Recombinant Production and Characterization of a Novel Esterase from a Hypersaline Lake, Acıgöl, by Metagenomic Approach

Overview

Journal Extremophiles

Publisher Springer

Date 2019 Jun 3

PMID 31154531

Citations 7

Authors

Havva Esra Tutuncu

Nurgul Balci

Melek Tuter

Nevin Gul Karaguler

Affiliations

Soon will be listed here.

Abstract

The aim of this study was to isolate a novel esterase from a hypersaline lake by sequence-based metagenomics. The metagenomic DNA was isolated from the enriched hypersaline lake sediment. Degenerate primers targeting the conserved regions of lipolytic enzymes of halophilic microorganisms were used for polymerase chain reaction (PCR) and a whole gene was identified by genome walking. The gene was composed of 783 bp, which corresponds to 260 amino acids with a molecular weight of 28.2 kDa. The deduced amino acid sequence best matched with the esterase from Halomonas gudaonensis with an identity of 91%. Recombinantly expressed enzyme exhibited maximum activity towards pNP-hexanoate with a k value of 12.30 s. The optimum pH and temperature of the enzyme were found as 9 and 30 °C, respectively. The effects of NaCl, solvents, metal ions, detergents and enzyme inhibitors were also studied. In conclusion, a novel enzyme, named as hypersaline lake "Acıgöl" esterase (hAGEst), was identified by sequence-based metagenomics. The high expression level, the ability to maintain activity at cold temperatures and tolerance to DMSO and metal ions are the most outstanding properties of the hAGEst.

Citing Articles

Pharmaceutical applications of halophilic enzymes.

Yavari-Bafghi M, Amoozegar M Heliyon. 2025; 11(4):e42754.

PMID: 40066035 PMC: 11891729. DOI: 10.1016/j.heliyon.2025.e42754.

Enzymatic Characterization of a Novel HSL Family IV Esterase EstD04 from sp. D01 in Mealworm Gut Microbiota.

Kuan J, Tsai C, Chou C, Wu C, Wu W Molecules. 2023; 28(14).

PMID: 37513282 PMC: 10385968. DOI: 10.3390/molecules28145410.

Molecular Cloning and Characterization of a New Family VI Esterase from an Activated Sludge Metagenome.

Liaw R, Chen J, Cheng M Microorganisms. 2022; 10(12).

PMID: 36557656 PMC: 9786865. DOI: 10.3390/microorganisms10122403.

Biotechnological potentials of halophilic microorganisms and their impact on mankind.

Dutta B, Bandopadhyay R Beni Suef Univ J Basic Appl Sci. 2022; 11(1):75.

PMID: 35669848 PMC: 9152817. DOI: 10.1186/s43088-022-00252-w.

Characterization of a Novel Esterase Est33 From an Antarctic Bacterium: A Representative of a New Esterase Family.

Liu X, Zhou M, Sun R, Xing S, Wu T, He H Front Microbiol. 2022; 13:855658.

PMID: 35655995 PMC: 9152352. DOI: 10.3389/fmicb.2022.855658.

References

Arpigny J, Jaeger K . Bacterial lipolytic enzymes: classification and properties. Biochem J. 1999; 343 Pt 1:177-83. PMC: 1220539. View

Huang X, Madan A . CAP3: A DNA sequence assembly program. Genome Res. 1999; 9(9):868-77. PMC: 310812. DOI: 10.1101/gr.9.9.868. View

Madern D, Ebel C, Zaccai G . Halophilic adaptation of enzymes. Extremophiles. 2000; 4(2):91-8. DOI: 10.1007/s007920050142. View

Stothard P . The sequence manipulation suite: JavaScript programs for analyzing and formatting protein and DNA sequences. Biotechniques. 2000; 28(6):1102, 1104. DOI: 10.2144/00286ir01. View

Linhart C, Shamir R . The degenerate primer design problem. Bioinformatics. 2002; 18 Suppl 1:S172-81. DOI: 10.1093/bioinformatics/18.suppl_1.s172. View

Bell P, Sunna A, Gibbs M, Curach N, Nevalainen H, Bergquist P . Prospecting for novel lipase genes using PCR. Microbiology (Reading). 2002; 148(Pt 8):2283-2291. DOI: 10.1099/00221287-148-8-2283. View

Fischer M, Pleiss J . The Lipase Engineering Database: a navigation and analysis tool for protein families. Nucleic Acids Res. 2003; 31(1):319-21. PMC: 165462. DOI: 10.1093/nar/gkg015. View

Zhao L, Han B, Huang Z, Miller M, Huang H, Malashock D . Epoxide hydrolase-catalyzed enantioselective synthesis of chiral 1,2-diols via desymmetrization of meso-epoxides. lzhao@diversa.com. J Am Chem Soc. 2004; 126(36):11156-7. DOI: 10.1021/ja0466210. View

Jiang Z, Wang H, Ma Y, Wei D . Characterization of two novel lipase genes isolated directly from environmental sample. Appl Microbiol Biotechnol. 2005; 70(3):327-32. DOI: 10.1007/s00253-005-0065-z. View

10.

Ferrer M, Golyshina O, Chernikova T, Khachane A, Martins Dos Santos V, Yakimov M . Microbial enzymes mined from the Urania deep-sea hypersaline anoxic basin. Chem Biol. 2005; 12(8):895-904. DOI: 10.1016/j.chembiol.2005.05.020. View

11.

Hanson C, Nishiyama Y, Paul S . Catalytic antibodies and their applications. Curr Opin Biotechnol. 2005; 16(6):631-6. DOI: 10.1016/j.copbio.2005.10.003. View

12.

Wei X, Kuhn D, Narasimhan G . Degenerate primer design via clustering. Proc IEEE Comput Soc Bioinform Conf. 2006; 2:75-83. View

13.

Tang K, Utairungsee T, Kanokratana P, Sriprang R, Champreda V, Eurwilaichitr L . Characterization of a novel cyclomaltodextrinase expressed from environmental DNA isolated from Bor Khleung hot spring in Thailand. FEMS Microbiol Lett. 2006; 260(1):91-9. DOI: 10.1111/j.1574-6968.2006.00308.x. View

14.

Labes A, Karlsson E, Fridjonsson O, Turner P, Hreggvidson G, Kristjansson J . Novel members of glycoside hydrolase family 13 derived from environmental DNA. Appl Environ Microbiol. 2008; 74(6):1914-21. PMC: 2268309. DOI: 10.1128/AEM.02102-07. View

15.

Oren A . Microbial life at high salt concentrations: phylogenetic and metabolic diversity. Saline Syst. 2008; 4:2. PMC: 2329653. DOI: 10.1186/1746-1448-4-2. View

16.

Ozcan B, Ozyilmaz G, Cokmus C, Caliskan M . Characterization of extracellular esterase and lipase activities from five halophilic archaeal strains. J Ind Microbiol Biotechnol. 2008; 36(1):105-10. DOI: 10.1007/s10295-008-0477-8. View

17.

Waterhouse A, Procter J, Martin D, Clamp M, Barton G . Jalview Version 2--a multiple sequence alignment editor and analysis workbench. Bioinformatics. 2009; 25(9):1189-91. PMC: 2672624. DOI: 10.1093/bioinformatics/btp033. View

18.

Contreras-Moreira B, Sachman-Ruiz B, Figueroa-Palacios I, Vinuesa P . primers4clades: a web server that uses phylogenetic trees to design lineage-specific PCR primers for metagenomic and diversity studies. Nucleic Acids Res. 2009; 37(Web Server issue):W95-W100. PMC: 2703966. DOI: 10.1093/nar/gkp377. View

19.

Hasan F, Shah A, Hameed A . Methods for detection and characterization of lipases: A comprehensive review. Biotechnol Adv. 2009; 27(6):782-798. DOI: 10.1016/j.biotechadv.2009.06.001. View

20.

Kotik M . Novel genes retrieved from environmental DNA by polymerase chain reaction: current genome-walking techniques for future metagenome applications. J Biotechnol. 2009; 144(2):75-82. DOI: 10.1016/j.jbiotec.2009.08.013. View