Immobilization of the Magnetic Nanoparticles with Alkaline Protease Enzyme Produced by Enterococcus hirae and Pseudomonas aeruginosa Isolated from Dairy Effluents

Abstract

Protease is an enzyme which has a wide range of applications in various fields. Extracellular protease was produced from Pseudomonas aeruginosa and Enterococcus hirae which were isolated from the effluents of diary industries. Protease immobilized with super paramagnetic nanoparticles was characterized by DLS, XRD and TEM methods in relation to their size and structure. The protease enzyme was bound to magnetic nanoparticles via surface transformation technique including Silica coated magnetic Nano composite, amine and cysteine functioned Nano composite formation. Successful binding of protease onto the particles was confirmed by TEM imaging. The maximal enzyme activity of immobilized protease was determined using universal protease assay and was found to be 105µg mL -1 & 290µg mL -1 for Pseudomonas sp. and Enterococcus sp. respectively. The immobilization capacity of protease onto nanoparticles was 6000µM/g. The stability of the immobilized enzyme increased in comparison with the free enzyme. Overall, this study showed that the stability and activity of the protease was enhanced by immobilization to the magnetic nanoparticles. This suggested that immobilized enzyme on the magnetic beads of nanoparticles could be used in an interesting range of applications, both in broader temperature and pH ranges, also permitting magnetic recovery of the enzyme for reuse or purification of the product.

Microbial proteases: ubiquitous enzymes with innumerable uses

“…Al-Dhabi-82 was reported by Al-Dhabi et al ( 2020 ). Similarly, Masi et al ( 2017 ) reported alkaline protease production from Enterococcus hirae and Pseudomonas aeruginosa isolated from dairy effluents. They further carried out formation of nanocomposite of the bacterial protease with super paramagnetic nanoparticles leading to protease stability enhancement.…”

Isolation, screening, characterization, and identification of alkaline protease-producing bacteria from leather industry effluent

“…The plates were observed for the zone of proteolysis around the inoculated wells. The enzymes which show a high zone of hydrolysis are selected and cultured in nutrient broth medium and also preserved in agar slants at 4°C for further studies (Sneha et al 2014, Masi et al 2017a).…”

“…Enzymes have better catalytic efficiency, adjustable activity, and high specificity when compared to catalysts of chemical or synthetic origin. These advantages have broadened the application of enzymes in various industries such as chemical, food, and pharmaceutical (Pires-Cabral et al 2010;Yucel 2012;Masi et al 2017a). This has generated a greater demand for enzyme production of high quality by cost-effective and commercial methods.…”

Impact of immobilization technology in industrial and pharmaceutical applications

“…The enzyme shows Brownian movement in aqueous solution when bounded with nanoparticles. Additionally, there is an additional advantage for magnetic nanoparticles as it can be separated easily using a magnetic field (Neda et al 2019;Masi et al 2017). The advantage of nanoparticles in immobilization is that it can improve stability and performance and reduce protein unfolding other advantages and disadvantages listed in Table 9.…”