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Sultan Qaboos University (2016)

Optimization and production of alkaline protease using fish waste as substrate

AishwaryaRamkuma

Titre : Optimization and production of alkaline protease using fish waste as substrate

Auteur : AishwaryaRamkumar

Université de soutenance : Sultan Qaboos University

Grade : Master of Science (MS) in Physics 2016

Résumé partiel
Proteases are protein hydrolysing enzymes. More than 60% of the industrial enzymes are proteases with a wide range of applications. Thermostable bacteria are capable of producing thermostable proteases that can resist denaturation due to extreme temperature, pH and chemicals. The guts from Sardinella longiceps were processed and used as substrate for low cost production of protease enzyme. Fish waste contained 61.6 % proteins, 21.8% lipids, 8.5% carbohydrates and trace elements such as K, Na+, Ca2+, Mg2+, Fe2+, Zn*, P3- and salts PO4, C12, SO42- and NO3. Soil samples were collected from Nakhi hot spring in Sultanate of Oman to isolate protease producing bacteria. From 101 isolates, one bacteria that produced the highest protease activity (51.8 U/ml) was identified as Bacillus licheniformis. The protease enzyme produced under optimized conditions was purified and characterized. Optimization of protease. production conditions was carried out using Placket Burman (PB) design and Response Surface Methodology (RSM). Positive factors for protease fermentation were studied using PB design with 13 runs using combinations of five factors. The factor ranges were temperature (40-80°C), pH (7-12), time (24-72 h), substrate concentration (0.5-2%), and inoculum size (1-5%). Based on PB results, factors and levels for RSM design were selected. The factors and levels were temperature (40 50°C), time (65-75 h), substrate concentration (1.5-3%), and inoculum size (4-5%) and the design generated 31 runs with different combinations. Model was significant (R2 – 82.92%) and production under optimized conditions produced an increase of 1.5-fold in activity resulting in 73.52 U/ml. Purified enzyme produced a final yield of 1.70% with a 6.46-fold purification.

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Page publiée le 17 avril 2022