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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2017, Vol. 37 Issue (12): 77-83    DOI: 10.13523/j.cb.20171211
Orginal Article     
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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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     
Received: 23 August 2017      Published: 16 December 2017
ZTFLH:  Q819  
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Shan XU
Ren-qiang LI
Ji-fu ZHANG
Yun ZHANG
Ai-jun SUN
Yun-feng Hu
Cite this article:

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.

URL:

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

Fig.1 Effect of different carriers on the immobilization of lipase
Fig.2 The optimization of conditions of immobilized lipase
(a) Effect of sodium alginate concentration on the activity of immobilized lipase (b) Effect of carboxymethyl cellulose sodium concentration on the activity of immobilized lipase (c) Effect of enzyme loading on lipase immobilization (d) Effect of CaCl2 concentration on the activity of immobilized lipase (e) Effect of buffer pH on the activity of immobilized lipase (f) Effect of immobilization time on the activity of immobilized lipase
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
Table 1 Results and analysis of orthogonal experiments for the optimization of immobilized condition of lipase
Fig.3 The optimization of conditions of crosslinking immobilized lipase
(a) Effect of crosslinking agent on the activity of immobilized lipase (b) Effect of crosslinking time with glutaraldehyde on the activity of immobilized lipase (c) Effect of crosslinking time with EGDE on the activity of immobilized lipase (d) The operation stability of immobilized lipase
Fig.4 The properties of immobilized and free lipase
(a) Optimal reaction pH of immobilized and free lipase (b) Optimal reaction temperature of immobilized and free lipase (c) Thermal stability of immobilized and free lipase
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