Optimized production of extracellular proteases by Bacillus subtilis from degraded abattoir waste

Proteases are ubiquitous in occurrence and are found in all living organisms. These are essential for cell growth and differentiation. The extracellular proteases are of a high commercial value and find multiple applications in various industrial sectors. The present study describes the screening of protease producing bacteria from a hitherto unexplored source i.e. degraded waste from abattoir. Three isolates were found namely yellow, white and orange coloured bacteria. Amongst them, white colored colony was found to be more suitable for protease production. The morphological, cultural, biochemical and 16S rRNA confirmed that the isolate was Bacillus subtilis. Physical and chemical parameters were optimized for maximum protease production and optimum temperature and pH was found to be 40oC at pH 14. Glucose as a carbon source and yeast extract as a nitrogen source further stimulated the production process giving maximum protease activity to be 20.74 U/ml and 20.67 U/ml. The applications of protease in detergent and solvent industry were tested and it was revealed that the purified enzyme can be used as an additive in detergent industry

Synthesis of glycinamides using protease immobilized magnetic nanoparticles

In the present investigation, Bacillus subtilis was isolated from slaughterhouse waste and screened for the production of protease enzyme. The purified protease was successfully immobilized on magnetic nanoparticles (MNPs) and used for the synthesis of series of glycinamides. The binding and thermal stability of protease on MNPs was confirmed by FTIR spectroscopy and TGA analysis. The surface morphology of MNPs before and after protease immobilization was carried out using SEM analysis. XRD pattern revealed no phase change in MNPs after enzyme immobilization. The processing parameters for glycinamides synthesis viz. temperature, pH, and time were optimized using Response Surface Methodology (RSM) by using Design Expert (9.0.6.2). The maximum yield of various amides 2 butyramidoacetic acid (AMD-1,83.4%), 2-benzamidoacetic acid (AMD-2,80.5%) and 2,2′((carboxymethyl) amino)-2-oxoethyl)-2-hydroxysuccinyl)bis(azanediyl))diacetic acid (AMD-3,80.8%) formed was observed at pH-8, 50 °C and 30 min. The synthesized immobilized protease retained 70% of the initial activity even after 8 cycles of reuse