使用国产环氧树脂LXEP-120固定化脂肪酶研究

doi:10.16088/j.issn.1001-6600.2018.04.014

Abstract

Abstract: The immobilization of lipase from procine pancrestic was performed by using domestic epoxy resin LXEP-120. The immobilization conditions were optimized, and the enzymatic properties of immobilized lipase and free lipase were also compared. The optimal immobilization conditions were as follows: using 1.0 mol/L pH6.0 potassium phosphate buffer, the quantity of epoxy resin being 5.0 g, the appropriate temperature being 22 ℃, and the immobilized time being 12 h. Under above optimized conditions, the activity of immobilized lipase was about 400 U/g, and the recovery rate of the enzymatic activity of the immobilized lipase reached 92.79%. The immobilized lipase retained 70% of its initial enzymatic activity after repeated usage for 10 times. The optimal reaction pH (pH 8.0) was the same as that of free lipase, and the optimal reaction temperature(45 ℃) was 5 ℃ higher than that of free lipase. The thermal-stability and operating-stability reuse rate of immobilized lipase were significantly better than those of free lipase. Additionally, immobilized lipase exhibited very good storage stability. The study shows that domestic epoxy resins LXEP-120 possesses great potential in the immobilization of industrial enzymes.

Key words: domestic epoxy resin LXEP-120, lipase, immobilization, enzymatic property

CLC Number:

Q814

XU Shan, LI Renqiang, ZHANG Jifu, ZHANG Yun, SUN Aijun, HU Yunfeng. Immobilization of Lipase Using Domestic Epoxy Resin LXEP-120[J].Journal of Guangxi Normal University(Natural Science Edition), 2018, 36(4): 108-118.

References

[1] 谢雪凤. 嗜热子囊菌产过氧化氢酶的生产、固定化及应用研究[D]. 杭州:浙江工业大学, 2009.
[2] KUMARI A, GUPTA R. Functional characterization of a novel aspartic acid rich lipase, talipc, from trichosporon asahii msr54: solvent-dependent enantio inversion during esterification of 1-phenylethanol[J]. Biotechnol Lett, 2015, 37(1): 121-130.
[3] CAO Y, ZHUANG Y, YAO C, et al. Purification and characterization of an organic solvent-stable lipase from pseudomonas stutzeri lc2-8 and its application for efficient resolution of (r, s)-1-phenylethanol[J]. Biochemical Engineering Journal, 2012, 64: 55-60.
[4] ROMDHANE I B, ROMDHANE Z B, BOUZID M, et al. Application of a chitosan-immobilized talaromyces thermophilus lipase to a batch biodiesel production from waste frying oils[J]. Appl Biochem Biotechnol, 2013, 171(8): 1986-2002.
[5] JAMIE A, ALSHAMI A S, MALIABARI Z O, et al. Immobilization and enhanced catalytic activity of lipase on modified MWCNT for oily wastewater treatment[J]. Environmental Progress and Sustainable Energy, 2016, 35(5): 1441-1449.
[6] HA S H, LAN M N, LEE S H, et al. Lipase-catalyzed biodiesel production from soybean oil in ionic liquids[J]. Enzyme and Microbial Technology, 2007, 41(4): 480-483.
[7] VAHEDI H. Evaluating the effects of different levels of lipase enzyme on the quality of baguette bread[J]. Acta Medica Mediterranea, 2015, 31: 1359-1363.
[8] LAUPRASERT P, CHANSIRIRATTANA J,PAENGJAN J. Effect of selected bacteria as bioremediation on the degradation of fats oils and greases in wastewater from cafeteria grease traps[J]. European Journal of Sustainable Development, 2017, 6(2): 180-185.
[9] 曾淑华, 杨江科, 闫云军. 固定化脂肪酶性质及其应用研究[J]. 生物加工过程, 2007, 5(1): 45-49.
[10] KIM H, CHOI N, OH S W, et al. Synthesis of alpha-linolenic acid-rich triacylglycerol using a newly prepared immobilized lipase[J]. Food Chemistry, 2017, 237: 654-658.
[11] MENDES A A, DE CASTRO H F, RODRIGUES D D, et al. Multipoint covalent immobilization of lipase on chitosan hybrid hydrogels: influence of the polyelectrolyte complex type and chemical modification on the catalytic properties of the biocatalysts[J]. Journal of Industrial Microbiology and Biotechnology, 2011, 38(8): 1055-1066.
[12] LEI M, HU D, YANG H, et al. Preparation and characterization of hollow magnetic composite nanoparticles for immobilized pectinase[J]. Surface and Coatings Technology, 2015, 271: 2-7.
[13] NUNES M A P, VILA-REAL H, FERNANDES P C B, et al. Immobilization of naringinase in pva-alginate matrix using an innovative technique[J]. Applied Biochemistry and Biotechnology, 2010, 160(7): 2129-2147.
[14] LOPEZ-GALLEGO F, BETANCOR L, MATEO C, et al. Enzyme stabilization by glutaraldehyde crosslinking of adsorbed proteins on aminated supports[J]. Journal of Biotechnology, 2005, 119(1): 70-75.
[15] TORRES P, BATISTA-VIERA F. Immobilization of β-galactosidase from bacillus circulans onto epoxy-activated acrylic supports[J]. Journal of Molecular Catalysis B: Enzymatic, 2012, 74(3/4): 230-235.
[16] 刘文涛, 段洪东, 王兴建, 等. 环氧基固定化酶载体的研究进展[J]. 山东轻工业学院学报, 2012, 26(3): 40-44.
[17] AGHABABAIE M, BEHESHTI M, RAZMJOU A, et al. Covalent immobilization of candida rugosa lipase on a novel functionalized Fe₃O₄@SiO₂ dip-coated nanocomposite membrane[J]. Food and Bioproducts Processing, 2016, 100: 351-360.
[18] XIANG X R, SUO H B, XU C, et al. Covalent immobilization of lipase onto chitosan-mesoporous silica hybrid nanomaterials by carboxyl functionalized ionic liquids as the coupling agent[J]. Colloids and Surfaces B, Biointerface, 2018, 165: 262-269.
[19] ZHAO J Q, WANG Y J, LUO G S, et al. Covalent immobilization of penicillin g acylase on aminopropyl-functionalized mesostructured cellular foams[J]. Bioresour Technol, 2010, 101(19): 7211-7217.
[20] 顾恺, 邹树平, 王志才, 等. 环氧树脂固定化卤醇脱卤酶的研究[J]. 现代化工, 2016, 36(11): 69-74.
[21] EPHRAIM K K, KRAEMER D M. Eupergit (r) c, a carrier for immobilization of enzymes of industrial potential[J]. Journal of Molecular Catalysis B-Enzymatic, 2000, 10(1-3): 157-176.
[22] 刘向前, 李道明, 王卫飞, 等. Lipozyme~CALB L固定化及催化合成EPA/DHA甘油酯的研究[J]. 中国油脂, 2016(11): 21-26.
[23] GONZALEZ-CORONEL L A, COBAS M, Rostro-Alanis M D, et al. Immobilization of laccase of pycnoporus sanguineus cs43[J]. New Biotechnology, 2017, 39: 141-149.
[24] KNEZEVIC-JUGOVIC Z D, GRBAVCIC S Z, JOVANOVIC J R, et al. Covalent immobilization of enzymes on Eupergit() supports: effect of the immobilization protocol[J]. Methods Mol Biol, 2017, 1504: 75-91.
[25] WANG J, LI W, NIU D D, et al. Improved synthesis of isomaltooligosaccharides using immobilized alpha-glucosidase in organic-aqueous media[J]. Food Science and Biotechnology, 2017, 26(3): 731-738.
[26] 侯爱军,徐冰斌,梁亮,等.改进铜皂-分光光度法测定脂肪酶活力[J].皮革科学与工程,2011,21(1):22-27.
[27] 嵇东情, 郑仁朝, 郑裕国. 环氧树脂固定化拟南芥腈水解酶NIT的研究[J]. 精细与专用化学品, 2015, 23(5): 12-16.
[28] 崔培梧, 黎继烈, 肖作为, 等. 环氧基树脂固定化青霉菌柚苷酶的工艺条件[J]. 食品与发酵工业, 2014, 40(5): 87-92.
[29] 侯丽云. 脂肪酶固定化及其在催化合成乙酸香茅酯中的应用[D]. 扬州: 扬州大学, 2013: 11.
[30] 刘自琴. 脂肪酶和胰蛋白酶的固定化及共固定化研究[D]. 广州: 华南理工大学, 2012: 4.
[31] MATEO C, GRAZú V, PESSELA B C, et al. Advances in the design of new epoxy supports for enzyme immobilization-stabilization[J]. Biochemical Society Transactions, 2007, 35(6):1593-1601.
[32] 雷生姣, 王可兴, 吕晓燕, 等. 聚乙烯醇-海藻酸钙固定化柚苷酶[J]. 食品科学,2011,32(3): 138-143.
[33] FERNANDES A I, GREGORIADIS G. Synthesic, characterization and properties of sialylated catalase[J]. Biochimica et Biophysica Acta (BBA) -Protein Structure and Molecular Enzymology, 1996, 1293(1): 90-96.
[34] AMIN H A, SECUNDO F, AMER H, et al. Improvement of aspergillus flavus saponin hydrolase thermal stability and productivity via immobilization on a novel carrier based on sugarcane bagasse[J]. Biotechnol Rep (Amst), 2018, 17: 55-62.
[35] DILEK A, BARIS B, DENIZ Y, et al. Improving stability of formate dehydrogenase from Candida methylica by immobilization onto Eupergit C 250 L[J]. Applied Biochemistry and Microbiology, 2016(1): 37-42.
[36] DRISS D, DRISS Z, CHAARI F, et al. Immobilized his-tagged recombinant xylanase from penicillium occitanis on nickel-chelate Eupergit C[J]. Bioengineered, 2014, 5(4): 274-279.

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Immobilization of Lipase Using Domestic Epoxy Resin LXEP-120

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