Preparation and Characterization of Glucose Oxidase Liposomes

doi:10.7506/spkx1002-6630-201613021

Abstract

Abstract:

Egg phosphatidylcholine and cholesterol were used as coating materials to prepare glucose oxidase (GOD)
liposomes by a thin film evaporation-freezing thawing method. Single factor experiments and orthogonal array design
methods were used to optimize the preparation conditions based on encapsulation efficiency. The shape, stability and
particle size distribution of liposome-encapsulated glucose oxidase were systematically investigated. The optimal process
parameters were obtained as follows: mass ratio between egg phosphatidylcholine and cholesterol of 4:1, 5 mg (500 U) of
GOD, hydration time of 15 min, and 10 freezing-thawing cycles. The encapsulation efficiency of GOD liposomes was up to
(87.7 ± 2.1)%, with an average diameter of approximately (164.75 ± 2.55) μm under the optimized conditions. The encapsulation
efficiency did not significant change at 4 ℃ and room temperature (23.0 ± 0.5) ℃, and the activity of GOD remained stable.

Key words: glucose oxidase, liposome, stability, film evaporation-freezing thawing method, characterization

CLC Number:

GUAN Huanan, HAN Bolin, NAO Amin, WANG Xin, XU Liping, SUN Lu. Preparation and Characterization of Glucose Oxidase Liposomes[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201613021.

References

[1] 周建芹, 陈韶华, 王剑文. 定向固定化葡萄糖氧化酶及其酶学性质

的研究[J]. 食品科学, 2009, 30(1): 186-189.

[2] 刘超, 袁建国, 王元秀, 等. 葡萄糖氧化酶的研究进展[J]. 食品与药

品, 2010, 12(7): 285-289.

[3] GULDEN Y, SEZGINTURK M K. A new methodology for the

determination of enzyme activity based on carbon nanotubes

and glucose oxidase[J]. Journal of Pharmaceutical and Biomedical

Analysis, 2015, 115(10): 254-259. DOI:10.1016/j.jpba.2015.07.037.

[4] PALOD P A, SINGH V. Improvement in glucose biosensing response

of electrochemically grown polypyrrole nanotubes by incorporating

crosslinked glucose oxidase[J]. Materials Science and Engineering: C,

2015, 55: 420-430. DOI:10.1016/j.msec.2015.05.038.

[5] 李艳, 李静. 葡萄糖氧化酶及其应用[J]. 食品工程, 2006, 9(3): 9-11.

[6] SUSANTO H, SAMSUDIN A M, ROKHATI N, et al. Immobilization

of glucose oxidase on chitosan-based porous composite

membranes and their potential use in biosensors[J]. Enzyme and

Microbial Technology, 2013, 52(6/7): 386-392. DOI:10.1016/

j.enzmictec.2013.02.005.

[7] 钱军民, 李旭祥. 纤维素固定化葡萄糖氧化酶的研究[J]. 西安交通

大学学报, 2001, 35(4): 416-420.

[8] ZHU Yanan, JIANG Yanjun, GAO Jing, et al. Immobilization of

glucose oxidase in liposome-templated biomimetic silica particles[J].

Chinese Journal of Catalysis, 2013, 34(1): 741-750. DOI:10.1016/

S1872-2067(11)60159-6.

[9] 熊伟, 李雄辉, 胡居吾, 等. 薄膜-超声法制备二氢杨梅素脂质体的

工艺研究[J]. 食品工业科技, 2012, 33(5): 254-257. DOI:10.13386/

j.issn1002-0306.2012.05.026.

[10] MAKOTO Y, NORRIYUKI T, MIKU Y. Catalase-conjugated

liposomes encapsulating glucose oxidase for controlled oxidation

of glucose with decomposition of hydrogen peroxide produced[J].

Colloids and Surfaces B: Biointerfaces, 2010, 79(2): 403-408.

DOI:10.1016/j.colsurfb.2010.05.006.

[11] 马宁, 濮少杰, 李峰, 等. 茶多酚脂质体的制备[J]. 食品科学, 2012,

33(20): 16-20.

[12] 徐云龙, 徐盛, 杨欣欣. 溶菌酶脂质体的制备工艺[J]. 华东理工

大学学报(自然科学版), 2006, 32(8): 939-942. DOI:10.14135/

j.cnki.1006-3080.2006.08.010.

[13] 宋佳佳, 刘宁. 薄膜蒸发-冷冻干燥法制备乳糖酶脂质体[J]. 食品工

业, 2011(9): 29-32.

[14] 王健, 李明轩. 冷冻干燥对提高脂质体稳定性的研究概况[J]. 中国

医药工业杂志, 2005, 36(9): 576-580.

[15] 吴亚妮, 徐云龙, 孙文晓. 木瓜蛋白酶纳米脂质体的制备及其粒度

控制[J]. 上海交通大学学报(农业科学版), 2007, 25(2): 1052-1091.

[16] 任文霞, 李建科. 茶多酚脂质体的制备[J]. 食品工业科技, 2008,

29(11): 186-189. DOI:10.13386/j.issn1002-0306.2008.11003.

[17] 胥传来, 乐国伟, 吴振国. 维生素C 新剂型脂质体制备

办法比较[J] . 中国饲料, 2002(10): 26-28. DOI:10.3969/

j.issn.1004-3314.2002.10.013.

[18] 熊伟, 李雄辉, 胡居吾, 等. 薄膜-超声法制备二氢杨梅素脂质体的

工艺研究[J]. 食品工业科技, 2012, 33(5): 254-257. DOI:10.13386/

j.issn1002-0306.2012.05.026.

[19] 孙维彤, 黄桂华, 叶杰胜, 等. 鱼精蛋白凝聚法测定脂质体和纳米脂

质体包封率[J]. 中国药学杂志, 2006, 41(22): 1717-1721.

[20] 曾德贵, 曾洪辉. 干扰素脂质体的制备与质量控制[J]. 制剂技术,

2010, 19(19): 37-38.

[21] 张一鸣, 刘靖新, 周颖琳, 等. 固相萃取-高效液相色谱法测定紫

杉醇脂质体的包封率[J]. 分析科学学报, 2014, 30(5): 705-708.

DOI:10.13526/j.issn.1006-6144.2014.05.021.

[22] 王妍, 张水华, 李云富, 等. 膜材性质及制备方法调控下的脂质体负

载干扰素的研究[J]. 食品科学, 2008, 29(2): 59-63.

[23] 张奇, 邓英杰. 冻融法制备5-氟尿嘧啶脂质体及其稳定性考察[J].

沈阳药科大学学报, 2000, 17(2): 87-89. DOI:10.14066/j.cnki.cn21-

1349/r.2000.02.003.

[24] 李红茹, 李淑芬. 脂质体中药物包封率的测定方法[J]. 药物分析杂

志, 2007, 27(11): 1844-1848. DOI:10.16155/j.0254-1793.2007.11.046.

[25] 叶兆伟, 承伟. 脂质体包封率测定方法及影响因素[ J ] . 中

国生物制品学杂志, 2007, 20(10): 789-792. DOI:10.3969/

j.issn.1004-5503.2007.10.022.

[26] 郑会娟, 刘成梅, 刘伟, 等. 中链脂肪酸脂质体的制备及其性质测定[J].

食品科学, 2010, 31(22): 170-175.

[27] YANG Shuibing, LIU Chengmei, LIU Wei, et al. Preparation and

characterization of nanoliposomes entrapping medium-chain fatty

acids and vitamin C by lyophilization[J]. International Journal of

Molecular Sciences, 2013, 14(10): 19763-19773. DOI:10.3390/

ijms141019763.