Covalent Immobilization of Marine Candida Rugosa Lipase Using Amino Carrier

doi:10.13523/j.cb.20190710

China Biotechnology

2019, Vol. 39

Issue (7): 71-78 DOI: 10.13523/j.cb.20190710

Covalent Immobilization of Marine Candida Rugosa Lipase Using Amino Carrier

Heng ZHU^1,²,Hai-jiao LIN^1,²,Ji-fu ZHANG³,Yun ZHANG¹,Ai-jun SUN¹,Yun-feng HU^1,^**(

)

¹ CAS Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301,China
² University of Chinese Academy of Sciences,Beijing 100049,China
³ Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510120,China

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Abstract

Covalent binding is one important immobilization method of industrial enzyme, which uses stable covalent bonds to immobilize industrial enzymes, creates multi-point covalent connections between carrier and enzyme, and prepares immobilized enzymes with good stability and possesses practical application value. Marine Candida rugosa lipase was immobilized using amino carrier through covalent binding method and relative inexpensive glutaraldehyde was used as the crosslinking reagent. By using single factor and orthogonal experiment, the optimal immobilization conditions were determined as follows:25℃, pH5.0,0.1% glutaraldehyde,0.25g carrier, crosslinked 0.5h,immobilization time 1h, enzyme loading 800U, with the final obtained enzyme activity being 83.01U/g. Compared with free enzyme, the optimal pH of immobilized lipase was shifted to alkaline direction, the optimal reaction temperature was increased by 10℃, the thermal stability and acid-alkali stability of immobilized enzyme were better than that of free enzyme, and the reusability and storage stability were better than that of free enzyme. Meanwhile, crosslinking agent was found to be an important factor in the preparation of immobilized lipase. Thus it is of great significance to explore new crosslinking agent to improve immobilization effect. A good foundation for the immobilization technique and industrial application of marine candida rugosa lipase were laid.

Key words： Amino carrier Lipase Glutaraldehyde Covalent crosslinking Immobilized enzyme

Received: 29 December 2018 Published: 05 August 2019

ZTFLH:

Q814.2

Corresponding Authors: Yun-feng HU E-mail: yunfeng.hu@scsio.ac.cn

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	Heng ZHU
	Hai-jiao LIN
	Ji-fu ZHANG
	Yun ZHANG
	Ai-jun SUN
	Yun-feng HU

Cite this article:

Heng ZHU,Hai-jiao LIN,Ji-fu ZHANG,Yun ZHANG,Ai-jun SUN,Yun-feng HU. Covalent Immobilization of Marine Candida Rugosa Lipase Using Amino Carrier. China Biotechnology, 2019, 39(7): 71-78.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20190710 OR https://manu60.magtech.com.cn/biotech/Y2019/V39/I7/71

Fig.1 Optimization of the immobilization conditions of lipase (a) Effect of carrier volume on immobilized enzyme activity (b) Effects of temperature on immobilized enzyme activity (c) Effect of pH on immobilized enzyme activity (d) Effect of glutaraldehyde volume fraction on immobilized enzyme activity (e) Effect of crosslinking time on immobilized enzyme activity (f) Effect of immobilized time on immobilized enzyme activity

Fig. 2 Characterization of enzymatic properties of immobilized lipase and free lipase (a) Comparison of the optimum reaction temperature of free enzymes and immobilized enzymes (b) Comparison of the optimum reaction pH of free enzymes and immobilized enzymes (c) Comparison of the thermal stability of free enzymes with immobilized enzymes (d) Comparison of the stability of free enzymes with immobilized enzymes (e) Effects of the number of immobilized enzyme uses on catalytic capacity (f) Changes in enzyme activity within one months of preservation at 4℃ and room temperature

Table 1 Results and analysis of orthogonal experiments for the condition optimization of immobilized lipase


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