Difference between revisions of "Part:BBa K258001"

Latest revision as of 11:05, 17 October 2009

Lipase ABC transporter recognition domain (LARD 1)

Lard 1 was designed for the secretion of fusion proteins.The LARD 1 included four glycine-rich repeats comprising a β-roll structure, and were added to the C-terminus of test proteins. Either Pro-Gly linker or Factor Xa site can be added between fusion proteins and LARD 1. Upon supplementation of E. coli with ABC transporter from E. chrysanthemi (ABC transporter PrtDEF), which function well in the phylogenetically-neighboring genus E.coli, EGF-LARDs were excreted into the culture supernatant. For LARD1, linkers may be engineered to have a Factor Xa cleavage site. LARD1 also contained residues 303–476 of TliA. The thermostable lipase, TliA, from which Jung Hoon Ahn designed LARD1, has four GGXGXD consensus sequences and EGVLIS as its final six C-terminal residues, showing similar organization of several hydrophobic residues preceded by an acidic residue, Glu. In our Wound Dressing project, EGF is one of tke key proteins although EGF has three disulfide bridges. Thus, exportation of EGF from E.coli was actually difficult. Happily with Jung Hoon Ahn's support, EGF can be exported in E. coli with LARD1 and also TliA, showing the possibility that the protein having disulfide bonds can be exported.

Sequence and Features