Characterization of an alkaline esterase from an enriched metagenomic library derived from an oil-spill area
| Autor: | Seung Cheol Baek, Jeong Min Jo, Soo-Mi Jeong, Jae Pil Lee, Hyun Woo Lee, Jungho Kim, Hoon Kim |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
0301 basic medicine chemistry.chemical_classification biology Molecular mass Esterase Gene 030106 microbiology Organic Chemistry Bioengineering biology.organism_classification 01 natural sciences Esterase Amino acid 03 medical and health sciences Carboxylesterase Enzyme chemistry Biochemistry 010608 biotechnology Catalytic triad Alcanivorax |
| Zdroj: | Journal of Applied Biological Chemistry. 62:73-79 |
| ISSN: | 2234-7941 1976-0442 |
| DOI: | 10.3839/jabc.2019.011 |
| Popis: | A novel esterase gene (est7S) was cloned from an enriched metagenomic library derived from an oil-spill area. The gene encoded a protein of 505 amino acids, and the molecular mass of the Est7S was estimated to be 54,512 Da with no signal peptide. Est7S showed the highest identity of 40% to an esterase from a sludge metagenome compared to the characterized enzymes with their properties, although it showed 99% identity to a carboxylesterase in the genome sequence of Alcanivorax borkumensis SK2. Est7S had catalytic triad residues, Ser183, Glu312, and His420, and the GESAG motif in most family VII lipolytic enzymes. Est7S was purified from the crude extract of clone SM7 using Sephacryl S-200 HR and HiTrap Q column chromatographies. The purified Est7S was optimally active at 50 ℃ and pH 10.0. Est7S showed a high specific activity of 366.7 U/mg protein. It preferred short length esters, particularly p-nitrophenyl acetate, efficiently hydrolyzed R- and S-enantiomers of methyl-3-hydroxy-2-methylpropionate, and glyceryl tributyrate. These properties of Est7S may provide potential merits in biotechnological applications such as detergent and paper processing under alkaline conditions. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|