Preparation and Properties of Purified Phytase from Oakbug Milkcap (<i>Lactarius Quietus</i>) Immobilised on Coated Chitosan with Iron Nano Particles and Investigation of Its Usability in Food Industry

Abstract

In this study, phytase enzyme was purified and characterized from Oakbug Milkcap mushrooms (Lactarius quietus) and the purified phytase enzyme was immobilized on the surface of modified chitosan with nano-Fe 3 O 4 nanoparticles. The phytase was purified from Oakbug Milkcap mushrooms using ammonium sulphate precipitation in the range of 40-80%. and DEAE-sephadex ion-exchange chromatography. The purification fold was calculated by determining the activity and amount of protein for each step. The purified phytase enzyme was then immobilized on modified chitosan support material with Fe 3 O 4 nanoparticles. The optimum pHs for the immobilized and free enzyme were 6.0 and the optimum temperatures were 60°C. The effects of some metal ions such as CuCl 2 , Hg 2 Cl 2 , FeCl 2 , MgCl 2 , ZnCl 2 , CaCl 2 were investigated on both the bound and free purified phytase. The molecular weight of the enzyme was determined using the SDS-PAGE electrophoresis method. Finally, we investigated whether or not the immobilized and free enzyme could hydrolyzed the phytic acid in green lentils, red lentils, peas, pinto beans, beans, brass, corn, dried corn, oat, rye, wheat, broad bean, chickpeas and peanuts. It was concluded that the immobilized phytase was quite resistant to temperature, pH and metal ions and it could be safely used in the hydrolysis of phytic acid in the legumes in the food industry.

Enhancement of ZnO Nanorods Properties Using Modified Chemical Bath Deposition Method: Effect of Precursor Concentration

“…The widening of E g is related to an increase in the transition tail width [83,84]. The increase in the compressive strain alongside the c-axis and decreases with an increase in tensile strain causes an increase in E g [85,86]. The investigating results show an increase in E g and a decrease in the tensile strain as an increase in the precursor concentration [52].…”

Immobilization of Phytase from Rice Bean (Vigna umbellata Thunb.) on Glutaraldehyde Activated Chitosan Microspheres

“…Though the optimum temperature is 40 °C, both the free and the immobilized enzymes are active at 80 °C with 36 and 44% activities, respectively. In some other cases also, there is no change in optimum temperatures of both the free, and the immobilized phytase (Celem & Onal, 2009a,b;Cho et al, 2011;Onem & Nadaroglu, 2014). At higher temperatures, the microspheres tend to become softer, as the incubation time is also 30 min.…”

“…It has been purified from some plant sources and characterized (Gibson & Ullah, 1988;Konietzny et al, 1995;Mahajan & Dua, 1997;Phillippy, 1998;Greiner et al, 1998Greiner et al, , 2001Greiner & Alminger 1999;Greiner, 2002;Andriotis & Ross, 2003;Belho et al, 2016;Belho & Ambasht, 2017). Immobilization of phytase has been pursued mostly from microbial sources like Aspergillus ficuum (Liu et al, 1999;Moraes et al, 2008), A. niger (Menezes-Blackburn et al, 2011;Trouillefou et al, 2015;Coutinho et al, 2020), A. heteromorphus (Lata et al, 2014), Escherichia coli (Menezes-Blackburn et al, 2011;Cho et al, 2011;El-Shora et al, 2019), Candida krusei (Quan et al 2003), and mushrooms (Onem & Nadaroglu, 2014;Onem et al, 2016). The literature on the enzyme immobilization, and the use of different matrices, has been extensively discussed (Mosbach, 1971;Mateo et al, 2007;Spahn & Minteer, 2008;Liu et al, 2018;Sastre et al, 2020).…”

Biosynthesis, purification, kinetic and thermodynamic analysis of phytase from Aspergillus oryzae

“…Fractional precipitation was carried out using different concentrations of ammonium sulfate (10-90%) for partial purification of the protein of interest and dialyzed to remove excessive salt (Onem and Nadaroglu, 2014 (Demir et al, 2017).…”