Phospholipase A1 is known as an effective catalyst for hydrolysis of various phospholipids in enzymatic vegetable oil degumming. Immobilization is one of the most efficient strategies to improve its activity, recovery and functional properties. In this study, chitosan-co-polyethylene oxide (90:10) nanofibrous mat was successfully fabricated and modified with atmospheric plasma at different times (2, 6 and 10 min) to interact with enzyme molecules. Scanning electron microscopy images revealed that the membranes retained uniform nanofibrous and open porous structures before and after the treatment. PLA1 was successfully immobilized onto the membrane surfaces via covalent bonds with the functional groups of chitosan nanofibrous mat. Response surface methodology was used to optimize the immobilization conditions for reaching the maximum immobilization efficiency. Enzyme concentration, pH, and immobilization time were found to be significant key factors. Under optimum conditions (5.03 h, pH 5.63, and enzyme dosage 654.36 UI), the atmospheric plasma surface modified chitosan nanofibers reached the highest immobilization efficiency (78.50%). Fourier transform infrared spectroscopy of the control and plasma surface-modified chitosan nanofibers revealed the functional groups of nanofibers and their reaction with the enzyme. The results indicated that surface modification by atmospheric plasma induced an increase in PLA1 loading on the membrane surfaces.
Richmond G, Smith T. Phospholipase A1. Int J Mol Sci. 2011; 12:588-612.
Yu D, Ma Y, Xue S. Jiang, L, Shi, L. Characterization of immobilized phospholipase A1 on magnetic nanoparticles for oil degumming application. LWT Food Sci Technol. 2012; 50:519-525.
Yang B, Zhou R, Yang JG, Wang YH, Wang WF. Insight into the enzymatic degumming process of soybean oil. J Am Oil Chem Soc. 2008; 85:421-425.
Manjula S, Jose A, Divakar S, Subramanian R. Degumming rice bran oil using phospholipase A1. Europ J Lipid Sci Technol. 2011; 113:658-664.
Jiang X, Chang M, Wang X, Jin Q, Wang X. The effect of ultrasound on enzymatic degumming process of rapeseed oil by the use of phospholipase A1. Ultrason Sonochem. 2014; 21:142-148.
Eslamipour F, Hejazi P. Effects of surface modification and activation of magnetic nanoparticles on the formation of amylase immobilization bonds under different ionic strength conditions. Mol J Catal B: Enzym. 2015; 119:1-11.
Soleimani M, Khani A, Najafzadeh K. α-Amylase immobilization on the silica nanoparticles for cleaning performance towards starch soils in laundry detergents. J Mol Catal B: Enzym. 2012; 74:1-5.
Huang XJ, Xu ZK, Wan LS, Innocent C, Seta P. Electrospun nanofibers modified with phospholipid moieties for enzyme immobilization. Macromol Rapid Commun. 2006; 27:1341-1345.
Park K, Ju YM, Son JS, Ahn K, Han DK. Surface modification of biodegradable electrospun nanofiber scaffolds and their interaction with fibroblasts. J Biomat Sci Polym E. 2007; 18:369-382.
Nickpour M, Pazouki M. Synthesis and characteristics of mesoporous sol-gels for lipase immobilization. Int J Eng. 2014; 27:1495-1502.
Zhu J, Sun G. Lipase immobilization on glutaraldehyde-activated nanofibrous membranes for improved enzyme stabilities and activities. React Funct Polym. 2012; 72: 839-845.
Lu P, Hsieh YL. Layer-by-layer self-assembly of Cibacron Blue F3GA and lipase on ultra-fine cellulose fibrous membrane. J Membrane Sci. 2010; 348:21-27.
Mourya VK, Inamdar NN. Chitosan-modifications and applications: Opportunities galore. React Funct Polym. 2008; 68:1013-1051.
Chang SH, Chian CH. Plasma surface modification effects on biodegradability and protein adsorption properties of chitosan films. Appl Surf Sci. 2013; 282:735-740.
Ni HC, Lin ZH, Hsu SH, Chiu IM. The use of air plasma in surface modification of peripheral nerve conduits. Acta Biomater. 2010; 6: 2066-2076.
Montogomery DC. Design and Analysis of Experiments, Fifth ed. John Wiley and Sons, New York, 1976; 427-511.
Zhou Y, Wang L, Wu L, Tang X, Pan S. Optimal immobilization of β-glucosidase into chitosan beads using response surface methodology. Electron. J Biotechnol. 2013; 16 (6):10.
Dwevedi A, Kayastha AM. Optimal immobilization of β-galactosidase from Pea (PsBGAL) onto Sephadex and chitosan beads using response surface methodology and its applications. Bioresour Technol. 2009; 100: 2667-2675.
Mirzaei E, Majidi RF, Shokrgozar MA, Paskiabi FA. Genipin cross-linked electrospun chitosan-based nanofibrous mat as tissue engineering scaffold. Nanomed J. 2014; 1:137-146.
Uygun A, Kiristi M, Oksuz L, Manolache S, Ulusoy S. RF hydrazine plasma modification of chitosan for antibacterial activity and nanofiber applications. Carbohydr Res. 2011; 346: 259-265.
Yang JG, Wang YH, Yang B, Mainda G, Guo Y. Degumming of vegetable oil by a new microbial lipase. Food Technol Biotechnol. 2006; 44: 101-104.
Bradford MM. A rapid and sensitive method for the quantitation of micro-gram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72: 248-254.
Huang XJ, Ge D, Xu ZK. Preparation and characterization of stable chitosan nanofibrous membrane for lipase immobilization. Europ Polym J. 2007; 43: 3710-3718.
Wanichapichart P, Sungkum R, Taweepreda W, Nisoa M. Characteristics of chitosan membranes modified by argon plasmas. Surf Coat Technol. 2009; 203: 2531-2535.
Arof AK, Osman Z. FTIR studies of chitosan acetate based polymer electrolytes. Electrochim Acta. 2003; 48: 993-999.
Bahrami A, Hejazi P. Electrostatic immobilization of pectinase on negatively charged AOT-Fe3O4 nanoparticles. J Mol Catal B: Enzym. 2013; 93: 1-7.
Xu Y, Du Y. Effect of molecular structure of chitosan on protein delivery properties of chitosan nanoparticles. Int J Pharm. 2003; 250: 215-226.
Boonsongrit Y, Mueller BW, Mitrevej A. Characterization of drug-chitosan: Interaction by H1 NMR, FTIR and isothermal titration calorimetry. Europ J Pharm Biopharm. 2008; 69: 388-395.
Yu D, Shi J, Jiang L, Xue J, Li Z, Kakuda Y. Immobilization of phospholipase A1 and its application in soybean oil degumming. J Am Oil Chem Soc. 2012; 89: 649-656.
Lee DG, Ponvel KM, Kim M, Hwang S, Ahn IS, Lee CH. Immobilization of lipase on hydrophobic nano-sized magnetite particles. J Mol Catal B: Enzym. 2009; 57: 62-66.
Sanjay G, Sugunan S. Enhanced pH and thermal stabilities of invertase immobilized on montmorillonite K-10. Food Chem. 2006; 94: 573-579.