Part:BBa_K4989010

GFP protein binded to Butyryl-CoA:Acetyl-CoA transferase

Application in the field of biology

Green Fluorescent Protein (GFP)

is a naturally occurring protein that exhibits remarkable fluorescence when exposed to ultraviolet or blue light. The fluorescence of GFP arises from a chemical reaction within the protein. When exposed to UV or blue light, GFP absorbs photons and undergoes a process called fluorescence, where it emits green light. The chromophore, a small molecular group within the protein, is responsible for this fluorescence.GFP revolutionized cell and molecular biology by allowing scientists to visualize and track specific proteins or cells within living organisms.

It has been used to study processes such as gene expression, protein localization, cell migration, and cell division. GFP fusions are created by attaching the GFP gene to the gene of interest, enabling researchers to monitor its expression and localization in real-time. Also, it is commonly used as a reporter gene in molecular biology to assess the activity of promoters or the effectiveness of gene expression. By fusing the GFP gene with a promoter of interest, researchers can easily monitor when and where a gene is active. GFP-tagged proteins have been instrumental in studying protein localization within cells and tracking their movement. This has provided insights into organelle dynamics, vesicle transport, and the functioning of cellular structures.
In genetic studies, GFP has been used to label specific cells or tissues, making it possible to trace the development of embryos, observe the migration of cells, or identify specific cell types. It has played a crucial role in understanding developmental biology and genetics.

Butyryl-CoA:Acetyl-CoA transferase:

The butcoat gene encodes for a transferase that catalyzes the final step of the metabolic pathway production: Butyryl-CoA:Acetyl-CoA transferase.The primary function of Butyryl-CoA:Acetyl-CoA transferase is to facilitate the transfer of the acetyl group from butyryl-CoA to coenzyme A (CoA)[Figure A]. In this way the transferase can utilize the butyryl-CoA an the acetate as substrates in order to produce butyrate.

This process results in the formation of acetyl-CoA as a second product, which is a crucial molecule in the citric acid cycle (Krebs cycle) and oxidative phosphorylation, two central pathways of energy production in the cell. By converting butyryl-CoA into acetyl-CoA, this enzyme allows the cell to harness the energy stored in fatty acids more efficiently.

Figure A.The transfer of the acetyl group from butyryl-CoA to coenzyme A (CoA) for the production of butyrate

The expression of genes encoding Butyryl-CoA:Acetyl-CoA transferase in bacteria is regulated in response to the availability of specific carbon sources. The presence of medium-chain fatty acids or other substrates can trigger the upregulation of genes encoding this enzyme, allowing bacteria to adapt to their nutritional environment.
Butyryl-CoA:Acetyl-CoA transferase, often referred to as Butyryl-CoA transferase, is an enzyme found in both bacteria and eukaryotes. Its role in bacteria is primarily associated with metabolic pathways related to fatty acid degradation, energy production, and the catabolism of various carbon sources. This enzyme is particularly significant for individuals on diets rich in medium-chain triglycerides and for those with certain metabolic disorders, as it helps ensure the efficient utilization of these fatty acids for energy. In the human gastrointestinal tract, the enzyme plays a role in the microbial fermentation of dietary fibers and complex carbohydrates. Certain bacteria in the gut produce short-chain fatty acids, such as butyrate, which have important implications for gut health and host physiology.

Biosafety

Our part is safe to be synthesized and utilized on an open bench. Also, the product of the gene does not include any biohazard and does not pose any threat even if by chance there is a leak.

Characterization

Our team, chose for the gold medal to create a new improved part and characterize it as well as possible for the competition standards and the other IGEM teams. So we did not characterize this specific part any further, as something that is crucial to be conducted in the future.

References

[1]Zimmer, Marc. “Green Fluorescent Protein (GFP):  Applications, Structure, and Related Photophysical Behavior.” Chemical Reviews, vol. 102, no. 3, Mar. 2002, pp. 759–82, https://doi.org/10.1021/cr010142r.
[2]Tsien, Roger Y. “THE GREEN FLUORESCENT PROTEIN.” Annual Review of Biochemistry, vol. 67, no. 1, June 1998, pp. 509–44, https://doi.org/10.1146/annurev.biochem.67.1.509.
[3]Chalfie, M. “Green Fluorescent Protein.” Photochemistry and Photobiology, vol. 62, no. 4, 1995, pp. 651–56, https://doi.org/10.1111/j.1751-1097.1995.tb08712.x.
[4]Duncan, Sylvia H., et al. “Acetate Utilization and Butyryl Coenzyme a (CoA):Acetate-CoA Transferase in Butyrate-Producing Bacteria from the Human Large Intestine.” Applied and Environmental Microbiology, vol. 68, no. 10, Oct. 2002, pp. 5186–90, https://doi.org/10.1128/AEM.68.10.5186-5190.2002. Accessed 20 Feb. 2021.
[5]Hippe, Berit, et al. “Quantification of Butyryl CoA:acetate CoA-Transferase Genes Reveals Different Butyrate Production Capacity in Individuals according to Diet and Age.” FEMS Microbiology Letters, vol. 316, no. 2, Jan. 2011, pp. 130–35, https://doi.org/10.1111/j.1574-6968.2010.02197.x. Accessed 6 June 2021.
[6]Petra Louis, Sheila I. McCrae, Cédric Charrier, Harry J. Flint, Organization of butyrate synthetic genes in human colonic bacteria: phylogenetic conservation and horizontal gene transfer, FEMS Microbiology Letters, Volume 269, Issue 2, April 2007, Pages 240–247,https://doi.org/10.1111/j.1574-6968.2006.00629.x
[7]“Roseburia Intestinalis L1-82 Chromosome 1, Complete Sequence.” NCBI Nucleotide, Mar. 2023, www.ncbi.nlm.nih.gov/nuccore/NZ_LR027880.1?report=genbank&from=430426&to=431769&strand=true.
[8]Louis, Petra, et al. “Diversity of Human Colonic Butyrate-Producing Bacteria Revealed by Analysis of the Butyryl-CoA:acetate CoA-Transferase Gene.” Environmental Microbiology, vol. 12, no. 2, Feb. 2010, pp. 304–14, https://doi.org/10.1111/j.1462-2920.2009.02066.x. Accessed 31 Oct. 2019.