Magnetic immobilisation of phospholipase C and its hydrolysis of phospholipids in crude soybean oil

Main Article Content

D. Yu
C. Yu
Y. Jiang
X. Zhang
T. Yuan
L. Wang
W. Elfalleh
L. Jiang

Keywords

phospholipase C, magnetic immobilisation, stability, oil degumming, 1,2-DAG

Abstract

In this study, free phospholipase C (PLC) was immobilised onto magnetic carriers, including sodium alginate (MPCSA), magnetic chitosan microparticles (MCM), Fe3O4/P (glycidyl methacrylate (GMA)-EDGM-St) and Fe3O4/SiOx-g-P (GMA). The magnetic immobilised PLC had a broader pH activity and temperature tolerance, which could be separated quickly and reused. The highest enzyme loading and enzymatic activity (135.64 mg/g and 8,560 U/g, respectively) were obtained using the magnetic immobilised PLC-Fe3O4/SiOx-g-P (GMA) system. Its optimum pH range was 5.5-7.0, and activity remained stable between 50 and 70 °C. After 6 cycles in soybean lecithin emulsion, the magnetic immobilised enzyme retained more than 80% of its initial activity. Enzymatic degumming with PLC-Fe3O4/SiOx-g-P (GMA) was carried out 2.0 h at 60 °C, resulting in 1,2-diacylglycerol (1,2-DAG) content of 1.07%, which improving the refining rate of soybean oil. Moreover, PLC-Fe3O4/SiOx-g-P (GMA) still possessed more than 80% of its initial activity after 5 cycles of degumming in crude soybean oil. The results show that magnetic immobilised PLC has a good industrial application.

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References

Bradford, M.M., 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248-254.
Brühl, L., 1997. Official methods and recommended practices of the American Oil Chemist’s Society. Physical and chemical characteristics of oils, fats and waxes. Lipid/Fett 99: 197-197.
Chang, M.Y. and Juang, R.S., 2005. Activities, stabilities, and reaction kinetics of three free and chitosan-clay composite immobilised enzymes. Enzyme and Microbial Technology 36: 75-82.
Dayton, C.L., Rosswurm, E.M. and Da Silva Galhardo, F., 2013. Enzymatic degumming utilizing a mixture of PLA and PLC phospholipases with reduced reaction time. U.S. Patent No. 8,460,905. U.S. Patent and Trademark Office, Washington, DC, USA.
Dijkstra, A.J., 2010. Enzymatic degumming. European Journal of Lipid Science and Technology 112: 1178-1189.
Elena, C., Ravasi, P., Cerminati, S., Peiru, S., Castelli, M.E. and Menzella, H.G., 2016. Pichia pastoris engineering for the production of a modified phospholipase C. Process Biochemistry 51: 1935-1944.
Garcia-Galan, C., Berenguer-Murcia, Á., Fernandez-Lafuente, R. and Rodrigues, R.C., 2011. Potential of different enzyme immobilisation strategies to improve enzyme performance. Advanced Synthesis and Catalysis 353: 2885-2904.
Guo, H., Tang, Y., Yu, Y., Xue, L. and Qian, J., 2016. Covalent immobilisation of ?-amylase on magnetic particles as catalyst for hydrolysis of high-amylose starch. International Journal of Biological Macromolecules 87: 537-544.
Ibrahim, A.S., Al-Salamah, A.A., El-Toni, A.M., El-Tayeb, M.A. and Elbadawi, Y.B., 2013. Immobilisation of cyclodextrin glucanotransferase on aminopropyl-functionalized silica-coated superparamagnetic nanoparticles. Electronic Journal of Biotechnology 16: 10.
Jiang, X., Chang, M., Jin, Q. and Wang, X., 2015a. Application of phospholipase A1 and phospholipase C in the degumming process of different kinds of crude oils. Process Biochemistry 50: 432-437.
Jiang, X., Chang, M., Jin, Q. and Wang, X., 2015b. Optimisation of the degumming process for camellia oil by the use of phospholipase C in pilot-scale system. Journal of Food Science and Technology 52: 3634-3644.
Jiang, X., Chang, M., Wang, X., Jin, Q. and Wang, X., 2014. A comparative study of phospholipase A 1 and phospholipase C on soybean oil degumming. Journal of the American Oil Chemists’ Society 91: 2125-2134.
Laatikainen, M., Srithammavut, W., Toukoniitty, B., Turunen, I. and Sainio, T., 2015. Phospholipid adsorption from vegetable oils on acid-activated sepiolite. Adsorption 21: 409-417.
Lamas, D.L., Constenla, D.T. and Raab, D., 2016. Effect of degumming process on physicochemical properties of sunflower oil. Biocatalysis and Agricultural Biotechnology 6: 138-143.
Lamas, D.L., Crapiste, G.H. and Constenla, D.T., 2014. Changes in quality and composition of sunflower oil during enzymatic degumming process. LWT – Food Science and Technology 58: 71-76.
Li, Z., Liu, H., Zhao, G., Wang, P., Wang, L., Wu, H., Fang, X., Sun, X., Wu, X. and Zheng, Z., 2016. Enhancing the performance of a phospholipase A1 for oil degumming by bio-imprinting and immobilisation. Journal of Molecular Catalysis B: Enzymatic 123: 122-131.
Lima, J.S., Araújo, P.H., Sayer, C., Souza, A.A., Viegas, A.C. and De Oliveira, D., 2017. Cellulase immobilisation on magnetic nanoparticles encapsulated in polymer nanospheres. Bioprocess and Biosystems Engineering 40: 511-518.
Lin, F. and Doong, R., 2011. Bifunctional Au-Fe3O4 heterostructures for magnetically recyclable catalysis of nitrophenol reduction. Journal of Physical Chemistry C115: 6591-6598.
Liu, Q., Hua, Y., Kong, X., Zhang, C. and Chen, Y., 2013. Covalent immobilisation of hydroperoxide lyase on chitosan hybrid hydrogels and production of C6 aldehydes by immobilised enzyme. Journal of Molecular Catalysis B: Enzymatic 95: 89-98.
Long, J., Jiao, A., Wei, B., Wu, Z., Zhang, Y., Xu, X. and Jin, Z., 2014. A novel method for pullulanase immobilised onto magnetic chitosan/Fe3O4 composite nanoparticles by in situ preparation and evaluation of the enzyme stability. Journal of Molecular Catalysis B: Enzymatic 109: 53-61.
Mei, L., Wang, L., Li, Q., Yu, J. and Xu, X., 2013. Comparison of acid degumming and enzymatic degumming process for Silybum marianum seed oil. Journal of the Science of Food and Agriculture 93: 2822-2828.
Milosavi?, N., Prodanovi?, R., Jovanovi?, S. and Vuj?i?, Z., 2007. Immobilisation of glucoamylase via its carbohydrate moiety on macroporous poly (GMA-co-EGDMA). Enzyme and Microbial Technology 40: 1422-1426.
Palvannan, T. and Boopathy, R., 2005. Phosphatidylinositol-specific phospholipase C production from Bacillus thuringiensis serovar. kurstaki using potato-based media. World Journal of Microbiology and Biotechnology 21: 1153-1155.
Piel, M.S., Peters, G.H. and Brask, J., 2017. Chemoenzymatic synthesis of fluorogenic phospholipids and evaluation in assays of phospholipases A, C and D. Chemistry and Physics of Lipids 202: 49-54.
Qu, Y., Sun, L., Li, X., Zhou, S., Zhang, Q., Sun, L., Yu, D., Jiang, L. and Tian, B., 2016. Enzymatic degumming of soybean oil with magnetic immobilised phospholipase A2. LWT – Food Science and Technology 73: 290-295.
Ravasi, P., Braia, M., Eberhardt, F., Elena, C., Cerminati, S., Peirú, S., Castelli, M.E. and Menzella, H.G., 2015. High-level production of Bacillus cereus phospholipase C in Corynebacterium glutamicum. Journal of Biotechnology 216: 142-148.
Subrahmanyam, C.V., Rao, M.V., Balasubrahmanyam, V. and Bhowmick, D.N., 2006. Membrane degumming of crude rice bran oil: pilot plant study. European Journal of Lipid Science and Technology 108: 746-752.
Xie, W. and Wang, J., 2012. Immobilised lipase on magnetic chitosan microspheres for transesterification of soybean oil. Biomass & Bioenergy 36: 373-380.
Xu, F.J., Cai, Q.J., Li, Y.L., Kang, E.T. and Neoh, K.G., 2005. Covalent immobilisation of glucose oxidase on well-defined poly (glycidyl methacrylate)-Si(111) hybrids from surface-initiated atom-transfer radical polymerisation. Biomacromolecules 6: 1012-1020.
Yang, L., Zou, P., Cao, J., Sun, Y., Han, D., Yang, S., Chen, G., Kong, X. and Yang, J., 2014. Facile synthesis and paramagnetic properties of [email protected] core-shell nanoparticles. Superlattices and Microstructures 76: 205-212.
Ye, L., Deng, K., Xu, F., Tian, L., Peng, T. and Zan, L., 2012. Increasing visible-light absorption for photocatalysis with black BiOCl. Physical Chemistry Chemical Physics 14: 82-85.
Yu, D., Jiang, L., Li, Z., Shi, J., Xue, J. and Kakuda, Y., 2012. Immobilisation of phospholipase A1 and its application in soybean oil degumming. Journal of the American Oil Chemists’ Society 89: 649-656.
Yu, D., Ma, Y., Jiang, L., Walid, E., He, S., He, Y., Xiaoyu, Z., Zhang, J. and Hu, L., 2014. Stability of soybean oil degumming using immobilised phospholipase A2. Journal of Oleo Science 63: 25-30.
Yu, D., Ma, Y., Xue, S.J., Jiang, L. and Shi, J., 2013. Characterisation of immobilised phospholipase A1 on magnetic nanoparticles for oil degumming application. LWT – Food Science and Technology 50: 519-525.
Zamora, R., Olmo, C., Navarro, J.L. and Hidalgo, F.J., 2004. Contribution of phospholipid pyrrolisation to the color reversion produced during deodorisation of poorly degummed vegetable oils. Journal of Agricultural and Food Chemistry 52: 4166-4171.