Physicochemical and Enzymatic Behavior Assessment of a Carbohydrate-Oxidizing Biocatalyst Extracted from Native Pseudomonas and Actinomyces Organisms

Abstract

Carbohydrate-oxidizing biocatalysts derived from microbial systems have gained significant attention due to their applications in biosensing, bioprocessing, and industrial biocatalysis. This study investigates the physicochemical stability, enzymatic behavior, and functional efficiency of a glucose-oxidizing biocatalyst extracted from native Pseudomonas and Actinomyces species. The research integrates structural-functional protein theory, enzyme kinetics, and disorder prediction frameworks to evaluate catalytic performance under variable environmental conditions.

Intrinsic protein disorder is increasingly recognized as a determinant of enzymatic flexibility and functional adaptability, particularly in microbial enzymes (Wright & Dyson, 1999; Dunker et al., 2002). Disorder-based functional plasticity is further supported by genome-wide analyses of protein flexibility patterns in microbial systems (Dunkor et al., 2000). In this context, computational and experimental approaches are combined to assess enzyme stability, substrate affinity, and thermodynamic efficiency.

The study builds upon established methodologies in protein disorder prediction (Ward et al., 2004) and enzymatic characterization protocols, aligning them with biochemical kinetic frameworks (McCormick, 1981). Furthermore, recent biochemical investigations into glucose oxidase systems demonstrate the relevance of microbial enzymatic variability in catalytic efficiency and thermodynamic stability under natural conditions (Singh, Modi, & Tiwari, 2019).

Results indicate that the extracted biocatalyst exhibits optimal catalytic activity under moderate pH and temperature conditions, with measurable shifts in enzymatic velocity corresponding to structural flexibility in protein conformation. Thermodynamic analysis suggests a balance between stability and reactivity, consistent with partially disordered protein regions contributing to catalytic adaptability.

The findings highlight the importance of integrating structural disorder theory with enzymatic kinetics to better understand microbial biocatalysts. This study contributes to the development of optimized bio-based oxidation systems with potential applications in industrial biotechnology, biosensor engineering, and metabolic pathway optimization.