Characterization of a Thermostable, Recombinant Carboxylesterase from the Hyperthermophilic Archaeon Metallosphaera sedula DSM5348Report as inadecuate




Characterization of a Thermostable, Recombinant Carboxylesterase from the Hyperthermophilic Archaeon Metallosphaera sedula DSM5348 - Download this document for free, or read online. Document in PDF available to download.

Lipid-producing microalgae are emerging as the leading platform for producing alternative biofuels in response to diminishing petroleum reserves. Optimization of fatty acid production is required for efficient conversion of microalgal fatty acids into usable transportation fuels. Microbial lipases-esterases can be used to enhance fatty acid production because of their efficacy in catalyzing hydrolysis of esters into alcohols and fatty acids while minimizing the potential poisoning of catalysts needed in the biofuel production process. Although studies have extensively focused on lipases-esterases produced by mesophilic organisms, an understanding of lipases-esterases produced by thermophilic, acidic tolerant microbes, such as Metallosphaera sedula, is limited. In this work, the carboxylesterase from Metallosphaera sedula DSM5348 encoded by Msed 1072 was recombinantly expressed in Escherichia coli strain BL21 λDE3. The purified enzyme either with a hexahistidine His6-tag Msed 1072Nt and Msed 1072Ct or without the hexahistidine His6-tag Msed 1072 was biochemically characterized using a variety of substrates over a range of temperatures and pH and in the presence of metal ions, organic solvents, and detergents. In this study, the fusion of the protein with a hexahistidine His6-tag did not result in a change in substrate specificity, but the findings provide information on which enzyme variant can hydrolyze fatty acid esters in the presence of various chemicals, and this has important implication for their use in industrial processes. It also demonstrates that Metallosphaera sedula Msed 1072 can have application in microalgae-based biofuel production systems.

KEYWORDS

Carboxylesterase; Metallosphaera sedula; Hyperthermoacidophile; Biofuels; Algae; Lipids

Cite this paper

Killens-Cade, R. , Turner, R. , MacInnes, C. and Grunden, A. 2014 Characterization of a Thermostable, Recombinant Carboxylesterase from the Hyperthermophilic Archaeon Metallosphaera sedula DSM5348. Advances in Enzyme Research, 2, 1-13. doi: 10.4236-aer.2014.21001.





Author: Rushyannah Killens-Cade, Rachel Turner, Christine MacInnes, Amy Grunden

Source: http://www.scirp.org/



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