Production of lipase enzyme using a mixed microbial vaccine technique from two local bacterial isolates, Kytococcus sedentarius and Pseudomonas oleovorans, utilizing solid medical waste

Document Type : Research Paper

Authors

Department of Biology, College of Education for Pure Sciences, University of Anbar, Anbar, Iraq

Abstract

Objective
This study aims to isolate and identify local bacterial strains capable of degrading medical waste while producing lipase enzymes. Key factors influencing enzyme production, including pH, temperature, and nutrient composition, were optimized to enhance yield. The findings contribute to sustainable waste management and industrial enzyme applications.
Materials and Methods
This study was conducted in the laboratories of the Department of Life Sciences, College of Education for Pure Sciences, University of Anbar. Local bacterial isolates capable of degrading solid medical waste were obtained and identified using standard diagnostic methods. Identification was confirmed using the Vitek system, which classified the isolates as Kytococcus sedentarius and Pseudomonas oleovorans. Lipase production was carried out using the submerged culture technique, with solid medical waste serving as the primary carbon and energy source. Various factors affecting enzyme production—including pH, temperature, inoculum size, nitrogen source, waste concentration, and incubation duration—were investigated to determine optimal conditions. Enzyme activity was measured, and the effect of nitrogen supplementation on lipase production was assessed.

Results
The bacterial isolates Kytococcus sedentarius and Pseudomonas oleovorans demonstrated high efficiency in degrading solid medical waste and producing lipase enzymes. Optimal conditions for maximum enzyme yield were determined to be pH 8, a temperature of 45°C, an inoculum volume of 6 mL per 100 mL of medium, and a waste concentration of 2 g per 100 mL of medium. Under these conditions, the highest recorded enzyme activity was 8.5 units/mL. The addition of a nitrogen source led to a decrease in enzyme activity from 8.5 to 8.1 units/mL, suggesting that the solid medical waste medium provided sufficient nitrogen, with additional supplementation acting as an inhibitor of enzyme production.
Conclusions
This study successfully isolated and identified two bacterial strains, Kytococcus sedentarius and Pseudomonas oleovorans, with high efficiency in degrading solid medical waste and producing lipase enzymes. The optimal conditions for maximum enzyme production were established, highlighting the potential of these bacteria for biotechnological applications. The findings suggest that solid medical waste can serve as a viable substrate for enzyme production, simultaneously reducing waste and generating valuable bioproducts. Furthermore, the observed inhibition of enzyme activity by nitrogen supplementation underscores the importance of optimizing nutrient composition. This research contributes to sustainable waste management and industrial enzyme production, paving the way for further studies on large-scale applications.

Keywords

References

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Agricultural Biotechnology Journal
Volume 17, Issue 1
March 2025
Pages 247-270

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