Zhang Xiaoyan, Lomora Mihai, Einfalt Tomaz, Meier Wolfgang, Klein Noreen, Schneider Dirk, Palivan Cornelia G
Chemistry Department, University of Basel, Klingelbergstrasse 80, Basel, Switzerland.
Biochemistry, Institute of Pharmacy & Biochemistry, University of Mainz, Germany.
Biomaterials. 2016 May;89:79-88. doi: 10.1016/j.biomaterials.2016.02.042. Epub 2016 Feb 26.
We introduce active surfaces generated by immobilizing protein-polymer nanoreactors on a solid support for sensitive sugar alcohols detection. First, such selective nanoreactors were engineered in solution by simultaneous encapsulation of specific enzymes in copolymer polymersomes, and insertion of membrane proteins for selective conduct of sugar alcohols. Despite the artificial surroundings, and the thickness of the copolymer membrane, functionality of reconstituted Escherichia coli glycerol facilitator (GlpF) was preserved, and allowed selective diffusion of sugar alcohols to the inner cavity of the polymersome, where encapsulated ribitol dehydrogenase (RDH) enzymes served as biosensing entities. Ribitol, selected as a model sugar alcohol, was detected quantitatively by the RDH-nanoreactors with GlpF-mediated permeability in a concentration range of 1.5-9 mM. To obtain "active surfaces" for detecting sugar alcohols, the nanoreactors optimized in solution were then immobilized on a solid support: aldehyde groups exposed at the compartment external surface reacted via an aldehyde-amino reaction with glass surfaces chemically modified with amino groups. The nanoreactors preserved their architecture and activity after immobilization on the glass surface, and represent active biosensing surfaces for selective detection of sugar alcohols, with high sensitivity.
我们介绍了通过将蛋白质-聚合物纳米反应器固定在固体支持物上而产生的活性表面,用于灵敏检测糖醇。首先,通过将特定酶同时封装在共聚物聚合物囊泡中,并插入用于糖醇选择性传导的膜蛋白,在溶液中构建了这种选择性纳米反应器。尽管存在人工环境以及共聚物膜的厚度,但重组大肠杆菌甘油转运蛋白(GlpF)的功能得以保留,并允许糖醇选择性扩散到聚合物囊泡的内腔,其中封装的核糖醇脱氢酶(RDH)酶作为生物传感实体。以核糖醇作为模型糖醇,RDH纳米反应器通过GlpF介导的渗透性在1.5-9 mM的浓度范围内对其进行了定量检测。为了获得用于检测糖醇的“活性表面”,将在溶液中优化的纳米反应器固定在固体支持物上:隔室外表面暴露的醛基通过醛-氨基反应与用氨基化学修饰的玻璃表面发生反应。纳米反应器固定在玻璃表面后保留了其结构和活性,代表了用于选择性检测糖醇的活性生物传感表面,具有高灵敏度。