Difference between revisions of "Part:BBa K1149003"

Revision as of 19:38, 15 September 2013

pelB-pueA expression

This part expresses a plastic degradation enzyme under the control of xylose inducible promoter.

The enzyme is the pueA from Pseudomonas chlororaphis, that encodes carboxylesterase that breaks down ester bonds in polyurethane. This protein is [http://onlinelibrary.wiley.com/doi/10.1111/j.1574-6976.2010.00231.x/full naturally extracellular], it has a C-terminal secretion signal located downstream of a domain containing three glycine-rich repeats. These glycine-rich repeats are nonapeptide motifs involved in Ca2+-binding found in proteins secreted via type I systems. It uses the RTX system. To ensure secretion in E.coli, we have also added a pelB seretion tag to the N-terminal. We also added a 6xHistidine tag for Western blotting and protein purification purposes.

PUR degradation by esterase:

references:

[http://onlinelibrary.wiley.com/doi/10.1111/j.1574-6976.2010.00231.x/full RTX proteins: a highly diverse family secreted by a common mechanism ]

Stern, R. V., and G. T. Howard. 2000. The polyester polyurethane gene (pueA) from Pseudomonas clororaphis encodes lipase. FEMS Microbiol. Lett. 185:163–168.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 99
Illegal BglII site found at 964
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 7: / Line 7: @@
 To ensure secretion in E.coli, we have also added a [https://parts.igem.org/Part:BBa_K1149023 pelB] seretion tag to the N-terminal. We also added a 6xHistidine tag for Western blotting and protein purification purposes.
-Theoretical pathway of PUR degradation by esterase:
+PUR degradation by esterase:
 https://static.igem.org/mediawiki/parts/3/31/PUR_esterase_theoretical_pathway_2.jpg