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Extremophiles

, Volume 14, Issue 3, pp 273–285 | Cite as

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

  • Rami Al Khudary
  • Ramprasath Venkatachalam
  • Moritz Katzer
  • Skander Elleuche
  • Garabed AntranikianEmail author
Original Paper

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).

Keywords

Esterase Psychrophilic enzyme Pseudoalteromonas arctica OsmC 

Notes

Acknowledgments

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

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Copyright information

© Springer 2010

Authors and Affiliations

  • Rami Al Khudary
    • 1
    • 2
  • Ramprasath Venkatachalam
    • 1
  • Moritz Katzer
    • 1
  • Skander Elleuche
    • 1
  • Garabed Antranikian
    • 1
    Email author
  1. 1.Institute of Technical MicrobiologyHamburg University of Technology (TUHH)HamburgGermany
  2. 2.American University of the Middle EastEgailaKuwait

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