Use of synthetic fusion gene to produce biodiesel from lignocellulosic biomass

ABSTRACT

In second generation biofuels physiochemical and structural compositions of lignocellulose make it recalcitrant to direct fermentation. The drawbacks in pretreatment and quantity of biodiesel produced created barriers in blooming this technology. This paper discusses a new pathway to resolve the hurdles using fusion gene to produce biodiesel from lignocellulosic biomass. A plasmid pCOLA Duet 1Z1 was constructed by cloning aft A gene of A. baylyi into multiple cloning site, followed by cloning of fusion genes, hex1 and pox1 into pCola Duet 1Z.

Co-expression of pCOLA Duet 1 Z1carrying hex 1, pox 1 & aft A gene with pLOI297 carrying pdc and adh genes from Z. mobilis helped in hydrolysis and fermentation of lignocellulose to biodiesel. While hex1 and pox1 helped in direct hydrolysis of lignocellulose including lignin, pdc and adh gene converted glucose to ethanol and aft A gene helped in esterifying ethanol to biodiesel.

Different lignocellulosic biomass from wastes of paddy such as husk and straw were hydrolyzed and fermented to produce biodiesel using traditional pretreatment (acid/enzyme, base/enzyme) and using fusion gene. FAEE (fatty acid ethyl ester) samples produced from various methods were analyzed through GC-MS and it was concluded that the sample produced using fusion gene tend to have better yield.

KEYWORDS:

• Biodiesel
• lignocellulose
• hydrolysis
• fusion gene
• fermentation technology
• metabolic engineering

Competing interests

Fusion gene referred in the manuscript has been protected by Universiti Brunei Darussalam under Brunei Patent Registration Number: BN/N/2014/0037

FadE Del E. coli was been gifted by University of California and might be protected by its inventor Prof. Jay D. Keasling.

Acknowledgements

I wish to express my deep sense of gratitude to Universiti Brunei Darussalam for accepting my research proposal and funding me with a Graduate Research Scholarship that made my PhD work possible.

Disclosure statement

No potential conflict of interest was reported by the authors.