Scholarlink

<p>Surface display of proteins on bacteria bears several advantages for binding studies, library screening or enzyme inhibitor testing. Here, we present a set of plasmids for autodisplay-based surface expression of recombinant proteins, named <b>A</b>utodisplay-<b>T</b>ool<b>B</b>ox (ATB). In this set, crucial parts as required for autodisplay, including promotor, SP, linker and β-barrel can be combined in varying permutations to find the best combination for a certain protein. The plasmids can be applied individually or in library approaches, enabling optimization of surface display in a single step. By such a library approach, the activity of surface-displayed β-glucosidase (β-Gluc) is increased by a factor of 4.9, the activity of a laccase (CotA) by a factor of 4.7 and the binding capacity of the surface-displayed nucleotide binding domain of human HCN2 channel (HCN2-CNBD) by a factor of 10.3. It is shown that the ATB can be used in different strains of <i>E. coli</i> as well as in <i>Pseudomonas putida</i>. The aim is to provide a selection of plasmids, accessible by Addgene, that every scientist can use for their own protein, either individually based on personal preferences, structural features, etc., or as a library, as shown here for three different examples.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

A plasmid toolbox for the easy autodisplay of recombinant proteins and its optimization

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年5月21日

查看全文
  • Christoph Furtmann,
  • Philip Röhe,
  • Katrin Gesing,
  • Hanna Kuss,
  • Florian Lenz,
  • Joachim Jose

摘要

Surface display of proteins on bacteria bears several advantages for binding studies, library screening or enzyme inhibitor testing. Here, we present a set of plasmids for autodisplay-based surface expression of recombinant proteins, named Autodisplay-ToolBox (ATB). In this set, crucial parts as required for autodisplay, including promotor, SP, linker and β-barrel can be combined in varying permutations to find the best combination for a certain protein. The plasmids can be applied individually or in library approaches, enabling optimization of surface display in a single step. By such a library approach, the activity of surface-displayed β-glucosidase (β-Gluc) is increased by a factor of 4.9, the activity of a laccase (CotA) by a factor of 4.7 and the binding capacity of the surface-displayed nucleotide binding domain of human HCN2 channel (HCN2-CNBD) by a factor of 10.3. It is shown that the ATB can be used in different strains of E. coli as well as in Pseudomonas putida. The aim is to provide a selection of plasmids, accessible by Addgene, that every scientist can use for their own protein, either individually based on personal preferences, structural features, etc., or as a library, as shown here for three different examples.