Difference between revisions of "Part:BBa K2052018"

Revision as of 15:48, 16 October 2016

Butyryl-CoA:acetate CoA-transferase

ButCoAT is an enzyme (Butyryl-CoA:acetate CoA-transferase) (EC 2.8.3.8) (Uniprot:C7GB37) which converts Acetyl-CoA (Acetyl-CoA is readily present in the cytosol and it can be directly converted into Butyrate.) into Butyrate and can be found in Roseburia intestinalis L1-82. The equation of the reaction (acyl-CoA:acetate CoA-transferase) (KEGG Reaction No. : R01179) is given down below :

Butanoyl-CoA + Acetate <=> Butanoic acid + Acetyl-CoA

Figure 1: Crystal Structures of Acetobacter aceti Succinyl(Butyryl)-CoA:Acetate CoA-Transferase Reveal Specificity Determinants and Illustrate the Mechanism Used by Class I CoA-Transferases.(Mullins, E.A. et al., 2012)

Butyrate

Butyrate is a short chain fatty acid which is formed by bacterial fermentation of carbohydrates (Sengupta et al., 2005).

Modeling

The figure above shows the increase in ButCoAT molecules over a time span of 10 seconds.

The figure above shows the molar increase in Butyrate concentration over a time span of 10 seconds.

Design Notes

E.coli is synthesized suitable for Codon Bias.

In the expression of pet28a (expression vector) , stop codon is removed for the addition of HisTag.

DNA Gel Analysis

Characterization

PCR Conformation of ButCoAT + pSB1C3

We have loaded the uncut version of K2052015 (RBS + ButCoAT) next to EcoRI single digested one. In the third lane, RBS ligated ButCoat (K2052018) was loaded with its single digested version. Digested ones gave sharp lane at 3600 bp as we expected.

Source

Roseburia intestinalis L1-82

Sequence and Features

References

Weinstock G., Sodergren E., Clifton S., Fulton L., Fulton B., Courtney L., Fronick C., Harrison M., Strong C., Farmer C., Delahaunty K., Markovic C., Hall O., Minx P., Tomlinson C., Mitreva M., Nelson J., Hou S. Wilson R.K. (2009 , August ). UniProtKB - C7GB37 (C7GB37_9FIRM). Retrieved from http://www.uniprot.org/uniprot/C7GB37?sort=score