Enhanced production of n-alkanes in Escherichia coli by spatial organization of biosynthetic pathway enzymes.
J Biotechnol
; 192 Pt A: 187-91, 2014 Dec 20.
Article
in En
| MEDLINE
| ID: mdl-25456061
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 ADOAAR binding site ratio on a DNA scaffold was 31, 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.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Oxidoreductases
/
Oxygenases
/
Bacterial Proteins
/
Alkanes
/
Escherichia coli
Language:
En
Journal:
J Biotechnol
Journal subject:
BIOTECNOLOGIA
Year:
2014
Document type:
Article