Part:BBa_K2052018

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.) to 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) , the stop codon is removed for the addition of HisTag.

DNA Gel Analysis

Characterization

Source

Roseburia intestinalis L1-82

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

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

http://pubs.acs.org/doi/abs/10.1021/bi300957f