新生牛血中Cu/Zn-SOD的提纯及肝素、壳聚糖的逐层自组装包裹

doi:10.16431/j.cnki.1671-7236.2016.08.015

《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (8): 2032-2037.doi: 10.16431/j.cnki.1671-7236.2016.08.015

新生牛血中Cu/Zn-SOD的提纯及肝素、壳聚糖的逐层自组装包裹

侯静¹, 林春丽², 白志军¹, 魏建民¹, 张永明², 张峰¹

1. 内蒙古农业大学生命科学学院, 呼和浩特 010018;
2. 内蒙古维克生生物科技有限公司, 呼和浩特 010015

收稿日期:2016-01-25 出版日期:2016-08-20 发布日期:2016-08-23
通讯作者: 张峰 E-mail:fengzhang1978@hotmail.com
作者简介:侯静(1990-),女,内蒙古巴彦淖尔人,硕士生,研究方向:逐层自组装微囊的制备,E-mail:13674852359@163.com
基金资助:
国家自然科学基金（21171086、81160213）；内蒙古自治区草原英才工程项目（108-108038）

Separation and Purification of Cu/Zn-SOD from Newborn Bovine Blood and Prepatation of HEP and CHI Using Layer-by-layer Self-assembly Technique

HOU Jing¹, LIN Chun-li², BAI Zhi-jun¹, WEI Jian-min¹, ZHANG Yong-ming², ZHANG Feng¹

1. College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. Inner Mongolia WeiKeSheng Biological Technology Co., Ltd., Hohhot 010015, China

Received:2016-01-25 Online:2016-08-20 Published:2016-08-23

摘要/Abstract

摘要：

试验旨在通过对超氧化物歧化酶（super oxide dismutase, SOD）改性，提高其稳定性。本试验采用了热变性提取方法从新生牛血中提取Cu/Zn-SOD，采用逐层自组装技术，以肝素与壳聚糖为材料，对从新生牛血中分离提纯的SOD进行物理包裹试验，并对包裹后Cu/Zn-SOD的稳定性进行研究。结果显示，在耐热稳定性上，包裹的SOD相对酶活性比天然SOD提高了3.3倍；在耐酸碱性方面，在pH分别为3.0、11.0的条件下，包裹的SOD相对酶活性较天然SOD提高了2.0倍；在抗胰蛋白酶水解的能力上，包裹的SOD相对酶活性较天然SOD提高了2.7倍。采用肝素、壳聚糖制成的外壳对SOD进行物理修饰可得到较好的效果，且其具有较强的机械强度、生物相容性较好，外表面的壳聚糖有助于帮助跨细胞膜转运，可应用于口服剂型SOD的开发。

关键词: SOD; 肝素; 壳聚糖; 逐层自组装技术; 相对酶活性

Abstract:

In order to improve the stability of super oxide dismutase (SOD) with some modification,we used thermal denaturation extraction method to extract Cu/Zn-SOD from newborn bovine blood,and used layer-by-layer self-assembly technique,heparin and chitosan as material to modificate the SOD which separated and purified from newborn bovine blood.The results showed that compared with the natural SOD,the modified SOD increased 3.3 times in the thermal stability;In acid and alkaline,under the condition of pH 3.0 and pH 11.0,the acid-base resistance property of the modified SOD raised 2.0 times;On the antitrypsin hydrolysis ability,the relative enzyme activity of modified SOD had increased 2.7 times than natural SOD.The modified SOD with shell outside made of heparin and chitosan by physical modification could get better effect,and had the strong mechanical strength,better biocompatible,outer surface of chitosan was helpful to cross the cell membrane transport,and it could be applied to the development of oral dosage forms of SOD.

Key words: SOD; heparin; chitosan; layer-by-layer self-assembly technique; relative enzyme activity

中图分类号:

S852.5

侯静, 林春丽, 白志军, 魏建民, 张永明, 张峰. 新生牛血中Cu/Zn-SOD的提纯及肝素、壳聚糖的逐层自组装包裹[J]. 《中国畜牧兽医》, 2016, 43(8): 2032-2037.

HOU Jing, LIN Chun-li, BAI Zhi-jun, WEI Jian-min, ZHANG Yong-ming, ZHANG Feng. Separation and Purification of Cu/Zn-SOD from Newborn Bovine Blood and Prepatation of HEP and CHI Using Layer-by-layer Self-assembly Technique[J]. , 2016, 43(8): 2032-2037.

参考文献

[1] 周轩,张天,任晓敏,等.SOD提取工艺研究进展[J].粮食与食品工业,2015,22(4):57-62.
[2] Celojevic D,Nisson S,Behndig A,et al.Superoxide dismutase gene polymorphisms in patients with age-related cataract[J].Ophthalmic Genet,2013,34(3):140-145.
[3] Kodydkova J,Vavrova L,Stankova B,et al.Antioxidant satus and oxidative stress markers in pancreatic cancer and chronic pancreatitis[J].Pancreas,2013,42(4):614-621.
[4] Ahmed M N,Zhang Y,Codipilly C,et al.Extracellular superoxide dismutase overexpression can reverse the course of hypoxia induced pulmonary hypertension[J].Mol Med,2012,18:38-46.
[5] 陈祥娥,凌沛学,张天民.超氧化物歧化酶的应用[J].食品与药品,2013,5(4):283-286.
[6] Veronese F M,Caliceti P,Schiavon O,et al.Polyethylene glycol-superoxide dismutase,a conjugate in search of exploitation[J].Adv Drug Deliv Rev,2002,54(4):587-606.
[7] 孔庆胜,迮玉官.聚乙二醇修饰超氧化物歧化酶的基础研究[J].济宁医学院学报,2006,29(1):16-17.
[8] Perez Y,Valdivia A,Gomez L,et al.Glycosidation of Cu,Zn-superoxide dismutase with end-group aminated dextran.Pharmacological and pharmacokinetics properties[J].Macromol Biosci,2005,5:1220-1225.
[9] 陈吉祥,袁勤生.β-环糊精对SOD的化学修饰[J].中国医药工业杂志,1998,29(2):59-61.
[10] Ghys R,Benoit J C,Paquette J C,et al.Radioisotope study of metabolic requirements of tubercle bacilli in vitro.Absorption of various trace elements by BCG[J].B World Health Organ,1966,35(4):569-580.
[11] Decher G,Ringsdorf H,Venzmer J,et al.Giant liposomes as model membranes for immunological studies:Spontaneous insertion of purified K1-antigen (poly-alpha-2,8-NeuAc) of Escherichia coli[J].Biochim Biophys Acta,1990,1023(3):357-364.
[12] Khopade A J,Caruso F.Stepwise self-assembled poly (amidoamine) dendrimer and poly (styrenesulfonate) microcapsules as sustained delivery vehicles[J].Biomacromolecules,2002,3(6):1154-1162.
[13] Mohanta V,Madras G,Patil S.Layer-by-layer assembled thin film of albumin nanoparticles for delivery of doxorubicin[J].J Phys Chem C,2012,116:5333-5341.
[14] Mahanta D,Manna U,Madras G,et al.Multilayer self-assembly of TiO₂ nanoparticles and polyaniline-grafted-chitosan copolymer (CPANI) for photocatalysis[J].ACS Appl Mater Inter,2011,3:84-92.
[15] Kharlampieva E,Kozlovskaya V,Sukhishvili S A.Layer-by-layer hydrogen-bonded polymer films:From fundamentals to applications[J].Adv Mater,2009,21:3053-3065.
[16] Rogach A,Susha A,Caruso F,et al.Nano-and microengineering:Three-dimensional colloidal photonic crystals prepared from submicrometer-sized polystyrene latex spheres pre-coated with luminescent polyelectrolyte/nanocrystal shells[J].Adv Mater,2000,12(5):333-339.
[17] Lee H,Jeong C,Ghafoor K,et al.Oral delivery of insulin using chitosan capsules cross-linked with phytic acid[J].Biomed Mater Eng,2011,21(1):25-36.
[18] Prego C,Torres D,Fernandez-Megia E,et al.Chitosan-PEG nanocapsules as new carriers for oralpeptide delivery——Effect of chitosan pegylation degree[J].J Control Release,2006,111(3):299-308.
[19] Lam P L,Lee K K H,Wong R S M,et al.Development of hydrocortisone succinic acid/and5-fluorouracil/chitosan microcapsules for oral and topical drug deliveries[J].Bioorg Med Chem Lett,2012,22(9):3213-3218.
[20] Gulbake A,Jain S K.Chitosan:A potential polymer for colon-specific drug delivery system[J].Expert Opin Drug Deliv,2012,9(6):713-729.

新生牛血中Cu/Zn-SOD的提纯及肝素、壳聚糖的逐层自组装包裹

Separation and Purification of Cu/Zn-SOD from Newborn Bovine Blood and Prepatation of HEP and CHI Using Layer-by-layer Self-assembly Technique

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	陈思宇, 隋卓君, 王占新, 李强, 陈艺, 王飞, 陈爽, 刘心怡, 严专强, 何诚. 新型滴鼻灭活鹦鹉热衣原体疫苗的免疫效力评价[J]. 中国畜牧兽医, 2022, 49(5): 1951-1959.
[2]	王思博, 崔红霞, 杨季, 许丽. 超氧化物歧化酶模拟物对氧化应激IPEC-J2细胞抗氧化性能的影响[J]. 中国畜牧兽医, 2021, 48(5): 1566-1573.
[3]	傅春妮, 李元辉, 李鹏程, 李留安, 秦顺义. 低聚壳聚糖硒抗玉米赤霉烯酮对IPEC-J2细胞紧密连接蛋白表达的影响[J]. 中国畜牧兽医, 2021, 48(11): 4311-4318.
[4]	范秋丽, 蒋守群, 苟钟勇, 李龙, 林厦菁, 王一冰, 陈芳. 枯草芽孢杆菌、低聚壳聚糖和丁酸钠对黄羽肉鸡生长性能、免疫功能和肉品质的影响[J]. 中国畜牧兽医, 2020, 47(4): 1080-1091.
[5]	邓小亮, 马文康, 洪玮, 付欣, 谭茵. 新西兰白兔SOD1基因编码区克隆及生物信息学分析[J]. 《中国畜牧兽医》, 2019, 46(8): 2236-2245.
[6]	崔红霞, 郭照宙, 赵旦华, 马渭青, 程皇座, 许丽. 超氧化物歧化酶模拟物对肉仔鸡肉品质和超氧化物歧化酶活性的影响[J]. 《中国畜牧兽医》, 2019, 46(5): 1379-1387.
[7]	刘莹莹, 任慧波, 张星, 邓缘, 罗璇, 陈晨, 胡雄贵, 朱吉, 彭英林. 母猪日粮中添加低聚壳聚糖对哺乳仔猪生长、抗氧化能力和免疫功能的影响[J]. 《中国畜牧兽医》, 2019, 46(1): 166-173.
[8]	安琪, 黄海峰, 张振兴, 李宝宝, 张萌萌, 郑义盈, 王成强, 章泸尹, 杨小健, 曹瑞勇, 聂鑫, 杜丽, 王凤阳. 羊源多杀性巴氏杆菌sodA基因的克隆、表达及其生物信息学分析[J]. 《中国畜牧兽医》, 2018, 45(8): 2067-2075.
[9]	夏威, 漆丹, 许晴, 叶思捷, 蒋涛. 不同收集方法及添加半胱氨酸、肝素钠对黄牛卵母细胞体外成熟及发育的影响[J]. 《中国畜牧兽医》, 2018, 45(5): 1297-1303.
[10]	张恬, 张龙超, 王立刚, 王立贤. 猪脂肪沉积候选基因AOC3、PPARG1及SOD3 DNA甲基化差异研究[J]. 《中国畜牧兽医》, 2016, 43(11): 2820-2825.
[11]	刘进，张峰，魏建民. 牛血Cu/Zn-SOD提取条件的研究[J]. 中国畜牧兽医, 2014, 41(2): 144-147.
[12]	李红, 周恩民, 易建中. 猪繁殖与呼吸综合征病毒细胞受体研究进展[J]. , 2013, (7): 77-81.
[13]	李贺, 郑世民. 转录反式作用催化蛋白－超氧化物歧化酶(TAT-SOD)对神经细胞凋亡的保护作用[J]. , 2012, 39(7): 151-154.
[14]	鲁京兰;李继鹏;崔成都. 肝素、咖啡因、丙酮酸钠、BSA对延边黄牛卵母细胞体外受精效果的影响[J]. , 2012, 39(1): 118-121.
[15]	王丹;赵洁. 日粮中添加不同水平壳聚糖对天山雪鹅生长性能和免疫器官的影响[J]. , 2010, 37(9): 13-16.