Part:BBa_K1498001
constitutive promoter with lipase (BBa_J23110, BBa_K258006)
Composite comprises of the constitutive promoter (BBa_J23110) with Thermostable lipase (tliA) of Pseudomonas fluorescens SIK W1 (codon optimised for E. coli version of BBa_K258006).
Four glycine-rich repeats (GGXGXD) in the tliA C-terminus, allow ATP binding cassette (ABC) transporter secretion of the lipase (Chung et al., 2009). ABC transportation results in the movement of protein through the inner and outer membrane of gram negative bacteria into the extracellular space.
The composite part must be transformed into cells which express ABC transporters. Here we coexpressed the part with BBa_K258008. TliA was successfully transported into culture supernatant, thus suggesting directed transport through the ABC transporter. This was quantified by measured lipase activity on Rhodamine B plates supplemented with olive oil.
References.
Chung, C. W., You, J., Kim, K., Moon, Y., Kim, H., & Ahn, J. H. (2009). Export of recombinant proteins in Escherichia coli using ABC transporter with an attached lipase ABC transporter recognition domain (LARD). Microbial cell factories, 8(1), 11. doi:10.1186/1475-2859-8-11
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 692
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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