Characterization of a Thermostable, Recombinant Carboxylesterase from the Hyperthermophilic Archaeon Metallosphaera sedula DSM5348
Open Access
- 1 January 2014
- journal article
- Published by Scientific Research Publishing, Inc. in Advances in Enzyme Research
- Vol. 02 (01), 1-13
- https://doi.org/10.4236/aer.2014.21001
Abstract
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
This publication has 30 references indexed in Scilit:
- Complete Genome Sequence of Metallosphaera cuprina, a Metal Sulfide-Oxidizing Archaeon from a Hot SpringJournal of Bacteriology, 2011
- Fatty acid production in genetically modified cyanobacteriaProceedings of the National Academy of Sciences of the United States of America, 2011
- Carboxylic ester hydrolases from hyperthermophilesExtremophiles, 2009
- Biogeography of theSulfolobus islandicuspan-genomeProceedings of the National Academy of Sciences of the United States of America, 2009
- Crystal Structure and Biochemical Properties of a Novel Thermostable Esterase Containing an Immunoglobulin-Like DomainJournal of Molecular Biology, 2009
- Microalgal triacylglycerols as feedstocks for biofuel production: perspectives and advancesThe Plant Journal, 2008
- Purification and Characterization of Two Highly Thermophilic Alkaline Lipases fromThermosyntropha lipolyticaApplied and Environmental Microbiology, 2007
- Protein production by auto-induction in high-density shaking culturesProtein Expression and Purification, 2005
- Role of the N Terminus in Enzyme Activity, Stability and Specificity in Thermophilic Esterases Belonging to the HSL FamilyJournal of Molecular Biology, 2005
- The crystal structure of a hyper-thermophilic carboxylesterase from the archaeon Archaeoglobus fulgidusJournal of Molecular Biology, 2001