimizing the translational and secretory pathways in microbial systems to enhance the expression of recombinant proteins, including industrially relevant enzymes like cellulases. By manipulating key factors such as signal peptide sequences, we are trying to improve protein yield and facilitate efficient secretion into the extracellular medium. These optimizations are essential for scalable protein production, ensuring that high-quality enzymes can be produced in sufficient quantities for biotechnological applications, including biofuel production, bioremediation, and enzymatic processing of biomass.