Purification and Characterization of a Novel Cholesterol Esterase from Pseudomonas aeruginosa, with Its Application to Cleaning Lipid-stained Contact Lenses
Purification and Characterization of a Novel Cholesterol Esterase from Pseudomonas aeruginosa, with Its Application to Cleaning Lipid-stained Contact Lenses
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铜绿假单胞菌新型胆固醇酯酶的纯化和表征及其在清洁脂质染色隐形眼镜中的应用
DOI：
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Y. Tominaga
中科院分区：
文献类型：
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作者：
A. Sugihara;Y. Shimada;A. Nomura;T. Terai;M. Imayasu;Y. Nagai;T. Nagao;Yomi Watanabe;Y. Tominaga
为了开发一种新的胆固醇酯酶来消除用过的隐形眼镜上的脂质，对微生物进行了酶活性筛选。从土壤中分离到一株铜绿假单胞菌，经鉴定能产生一种理想的酶。该酶的等电点为3.2，分子量为58 kDa。最适温度为53°C左右，pH为7.0，最适pH为5.5至9.5。该酶在25°C下在pH 5和10之间稳定19小时，并且在pH 7.0下孵育30分钟后在高达53°C下保持其活性。不同脂肪酸胆固醇酯的水解速率顺序为：亚油酸酯>油酸酯>硬脂酸酯>棕榈酸酯>辛酸酯>肉豆蔻酸酯>月桂酸酯、癸酸酯>己酸酯>丁酸酯、乙酸酯。添加（牛磺）胆酸盐至100 mM的最终浓度显着促进短链，中链和长链脂肪酸的甘油三酯的水解。当与牛磺胆酸盐一起使用时，该酶可作为一种有效的清洁剂，用于清洁被由油酸胆固醇酯、三棕榈酸甘油酯和硬脂酸硬脂酯组成的脂质染色的隐形眼镜。
With the aim of developing a new cholesterol esterase for eliminating lipids on used contact lenses, microorganisms were screened for the enzyme activity. A Pseudomonas aeruginosa isolated from soil was found to produce a desirable enzyme. The enzyme had an isoelectric point of 3.2, and molecular mass of 58 kDa. The optimal temperature was around 53°C at pH 7.0, and the optimal pH was from 5.5 to 9.5. The enzyme was stable between pH 5 and 10 for 19 h at 25°C, and retained its activity up to 53°C on 30 min of incubation at pH 7.0. The rates of hydrolysis of cholesteryl esters of different fatty acids were in the following order: linoleate>oleate>stearate>palmitate>caprylate>myristate> laurate, caprate>caproate>butyrate, acetate. Addition of (tauro)cholate to a final concentration of 100 mM markedly promoted the hydrolysis of triglycerides of short-, medium-, and long-chain fatty acids. When used with taurocholate, the enzyme acted as an effective cleaner for contact lenses stained with lipids consisting of cholesteryl oleate, tripalmitin, and stearyl stearate.
影响因子：
2.9
作者：
Chen, JCH;Miercke, LJW;Stroud, RM
通讯作者：
Stroud, RM