Improved N-alkanes Production in Escherichia coli By Spatial Organization of Alkane Biosynthetic Pathway Enzymes

Linear
hydrocarbons, especially n-alkanes, are the major constituents of
gasoline and jet fuel with ever increasing demand as alternative biofuels. However,
the conventional n-alkanes production process based on expensive raw
materials (coal, hydrogen and cobalt) increases the overall production cost of n-alkanes.
To meet the growing demand, efforts have been made to engineer microbial systems
for the economical production of n-alkanes. Nevertheless, the microbial
productions of n-alkanes are far below a commercial threshold. color:black;background:white'>Biologically, n-alkanes are produced from
fatty acyl-ACPs with the help of acyl-ACP reductases (AAR) and aldehyde
deformylating oxygenases (ADO). One of the major challenges in the biological n-alkanes
color:black;background:white'>production line-height:200%;font-family:"Times New Roman","serif";color:black;background:
white'> process is a slow catalytic turnover rate of ADO. color:black;background:white'>T 200%;font-family:"Times New Roman","serif";color:black;background:white'>o
increase the n-alkanes production, we controlled the spatial arrangement
and stoichiometric ratio of enzymes. First, a chimeric protein of color:black;background:white'>AAR and ADO was synthesized. Second, color:black;background:white'>the enzymes were arranged on a DNA scaffold with
various ratios. As the result, p line-height:200%;font-family:"Times New Roman","serif";color:black;background:
white'>roduction of font-family:"Times New Roman","serif";color:black;background:white'> n color:black;background:white'>-alkanes line-height:200%;font-family:"Times New Roman","serif";color:black;background:
white'>was increased (4.4-fold) by the chimeric fusion of color:black'>ADO-AAR font-family:"Times New Roman","serif";color:black;background:white'>compared to
a control strain expressing wild type AAR and ADO. Furthermore, when the ratio
of ADO to AAR was 3 to 1, 200%;font-family:"Times New Roman","serif";color:black;background:white'>n color:black;background:white'>-alkanes line-height:200%;font-family:"Times New Roman","serif";color:black;background:
white'>production was increased (8.8-fold) compared to the control strain. color:black;background:white'> Our results showed that the color:black;background:white'> spatial organizations of enzymes color:black;background:white'> 200%;font-family:"Times New Roman","serif";color:black;background:white'>using
protein color:black;background:white'>chimera line-height:200%;font-family:"Times New Roman","serif";color:black;background:
white'>and "Times New Roman","serif";color:black;background:white'> DNA scaffolds color:black;background:white'> are applicable for establishing an efficient n-alkanes
biosynthetic pathway font-family:"Times New Roman","serif";color:black;background:white'>in Escherichia
coli. "Times New Roman","serif";color:black;background:white'>