title: Enhanced production of n-alkanes in Escherichia coli by spatial organization of biosynthetic pathway enzymes.
abstract: Alkanes chemically mimic hydrocarbons found in petroleum, and their demand as biofuels is steadily increasing. Biologically, n-alkanes are produced from fatty acyl-ACPs by acyl-ACP reductases (AARs) and aldehyde deformylating oxygenases (ADOs). One of the major impediments in n-alkane biosynthesis is the low catalytic turnover rates of ADOs. Here, we studied n-alkane biosynthesis in Escherichia coli using a chimeric ADO-AAR fusion protein or zinc finger protein-guided ADO/AAR assembly on DNA scaffolds to control their stoichiometric ratios and spatial arrangements. Bacterial production of n-alkanes with the ADO-AAR fusion protein was increased 4.8-fold (24 mg/L) over a control strain expressing ADO and AAR separately. Optimal n-alkane biosynthesis was achieved when the ADO:AAR binding site ratio on a DNA scaffold was 3:1, yielding an 8.8-fold increase (44 mg/L) over the control strain. Our findings indicate that the spatial organization of alkane-producing enzymes is critical for efficient n-alkane biosynthesis in E. coli.
authors.forename: Ziaur
authors.initials: Z
authors.lastname: Rahmana
authors.forename: Bong Hyun
authors.initials: BH
authors.lastname: Sung
authors.forename: Ji-Yeun
authors.initials: JY
authors.lastname: Yi
authors.forename: Le Minh
authors.initials: le M
authors.lastname: Bui
authors.forename: Jun Hyoung
authors.initials: JH
authors.lastname: Lee
authors.forename: Sun Chang
authors.initials: SC
authors.lastname: Kim
mesh_terms: D000473:Alkanes / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids
chemical_list: D000473:Alkanes; D001426:Bacterial Proteins; D004247:DNA; D010088:Oxidoreductases; D010105:Oxygenases