Difference between revisions of "Part:BBa K2042000"

Revision as of 20:57, 29 August 2016

Propionate CoA transferase for P. putida

This is an evolved coding sequence for the enzyme Propionate CoA transferase (Pct) EC 2.8.3.1. The wild type enzyme come from Clostridium propionicum and normally catalyzes the reaction: acetyl-CoA + propanoate <--> acetate + propanoyl- CoA. This mutated sequence has one amino acid substitution: A243T allowing lactate to act as acceptor and convert it efficiently to lactyl-CoA. Moreover, this sequence is codon optimized for Pseudomonas putida using Codon Optimization Tool from IDT in order to produce Polylactic acid in P. putida. This enzyme is a variant of Yale 2013 BBa_K1211001 biobrick but codon optimized for Pseudomonas putida and with one different amino acid mutation (A243T instead of V193A) which led to the increase of both polymer content and lactate mole fraction in the copolymer.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 675
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1081
Illegal BsaI site found at 1552

Revision as of 20:54, 29 August 2016 (view source) RATOVOMANANA (Talk \| contribs) ← Older edit		Revision as of 20:57, 29 August 2016 (view source) RATOVOMANANA (Talk \| contribs) Newer edit →
Line 4:		Line 4:


−	This is an evolved coding sequence for the enzyme Propionate CoA transferase (Pct) EC 2.8.3.1. The wild type enzyme come from Clostridium propionicum and normally catalyzes the reaction: acetyl-CoA + propanoate <--> acetate + propanoyl- CoA. This mutated sequence has one amino acid substitution: A243T allowing lactate to act as acceptor and convert it efficiently to lactyl-CoA. Moreover, this sequence is codon optimized for Pseudomonas putida using Codon Optimization Tool from IDT in order to produce ~~(D)-Lactyl- CoA~~.	+	This is an evolved coding sequence for the enzyme Propionate CoA transferase (Pct) EC 2.8.3.1. The wild type enzyme come from Clostridium propionicum and normally catalyzes the reaction: acetyl-CoA + propanoate <--> acetate + propanoyl- CoA. This mutated sequence has one amino acid substitution: A243T allowing lactate to act as acceptor and convert it efficiently to lactyl-CoA. Moreover, this sequence is codon optimized for Pseudomonas putida using Codon Optimization Tool from IDT in order to produce Polylactic acid in P. putida.
	This enzyme is a variant of Yale 2013 <html><a href="/Part:BBa_K1211001">BBa_K1211001</a></html> biobrick but codon optimized for Pseudomonas putida and with one different amino acid mutation (A243T instead of V193A) which led to the increase of both polymer content and lactate mole fraction in the copolymer.		This enzyme is a variant of Yale 2013 <html><a href="/Part:BBa_K1211001">BBa_K1211001</a></html> biobrick but codon optimized for Pseudomonas putida and with one different amino acid mutation (A243T instead of V193A) which led to the increase of both polymer content and lactate mole fraction in the copolymer.