Purification and Characterization of Agarase PV-1 from Pseudomonas vesicularis MA103

Pseudomonas vesicularis MA103所產洋菜酶（Agarase）PV-1之純化與特性

10.29822/JFST.201109.0003

吳紹祺(Shao-Chi Wu)；曾國政(Kuo-Chang Zen)；吳欣蕙(Xin-Hui Wu)；潘崇良(Chorng-Liang Pan)

β-洋菜酶 ； 新洋菜二糖 ； 新洋菜四糖 ； 新洋菜六糖 ； β-agarase ； Neoagarobiose ； Neoagarohexaose ； Neoagarotetraose

臺灣水產學會刊

38卷3期（2011 / 09 / 01）

201 - 215

英文

液化洋菜菌株Pseudomonas vesicularis所產MA103-agarases經超濃縮、DE-52離子交換樹脂、與Bio-Gel P-100層析後可得純化洋菜黴（Agarase）PV-1。Agarase PV-1最適pH和溫度分別為6.0和40℃；然而其活性在50℃下15分鐘後減少80.0%。Agarase PV-1的分子量為62.9 kDa，且在鈉離子環境中可以提高20%的相對活性。Agarase PV-1以HIMEDIA（上標 TM） agar為基質時有最佳之基質特異性活性，相對agarose為基質分解活性分別為1.4倍。Agarase PV-1水解中性洋菜之水解產物經HPLC分析，所得產物為一群介於新洋菜二糖（neoagarobiose）與新洋菜六糖（neoagarohexose）間之寡醣。利用Agarases PV-1與新洋菜六糖混合反應，經HPLC分析，所得產物為數個大於新洋菜六糖之寡醣。

The agarases producing strain Pseudomonas (P.) vesicularis MA103 is an agar-liquefying strain. The crude MA103-agarases is concentrated by ultrafiltration, chromatography by ion-exchange, and Bio-Gel P-100 to obtained Agarase PV-1. The optimum pH and temperature for Agarase PV-1 were 6.0 and 40℃; however 80.0％ of the enzyme activity was inactivated in 15 min under 50℃. The molecular mass of Agarase PV-1 was 62.9 kDa and under sodium ion could enhance 20％ relative agarase activity. The best substrate specific activities of Agarase PV-1 toward brand Himedia(superscript TM) agar were 1.4-time digestion activity compare to agarose, respectively. Hydrolytic products of agarose digested by Agarase PV-1 were analyzed by HPLC, and the results showed that they were oligosaccharides, withe molecular size between neoagarohexaose and neoagarobiose. Products of neoagarohexaose reacted with Agarase PV-1 analyzed by HPLC were oligosaccharides, with molecular size larger than neoagarohexaose.