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http://dx.doi.org/10.5657/FAS.2013.0311

Molecular Cloning, Purification, and Characterization of a Cold-Adapted Esterase from Photobacterium sp. MA1-3  

Kim, Young-Ok (Biotechnology Research Division, National Fisheries Research and Development Institute)
Heo, Yu Li (Biotechnology Research Division, National Fisheries Research and Development Institute)
Nam, Bo-Hye (Biotechnology Research Division, National Fisheries Research and Development Institute)
Kim, Dong-Gyun (Biotechnology Research Division, National Fisheries Research and Development Institute)
Jee, Young-Ju (Biotechnology Research Division, National Fisheries Research and Development Institute)
Lee, Sang-Jun (Biotechnology Research Division, National Fisheries Research and Development Institute)
An, Cheul-Min (Biotechnology Research Division, National Fisheries Research and Development Institute)
Publication Information
Fisheries and Aquatic Sciences / v.16, no.4, 2013 , pp. 311-318 More about this Journal
Abstract
The gene encoding an esterase from Photobacterium sp. MA1-3 was cloned in Escherichia coli using the shotgun method. The amino acid sequence deduced from the nucleotide sequence (948 bp) corresponded to a protein of 315 amino acid residues with a molecular weight of 35 kDa and a pI of 6.06. The deduced protein showed 74% and 68% amino acid sequence identities with the putative esterases from Photobacterium profundum SS9 and Photobacterium damselae, respectively. Absence of a signal peptide indicated that it was a cell-bound protein. Sequence analysis showed that the protein contained the signature G-X-S-X-G included in most serine-esterases and lipases. The MA1-3 esterase was produced in both soluble and insoluble forms when E. coli cells harboring the gene were cultured at $18^{\circ}C$. The enzyme was a serine-esterase and was active against $C_2$, $C_4$, $C_8$ and $C_{10}$ p-nitrophenyl esters. The optimum pH and temperature for enzyme activity were pH 8.0 and $30^{\circ}C$, respectively. Relative activity remained up to 45% even at $5^{\circ}C$ with an activation energy of 7.69 kcal/mol, which indicated that it was a cold-adapted enzyme. Enzyme activity was inhibited by $Cd^{2+}$, $Cu^{2+}$, $Zn^{2+}$, and $Hg^{2+}$ ions.
Keywords
Photobacterium sp.; Cold-adapted esterase; Gene expression; Substrate specificity;
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