Production of keratinase enzyme from a local isolate of Kocuria rosea using environmental waste

Document Type : Research Paper

Authors

1 MSc Student, Department of Biology, College of Education for Pure Sciences, University of Anbar, Anbar, Iraq.

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

10.22103/jab.2025.24585.1642

Abstract

Objective
Keratinase is an enzyme that belongs to the metalloprotease group, along with many other protein-degrading enzymes. Keratinase is a specialized enzyme that acts on the substrate (keratin). It breaks the strong chemical bonds of keratin. This study aimed to produce the enzyme keratinase using local bacterial isolates obtained from soil samples and poultry waste from different areas in Al-Anbar province, using various waste materials such as hooves, horns, and hides.

Materials and methods
Seventeen bacterial isolates were obtained, which demonstrated a high ability to grow on feather agar medium used for isolation, from 20 soil and poultry waste samples. The five most efficient bacterial isolates were selected based on the diameter of the colonies growing on the feather agar medium. Among them, the bacterial isolate designated with the local code A2 was chosen as the most efficient in keratin degradation after culturing the five selected isolates on pure keratin medium. The bacterial isolate was identified based on morphological, cultural, microscopic characteristics, and biochemical tests. The VITEK 2 Compact device was used to confirm the identification.

Results
The results indicated that the isolate was Kocuria rosea. The optimal conditions for enzyme production from Kocuria rosea, including temperature, pH, inoculum size, carbon source and its concentration, nitrogen source and its concentration, and incubation time were studied. Based on the experiments and their results, the medium prepared using sheep hooves was selected for the growth of Kocuria rosea and the production of keratinase, particularly since it is an environmentally available and inexpensive waste in the local environment. The results of the optimal conditions study showed that the best production of keratinase enzyme was at a temperature of 30°C, a pH of 8.0, shaking speed of 150 rpm, with 3g/100mL of sheep hooves in the medium, 5mL/100mL inoculum, using urea as the nitrogen source at a concentration of 0.1g/100mL, and an incubation time of 72 hours. The enzymatic activity reached 0.301 U/mL.

Conclusions
These findings underscore the potential of Kocuria rosea in bioindustrial applications, particularly in processes involving keratin waste recycling and sustainable waste management.

Keywords

References

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Agricultural Biotechnology Journal
Volume 16, Issue 4
January 2025
Pages 391-412

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