Hughes_2011_J.Lab.Autom_16_17

Reference

Title : Production of Candida antarctica lipase B gene open reading frame using automated PCR gene assembly protocol on robotic workcell and expression in an ethanologenic yeast for use as resin-bound biocatalyst in biodiesel production - Hughes_2011_J.Lab.Autom_16_17
Author(s) : Hughes SR , Moser BR , Harmsen AJ , Bischoff KM , Jones MA , Pinkelman R , Bang SS , Tasaki K , Doll KM , Qureshi N , Saha BC , Liu S , Jackson JS , Robinson S , Cotta MC , Rich JO , Caimi P
Ref : J Lab Autom , 16 :17 , 2011
Abstract :

A synthetic Candida antarctica lipase B (CALB) gene open reading frame (ORF) for expression in yeast was constructed, and the lycotoxin-1 (Lyt-1) C3 variant gene ORF, potentially to improve the availability of the active enzyme at the surface of the yeast cell, was added in frame with the CALB ORF using an automated PCR assembly and DNA purification protocol on an integrated robotic workcell. Saccharomyces cerevisiae strains expressing CALB protein or CALB Lyt-1 fusion protein were first grown on 2% (w/v) glucose, producing 9.3 g/L ethanol during fermentation. The carbon source was switched to galactose for GAL1-driven expression, and the CALB and CALB Lyt-1 enzymes expressed were tested for fatty acid ethyl ester (biodiesel) production. The synthetic enzymes catalyzed the formation of fatty acid ethyl esters from ethanol and either corn or soybean oil. It was further demonstrated that a one-step-charging resin, specifically selected for binding to lipase, was capable of covalent attachment of the CALB Lyt-1 enzyme, and that the resin-bound enzyme catalyzed the production of biodiesel. High-level expression of lipase in an ethanologenic yeast strain has the potential to increase the profitability of an integrated biorefinery by combining bioethanol production with coproduction of a low-cost biocatalyst that converts corn oil to biodiesel.

PubMedSearch : Hughes_2011_J.Lab.Autom_16_17
PubMedID: 21609683

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Citations formats

Hughes SR, Moser BR, Harmsen AJ, Bischoff KM, Jones MA, Pinkelman R, Bang SS, Tasaki K, Doll KM, Qureshi N, Saha BC, Liu S, Jackson JS, Robinson S, Cotta MC, Rich JO, Caimi P (2011)
Production of Candida antarctica lipase B gene open reading frame using automated PCR gene assembly protocol on robotic workcell and expression in an ethanologenic yeast for use as resin-bound biocatalyst in biodiesel production
J Lab Autom 16 :17

Hughes SR, Moser BR, Harmsen AJ, Bischoff KM, Jones MA, Pinkelman R, Bang SS, Tasaki K, Doll KM, Qureshi N, Saha BC, Liu S, Jackson JS, Robinson S, Cotta MC, Rich JO, Caimi P (2011)
J Lab Autom 16 :17