Parthasarathy Ranganath, Bajaj Jitin, Boder Eric T
Department of Chemical and Biomolecular Engineering and Laboratory for Research on the Structure of Matter, University of Pennsylvania, 220 South 33rd Street, Philadelphia, 19104, USA.
Biotechnol Prog. 2005 Nov-Dec;21(6):1627-31. doi: 10.1021/bp050279t.
The Escherichia coli biotin ligase enzyme BirA has been extensively used in recent years to generate site-specifically biotinylated proteins via a biotin acceptor peptide tag. In the present study, BirA was displayed for the first time on the yeast Saccharomyces cerevisiae using the Aga1p-Aga2p platform and assayed using a peptide-tagged protein as the substrate. The enzyme is fully functional and resembles the soluble form in many of its properties, but the yeast-displayed enzyme demonstrates stability and reusability on the time scale of weeks. Thus, the yeast-displayed BirA system represents a facile and highly economical alternative for producing site-specifically biotinylated proteins.
近年来，大肠杆菌生物素连接酶BirA已被广泛用于通过生物素受体肽标签生成位点特异性生物素化蛋白。在本研究中，BirA首次使用Aga1p-Aga2p平台展示在酿酒酵母上，并以肽标签蛋白作为底物进行检测。该酶功能完全，在许多特性上类似于可溶形式，但酵母展示的酶在数周的时间尺度上表现出稳定性和可重复使用性。因此，酵母展示的BirA系统是生产位点特异性生物素化蛋白的一种简便且经济高效的替代方法。
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