Separation and Purification of Agarase and Study on Its Properties

doi:10.13523/j.cb.20170107

China Biotechnology

2017, Vol. 37

Issue (1): 46-52 DOI: 10.13523/j.cb.20170107

Separation and Purification of Agarase and Study on Its Properties

XIE Xi-zhen, LIN Juan, XIE Yong, YE Xiu-yun

Fujian Provincial Key Laboratory of Marine Enzyme Engineering, Fuzhou University, Fuzhou 350116, China

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Abstract

Agarase was fermented with Vibrio sp. ZC-1, and separated through hollow fiber concentration, ammonium sulfate precipitation and DEAE-anion exchange chromatography. A pure AgaZC-1 detected by SDS-PAGE was obtained. Its relative molecular mass and specific activity was about 45kDa and 114.613U/mg, respectively. Furthermore, the enzymatic properties of AgaZC-1 were studied. The results showed that the optimum pH was 7. It was stable in the range of 5.0 to 9.0 and could maintain more than 80% of its relative enzyme activity after incubation for 1 hour. The optimal reaction temperature was 50℃ and 60% of enzyme activity was remained after incubation for 1 hour under 45℃. At high concentrations (5mmol/L), Fe³⁺, Cu²⁺, Sn²⁺ and Zn²⁺ could completely inhibit the enzyme activity. While at low concentrations (1mmol/L), Cu²⁺, Ba²⁺, Na⁺, Zn²⁺, Ag⁺, Sr³⁺ and K⁺ had obvious inhibition on enzyme activity. The K_m and V_max of AgaZC-1 was 0.538mg/ml and 6.33μmol/(L·min), respectively. It was highly specific for agar and the hydrolysates were neoagarotetraose and neoagarohexaose analyzed by TLC.

Key words： Agarase Vibiro sp. Separation and purification Enzymatic properties

Received: 18 July 2016 Published: 25 January 2017

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Cite this article:

XIE Xi-zhen, LIN Juan, XIE Yong, YE Xiu-yun. Separation and Purification of Agarase and Study on Its Properties. China Biotechnology, 2017, 37(1): 46-52.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20170107 OR https://manu60.magtech.com.cn/biotech/Y2017/V37/I1/46

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