Coding

Part:BBa_K4879001

Designed by: Rohaneshwar Manikandan   Group: iGEM23_IISER-Pune-India   (2023-10-08)


JcFATB

The fatty acyl-ACP thioesterase B, from the plant Jatropha curcas. [1]

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Usage and Biology

Plant acyl-ACP thioesterases are enzymes that play a crucial role in various metabolic processes in plants. They function by catalyzing the hydrolysis of thioester bonds specifically between an acyl group and an acyl carrier protein (ACP). This hydrolysis reaction results in the release of the acyl group from ACP, producing a free fatty acid (FFA). Thus, acyl-ACP thioesterases are pivotal in regulating the chain length of fatty acids in de novo fatty acid biosynthesis by cleaving them from ACP.

This regulatory function of thioesterases in fatty acids' chain lengths significantly affects the free fatty acid as well as the triglyceride compositions in plant seed oils, affecting their value in terms of nutrition or biofuel potential. This has made thioesterases a focus in industries, with research aimed at manipulating them to give favourable lipid compositions for further processing.

Thioesterase B, in Jatropha curcas, acts upon palmitoyl-ACP and stearoyl-ACP to give palmitic acid and stearic acid. The production of these saturated fatty acids would act as further substrates for the desaturases in Y. lipolytica to act upon and produce the more ideal unsaturated FFAs, which is why we cloned thioesterase B in our chassis.


The protein structure of JcFATB, as predicted by us on AlphaFold


Design

The transcriptional unit for JcFATB (BBa_K4879007) is designed with the coding sequence flanked by the TEF1 promoter (BBa_K3629001) upstream and the XPR2 terminator (BBa_K3629004) downstream.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 831
    Illegal PstI site found at 50
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 831
    Illegal PstI site found at 50
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 831
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 831
    Illegal PstI site found at 50
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 831
    Illegal PstI site found at 50
  • 1000
    COMPATIBLE WITH RFC[1000]


References

1. Liu, Y.; Han, J.; Li, Z.; Jiang, Z.; Luo, L.; Zhang, Y.; Chen, M.; Yang, Y.; Liu, Z. Heterologous Expression of Jatropha curcas Fatty Acyl-ACP Thioesterase A (JcFATA) and B (JcFATB) Affects Fatty Acid Accumulation and Promotes Plant Growth and Development in Arabidopsis. Int. J. Mol. Sci. 2022, 23, 4209. https://doi.org/10.3390/ijms23084209

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