Abstract
Laccases have become desirable enzymes for application in many industrial processes. Nowadays, most of these enzymes are obtained from fungi. Among prospective studies for bacterial laccase genes, some have included actinomycetes, but only a few studies have characterized the enzyme produced. Thus, we have isolated a laccase-producing actinomycete from forest soil under restoration process and further aimed to characterize its produced enzyme. The isolate SB086 was assigned to the Streptomyces genus by a combination of phenotypical, chemical and phylogenetic properties. Our data indicate that the bacterium produces a thermotolerant laccase. The maximum activity was obtained in the pH range 4.0–5.0 and at 50 °C in reaction mixture containing 5 mM CuSO4; thermal stability was noted at 60 °C and 70 °C—a well-desired characteristic for industry. The active enzyme presented a high molecular mass (over 100 kDa) and was less sensitive to inhibition by metal ions than generally described for bacterial laccases. Our findings support in silico data of bacterial laccase secretion, and reinforce the view that actinomycetes may be a rich source of laccase for industrial application.
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References
Alexandre G, Zhulin IB (2000) Laccases are widespread in bacteria. Trends Biotechnol 18:41–42
Anderson AS, Wellington EMH (2001) The taxonomy of Streptomyces and related genera. Int J Syst Evol Microbiol 51:797–814
Arias ME, Arenas M, Rodríguez J, Soliveri J, Ball SA, Hernández M (2003) Kraft pulp biobleaching and mediated oxidation of a nonphenolic substrate by laccase from Streptomyces cyaneus CECT 3335. Appl Environ Microbiol 69:1953–1958
Ausec L, Zakrzewski M, Goesmann A, Schlüter A, Mandic-Mulec I (2011) Bioinformatic analysis reveals high diversity of bacterial genes for laccase-like enzymes. PLoS ONE 6(10):e25724
Baldrian P (2004) Purification and characterization of laccase from the white-rot fungus Daedalea quercina and decolorization of synthetic dyes by the enzyme. Appl Microbiol Biotechnol 63:560–563
Bugg DH, Ahmad M, Hardiman EM, Singh R (2011) The emerging role for bacteria in lignin degradation and bio-product formation. Curr Opin Biotechnol 22:394–400
Claus H (2004) Laccases: structure, reactions, distribution. Micron 35:93–96
Coll PM, Fernández-Abalos JM, Villanueva JR, Santamaría R, Pérez P (1993) Purification and characterization of a phenoloxidase (laccase) from the lignin-degrading basidiomycete PM1 (CECT 2971). Appl Environ Microbiol 59(8):2607–2613
Crawford DL (1978) Lignocellulosic decomposition by selected Streptomyces strains. Appl Environ Microbiol 35:1041–1045
Endo K, Hosono K, Beppu T, Ueda K (2002) A novel extracytoplasmic phenol oxidase of Streptomyces: its possible involvement in the onset of morphogenesis. Microbiology 148:1767–1776
Endo K, Hayashi Y, Hibi T, Hosono K, Beppu T, Ueda K (2003) Enzymological Characterization of EpoA, a Laccase-Like Phenol Oxidase Produced by Streptomyces griseus. J Biochem 133(5):671–677
Gonzalez JM, Saiz-Jimenez C (2002) A fluorimetric method for the estimation of G + C mol% content in microorganisms by thermal denaturation temperature. Environ Microbiol 4:770–773
Gordon RE, Mihm JM (1962) Identification of Nocardia caviae (Erikson) nov.comb. Ann NY Acad Sci 98:628–636
Gunne M, Urlacher VB (2012) Characterization of the Alkaline Laccase Ssl1 from Streptomyces sviceus with Unusual Properties Discovered by Genome Mining. PLoS ONE 7(12):1–8
Hatakka A (1994) Lignin-modifying enzyme from selected white-rot fungi: production and role in lignin degradation. FEMS Microbiol Rev 13:125–135
Hayakawa M, Nonumura H (1987) Humic acid – vitamin agar, a new medium for the selective isolation of soil actinomycetes. J Ferment Technol 65:501–509
Howard RL, Abotsi E, Jansen van Rensburg EL, Howard S (2003) Lignocellulose biotechnology: issues of bioconversion and enzyme production. Afr J Biotechnol 2(12):602–619
Kämpfer P (2012) Genus I. Streptomyces Waksman and Henrici 1943, 339 emend. Witt and Stackebrandt 1990, 370 emend. Wellington, Stackebrandt, Sanders, Wolstrup and Jorgensen 1992, 159. In: Goodfellow M, Kämpfer P, Busse H-J et al (eds) Bergey’s manual of systematic bacteriology, Part B, vol 5, 2nd edn. Springer, New York, pp 1455–1767
Kellner H, Luis P, Zimdars B, Kiesel B, Buscot F (2008) Diversity of bacterial laccase-like multicopper oxidase genes in forest and grassland Cambisol soil samples. Soil Biol Biochem 40:638–648
Kim OS, Cho YJ, Lee K, Yoon SH, Kim M, Na H, Park SC, Jeon YS, Lee JH, Yi H, Won S, Chun J (2012) Introducing EzTaxon-e: a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured species. Int J Syst Evol Microbiol 62:716–721
Luis P, Walther G, Kellner H, Martin F, Buscot F (2004) Diversity of laccase genes from basidiomycetes in a forest soil. Soil Biol Biochem 36:1025–1036
Machczynski MC, Vijgenboom E, Samyn B, Canter GW (2004) Characterization of SLAC: a small laccase from Streptomyces coelicolor with unprecedented activity. Protein Sci 13:2388–2397
Madhavi V, Lele SS (2009) Laccase: properties and applications. Bioresources 4(4):1694–1717
Manfio GP, Zakrzewska-Czerwinska J, Atalan E, Goodfellow M (1995) Towards minimal standards for the description of Streptomyces species. Biotechnologia 8:228–237
Martins LO, Soares CM, Pereira MM, Teixeira M, Costa T, Jones GH, Henriques AO (2002) Molecular and biochemical characterization of a highly stable bacterial laccase that occurs as a structural component of the Bacillus subtilis endospore coat. J Biol Chem 277:18849–18859
Molina-Guijarro JM, Pérez J, Muñoz-Dorado J, Guillén F, Moya R, Hernández M, Arias ME (2009) Detoxification of azo dyes by a novel pH-versatile, salt-resistant laccase from Streptomyces ipomoea. Int Microbiol 12:13–21
Piscitelli A, Pezzella C, Giardina P, Faraco V, Giovanni S (2010) Heterologous laccase production and its role in industrial applications. Bioeng Bugs 1(4):252–262
Sasser M (1990) Identification of bacteria by gas chromatography of cellular fatty acids. MIDI Technical Note 101. MIDI, Newark, DE
Schroeder M, Pöllinger-Zierler B, Aichernig N, Siegmund B, Guebitz GM (2008) Enzymatic removal of off-flavors from apple juice. J Agric Food Chem 56(7):2485–2489
Sharma P, Goel R, Capalash N (2007) Bacterial laccases. World J Microbiol Biotechnol 23(6):823–832
Shirling EB, Gottlieb D (1966) Methods for characterization of Streptomyces species. Int J Syst Bacteriol 16:313–340
Strong PJ, Claus H (2011) Laccase: A Review of Its Past and Its Future in Bioremediation. Crit Rev Environ Sci Technol 41(4):373–434
Suzuki T, Endo K, Ito M, Tsujibo H, Miyamoto K, Inamori Y (2003) A thermostable laccase from Streptomyces lavendulae REN-7: purification, characterization, nucleotide sequence, and expression. Biosci Biotechnol Biochem 67:2167–2175
Taherzadeh MJ, Karimi K (2007) Enzyme-based hydrolysis processes for ethanol from lignocellulosic materials: a review. Bioresources 2(4):707–738
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA 5: molecular evolutionary genetics analysis using maximum-likelihood, evolutionary distance, and maximum-parsimony methods. Mol Biol Evol 28:2731–2739
Tejirian A, Xu F (2010) Inhibition of cellulase-catalyzed lignocellulosic hydrolysis by iron and oxidative metal ions and complexes. Appl Environ Microbiol 76(23):7673–7682
Williams ST, Goodfellow M, Alderson G, Wellington EMH, Sneath PHA, Sackin MJ (1983) Numerical classification of Streptomyces and related genera. J Gen Microbiol 129:1743–1813
Zucchi TD, Almeida LG, Cônsoli FL (2011) Culturable bacterial diversity associated with cysts of Eurhizococcus brasiliensis (Hempel) (Hemiptera: Margarodidae). World J Microbiol Biotechnol 27:791–797
Zucchi TD, Kim B-Y, Bonda ANV, Goodfellow M (2013) Actinomadura xylanilytica sp. nov., a novel actinomycete isolate from soil. Int J Syst Evol Microbiol 63(2):576–580
Acknowledgment
The authors are indebted to FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo) for providing the funding for developing this research (grants 11/50243-1 and 11/14333-6).
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Fernandes, T.A.R., da Silveira, W.B., Passos, F.M.L. et al. Characterization of a thermotolerant laccase produced by Streptomyces sp. SB086. Ann Microbiol 64, 1363–1369 (2014). https://doi.org/10.1007/s13213-013-0781-z
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DOI: https://doi.org/10.1007/s13213-013-0781-z