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A cold-adapted esterase of a novel marine isolate, Pseudoalteromonas arctica: gene cloning, enzyme purification and characterization

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Abstract

A gene encoding an esterase (estO) was identified and sequenced from a gene library screen of the psychrotolerant bacterium Pseudoalteromonas arctica. Analysis of the 1,203 bp coding region revealed that the deduced peptide sequence is composed of 400 amino acids with a predicted molecular mass of 44.1 kDa. EstO contains a N-terminal esterase domain and an additional OsmC domain at the C-terminus (osmotically induced family of proteins). The highly conserved five-residue motif typical for all α/β hydrolases (G × S × G) was detected from position 104 to 108 together with a putative catalytic triad consisting of Ser106, Asp196, and His225. Sequence comparison showed that EstO exhibits 90% amino acid identity with hypothetical proteins containing similar esterase and OsmC domains but only around 10% identity to the amino acid sequences of known esterases. EstO variants with and without the OsmC domain were produced and purified as His-tag fusion proteins in E. coli. EstO displayed an optimum pH of 7.5 and optimum temperature of 25°C with more than 50% retained activity at the freezing point of water. The thermostability of EstO (50% activity after 5 h at 40°C) dramatically increased in the truncated variant (50% activity after 2.5 h at 90°C). Furthermore, the esterase displays broad substrate specificity for esters of short-chain fatty acids (C2–C8).

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Acknowledgments

R. A. K. received a scholarship from the DAAD (Deutscher Akademischer Austausch Dienst).

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  1. Rami Al Khudary

    Present address: American University of the Middle East, Egaila, Kuwait

Authors and Affiliations

  1. Institute of Technical Microbiology, Hamburg University of Technology (TUHH), Kasernenstr. 12, 21073, Hamburg, Germany

    Rami Al Khudary, Ramprasath Venkatachalam, Moritz Katzer, Skander Elleuche & Garabed Antranikian

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  1. Rami Al Khudary
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  2. Ramprasath Venkatachalam
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  3. Moritz Katzer
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Correspondence to Garabed Antranikian.

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Communicated by H. Santos.

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Al Khudary, R., Venkatachalam, R., Katzer, M. et al. A cold-adapted esterase of a novel marine isolate, Pseudoalteromonas arctica: gene cloning, enzyme purification and characterization. Extremophiles 14, 273–285 (2010). https://doi.org/10.1007/s00792-010-0306-7

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  • DOI: https://doi.org/10.1007/s00792-010-0306-7

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