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| Isolation, Identification of the Antimicrobial Protein on Staphyloccus aureus and Optimization of Fermentation Process from Marine Bacillus subtilis UBMR1027 |
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Abstract It was all known that Staphylococcus aureus was an infectious pathogen to human and animal. In order to study the protein of inhibitory activities to S. aureus, the protein from Bacillus subtilis UMBR1027 was extracted by ammonium sulfate precipitation, and separated by Sephadex LH-20 column chromatography as well as sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and was also identified by ABI 4800 plus MALDI-TOF. Furthermore, the conditions of fermentation were optimized by the response surface method to increase the quantity of the bioactive protein from B. subtilis UMBR1027. The bacteriostasis activity against S. aureus was assessed by the filter paper diffusion method. The 3 key factors including the pH value of the buffer, the ionic strength in the solution and temperature of culture were evaluated respectively to optimize the conditions of fermentation by design Box-Behnken experiments. The results showed that the alkaline serine protease, subtilisin, endo-β-1,3-1,4 glucanase and alaph amylase were identified in the bioactive protein from B. subtilis UMBR1027. The optimization experiments were achieved by systematically adjusting the pH of the buffer, the ionic strength in the solution and temperature. The optimal culture conditions were the medium pH 6.95, salinity 10.38% and the temperature 34.95 ℃, respectively. The selected experiment under the culture conditions were that pH was 6.95, salinity was 10.38% and the temperature was 35 ℃, and the bacteriostatic circle diameter was 20.4 mm which was close to the predicted values. This paper identified antibactial protein from B. subtilis UMBR1027 and optimized ferment process by the response surface method. The results will provide theoretical basis to discover new natural drug molecules from marine.
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Received: 10 March 2016
Published: 22 July 2016
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