Chitosanase displayed on liposome can increase its activity and stability
Chitosanase displayed on liposome can increase its activity and stability
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DOI：
10.1016/j.jbiotec.2010.01.014
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发表时间：
2010-04-01
影响因子：
4.1
通讯作者：
Kuboi, Ryoichi
中科院分区：
文献类型：
--
作者：
Ngo, Kien Xuan;Umakoshi, Hiroshi;Kuboi, Ryoichi
基于脂质体与灰色链霉菌细胞膜的直接相互作用，采用脂质体固定化壳聚糖酶（ICL）制备新型生物催化剂的策略。对ICL进行了表征，包括蛋白质的分子量、壳聚糖酶活性、各种脂质体表面水化对固定化壳聚糖酶水解活性的影响以及ICL在各种极端条件下的稳定性。纯化的 ICL 样品的 SOS-PAGE 分析显示存在约 39 kDa 的蛋白质，相当于成熟壳聚糖酶及其信号肽 (38.8 kDa) 的重量总和。 ICL的上述蛋白还表达了显着高于常规壳聚糖酶的壳聚糖酶活性。此外，用于制备ICL的脂质体的表面水合影响固定化壳聚糖酶的活性，证实了脂质体表面的重要性。事实上，通过测量壳聚糖酶活性测定的 ICL 在各种温度和 pH 条件下的稳定性明显高于传统壳聚糖酶。 ICL的这些特性表明，通过将酶适当地固定在脂质体囊泡上来制备生物催化剂是可能的。 (C) 2010 Elsevier B.V. 保留所有权利。
The strategy to prepare a novel biocatalyst by the immobilization of chitosanase onto liposome (ICL) was carried out based on the direct interaction of liposomes with cell membrane of Streptomyces griseus cell. The ICL was characterized in relation to the molecular weight of protein, the chitosanase activity, the effect of the surface hydration of various liposomes on hydrolysis activity of immobilized chitosanase and the stability of ICL under various extreme conditions. The SOS-PAGE analysis of the purified ICL sample shows the existence of a protein with approximately 39 kDa that corresponded to the sum of weight of the mature chitosanase and its signal peptide (38.8 kDa). The above protein of ICL also expresses the chitosanase activity that is significantly higher than that of the conventional chitosanase. Furthermore, the surface hydration of liposomes used to prepare ICL that affected the activity of immobilized chitosanase verified the importance of liposome surfaces. Indeed, the stability of ICL assayed by measuring the chitosanase activity is significantly higher than that of conventional chitosanase under various temperatures and pH conditions. These characteristics of ICL show the possible preparation of the biocatalysts that can be prepared by immobilizing enzymes onto liposome vesicles properly. (C) 2010 Elsevier B.V. All rights reserved.