Isolation and characterization of tyrosinase produced by marine actinobacteria and its application in the removal of phenol from aqueous environment

Abstract

The present study was focused on screening and characterization of tyrosinase enzyme produced by marine actinobacteria and its application in phenolic compounds removal from aqueous solution. A total of 20 strains were isolated from marine sediment sample and screened for tyrosinase production by using skimmed milk agar medium. Among 20 isolates, two isolates LK-4 and LK-20 showed zone of hydrolysis and these were taken for secondary screening by using tyrosine agar medium. Based on the result of secondary screening LK-4 was selected for further analysis, such as tyrosinase assay, protein content and specific activity of the enzyme. The tyrosinase enzyme was produced in a SS medium and was partially purified by ammonium sulfate precipitation, dialysis and SDS PAGE. The isolate (LK-4) was identified as Streptomyces espinosus using 16S rRNA gene sequencing and named as “Streptomyces espinosus strain LK4 (KF806735)”. The tyrosinase enzyme was immobilized in sodium alginate which was applied to remove phenolic compounds from water. The enzyme efficiently removed the phenolic compounds from aqueous solution within few hours which indicated that tyrosinase enzyme produced by Streptomyces espinosus strain LK-4 can be potently used for the removal of phenol and phenolic compounds from wastewater in industries.

Key words： actinobacteria; phenolic compounds; immobilization; tyrosinase

Acknowledgements

We are very thankful to the management of VIT University for providing laboratory and facilities to carry out our project work. We are also thankful to School of Advanced Science (SAS) and Chemistry Division for providing HPLC facility.

References

1 Adeyemi O, Oginni O, Osubor C C, Adeyemi O, Oloyede O B, Oladiji A T, Adebayo E A (2009). Effect of water contaminated with phthalate, benzene and cyclohexane on Clarias gariepinus’ cellular system. Food Chem Toxicol, 47(8): 1941–1944

PMID

2 Anwar A, Qader S A, Raiz A, Iqbal S, Azhar A (2009). Calcium alginate: a support material for immobilization of proteases from newly isolated strain of Bacillus subtilis KIBGE-HAS. World Appl Sci J, 7: 1281–1286

3 Bevilaqua J V, Freire M C, Anna S (2002). Phenol removal through combined biological and enzymatic treatments. Braz J Chem Eng, 19(2): 151–158

4 Bradford M M (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 72(1–2): 248–254

PMID

5 Chung T P, Tseng H Y, Juang R S (2003). Mass transfer effect and intermediate detection for phenol degradation in immobilized Pseudomonas putida systems. Process Biochem, 38(10): 1497–1507

6 Crecchio C, Ruggiero P, Pizzigallo M D R (1995). Polyphenoloxidases immobilized in organic gels: Properties and applications in the detoxification of aromatic compounds. Biotechnol Bioeng, 48(6): 585–591

PMID

7 Cuypers R, Sudhölter E J, Zuilhof H (2010). Hydrogen bonding in phosphine oxide/phosphate-phenol complexes. ChemPhysChem, 11(10): 2230–2240

PMID

8 Dajanta K, Wongkham S, Thirach P, Baophoeng P, Apichartsrangkoon A, Santithum P, Chukeatirote E (2009). Comparative study of proteolytic activity of protease-producing bacteria isolated from Thua nao. Maejo Int J Sci Technol, 3: 269–276

9 Dalfard B, Khajeh K, Soudi M R, Manesh H N, Ranjbar B, Sajedi R H (2006). Isolation and biochemical characterization of laccase and tyrosinase activities in a novel melanogenic soil bacterium. Enzyme Microb Technol, 39(7): 1409–1416

10 Decker H, Tuczek F (2000). Tyrosinase/catecholoxidase activity of hemocyanins: structural basis and molecular mechanism. Trends Biochem Sci, 25(8): 392–397

PMID

11 Della-Cioppa G, Garger S J, Holtz R B, McCulloch M J, Sverlow G G (1998a). Method for making stable extracellular tyrosinase and synthesis of polyphenolic polymers therefrom. US Patent 5801047

12 Della-Cioppa G, Garger S J, Sverlow G G, Turpen T H, Grill L K, Chedekal M R (1998b). Melanin production by Streptomyces. US Patent 5814495

13 Dolashki A, Gushterova A (2009). Identification and characterization of tyrosinase from Streptomyces albus by mass spectrometry. Biotechnol & Biotechnol, 23: 946–950

14 Dura’n N, Rosa M A, D’Annibale A, Gianfreda L (2002). Applications of laccases and tyrosinases (phenoloxidases) immobilized on different supports: a review. Enzyme Microb Technol, 31(7): 907–931

15 Escribano J, Cabanes J, Chazarra S, Garcı’a-Carmona F (1997). Characterization of monophenolase activity of table beet polyphenol oxidase. Determination of kinetic parameters on the tyramine/dopamine pair. J Agric Food Chem, 45(11): 4209–4214

16 Fairhead M, Thöny-Meyer L (2012). Bacterial tyrosinases: old enzymes with new relevance to biotechnology. New Biotechnol, 29(2): 183–191

PMID

17 Gernjak W, Krutzler T, Glaser A, Malato S, Caceres J, Bauer R, Fernández-Alba A R (2003). Photo-Fenton treatment of water containing natural phenolic pollutants. Chemosphere, 50(1): 71–78

PMID

18 Grady C P L Jr (1990). Biodegradation of toxic organics: status and potential. J Environ Eng, 116(5): 805–828

19 Ha S R, Vinitnantharat S, Ozaki H (2000). Bioregeneration by mixed microorganisms of granular activated carbon loaded with a mixture of phenols. Biotechnol Lett, 22(13): 1093–1096

20 Haghbeen K, Jazii F R, Karkhane A A, Borojerdi S S (2004). Purification of tyrosinase from edible mushroom. Iranian J Biotechnol, 2: 189–194

21 Halaouli S, Asther M, Sigoillot J C, Hamdi M, Lomascolo A (2006). Fungal tyrosinases: new prospects in molecular characteristics, bioengineering and biotechnological applications. J Appl Microbiol, 100(2): 219–232

PMID

22 Jones B V, Sun F, Marchesi J R (2007). Using skimmed milk agar to functionally screen a gut metagenomic library for proteases may lead to false positives. Lett Appl Microbiol, 45(4): 418–420

PMID

23 Kameda E, Langone M A, Coelho M A (2006). Tyrosinase extract from Agaricus bisporus mushroom and its in natura tissue for specific phenol removal. Environ Technol, 27(11): 1209–1215

PMID

24 Karthik L, Kumar G, Bhaskara Rao K V (2010). Diversity of marine actinomycetes from Nicobar marine sediments and its antifungal activity. Int J Pharm Pharm Sci, 2: 199–203

25 Kathiresan K, Balagurunathan R, Manilamani M, Selvan (2005). Fungicidal effect of marine actinomycetes against phytopathogenic fungi. Indian J Biotechnol, 4: 271–276

26 Katz E, Betancourt A (1988). Induction of tyrosinase by L-methionine in Streptomyces antibioticus. Can J Microbiol, 34(12): 1297–1303

PMID

27 Klibanov A M, Alberti B N, Morris E D, Felshin L M (1980). Enzymatic removal of toxic phenols and anilines from waste waters. J Appl Biochem, 2: 414–421

28 Kruger N J (1994). The Bradford method for protein quantitation. Methods Mol Biol, 32: 9–15

PMID

29 Matoba Y, Kumagai T, Yamamoto A, Yoshitsu H, Sugiyama M (2006). Crystallographic evidence that the dinuclear copper center of tyrosinase is flexible during catalysis. J Biol Chem, 281(13): 8981–8990

PMID

30 Maurya S, Singh D (2010). Quantitative Analysis of Total Phenolic Content in Adhatoda vasica Nees Extracts. Int J Pharm Tech Res, 2: 2403–2406

31 Mayer A M, Staples R C (2002). Laccase: new functions for an old enzyme. Phytochemistry, 60(6): 551–565

PMID

32 Molina L P, Hiner A N P, Tudela J, Garcı’a-Ca’novas F, Rodrı’guez-Lo’pez J N (2003). Enzymatic removal of phenolsfrom aqueous solution by artichoke (Cynara scolymus L.)extracts. Enzyme Microb Technol, 33(5): 738–742

33 Nambudiri A M D, Bhat J V (1972). Conversion of p-cumarate into caffeate by Streptomyces nigrifaciens. Biochem J, 130: 425–433

PMID

34 Peralta-Zamora P, Pereira C M, Tiburtius E R L, Moraes S G, Rosa M A, Minussi R C, Dura’n N (2003). Decolorization of reactive dyes by immobilized laccase. Appl Catal B, 42(2): 131–144

35 Philipp S, Held T, Kutzner H J (1991). Purifi cation and characterization of the tyrosinase of Streptomyces michiganensis DSM 40015. J Basic Microbiol, 31(4): 293–300

36 Popa C, Bahrim G (2011). Streptomyces tyrosinase: production and practical applications. Innov Rom Food Biotechnol, 8: 1–7

37 Raval K M, Vaswani P S, Majumder D R (2012). Biotransformation of a single amino acid L tyrosine into a bioactive molecule L-DOPA. Int J Sci Res, 2: 2250–3153

38 Rice R H, Cohen D E (1996). The basic science of poisons. McGraw-Hill, New York

39 Robb D A (1995). Exploiting tyrosinase activity in aqueous and nonaqueous media. Acs Sym Ser, 600: 159–165

40 Saboury A A, Zolghadri S, Haghbeen K, Moosavi-Movahedi A A (2006). The inhibitory effect of benzenethiol on the cresolase and catecholase activities of mushroom tyrosinase. J Enzyme Inhib Med Chem, 21(6): 711–717

PMID

41 Saiki R K, Scharf S, Faloona F, Mullis K B, Horn G T, Erlich H A, Arnheim N (1985). Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science, 230(4732): 1350–1354

PMID

42 Saitou N, Nei M (1987). The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol, 4(4): 406–425

PMID

43 Saiyood S, Vangnai A S, Thiravetyan P, Inthorn D (2010). Bisphenol A removal by the Dracaena plant and the role of plant-associating bacteria. J Hazard Mater, 178(1–3): 777–785

PMID

44 Seo S Y, Sharma V K, Sharma N (2003). Mushroom tyrosinase: recent prospects. J Agric Food Chem, 51(10): 2837–2853

PMID

45 Shesterenko Y A, Sevastyanov O V, Romanoyskaya L L (2012). Removal of phenols from aqueous solutions using Tyrosinase immobilized on polymer carriers and inorganic coagulants. J Water Chem Technol, 34(2): 107–111

46 Shi J, Bian W, Yin X (2009). Organic contaminants removal by the technique of pulsed high-voltage discharge in water. J Hazard Mater, 171(1–3): 924–931

PMID

47 Shubhrasekhar C, Supriya M, Karthik L, Gaurav K, Bhaskara Rao K V (2013). Isolation, characterization and application of biosurfactant produced by marine actinobacteria isolated from Saltpan soil from coastal area of Andhra Pradesh, India. Res J Biotechnol, 8: 18–25

48 Vermelho A B, Meirelles M N L, Lopes A, Petinate S D G, Chaia A A, Branquinha M H (1996). Detection of extracellular proteases from microorganisms on agar plates. Mem Inst Oswaldo Cruz, 91(6): 755–760

PMID

49 Wang Q, Fang X, Bai B, Liang X, Shuler P J, Goddard W A 3rd, Tang Y (2007). Engineering bacteria for production of rhamnolipid as an agent for enhanced oil recovery. Biotechnol Bioeng, 98(4): 842–853

PMID

50 Xu D Y, Yang Y, Yang Z (2011). Activity and stability of cross-linked tyrosinase aggregates in aqueous and nonaqueous media. J Biotechnol, 152(1–2): 30–36

PMID

51 Zhao J, Li Y, Zhang C, Zeng Q, Zhou Q (2008). Sorption and degradation of bisphenol A by aerobic activated sludge. J Hazard Mater, 155(1–2): 305–311

PMID