Isolation and Characterization of Gelatinase Enzyme-Producing Microorganisms from Soil for Industrial Applications

Abstract

The quest for novel enzymes with industrial applications has led to the exploration of diverse microbial ecosystems, including soil environments rich in microbial diversity. This study aimed to isolate and characterize gelatinase-producing microorganisms from soil samples for potential industrial applications. Soil samples were collected from various environments, including agricultural fields, forests, industrial sites, and natural habitats, to capture a wide range of microbial diversity. A total of 250 isolates were obtained through serial dilution and plating on gelatin agar, among which 58 isolates exhibited clear zones of hydrolysis, indicating potential gelatinase activity. Morphological and biochemical characterization revealed the presence of Gram-positive, Gram-negative, and spore-forming bacteria, along with fungal isolates. Molecular identification using 16S rRNA gene sequencing for bacteria and ITS sequencing for fungi identified Bacillus, Pseudomonas, and Aspergillus species as predominant gelatinase producers. Enzyme activity assays demonstrated significant gelatinase activity in selected isolates, including Bacillus subtilis, Pseudomonas aeruginosa, and Aspergillus niger. Optimization studies revealed the optimal pH, temperature, and substrate concentration for maximal enzyme activity, along with the influence of metal ions and inhibitors on enzyme stability. Stability studies indicated the robustness of the isolated gelatinases under various industrial conditions, including temperature and pH fluctuations. Application testing showcased the versatility of the enzymes in food processing, pharmaceuticals, and environmental remediation. Comparative analysis with commercial enzymes highlighted the competitiveness of the isolated enzymes in terms of activity, stability, and cost-effectiveness. This study demonstrates the potential of soil microorganisms as a source of industrially relevant enzymes, particularly gelatinases, for various applications. The isolation and characterization of potent gelatinase producers from soil underscore the importance of microbial diversity in bioprospecting efforts. These findings contribute to the development of sustainable and efficient industrial processes by harnessing the enzymatic capabilities of soil microorganisms. Future research directions may include scaling up production, genetic engineering for enhanced enzyme properties, and further exploration of microbial diversity for novel enzyme discovery.