PDANPs(polydopamine nanoparticles) were prepared by oxidative self-polymerization of dopamine and used as a support for Lecitase® Ultra immobilization. Enzyme activity recovery(71.2%)and immobilization efficiency (78.2%) were obtained with immobilization time of 9 h and enzyme dosage of 0.15 mL/mg. The characterizations of PDANPs and immobilized enzyme (PDANPs@Lecitase® Ultra) by transmission electron microscopy, Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy showed that nano-sized PDANPs were prepared, and Lecitase® Ultra was successfully immobilized on PDANPs. Compared with free Lecitase® Ultra, the optimum pH of PDANPs@Lecitase® Ultra increased by one unit, and optimum temperature was 5 ℃ higher. In addition, the thermal stability and storage stability were significantly improved. When PDANPs@Lecitase® Ultra was applied in camellia oil degumming, phosphorus content of the degummed oil can be reduced to less than 5 mg/kg under optimal conditions. In addition, PDANPs@Lecitase® Ultra still retained 57.6% of initial enzyme activity after 10 cycles, indicating that PDANPs@Lecitase® Ultra has a good industrial application prospect in oil degumming.
[1] LAMAS D L, CONSTENLA D T, RAAB D.Effect of degumming process on physicochemical properties of sunflower oil[J].Biocatalysis and Agricultural Biotechnology, 2016, 6:138-143.
[2] CHEW S C, NYAM K L.Refining of Edible Oils[M]//Lipids and Edible Oils.Amsterdam:Elsevier, 2020:213-241.
[3] 徐赢华, 王国敬, 李春, 等.酶法脱胶在植物油脂精炼中的应用进展[J].农业工程学报, 2015, 31(23):269-276.
XU Y H, WANG G J, LI C, et al.Application of enzymatic degumming on vegetable oils refining[J].Transactions of the Chinese society of Agricultural Engineering, 2015, 31(23):269-276.
[4] JIANG X F, CHANG M, WANG X S, et al.The effect of ultrasound on enzymatic degumming process of rapeseed oil by the use of phospholipase A1 [J].Ultrasonics Sonochemistry, 2014, 21(1):142-148.
[5] GOFFERJÉ G, MOTULEWICZ J, STÄBLER A, et al.Enzymatic degumming of crude Jatropha oil:Evaluation of impact factors on the removal of phospholipids[J].Journal of the American Oil Chemists' Society, 2014, 91(12):2 135-2 141.
[6] VIRGEN-ORTĺZ J J, DOS SANTOS J C S, ORTIZ C, et al.Lecitase ultra:A phospholipase with great potential in biocatalysis [J].Molecular Catalysis, 2019, 473:110405.
[7] YU D Y, MA Y, XUE S J, et al.Characterization of immobilized phospholipase A1 on magnetic nanoparticles for oil degumming application[J].LWT-Food Science and Technology, 2013, 50(2):519-525.
[8] DENG X, CAO S L, LI N, et al.A magnetic biocatalyst based on mussel-inspired polydopamine and its acylation of dihydromyricetin[J].Chinese Journal of Catalysis, 2016, 37(4):584-595.
[9] CHAO C, LIU J D, WANG J T, et al.Surface modification of halloysite nanotubes with dopamine for enzyme immobilization [J].ACS Applied Materials & Interfaces, 2013, 5(21):10 559-10 564.
[10] ZHAO J X, MA M M, YAN X H, et al.Green synthesis of polydopamine functionalized magnetic mesoporous biochar for lipase immobilization and its application in interesterification for novel structured lipids production[J].Food Chemistry, 2022, 379:132148.
[11] DING S W, CARGILL A A, MEDINTZ I L, et al.Increasing the activity of immobilized enzymes with nanoparticle conjugation [J].Current Opinion in Biotechnology, 2015, 34:242-250.
[12] YE Y W, ZHENG L B, WU T T, et al.Size-dependent modulation of polydopamine nanospheres on smart nanoprobes for detection of pathogenic bacteria at single-cell level and imaging-guided photothermal bactericidal activity[J].Acs Applied Materials & Interfaces, 2020, 12(31):35 626-35 637.
[13] CAO S L, DENG X, XU P, et al.Highly efficient enzymatic acylation of dihydromyricetin by the immobilized lipase with deep eutectic solvents as cosolvent[J].Journal of Agricultural and Food Chemistry, 2017, 65(10):2 084-2 088.
[14] BRADFORD M M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J].Analytical Biochemistry, 1976, 72(1-2):248-254.
[15] ZHAN J F, JIANG S T, PAN L J.Immobilization of phospholipase A1 using a polyvinyl alcohol-alginate matrix and evaluation of the effects of immobilization[J].Brazilian Journal of Chemical Engineering, 2013, 30(4):721-728.
[16] CAO D L, CHENG W J, TAO K, et al.Preparation of polydopamine-modified 3D interconnected macroporous silica for laccase immobilization[J].Macromolecular Research, 2018, 26(7):616-622.
[17] HO C C, DING S J.The pH-controlled nanoparticles size of polydopamine for anti-cancer drug delivery[J].Journal of Materials Science:Materials in Medicine, 2013, 24(10):2 381-2 390.
[18] DELFINO I, PORTACCIO M, DELLA VENTURA B D, et al.Enzyme distribution and secondary structure of sol-gel immobilized glucose oxidase by micro-attenuated total reflection FT-IR spectroscopy[J].Materials Science & Engineering C, 2013, 33(1):304-310.
[19] ZHENG G L, ZHANG D, ZHENG C, et al.Facile fabrication of biomimetic superoleophobic composite coating via Schiff base reaction and self-assembly[J].Progress in Organic Coatings, 2020, 142:105568.
[20] LOU W Y, ZONG M H, SMITH T J, et al.Impact of ionic liquids on papain:An investigation of structure-function relationships [J].Green Chemistry, 2006, 8(6):509-512.
[21] 周颖. 异源果聚糖蔗糖酶在B.subtilis中的分泌表达及固定化研究 [D].合肥:合肥工业大学, 2021.
ZHOU Y.Secretotry expression of heterologous levansucrase in B.subtilis and its immobilization [D].Hefei:Hefei University of Technology, 2021.
[22] SPINELLI D, FATARELLA E, DI MICHELE A, et al.Immobilization of fungal (Trametes versicolor) laccase onto amber lite IR-120 H beads:Optimization and characterization[J].Process Biochemistry, 2013, 48(2):218-223.
[23] YU D Y, ZHANG X, WANG T, et al.Immobilized Candida antarctica lipase B (CALB) on functionalized MCM-41:Stability and catalysis of transesterification of soybean oil and phytosterol [J].Food Bioscience, 2021, 40:100906.
[24] SATHISHKUMAR P, CHAE J C, UNNITHAN A R, et al.Laccase-poly (lactic-co-glycolic acid)(PLGA) nanofiber:Highly stable, reusable, and efficacious for the transformation of diclofenac[J].Enzyme and Microbial Technology, 2012, 51(2):113-118.
[25] XU M Q, LI F L, YU W Q, et al.Combined cross-linked enzyme aggregates of glycerol dehydrogenase and NADH oxidase for high efficiency in situ NAD(+) regeneration [J].International Journal of Biological Macromolecules, 2020, 144:1 013-1 021.
[26] 水龙龙, 操丽丽, 庞敏, 等.大孔树脂固定化磷脂酶 A1及其用于菜籽油脱胶工艺的优化[J].食品科学, 2018, 39(4):241-247.
SHUI L L, CAO L L, PANG M, et al.Optimization of process parameters for immobilization of phospholipase A1 onto macroporous resin and its application for degumming rapeseed oil[J].Food Science, 2018, 39(4):241-247.
