Isolation, screening and identification of haloarchaea with chitinolytic activity from hypersaline lakes of Iran

Maryam Yavari-Bafghi, Hamid Babavalian, Mohammad Ali Amoozegar

Abstract


Paper description:

  • The industrial application of enzymes which can withstand harsh conditions has greatly increased in recent years and extremozymes are good alternatives to mesophilic enzymes in industries. Chitinase is one of the most important hydrolytic enzymes with a variety of biotechnological applications.
  • We describe chitinase from a haloarchaeal strain belong to the genus Natrinema isolated from a hypersaline lake in Iran. To our knowledge, our investigation is among the first studies on chitinolytic activity of halophilic archaea.
  • This study revealed the strong potential of the genus Natrinema to produce chitinase at high salt concentrations without Mg2+ requirement.

Abstract: Halophiles produce stable enzymes under extreme conditions. The scant information about chitinolytic haloarchaea led us to conduct the present study in order to isolate and screen native halophilic archaea with chitinolytic activity and to optimize the enzyme production conditions. Among 500 haloarchaeal strains isolated from water samples from different hypersaline lakes of Iran, five strains showed chitinolytic activity. Based on biochemical, morphological and molecular analyses, we established that all five potent strains belonged to the genus Natrinema. Besides, observing chitinase function in culture media, through an additional molecular test the presence of the chitinase gene in chitinase-producing strains was also confirmed by PCR amplification. Compared with other potent strains, Natrinema sp. strain BS5 showed significant chitinase production. The production of chitinase in strain BS5 accompanied growth, started at the logarithmic phase and increased to its maximum level at the beginning of the stationary phase. Maximum chitinase production was obtained at 37˚C, pH 7.5, 3 M NaCl and 1% colloidal chitin. The strain BS5 showed 38%, 30%, 24% and 28% decreases in enzyme production at 40˚C, pH 8, 3.5 M NaCl and 0.5% substrate, respectively. This strain was able to produce the enzyme in NaCl 4 M and in the absence of MgCl2 and MgSO4. This study revealed the strong potential of the genus Natrinema to produce chitinase at high salt concentrations without Mg2+ requirement.

https://doi.org/10.2298/ABS180525049Y

Received: May 25, 2018; Revised: September 27, 2018; Accepted: October 4, 2018; Published online: October 30, 2018

How to cite this article: Yavari-Bafghi M, Babavalian H, Amoozegar MA. Isolation, screening and identification of haloarchaea with chitinolytic activity from hypersaline lakes of Iran. Arch Biol Sci. 2019;71(1):71-81.


Keywords


chitinase, halophile, archaea, Natrinema, chitin

Full Text:

PDF

References


Ara I, Daram D, Baljinova T, Yamamura H, Bakir M a, Suto M, Ando K. Isolation , classification , phylogenetic analysis and scanning electron microscopy of halophilic , halotolerant and alkaliphilic actinomycetes isolated from hypersaline soil. African J Microbiol Res. 2013;7(4):298-308.

Ventosa A. Unusual micro-organisms from unusual habitats: hypersaline environments. In: Logan NA, Lappin-Scott HM, Oyston PCF, editors. Prokaryotic diversity. Cambridge: Cambridge University Press; 2006. p. 223-54.

Yin J, Chen JC, Wu Q, Chen GQ. Halophiles, coming stars for industrial biotechnology. Vol. 33, Biotechnology Advances. 2015. p. 1433-42.

Kakhki AM, Amoozegar MA, Khaledi EM. Diversity of hydrolytic enzymes in haloarchaeal strains isolated from salt lake. Int J Environ Sci Technol. 2011;8(4):705-14.

Litchfield CD. Potential for industrial products from the halophilic Archaea. J Ind Microbiol Biotechnol. 2011;38(10):1635–47.

Amoozegar MA, Siroosi M, Atashgahi S, Smidt H, Ventosa A. Systematics of haloarchaea and biotechnological potential of their hydrolytic enzymes. Microbiology. 2017;163(5):623-45.

Islam R, Datta B. Diversity of chitinases and their industrial potential. Int J Appl Res. 2015;1:55-60.

Stoykov YM, Pavlov AI, Krastanov AI. Chitinase biotechnology: Production, purification, and application. Vol. 15, Engineering in Life Sciences. 2015. p. 30-8.

Adrangi S, Faramarzi M, Shahverdi A, Sepehrizadeh Z. Purification and characterization of two extracellular endochitinases from Massilia timonae. Carbohydr Res. 2010;345(3):402-7.

Hashimoto M, Ikegami T, Seino S, Ohuchi N, Fukada H, Sugiyama J, Shirakawa M, Watanabe T.. Expression and Characterization of the Chitin-Binding Domain of Chitinase A1 from Bacillus circulans WL-12. J Bacteriol. 2000;182(11):3045-54.

Ahmadian G, Degrassi G, Venturi V, Zeigler DR, Soudi M, Zanguinejad P. Bacillus pumilus SG2 isolated from saline conditions produces and secretes two chitinases. J Appl Microbiol. 2007;103(4):1081-9.

Delgado-García M, Aguilar CN, Contreras-Esquivel JC, Rodríguez-Herrera R. Screening for extracellular hydrolytic enzymes production by different halophilic bacteria. Mycopath. 2014;12(1):17-23.

Swiontek Brzezinska M, Jankiewicz U, Burkowska A, Walczak M. Chitinolytic Microorganisms and Their Possible Application in Environmental Protection. Curr Microbiol. 2014;68(1):71-81.

Li H, Greene LH. Sequence and Structural Analysis of the Chitinase Insertion Domain Reveals Two Conserved Motifs Involved in Chitin-Binding. Yang H, editor. PLoS One. 2010 Jan;5(1):e8654.

Adrangi S, Faramarzi MA. From bacteria to human: A journey into the world of chitinases. Biotechnol Adv. 2013 Dec;31(8):1786-95.

Revah-Moiseev S, Carroad PA. Conversion of the enzymatic hydrolysate of shellfish waste chitin to single-cell protein. Biotechnol Bioeng. 1981;23(5):1067-78.

García-Fraga B, da Silva AF, López-Seijas J, Sieiro C. Functional expression and characterization of a chitinase from the marine archaeon Halobacterium salinarum CECT 395 in Escherichia coli. Appl Microbiol Biotechnol. 2014 Mar 30;98(5):2133-43.

Gao J, Bauer MW, Shockley KR, Pysz MA, Kelly RM. Growth of Hyperthermophilic Archaeon Pyrococcus furiosus on Chitin Involves Two Family 18 Chitinases. Appl Environ Microbiol. 2003;69(6):3119-28.

Tanaka T, Fukui T, Imanaka T. Different Cleavage Specificities of the Dual Catalytic Domains in Chitinase from the Hyperthermophilic Archaeon Thermococcus kodakaraensis KOD1. J Biol Chem. 2001;276(38):35629-35.

Staufenberger T, Imhoff JF, Labes A. First crenarchaeal chitinase found in Sulfolobus tokodaii. Microbiol Res. 2012;167:262-9.

Amoozegar MA, Makhdoumi-Kakhki A, Shahzadeh Fazeli SA, Azarbaijani R, Ventosa A. Halopenitus persicus gen. nov., sp. nov., an archaeon from an inland salt lake. Int J Syst Evol Microbiol. 2012;62(Pt8):1932-6.

Pecher T, Böck A. In vivo susceptibility of halophilic and methanogenic organisms to protein synthesis inhibitors. FEMS Microbiol Lett. 1981;10(3):295-7.

Kaya M, Baran T, Karaarslan M. A new method for fast chitin extraction from shells of crab, crayfish and shrimp. Nat Prod Res. 2015;29(15):1477-80.

Dassault H. P. An improved technique for staining red halophilic bacteria. J Bacteriol. 1955;70(4):484-5.

Heimbrook ME, Wang WL, Campbell G. Staining bacterial flagella easily. J Clin Microbiol. 1989;27(11):2612-5.

Garrity GM, Holt JG. The Road Map to the Manual. In: Bergey’s Manual® of Systematic Bacteriology. 2001. p. 119-66.

Marmur J. A procedure for the isolation of deoxyribonucleic acid from micro-organisms. J Mol Biol. 1961;3(2):208.

Spencer JFT, Ragout de Spencer AL, editors. Environmental Microbiology. Totowa: Humana Press; 2004. 439 p. (Methods in Biotechnology; 16).

Klatte T, Evans L, Whitehead RN, Cole JA. Four PCR primers necessary for the detection of periplasmic nitrate reductase genes in all groups of Proteobacteria and in environmental DNA. Biochem Soc Trans. 2011;39(1):321-6.

Kim O-S, Cho Y-J, Lee K, Yoon S-H, Kim M, Na H, Park SC, Jeon Y, Lee JH, Yi H, Won S, Chun J. Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int J Syst Evol Microbiol. 2012;62(Pt 3):716-21.

Miller GL. Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar. Anal Chem. 1959;31(3):426-8.

Nawani NN, Kapadnis BP. Optimization of chitinase production using statistics based experimental designs. Process Biochem. 2005;40(2):651-60.

Xu XW, Ren PG, Liu SJ, Wu M, Zhou PJ. Natrinema altunense sp. nov., an extremely halophilic archaeon isolated from a salt lake in Altun Mountain in Xinjiang, China. Int J Syst Evol Microbiol. 2005;55(3):1311-4.

Ali I, Prasongsuk S, Akbar A, Aslam M, Lotrakul P, Punnapayak H, Rakshit S. Hypersaline habitats and halophilic microorganisms. Maejo Int J Sci Technol. 2016;10(3):330-45.

Margesin R, Schinner F. Potential of halotolerant and halophilic microorganisms for biotechnology. Extremophiles. 2001;5(2):73-83.

Delgado-García M, Valdivia-Urdiales B, Aguilar-González CN, Contreras-Esquivel JC, Rodríguez-Herrera R. Halophilic hydrolases as a new tool for the biotechnological industries. J Sci Food Agric. 2012;92(13):2575-80.

Saima, Kuddus M, Roohi, Ahmad IZ. Isolation of novel chitinolytic bacteria and production optimization of extracellular chitinase. J Genet Eng Biotechnol. 2013;11(1):39-46.

Monreal J, Reese ET. The chitinase of Serratia marcescens. Can J Microbiol. 1969;15(7):689-96.

Berger LR RD. The chitinase system of a strain of Streptomyces griseus. Biochem Biophys Acta. 1958;29:522-34.

Karthik N, Akanksha K PA. Production, purification and properties of fungal chitinases. Indian J Exp Biol. 2014;11(1025–35.):1025-35.

Percot A, Viton C, Domard A. Optimization of chitin extraction from shrimp shells. Biomacromolecules. 2003;4(1):12-8.

Sorokin DY, Toshchakov S V, Kolganova T V, Kublanov I V. Halo(natrono)archaea isolated from hypersaline lakes utilize cellulose and chitin as growth substrates. Front Microbiol. 2015;6:942.

Deeba F, Shakir HA, Irfan M, Qazi JI. Chitinase production in organisms : a review. Punjab Univ J Zoo. 2016;31(1):101-6.

Hou J, Han J, Cai L, Zhou J, Lü Y, Jin C, Liu J, Xiang H. Characterization of genes for chitin catabolism in Haloferax mediterranei. Appl Microbiol Biotechnol. 2014;98(3):1185-94.

Patil J, Bajekal S. Diversity of hydrolytic enzymes in Haloalkaliphilic archaea isolated from Lonar Lake. Microbiology. 2013;2(7).

Shi W, Tang X-F, Huang Y, Gan F, Tang B, Shen P. An extracellular halophilic protease SptA from a halophilic archaeon Natrinema sp. J7: gene cloning, expression and characterization. Extremophiles. 2006;10(6):599-606.


Refbacks

  • There are currently no refbacks.


Copyright (c) 2019 ARCHIVES OF BIOLOGICAL SCIENCES

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.