Extremophiles

, Volume 20, Issue 1, pp 45–55 | Cite as

Cloning, expression and characterization of a novel cold-adapted GDSL family esterase from Photobacterium sp. strain J15

  • Mehrnoush Hadaddzadeh Shakiba
  • Mohd Shukuri Mohamad Ali
  • Raja Noor Zaliha Raja Abd Rahman
  • Abu Bakar Salleh
  • Thean Chor Leow
Original Paper
First Online:
Received:
Accepted:

Abstract

The gene encoding for a novel cold-adapted enzyme from family II of bacterial classification (GDSL family) was cloned from the genomic DNA of Photobacterium sp. strain J15 in an Escherichia coli system, yielding a recombinant 36 kDa J15 GDSL esterase which was purified in two steps with a final yield and purification of 38.6 and 15.3 respectively. Characterization of the biochemical properties showed the J15 GDSL esterase had maximum activity at 20 °C and pH 8.0, was stable at 10 °C for 3 h and retained 50 % of its activity after a 6 h incubation at 10 °C. The enzyme was activated by Tween-20, -60 and Triton-X100 and inhibited by 1 mM Sodium dodecyl sulphate (SDS), while β-mercaptoethanol and Dithiothreitol (DTT) enhanced activity by 4.3 and 5.4 fold respectively. These results showed the J15 GDSL esterase was a novel cold-adapted enzyme from family II of lipolytic enzymes. A structural model constructed using autotransporter EstA from Pseudomonas aeruginosa as a template revealed the presence of a typical catalytic triad consisting of a serine, aspartate, and histidine which was verified with site directed mutagenesis on active serine.

Keywords

Cold-adapted GDSL family Esterase Cloning Characterization Photobacterium 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

This work was supported by Science Fund (02-01-04-SF1321) Ministry of Science Technology and Innovation, Malaysia.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interests regarding the publication of this paper.

Supplementary material

792_2015_796_MOESM1_ESM.docx (33 kb)
Supplementary material 1 (DOCX 32 kb)
792_2015_796_MOESM2_ESM.docx (18 kb)
Supplementary material 2 (DOCX 17 kb)
792_2015_796_MOESM3_ESM.docx (25 kb)
Supplementary material 3 (DOCX 24 kb)

References

  1. Akoh CC, Lee GC, Liaw YC, Huang TH, Shaw JF (2004) GDSL family of serine esterases/lipases. Prog Lipid Res 43(6):534–552PubMedCrossRefGoogle Scholar
  2. Arpigny JL, Jaeger KE (1999) Bacterial lipolytic enzymes: classification and properties. Biochem J 343(Pt 1):177PubMedPubMedCentralCrossRefGoogle Scholar
  3. Bradford MM (1976) Purification and characterization of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72(1):248–254PubMedCrossRefGoogle Scholar
  4. Cavicchioli R, Siddiqui KS, Andrews D, Sowers KR (2002) Low-temperature extremophiles and their applications. Curr Opin Biotechnol 13(3):253–261. doi: 10.1016/S0958-1669(02)00317-8 PubMedCrossRefGoogle Scholar
  5. Cieśliński H, Białkowska AM, Długołęcka A, Daroch M, Tkaczuk KL, Kalinowska H, Kur J, Turkiewicz M (2007) A cold-adapted esterase from psychrotrophic Pseudoalteromas sp. strain 643A. Arch Microbiol 188(1):27–36PubMedCrossRefGoogle Scholar
  6. Dalrymple BP, Cybinski DH, Layton I, McSweeney CS, Xue GP, Swadling YJ, Lowry JB (1997) Three Neocallimastix patriciarum esterases associated with the degradation of complex polysaccharides are members of a new family of hydrolases. Microbiology 143(8):2605PubMedCrossRefGoogle Scholar
  7. D’Amico S, Collins T, Marx J-C, Feller G, Gerday C (2006) Psychrophilic microorganisms: challenges for life. EMBO Rep 7(4):385–389PubMedPubMedCentralCrossRefGoogle Scholar
  8. Gerday C, Aittaleb M, Bentahir M, Chessa J-P, Claverie P, Collins T, D’Amico S, Dumont J, Garsoux G, Georlette D, Hoyoux A, Lonhienne T, Meuwis M-A, Feller G (2000) Cold-adapted enzymes: from fundamentals to biotechnology. Trends Biotechnol 18(3):103–107. doi: 10.1016/S0167-7799(99)01413-4 PubMedCrossRefGoogle Scholar
  9. Gianese G, Argos P, Pascarella S (2001) Structural adaptation of enzymes to low temperatures. Protein Eng 14(3):141–148PubMedCrossRefGoogle Scholar
  10. Hu XP, Heath C, Taylor MP, Tuffin M, Cowan D (2012) A novel, extremely alkaliphilic and cold-active esterase from Antarctic desert soil. Extremophiles 16(1):79–86PubMedCrossRefGoogle Scholar
  11. Jaeger K, Dijkstra B, Reetz M (1999) Bacterial biocatalysts: molecular biology, three-dimensional structures, and biotechnological applications of lipases. Annu Rev Microbiol 53(1):315–351PubMedCrossRefGoogle Scholar
  12. Kelley LA, Sternberg MJE (2009) Protein structure prediction on the Web: a case study using the Phyre server. Nat Protocols 4(3):363–371PubMedCrossRefGoogle Scholar
  13. Kulakova L, Galkin A, Nakayama T, Nishino T, Esaki N (2004) Cold-active esterase from Psychrobacter sp. Ant300: gene cloning, characterization, and the effects of Gly→Pro substitution near the active site on its catalytic activity and stability. Biochim Biophys Acta (BBA)-Proteins Proteomics 1696(1):59–65CrossRefGoogle Scholar
  14. Laemmli U (1970) Most commonly used discontinuous buffer system for SDS electrophoresis. Nature 227:680–685PubMedCrossRefGoogle Scholar
  15. Li J, Derewenda U, Dauter Z, Smith S, Derewenda ZS (2000) Crystal structure of the Escherichia coli thioesterase II, a homolog of the human Nef binding enzyme. Nat Struct Biol 7(7):555–559PubMedCrossRefGoogle Scholar
  16. Ling H (2008) Sequence analysis of GDSL lipase gene family in Arabidopsis thaliana. Pak J Biol Sci 11(5):763PubMedCrossRefGoogle Scholar
  17. Ling H, Zhao J, Zuo K, Qiu C, Yao H, Qin J, Sun X, Tang K (2006) Isolation and expression analysis of a GDSL-like lipase gene from Brassica napus L. J Biochem Mol Biol 39(3):297PubMedCrossRefGoogle Scholar
  18. Lyu PC, Sherman JC, Chen A, Kallenbach NR (1991) Alpha-helix stabilization by natural and unnatural amino acids with alkyl side chains. Proc Natl Acad Sci 88(12):5317–5320PubMedPubMedCentralCrossRefGoogle Scholar
  19. Marshall CJ (1997) Cold-adapted enzymes. Trends Biotechnol 15(9):359–364PubMedCrossRefGoogle Scholar
  20. Messaoudi A, Belguith H, Gram I, Hamida JB (2010) Classification of EC 3.1. 1.3 bacterial true lipases using phylogenetic analysis. Afr J Biotechnol 9(48):8243–8247Google Scholar
  21. Mølgaard A, Kauppinen S, Larsen S (2000) Rhamnogalacturonan acetylesterase elucidates the structure and function of a new family of hydrolases. Structure 8(4):373–383PubMedCrossRefGoogle Scholar
  22. Panda T, Gowrishankar BS (2005) Production and applications of esterases. Appl Microbiol Biotechnol 67(2):160–169. doi: 10.1007/s00253-004-1840-y PubMedCrossRefGoogle Scholar
  23. Schandl A, Pittner F (1984) The role of Na+ and Ca+ ions on the action of pancreatic lipase studied with the help of immobilisation techniques. Eur J Biochem 140(3):547–551PubMedCrossRefGoogle Scholar
  24. Suzuki T, Nakayama T, Choo DW, Hirano Y, Kurihara T, Nishino T, Esaki N (2003) Cloning, heterologous expression, renaturation, and characterization of a cold-adapted esterase with unique primary structure from a psychrotroph Pseudomonas sp. strain B11-1. Protein Expr Purif 30(2):171–178PubMedCrossRefGoogle Scholar
  25. Tokunaga H, Arakawa T, Tokunaga M (2008) Engineering of halophilic enzymes: two acidic amino acid residues at the carboxy-terminal region confer halophilic characteristics to Halomonas and Pseudomonas nucleoside diphosphate kinases. Protein Sci 17(9):1603–1610PubMedPubMedCentralCrossRefGoogle Scholar
  26. Upton C, Buckley JT (1995) A new family of lipolytic enzymes? Trends Biochem Sci 20(5):178PubMedCrossRefGoogle Scholar
  27. van den Berg B (2010) Crystal structure of a full-length autotransporter. J Mol Biol 396(3):627–633. doi: 10.1016/j.jmb.2009.12.061 PubMedCrossRefGoogle Scholar
  28. Xu Y, Zhang Y, Liang Z, Van de Casteele M, Legrain C, Glansdorff N (1998) Aspartate carbamoyltransferase from a psychrophilic deep-sea bacterium, Vibrio strain 2693: properties of the enzyme, genetic organization and synthesis in Escherichia coli. Microbiology 144(5):1435–1441PubMedCrossRefGoogle Scholar
  29. Yang Z, Zhang Y, Shen T, Xie Y, Mao Y, Ji C (2013) Cloning, expression and biochemical characterization of a novel, moderately thermostable GDSL family esterase from Geobacillus thermodenitrificans T2. J J Biosci Bioeng 115(2):133–137PubMedCrossRefGoogle Scholar
  30. Yu Shanshan ZB, Zhao Xinyu, Feng Yan (2010) Gene cloning and characterization of a novel thermophilic esterase from Fervidobacterium nodosum Rt17-B1. Acta Biochim Biophys Sin: 288–295Google Scholar
  31. Zuo K, Zhang L, Yao H, Wang J (2010) Isolation and functional expression of a novel lipase gene isolated directly from oil-contaminated soil. Acta Biochim Pol 57(3):305PubMedGoogle Scholar

Copyright information

© Springer Japan 2015

Authors and Affiliations

  1. 1.Enzyme and Microbial Technology Research CentreUniversiti Putra MalaysiaSerdangMalaysia
  2. 2.Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular SciencesUniversiti Putra MalaysiaSerdangMalaysia
  3. 3.Department of BiochemistryUniversiti Putra MalaysiaSerdangMalaysia
  4. 4.Department of Microbiology, Faculty of Biotechnology and Biomolecular ScienceUniversiti Putra MalaysiaSerdangMalaysia

Personalised recommendations