Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Jung Hoon Ahn from Korea Advanced Institute of Science and Technology
Date created: 2009-10-08 11:00:00
Date modified: 2015-05-08 01:11:42
Thermostable lipase (TliA) of Pseudomonas fluorescens SIK W1
| Types | DnaRegion |
| Roles | tag Tag |
| Sequences | BBa_K258006_sequence (Version 1) |
Description
TliA has four glycinerichrepeats (GGXGXD) in its C-terminus, which appear in many ABC transporter-secreted
proteins.Export of fusion proteins with the whole TliA through the ABC transporter
was evident on the basis of lipase enzymatic activity. Upon supplementation of E. coli with ABC
transporter, EGF-TliA was excreted into the culture supernatant.Whole TliA were attached to C-termini of model proteins and enabled
the export of the model proteins such as GFP and EGF which has 3 disulfide bonds in E. coli supplemented with ABC
transporter. Activity domain (residues 1???268) and
secretion/chaperon domain (residues 279???476). In our experiment, we observed that TliA fused proteins were excreted to supernatant culture succesfully by detecting lipase activity with tributyrin and spectrophotometric detection with the substrate p-nitrophenyl phosphate at 420 nm. At the experiments, Tlia fused proteins were excreted ten-fold more with ABC transporter PrtDEF of Erwinia chrysanthemi.
Notes
The sequence was optimized for E.coli with Biobrick restriction sitesSource
The thermostable lipase (TliA) of P. fluorescens SIK W1 iscomprised of 476 amino acids and has the characteristic
C-terminal signal sequence recognized by the ABC transporter.