Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Marie Burghard, Henrik Cordes, Jascha Diemer, Adrian Eilingsfeld, Sven Jager, Philipp Rottmann, Rene Sahm, Arne Wehling
Date created: 2012-09-01 11:00:00
Date modified: 2015-05-08 01:13:26
EstA : inactive fusion protein for bacterial surface expression
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K808027_sequence (Version 1) |
Description
This Esterase A is a inactivated membrane bound esterase from Pseudomonas putida. Its active site is mutated so it shows no more hydrolytic activity. It can be used for cell surface display like its shown in the following paper:Taek Ho Yang, Jae Gu Pan, Yeon Soo Seo, Joon Shick Rhee
Use of Pseudomonas putida EstA as an Anchoring Motif for Display of a Periplasmic Enzyme on the Surface of Escherichia coli
Appl Environ Microbiol. 2004 December; 70(12): 6968???6976. doi: 10.1128/AEM.70.12.6968-6976.2004
Notes
This certain variant of EstA should be used for cell surface expression and displaying of almost every kind of enzyme. There is no ATG at its 5' because its an C-terminal fusion protein. If you want to use you need to design it in the following way:N - Peri plasmatic signal sequence - Protein of interest - EstA - C
We did not use any kind of BioBrick Assembly due to the generation of stop codons. If you would like to fuse EstA to any kind of protein, please use SOE PCR or an alternative kind of assembly strategy.
Source
Our parts derives from an established inactive EstA mutant on pEST100 vector, which was used in the following paper:Becker, S., Theile, S., Heppeler, N., Michalczyk, A., Wentzel, A., Wilhelm, S., Jaeger, K.-E., et al. (2005). A generic system for the Escherichia coli cell-surface display of lipolytic enzymes. FEBS letters, 579(5), 1177???82. doi:10.1016/j.febslet.2004.12.087