Part:BBa_K4788006

IsmA-BCoAT

Transforming cholesterol into 4-cholesten-3-one while producing short chain fatty acids.

Usage and Biology

It is derived from the combination of the IsmA gene and the BCoAT gene, which conforms to the part. It can directly degrade cholesterol while producing short chain fatty acids to indirectly regulate cholesterol.

Characterization

In order to characterize the corresponding components, we need to develop different characterization schemes for different genes. For the BCoAT gene, We use gas chromatography to examine the production of short chain fatty acids in strains containing the BCoAT gene to characterize the gene, and for the IsmA gene, we used the OPA method to detect cholesterol residue after cholesterol culture medium cultivation to characterize it. Meanwhile, as short chain fatty acids are also important cholesterol regulatory substances, we can also construct the IsmA-BSH-BCoAT triplet gene to achieve more complex cholesterol regulation. For more information on triplets, please refer to BBa_K4788007. In the characterization of the triad, as shown in Figure 1, we can see that the IsmA gene in the I-BC gene has been successfully characterized. As shown in Figure 2,We can also see the characterization of the BCoAT gene from its GC data.However, the characterization of BCoAT in the binary system is not significant

Figure1: a) Cholesterol residue in the culture medium after cultivation of engineering bacteria on basic cholesterol medium b) Percentage of cholesterol degradation in the culture medium after cultivation of engineering bacteria on basic cholesterol medium

Figure1: GC detection of short chain fatty acid production by BCoAT containing strains

Reference

Jia B, Zou Y, Han X, Bae JW, Jeon CO. Gut microbiome-mediated mechanisms for reducing cholesterol levels: implications for ameliorating cardiovascular disease. Trends Microbiol. 2023 Jan;31(1):76-91. doi: 10.1016/j.tim.2022.08.003. Epub 2022 Aug 22. PMID: 36008191. Kenny DJ, Plichta DR, Shungin D, Koppel N, Hall AB, Fu B, Vasan RS, Shaw SY, Vlamakis H, Balskus EP, Xavier RJ. Cholesterol Metabolism by Uncultured Human Gut Bacteria Influences Host Cholesterol Level. Cell Host Microbe. 2020 Aug 12;28(2):245-257.e6. doi: 10.1016/j.chom.2020.05.013. Epub 2020 Jun 15. PMID: 32544460; PMCID: PMC7435688.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1861
  Illegal AgeI site found at 58
  
• 1000
  COMPATIBLE WITH RFC[1000]