Team:CityU HK/project/module fad

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Module description


FadD & FadL Module

The ultimate goal of our project is to genetically engineer an E. coli strain to increase its fatty acid uptake efficiency and ability to convert the absorbed fatty acid into α-linolenic acid (ALA).

In order to achieve this, we have sub-divided our project into three parts:
a. Enhanced uptake of long chain fatty acids (LCFAs) by E. coli
b. Increased conversion of fatty acyl-CoA into free fatty acid
c. Enhanced conversion of free fatty acid into ALA

The uptake of exogenous long chain fatty acids is controlled by the fadL and fadD genes.


Figure 1. Importation of long-chain fatty acids by FadL and FadD.


Long chain fatty acids (LCFAs) are protonated in the periplasmic space and FadD facilitates its transport into the cytosol.

The fadL gene codes for an outer membrane-bound fatty acids transport protein that facilitates the import of long chain fatty acids into the periplasmic space where they are protonated and then partitioned at the inner membrane (with the ionic head pointing towards the periplasmic space). Protonated fatty acids are unable to pass through the inner membrane into the cytosol and requires the assistance of the FadD protein. The fadD gene codes for an inner membrane-associated long chain acyl-CoA synthetase enzyme in E. coli which catalyses the addition of a CoA moiety to the fatty acid and its subsequent transport across the inner membrane into the cytosol. FadD is therefore required both for the transport of LCFAs through the inner membrane into the cytosol and their activation for metabolism via the β-oxidation pathway. By overexpressing FadL and FadD, we aim to enhance LCFA uptake to boost the subsequent conversion of long chain acyl-CoA to ALA.


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