Enhancement of the hyaluronidase production from isolated Staphylococcus aureus using factorial design technique and partial purification

• An isolated bacterial strain showed considerable ability to produce hyaluronidase enzyme.
• New medium contains waste milk whey supported hyaluronidase production (189 U/ml).
• Optimization with factorial design model increased the enzyme activity to 492 U/ml.
• A trial to improve the enzyme activity was made through fractional precipitation with organic solvents and ammonium sulphate.

Hyaluronidase is an enzyme has a great potential in diverse medical fields. A group of bacterial isolates was screened for the production of hyaluronidase. The most potent isolate was identified depending on its morphological, biochemical and molecular characteristics. Thereafter, the new powerful enzyme producer was identified as Staphylococcus aureus. Optimization strategy was conducted to enhance the enzyme production by S. aureus in a medium containing a waste material (milk whey). The first step of the optimization process included choosing of the most significant factors after screening with Plackett–Burman model. Eleven factors, medium composition and some cultural conditions, were screened. Hence, the most significant factors were subjected to further optimization step using Box–Behnken model. Box–Behnken model had the ability to suggest the optimum concentrations of the significant parameters and to predict the maximal enzyme activity. The predicted enzyme activity was 500 U/ml. The predicted value was verified experimentally, and the maximum enzyme activity reached 492 U/ml. Overall, the final enzyme outcome 492 U/ml is considered absolutely high if it compared with all previous findings with free cells. In addition to using of competitive cheaper media, attempts were made to increase the effectiveness of the enzyme through partial purification.

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