BBa_K258006 1 BBa_K258006 Thermostable lipase (TliA) of Pseudomonas fluorescens SIK W1 2009-10-08T11:00:00Z 2015-05-08T01:11:42Z The thermostable lipase (TliA) of P. fluorescens SIK W1 is comprised of 476 amino acids and has the characteristic C-terminal signal sequence recognized by the ABC transporter. TliA has four glycinerich repeats (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. false false _358_ 0 4260 9 Not in stock true The sequence was optimized for E.coli with Biobrick restriction sites false Jung Hoon Ahn from Korea Advanced Institute of Science and Technology BBa_K258006_sequence 1 atgggtgtgttcgattacaagaacctgggtacggaagcctctaaaaccctgttcgctgatgcgaccgcgatcaccctgtatacctaccacaacctggataacggtttcgctgtaggttatcagcagcacggcctgggtctgggcctgccagcgaccctggtcggcgcgctgctgggttctaccgactctcaaggcgtgatcccgggcatcccatggaacccggactctgaaaaagcagctctggacgcggtacatgcagctggctggactccgatctccgcttctgcgctgggctacggtggcaaagtcgacgctcgtggcaccttcttcggcgaaaaagcaggctataccacggcacaggccgaagtactgggcaagtacgacgatgctggtaaactgctggaaatcggcatcggtttccgtggcaccagcggtccgcgtgaatccctgatcactgactccatcggtgatctggtaagcgatctgctggcggctctgggtccgaaggactacgcgaaaaactacgcaggtgaagccttcggtggtctgctgaagactgttgcagactacgccggtgcccacggtctgtccggtaaggacgtcctggtgtccggtcatagcctgggtggtctggcggtgaactctatggcagatctgagcaccagcaagtgggcgggcttttacaaagatgcaaactacctggcttacgcctctccaacgcagtctgctggtgataaagtcctgaacatcggttacgaaaacgacccggtctttcgtgctctggacggttccactttcaacctgtcctctctgggtgtacacgacaaagcccacgaatccactacggacaacattgtcagctttaacgaccactatgcgtctactctgtggaacgtcctgccgttctctattgccaacctgtccacttgggtgtctcacctgccgtctgcttacggtgacggcatgacccgtgttctggaaagcggcttctacgaacagatgacccgtgattccactattattgtggctaacctgtccgacccggcgcgcgctaatacgtgggttcaggatctgaaccgtaacgcagaaccgcataccggcaatactttcatcatcggttctgatggtaatgacctgattcagggcggtaaaggtgctgatttcatcgagggcggcaaaggtaatgacactatccgtgataattccggccacaatacgttcctgttttccggtcacttcggccaggaccgtattatcggctatcagccgaccgaccgtctggtgttccagggtgcggacggtagcaccgacctgcgtgaccacgcgaaagccgtgggcgctgacaccgttctgtctttcggcgcagactctgtgactctggtcggcgtcggcctgggtggcctgtggtctgaaggcgttctgatttct igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 Chris J. Myers James Alastair McLaughlin 2017-03-06T15:00:00.000Z