Part:BBa_K2924022
accA
This part contains the coding sequence for the Acetyl-coenzyme A carboxylase carboxyl transferase subunit alpha. It is a component of the Acetyl-CoA carboxylase complex (ACC) from Escherichia coli (strain K12), involved in the fatty acid biosynthesis. Cloned into an inducible pET21a (Novagen) vector carrying an ampicillin resistance and containing strong T7 promoter and C-terminal histidine tag 1. ACC is allosterically inhibited by the accumulated fatty acyl-CoAs 1.
Usage and Biology
The accA is one of four genes in the composite part AccABCD. The composite part encodes for ACC and catalyzes the first committed step in fatty acid biosynthesis - the ATP-dependent formation of malonyl-CoA from acetyl-CoA and bicarbonate (HCO3-) 1 [Fig. 1]. Fatty acids are carboxylic acids and have a chain length of 4 up to 36 carbon atoms.1
AccA encodes the alpha subunit of the Acetyl-coenzyme A carboxylase carboxyl transferase (CT). Together with the beta subunit AccD, they coordinate the transfer of a carboxyl group onto the Acetyl-CoA to convert Malonyl-CoA [Fig. 2]. The gene was replicated and overexpressed in E. coli BL21 to increase carbon flux towards the precursor malonyl-CoA. An improved cellular concentration of malonyl-CoA contributes to increased production of malonyl-CoA derived compounds like fatty acids 1
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 579
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 517
Illegal SapI.rc site found at 714
References
[1] Faergeman, Nils Joakim, and Jens Knudsen. "Role of long-chain fatty acyl-CoA esters in the regulation of metabolism and in cell signalling." Biochemical Journal 323.Pt 1 (1997): 1.
[2] Fujita, Yasutaro, Hiroshi Matsuoka, and Kazutake Hirooka. "Regulation of fatty acid metabolism in bacteria." Molecular microbiology 66.4 (2007): 829-839.
[3] Lehninger, A. L., Nelson, D. L., Cox, M. M., & Osgood, M. (2005). Lehninger principles of biochemistry. New York: W.H. Freeman.
[4] Xu, Peng, et al. "Modular optimization of multi-gene pathways for fatty acids production in E. coli." Nature communications 4 (2013): 1409.
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