Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

A Novel Esterase from Paenibacillus sp. PBS-2 Is a New Member of the ${\beta}$-Lactamase Belonging to the Family VIII Lipases/Esterases

  • Kim, Young-Ok (Biotechnology Research Division, National Fisheries Research and Development Institute) ;
  • Park, In-Suk (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)
  • Received : 2014.05.20
  • Accepted : 2014.06.20
  • Published : 2014.09.28
Download PDF
⟨ Previous Next ⟩

Abstract

Screening of a gene library from Paenibacillus sp. PBS-2 generated in Escherichia coli led to the identification of a clone with lipolytic activity. Sequence analysis showed an open reading frame encoding a polypeptide of 378 amino acid residues with a predicted molecular mass of 42 kDa. The esterase displayed 69% and 42% identity with the putative ${\beta}$-lactamases from Paenibacillus sp. JDR-2 and Clostridium sp. BNL1100, respectively. The esterase contained a Ser-x-x-Lys motif that is conserved among all ${\beta}$-lactamases found to date. The protein PBS-2 was produced in both soluble and insoluble forms when E. coli cells harboring the gene were cultured at $18^{\circ}C$. The enzyme is a serine protein and was active against p-nitrophenyl esters of $C_2$, $C_4$, $C_8$, and $C_{10}$. The optimum pH and temperature for enzyme activity were pH 9.0 and $30^{\circ}C$, respectively. Relative activity of 55% remained at up to $5^{\circ}C$ with an activation energy of 5.84 kcal/mol, which indicates that the enzyme is cold-adapted. Enzyme activity was inhibited by $Cd^{2+}$, $Cu^{2+}$, and $Hg^{2+}$ ions. As expected for a serine esterase, activity was inhibited by phenylmethylsulfonyl fluoride. The enzyme was remarkably active and stable in the presence of commercial detergents and organic solvents. This cold-adapted esterase has potential as a biocatalyst and detergent additive for use at low temperatures.

Keywords

References

  1. Arpigny JL, Jaeger KE. 1999. Bacterial lipolytic enzymes: classification and properties. Biochem. J. 343: 177-183. https://doi.org/10.1042/0264-6021:3430177
  2. Bornscheuer UT, Kazlauskas RJ. 1999. Hydrolases in Organic Synthesis - Regio- and Stereoselective Biotransformations. Wiley- VCH, Weinheim.
  3. Bornscheuer UT. 2002. Microbial carboxylesterases: classification, properties and applications in biocatalysis. FEMS Microbiol. Rev. 26: 73-81. https://doi.org/10.1111/j.1574-6976.2002.tb00599.x
  4. Brenner S. 1988. The molecular evolution of genes and proteins: a tale of two serines. Nature 334: 528-530. https://doi.org/10.1038/334528a0
  5. Choo DW, Kurihara T, Suzuki T, Soda K, Esaki N. 1998. A cold-adapted lipase of an Alaskan psychrotroph, Pseudomonas sp. strain B11-1: gene cloning and enzyme purification and characterization. Appl. Environ. Microbiol. 64: 486-491.
  6. Dale JW, Godwin D, Mossakowska D, Stephenson P, Wall S. 1985. Sequence of OXA2 $\beta$-lactamase: comparion with other penicillin-reactive enzymes. FEMS Lett. 191: 39-44.
  7. Drauz K, Waldmann H. 1995. Enzyme Catalysis in Organic Synthesis. Vol. 1 and 2. Wiley-VCH, Weinheim.
  8. Elend C, Schmeisser C, Leggewise C, Babiak P, Carballeira JD, Steele HL, et al. 2006. Isolation and biochemical characterization of two novel metagenome-derived esterases. Appl. Environ. Microbiol. 72: 3637-3645. https://doi.org/10.1128/AEM.72.5.3637-3645.2006
  9. Feller G, Thiry M, Arpigny JL, Gerday C. 1991. Nucleotide sequence of the lipase gene lip2 from the antarctic psychrotrophic Moraxella TA144 and site-specific mutagenesis of the conserved serine and histidine resides. DNA Cell Biol. 10: 381-388. https://doi.org/10.1089/dna.1991.10.381
  10. Hasan F, Shah AA, Hameed A. 2006. Industrial applications of microbial lipases. Enzyme Microb. Technol. 39: 234-251.
  11. Jaeger KE, Dijkstra BW, Reetz MT. 1999. Bacterial biocatalysis: molecular biology, three-dimensional structures, and biotechnological applications of lipases. Annu. Rev. Microbiol. 53: 315-351. https://doi.org/10.1146/annurev.micro.53.1.315
  12. Jaeger KE, Ransac S, Kock BH, Feratto FG, Dijkstra BW. 1993. Topology characterization and modeling of the 3D structure of lipase from Pseudomonas aeruginosa. FEBS Lett. 332: 143-149. https://doi.org/10.1016/0014-5793(93)80501-K
  13. Joris B , Ghuysen J M, D ive G, R enard A, Did eberg O , Charlier P, et al. 1988. The active-site-serine penicillinrecognizing enzymes as members of the Streptomyces R61 DD-peptidase family. Biochem. J. 250: 313-324. https://doi.org/10.1042/bj2500313
  14. Jung YJ, Kim HK, Kim JH, Park SH, Oh TK, Lee JK. 2005. A direct approach for finding functional lipolytic enzymes from the Paenibacillus polymyxa genome. J. Microbiol. Biotechnol. 15: 155-160.
  15. Kim HE, Kim KR. 2002. Purification and characterization of an esterase from Acinetobacter lwoffii 16C-1. Curr. Microbiol. 44: 401-405. https://doi.org/10.1007/s00284-001-0008-6
  16. Kim HK, Choi HJ, Kim MH, Sohn CB, Oh TK. 2002. Expression and characterization of $Ca^{2+}$-independent lipase from Bacillus pumilus B26. Biochim. Biophys. Acta 1583: 205-212. https://doi.org/10.1016/S1388-1981(02)00214-7
  17. Kim YO, Heo YL, Nam BH, Jee YJ, Lee SJ, An CM. 2013. Molecular cloning, purification, and characterization of a cold-adapted esterase from Photobacterium sp. MA1-3. Fish. Aquat. Sci. 16: 311-318. https://doi.org/10.5657/FAS.2013.0311
  18. Kim YO, Park EM, Seo JS, Nam BH, Kong HJ, Kim WJ, et al. 2013. Shewanella sp. Ke75 esterase with specificity for pnitrophenyl butyrate: gene cloning and characterization. J. Korean Soc. Appl. Biol. Chem. 56: 55-62. https://doi.org/10.1007/s13765-012-2089-2
  19. Kim YO, Park IS, Kim HK, Nam BH, Kong HJ, Kim WJ, et al. 2011. A novel cold-adapted esterase from Salinisphaera sp. P7-4: gene cloning, overexpression, and characterization. J. Gen. Appl. Microbiol. 57: 357-364. https://doi.org/10.2323/jgam.57.357
  20. 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 GlyPro substitution near the active site on its catalytic activity and stability. Biochim. Biophys. Acta 1696: 59-65. https://doi.org/10.1016/j.bbapap.2003.09.008
  21. Lee HK, Min JA, Sung HK, Won HS, Byeong CJ. 2003. Purification and characterization of cold active lipase from psychrotrophic Aeromonas sp. LPB4. J. Microbiol. 41: 22-27.
  22. Lee SJ, Park EH, Han YH, Kim YO, Park SW. 2013. Isolation of a marine bacterium capable of biodegrading poly (butylenes succinate). Fish. Aquat. Sci. 16: 41-44. https://doi.org/10.5657/FAS.2013.0041
  23. Okuda H. 1991. Esterases, pp. 563-577. In Kuby SA (ed.). A Study of Enzymes. CRC Press, Boca Raton, FL.
  24. Panda T, Gowrishankar BS. 2005. Production and applications of esterase. Appl. Microbiol. Biotechnol. 67: 160-169. https://doi.org/10.1007/s00253-004-1840-y
  25. Park HJ, Jeong JH, Kang SG, Lee JH, Lee SA, Kim HK. 2007. Functional expression and refolding of new alkaline esterase, EM2L8 from deep-sea sediment metagenome. Protein Expr. Purif. 52: 340-347. https://doi.org/10.1016/j.pep.2006.10.010
  26. Patel RN. 2000. Stereoselective Biocatalysis. Marcel Dekker, New York.
  27. Petersen SB, Drablos F. 1994. A sequence analysis of lipases, esterases and related proteins, pp. 23-48. In Wooley P, Petersen SB (eds.). Lipases: Their Structure, Biochemistry and Applications. Press Syndicate of the University of Cambridge, New York.
  28. Petersen EI, Valinger G, Sölkner B, Stubenrauch G, Schwab H. 2001. A novel esterase from Burkholderia gladioli that shows high deacetylation activity on cephalosporins is related to $\beta$-lactamases and DD-peptidases. J. Biotechnol. 89: 11-25. https://doi.org/10.1016/S0168-1656(01)00284-X
  29. Prim N, Blanco A, Martinez J, Pastor FIJ, Diaz P. 2000. EstA, a gene coding for a cell-bound esterase from Paenibacillus sp. BP-23, is a new member of the bacterial subclass of type B carboxylesterases. Res. Microbiol. 151: 303-312. https://doi.org/10.1016/S0923-2508(00)00150-9
  30. Ryu HS, Kim HK, Choi WC, Kim MH, Park SY, Han NS, et al. 2006. New cold-adapted lipase from Photobacterium lipolyticum sp. nov. that is closely related to filamentous fungal lipases. Appl. Microbiol. Biotechnol. 70: 321-326. https://doi.org/10.1007/s00253-005-0058-y
  31. Saito H, Miura KI. 1963. Preparation of transforming deoxyribonucleic acid by phenol treatment. Biochim. Biophys. Acta 72: 619-629. https://doi.org/10.1016/0926-6550(63)90386-4
  32. 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: 171-178. https://doi.org/10.1016/S1046-5928(03)00128-1
  33. Suzuki T, Nakayama T, Kurihara T, Nishino T, Esaki N. 2002. A cold active esterase with a substrate preference for vinyl esters from a psychrotroph, Acinetobacter sp. strain no. 6: gene cloning, purification, and characterization. J. Mol. Catal. B Enzym. 16: 255-263. https://doi.org/10.1016/S1381-1177(01)00070-4
  34. Thompson JD, Higgins DG, Gibson TJ. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22: 4673-4680. https://doi.org/10.1093/nar/22.22.4673
  35. Tsujita T, Shirai K, Saito Y, Okuda H. 1990. Relationship between lipase and esterase. Isozymes: structure, function, and use in biology and medicine. Prog. Clin. Biol. Res. 344: 915-933.
  36. Upton C, Buckley JT. 1995. A new family of lipolytic enzymes. Trends Biochem. Sci. 20: 178-179. https://doi.org/10.1016/S0968-0004(00)89002-7
  37. Wei P, Bai L, Song W, Hao G. 2009. Characterization of two soil metagenome-derived lipases with high specificity for pnitrophenyl palmitate. Arch. Microbiol. 191: 233-240. https://doi.org/10.1007/s00203-008-0448-5
  38. Wei Y, Schottel JL, Derewenda U, Swenson L, Patkar S, Derewenda ZS. 1995. A novel variant of the catalytic triad in the Streptomyces scabies esterase. Nat. Struct. Biol. 2: 218-223. https://doi.org/10.1038/nsb0395-218
  39. Wong CH, Whitesids GM. 1994. Enzymes in Synthetic Organic Chemistry. Pergamon Press, Oxford.

Cited by

  1. Metagenomics of an Alkaline Hot Spring in Galicia (Spain): Microbial Diversity Analysis and Screening for Novel Lipolytic Enzymes vol.6, pp.None, 2015, https://doi.org/10.3389/fmicb.2015.01291
  2. Characterization of a Novel Alkaline Family VIII Esterase with S-Enantiomer Preference from a Compost Metagenomic Library vol.26, pp.2, 2014, https://doi.org/10.4014/jmb.1509.09081
  3. Cloning, Expression, and Characterization of a Cold-Adapted Shikimate Kinase from the Psychrophilic Bacterium Colwellia psychrerythraea 34H vol.26, pp.12, 2014, https://doi.org/10.4014/jmb.1608.08049
  4. Molecular characterization, overexpression and comparison of esterases-encoding LipRT, Lip4 and Lip20 from moderately thermophilic and mesophilic bacteria vol.169, pp.None, 2014, https://doi.org/10.1051/matecconf/201816901018
  5. Enhancement of the efficiency of Cd phytoextraction using bacterial endophytes isolated fromChromolaena odorata, a Cd hyperaccumulator vol.20, pp.11, 2018, https://doi.org/10.1080/15226514.2017.1365338
  6. Molecular Characterization of Novel Family IV and VIII Esterases from a Compost Metagenomic Library vol.9, pp.8, 2014, https://doi.org/10.3390/microorganisms9081614