Cloning, molecular modelling, and docking analysis of GH-13 alpha-amylase from rumen metagenome for saccharification of starch rich biomass for greener future Jasmine Mansuri, Tripti Dadheech, Prakram Singh Chauhan, Anjali B. Thakkar, Dharamshibhai Rank, Chaitanya G. Joshi, Hiren K. Patel, Anju P. Kunjadiya Biocatalysis and Biotransformation, 2026 α-Amylases are used in various biotechnological processes including the textile, paper, food, biofuels, detergents, and pharmaceutical industries. In this study, a unique starch-degrading enzyme (Alpha-Amy4) was isolated from the microbial diversity present in cattle rumen using functional metagenomic approach. A metagenomic library was constructed using fosmid vector (pCC1FOS) for high-throughput screening of enzymes through a multistage functional screening pipeline. Sequence analysis revealed an open reading frame of 1488 bp encoding a 496 amino acid long Alpha-Amy4 (GH13 family), sharing 92.73% similarity with an alpha amylase <i>Escherichia coli</i> (NCBI Reference Sequence: WP 001245695.1). The recombinant purified protein has a molecular weight of ∼54.0 kDa on SDS-PAGE and exhibited optimal activity at 45 °C and pH 5.5. The apparent K<sub>m</sub> and V<sub>max</sub> values were 0.368 mol mL<sup>−1 </sup>min<sup>−1</sup> and 1092 µmol mL<sup>−1 </sup>min<sup>−1</sup>, respectively, according to the plot data in the absence of calcium activator. Homology modelling and molecular docking identified conserved catalytic triad residues (D172, E203, and D289) within the TIM (β/α)8-barrel motif involved in substrate hydrolysis. Finally, potato peel, a low-cost and high-starch agricultural waste was used to confirm the activity of Alpha-amy4. The enzyme demonstrated efficiency at a lower dosage (20 IU), resulting in enhanced saccharification (78.7%) of potato peels in a shorter time. Thus, Alpha-amy4 represents a novel member of the GH-13 glycoside hydrolase (GHs) family, distinguished by its enhanced enzymatic activity on raw agro-wastes, highlighting its potential for efficient and sustainable biomass valorisation.
Microbial Production of Human Milk Oligosaccharides (HMOs) Prakram Singh Chauhan, Tripti Dadheech, Arunika Saxena Microbial Nutraceuticals Products and Processes, 2025 Human milk oligosaccharides (HMOs) have been the subject of widespread interest in recent years due to their beneficial effects on neonatal health. In light of the availability, safety, and affordability of host strains, an engineered microbial method has been investigated for making HMOs on a large scale. The production of HMOs is becoming more efficient due to advances in molecular biology and metabolic engineering. A number of fucosylated HMOs with complex structures, such as 2′-fucosyllactose (2′-FL), 3′-fucosyllactose (3′-FL), lacto- N -tetraose (LNT), lacto- N -neotetraose (LNnT), 3′-sialyllactose (3′-SL), 6′-sialyllactose (6′-SL), and 3′-SL, have been produced via the engineered microbial route, with 2′-FL being the most produced. It is difficult to select a host strain due to the ambiguity of metabolic processes. Additionally, various HMOs are synthesized in microorganisms by expressing glycosyltransferases (GTs). In order to employ designed microbial pathways effectively, it is necessary to develop a GTs that is efficient and safe. An overview of most recent studies on HMO generation by engineered microbial pathways, purification methods, market analyses, and challenges for scaling up is presented in this chapter.
Preface Management and Mitigation of Emerging Pollutants, 2023
Management and mitigation of emerging pollutants Nancy George, Vagish Dwibedi, Santosh Kumar Rath, Prakram Singh Chauhan Management and Mitigation of Emerging Pollutants, 2023 This book covers the latest advances in the management and mitigation of emerging pollutants, including microbial functional applications.
Statistical optimization of process parameters for production of alkaline protease from Vibrio metschnikovii NG155 having application in leather industry International Journal of Pharma and Bio Sciences, 2014
A mannanase from Bacillus nealsonii PN-11: Statistical optimization of production and application in biobleaching of pulp in combination with xylanase International Journal of Pharma and Bio Sciences, 2014
An overview of purification strategies for microbial mannanases International Journal of Pharma and Bio Sciences, 2014
