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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2017, Vol. 37 Issue (12): 77-83    DOI: 10.13523/j.cb.20171211
技术与方法     
乙二醇缩水甘油醚交联海藻酸钠-羧甲基纤维素钠固定化脂肪酶 *
徐珊1,李任强1,张继福4,张云2,3,孙爱君2,3,胡云峰2,3*()
1 暨南大学生物工程学系 广州 510632
2 中国科学院南海海洋研究所 广东省海洋药物重点实验室 广州 510301
3 中国科学院南海海洋研究所 中国科学院热带海洋生物资源与生态重点实验室 广州 510301
4 广东省中医院 广州 510120
Ethylene Glycol Diglycidyl Ether Cross-linked with Sodium Alginate- carboxymethyl Cellulose to Immobilize Lipase
Shan XU1,Ren-qiang LI1,Ji-fu ZHANG4,Yun ZHANG2,3,Ai-jun SUN2,3,Yun-feng Hu2,3*()
1 Department of Biotechnology, Jinan University, Guangzhou 510632, China
2 Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology,Chinese Academy of Sciences, Guangzhou 510301, China
3 Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology,Chinese Academy of Sciences, Guangzhou 510301, China
4 Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510120, China
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摘要：

以海藻酸钠、羧甲基纤维素钠(CMC)为载体,分别以乙二醇缩水甘油醚(EGDE)和戊二醛为交联剂,采用包埋交联法对脂肪酶进行固定化,结果显示EGDE的交联效果要优于戊二醛,添加EGDE的固定化酶酶活最好。得到制备固定化酶的最优方案为海藻酸钠2.5%,CMC浓度1.5%,给酶量800U/ml复配载体,氯化钙5%,以0.02%的EGDE交联固定30min,由此制备得到酶活约为380 U/g的固定化酶,酶活收率约为50.09%。固定化酶的最适反应pH为8.5,比游离酶增大0.5个单位;最适反应温度是45℃,比游离酶提高5℃;耐热性能变好,且重复使用7次后仍能保持60%左右的相对酶酶活。
关键词： 海藻酸钠;  羧甲基纤维素钠;  乙二醇缩水甘油醚;  固定化酶    
Abstract:

Sodium alginate and carboxymethyl cellulose sodium were used as the carriers to immobilize lipase, using ethylene glycol diglycidyl ether (EGDE) and glutaraldehyde as the cross-linking agent, respectively. The results showed that EGDE was superior to glutaraldehyde and was the most effective cross-linking agent. The optimal conditions for lipase immobilized were as follows: sodium alginate 2.5%, carboxymethyl cellulose sodium 1.5%, enzyme loading 800U/ml, CaCl2 5%, EGDE 0.02%, and immobilization time 30 min. Under above optimized conditions, the activity of immobilized lipase was about 380 U/g, and the recovery of the enzyme activity of the immobilized lipase reached about 50.09%. Compared with those from free lipase, the optimal pH (8.0)and temperature (45℃) were higher. The thermal stability of immobilized lipase was better than that of free lipase, and the immobilized lipase could be reused for seven times and still retained about 60% activity.
Key words: Sodium alginate    Carboxymethyl cellulose sodium    Ethylene glycol diglycidyl ether    Immobilized enzyme
收稿日期: 2017-08-23 出版日期: 2017-12-16
ZTFLH:  Q819  
基金资助: 中国科学院战略性先导科技专项(XDA11030404)、广东省海洋渔业科技攻关与研发方向项目和广州市科技计划(201510010012)资助项目
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徐珊
李任强
张继福
张云
孙爱君
胡云峰

引用本文:

徐珊,李任强,张继福,张云,孙爱君,胡云峰. 乙二醇缩水甘油醚交联海藻酸钠-羧甲基纤维素钠固定化脂肪酶 *[J]. 中国生物工程杂志, 2017, 37(12): 77-83.

Shan XU,Ren-qiang LI,Ji-fu ZHANG,Yun ZHANG,Ai-jun SUN,Yun-feng Hu. Ethylene Glycol Diglycidyl Ether Cross-linked with Sodium Alginate- carboxymethyl Cellulose to Immobilize Lipase. China Biotechnology, 2017, 37(12): 77-83.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20171211    或    https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I12/77

图1  不同载体对脂肪酶固定化的影响
图2  脂肪酶固定化条件的优化
试验号 A(海藻酸钠浓度%) B (CMC浓度%) C(氯化钙浓度%) D(固定化时间min) 固定效率%
1 1(1.5) 1(0.75) 1(4) 1(30) 24.58
2 1 2(1) 2(5) 2(40) 35.61
3 1 3(1.5) 3(6) 3(50) 38.18
4 2(2) 1 2 3 32.09
5 2 2 3 1 33.77
6 2 3 1 2 36.74
7 3(2.5) 1 3 2 30.45
8 3 2 1 3 35.22
9 3 3 2 1 39.34
k1 32.79 29.04 32.18 32.56
k2 34.20 34.87 35.68 34.27
k3 35.00 38.09 34.13 35.16
R 2.21 9.05 3.50 2.60
表1  脂肪酶固定化条件优化正交试验结果与分析
图3  固定化酶交联条件的优化
图4  固定化酶与游离酶酶学性质比较
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