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Low-Scale expression and purification of an active putative iduronate 2-sulfate sulfatase-Like enzyme from Escherichia coli K12

  • Systems and Synthetic Microbiology and Bioinformatics
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Abstract

The sulfatase family involves a group of enzymes with a large degree of similarity. Until now, sixteen human sulfatases have been identified, most of them found in lysosomes. Human deficiency of sulfatases generates various genetic disorders characterized by abnormal accumulation of sulfated intermediate compounds. Mucopolysaccharidosis type II is characterized by the deficiency of iduronate 2-sulfate sulfatase (IDS), causing the lysosomal accumulation of heparan and dermatan sulfates. Currently, there are several cases of genetic diseases treated with enzyme replacement therapy, which have generated a great interest in the development of systems for recombinant protein expression. In this work we expressed the human recombinant IDS-Like enzyme (hrIDS-Like) in Escherichia coli DH5α. The enzyme concentration revealed by ELISA varied from 78.13 to 94.35 ng/ml and the specific activity varied from 34.20 to 25.97 nmol/h/mg. Western blotting done after affinity chromatography purification showed a single band of approximately 40 kDa, which was recognized by an IgY polyclonal antibody that was developed against the specific peptide of the native protein. Our 100 ml-shake-flask assays allowed us to improve the enzyme activity seven fold, compared to the E. coli JM109/pUC13-hrIDS-Like system. Additionally, the results obtained in the present study were equal to those obtained with the Pichia pastoris GS1115/pPIC-9-hrIDS-Like system (3 L bioreactor scale). The system used in this work (E. coli DH5α/pGEX-3X-hrIDS-Like) emerges as a strategy for improving protein expression and purification, aimed at recombinant protein chemical characterization, future laboratory assays for enzyme replacement therapy, and as new evidence of active putative sulfatase production in E. coli.

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References

  • Altschul, S.F., Madden, T.L., Schäffer, A.A., Zhang, J., Zhang, Z., Miller, W.J., and Lipman, D. 1997. Gapped BLAST and PSI-LAST: A new generation of protein database search programs. Nucleic Acid Res. 25, 3389–3402.

    Article  PubMed  CAS  Google Scholar 

  • Archer, I.M., Harper, P.S., and Wusteman, F.S. 1982. Multiple forms of Iduronate 2-sulphate sulphatase in human tissues and body fluids. Biochem. Biophys. Acta. 708, 134–140.

    Article  PubMed  CAS  Google Scholar 

  • Bader, R. and Leisinger, T. 1994. Isolation and characterization of the Methylophilus sp. strain DMll gene encoding dichloromethane dehalogenase/glutathione S-transferase. J. Bacteriol. 176, 3466–3473.

    PubMed  CAS  Google Scholar 

  • Baneyx, F. and Mujacic, M. 2004. Recombinant protein folding and misfolding in Escherichia coli. Nat. Biotechnol. 22, 1399–1408.

    Article  PubMed  CAS  Google Scholar 

  • Barrera, L.A. 1990. Enfermedades genéticas de origen metabólico. Pediatría 25, 64–68.

    Google Scholar 

  • Barrera, L.A. 1993. Errores innatos del metabolismo. Acta Méd. Colomb. 18, 31–40.

    Google Scholar 

  • Beaudet, A.L., Scriver, C.R., Sly, W.S., and Valle, D. 2001. The metabolic and molecular bases of inherited disease. McGraw-Hill.

    Google Scholar 

  • Benjdia, A., Deho, G., Rabot, S., and Berteau, O. 2007. First evidences for a third sulfatase maturation system in prokaryotes from E. coli aslB and ydeM deletion mutants. FEBS Lett. 581, 1009–1014.

    Article  PubMed  CAS  Google Scholar 

  • Bielicki, J., Freeman, C., Clements, P.R., and Hopwood, J.J. 1990. Human liver iduronate 2-sulfatase. Purification, characterization and catalytic properties. Biochem. J. 271, 75–86.

    PubMed  CAS  Google Scholar 

  • Bielicki, J., Hopwood, J., Wilson, P.J., and Anson, D.S. 1993. Recombinant human iduronate -2-sulphatase: correction of mucopolysaccharidosis — type II fibroblasts and characterization of the purified enzyme. J. Biochem. 289, 241–246.

    CAS  Google Scholar 

  • Bond, C.S., Clements, P.R., Ashby, S.J., Collyer, C.A., Harrop, S., Hopwood, J.J., and Guss, M. 1997. Structure of human lysosomal sulfatase. Struct. 5, 277–289.

    Article  CAS  Google Scholar 

  • Córdoba-Ruiz, H.A., Poutou-Piñales, R.A., Echeverri-Peña, O.Y., Algecira-Enciso, N.A., Landázuri, P., Sáenz, H., and Barrera-Avellaneda, L.A. 2009. Laboratory scale production of the human recombinant iduronate 2-sulfate sulfatase-Like from Pichia pastoris. Afr. J. Biotechnol. 8, 1786–1792.

    Google Scholar 

  • Dakterzada, F., Mobarez, A.M., Roudkenar, M.H., and Forouzandeh, M. 2012. Production of pentameric cholera toxin B subunit in Escherichia coli. Avicenna J. Med. Biotechnol. 4, 89–94.

    PubMed  CAS  Google Scholar 

  • Di Natale, P. and Ronsisville, L. 1981. Identification and partial characterization of two enzymes forms of iduronate sulfatase from human placenta. Biochem. Biophys. Acta. 661, 106–111.

    Article  PubMed  Google Scholar 

  • Dierks, T., Lecca, M.R., Schlotterhose, P., Schmidt, B., and von Figura, K. 1999. Sequence determinants directing conversion of cysteine to formylglycine in eukariotic sulfatases. Embo. J. 18, 2084–2091.

    Article  PubMed  CAS  Google Scholar 

  • Dierks, T., Lecca, M.R., Schmidt, B., and von Figura, K. 1998a. Conversion of cysteine to formylglycine in eukaryotic sulfatases occurs by a common mechanism in the endoplasmic reticulum. FEBS Lett. 423, 61–65.

    Article  PubMed  CAS  Google Scholar 

  • Dierks, T., Miech, C., Hummerjohann, J., Schmidt, B., Kertesz, M.A., and von Figura, K. 1998b. Posttranslational formation of formylglycine in prokaryotic sulfatases by modification of either cysteine or serine. J. Biol. Chem. 273, 25560–25564.

    Article  PubMed  CAS  Google Scholar 

  • Dong, X., Stothard, P., Forsythe, I., and Wishart, D. 2004. Plas-Mapper: a web server for drawing and auto-annotating plasmid maps. Nucleic Acids Res. 32, W660–664.

    Article  PubMed  CAS  Google Scholar 

  • Friedman, B., Vaddi, K., Preston, C., Mahon, E., Cataldo, J.R., and Macpherson, J.M. 1999. A comparison of pharmacological properties of carbohydrate remodeled recombinant and placental-derived β-glucocerebrosidase: Implications for clinical efficacy in treatment of Gaucher disease. Blood 93, 2807–2816.

    PubMed  CAS  Google Scholar 

  • Froissart, R., Millat, G., Mathiu, M., Bozon, D., and Maire, I. 1995. Processing of iduronate 2-sulfatase in human fibroblasts. Biochem. J. 309, 425–430.

    PubMed  CAS  Google Scholar 

  • García, A.R., DaCosta, J.M., Pan, J., Muenzer, J., and Lamsa, J.C. 2007. Preclinical dose ranging studies for enzyme replacement therapy with idursulfase in a knock-out mouse model of MPS II. Mol. Gen. Metabol. 91, 183–190.

    Article  Google Scholar 

  • Gasteiger, E., Hoogland, C., Gattiker, A., Duvaud, S., Wilkins, M.R., Appel, R.D., and Bairoch, A. 2005. Protein identification and analysis tools on the ExPASy server, pp. 571–660. Humana Press, New York, USA.

    Google Scholar 

  • Guez, N. and Peitisch, M.C. 1997. SWISS-MODEL and the Swiss-PdbViewer: An environment for comparative protein modeling. Electroph. 18, 2714–2723.

    Article  Google Scholar 

  • Hilman, B.C. and Sorensen, R.U. 1994. Management options: SCIDS with adenosine deaminase deficiency. Ann. Aller. 72, 395–402.

    CAS  Google Scholar 

  • Hopwood, J.J. and Ballabio, A. 2001. Multiple sulfatase deficiency and the nature of sulfatae family. In Scriver, C.R., Beaudet, A., Sly, W., and Valle, D. (eds.), pp. 3725–3732. The metabolic and molecular bases of inherited disease. Mc Graw-Hill, New York, N.Y., USA.

    Google Scholar 

  • Invitrogen. 2001. Subcloning efficiency DH5α. Catalog no. 18265-017.

    Google Scholar 

  • Jacobs, P.P., Geysens, S., Vervecken, W., Contreras, R., and Callewaert, N. 2008. Engineering complex-type N-glycosylation in Pichia pastoris using GlycoSwitch technology. Nat. Protocols 4, 58–70.

    Article  Google Scholar 

  • Kakkis, E.D., Muenzer, J., Tiller, G.E., Waber, L., Belmont, J., Passage, M., Izykowki, B., Philips, J., Doroshow, R., Walot, I., and et al. 2001. Enzyme-replacement therapy in mucopolysaccharidosis I. New Eng. J. Med. 344, 182–188.

    Article  PubMed  CAS  Google Scholar 

  • Kaplan, W., Husler, P., Klump, H., Erhardt, J., Sluis-Cremer, N., and Dirr, H. 1997. Conformational stability of pGEX-expressed Schistosoma japonicum glutathione S-transferase: A detoxification enzyme and fusion-protein affinity tag. Prot. Sci. 6, 399–406.

    Article  CAS  Google Scholar 

  • La Vallie, E.R. and McCoy, J.M. 1994. Enzymatic cleavage of fusion proteins with factor Xa. John Wiley & Sons, pp. 16.14.16–16.14.17. New York, N.Y., USA.

    Google Scholar 

  • Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.

    Article  PubMed  CAS  Google Scholar 

  • Landázuri, P. 2002. Clonación Del cDNA de la Iduronato 2-Sulfato Sulfatasa Humana. Expresión de la Enzima en Escherichia coli y Pichia pastoris, p. 134. Tesis Doctoral Thesis, Pontificia Universidad Javeriana, Bogotá, Columbia.

    Google Scholar 

  • Landázuri, P., Poutou-Piñales, R.A., Acero-Godoy, J., Córdoba-Ruiz, H.A., Echeverri-Peña, O.Y., Sáenz, H., Delgado-Boada, J.M., and Barrera-Avellaneda, L.A. 2009. Cloning and shake flask expression of hrIDS-Like in Pichia pastoris. Afr. J. Biotechnol. 8, 2871–2877.

    Google Scholar 

  • Larkin, M.A., Blackshields, G., Brown, N.P., Chenna, R., McGettigan, P.A., McWilliam, H., Valentin, F., Wallace, I.M., Wilm, A., López, R., Thompson, J.D., Gibson, T.J., and Higgins, D.G. 2007. Clustal W and Clustal X version 2.0. Bioinformatics 23, 2947–2948.

    Article  PubMed  CAS  Google Scholar 

  • Lili, W., Chaozhan, W., and Xindu, G. 2006. Expression, renaturation and simultaneous purification of recombinant human stem cell factor in Escherichia coli. Biotechnol. Lett. 28, 993–997.

    Article  PubMed  Google Scholar 

  • Lissens, W., Zenati, A., and Liebaers, I. 1984. Polyclonal antibodies against iduronate 2-sulfatase from human urine. Biochem. Biophys. Acta. 801, 365–371.

    Article  PubMed  CAS  Google Scholar 

  • Majidzadeh-A, K., Mahboudi, F., Hemayatkar, M., Davami, F., Barkhordary, F., Adeli, A., Soleimani, M., Davoudi, N., and Khalaj, V. 2010. Human tissue plasminogen activator expression in Escherichia coli using cytoplasmic and periplasmic cumulative power. Avicenna J. Med. Biotechnol. 2, 131–136.

    PubMed  CAS  Google Scholar 

  • Maleki, A., Roohvand, F., Tajerzadeh, H., Khanahmad, H., Nobari, M.B., Beiruti, A., and Najafabadi, A.R. 2010. High expression of methylotrophic yeast-derived recombinant human erythropoietin in a pH-controlled batch system. Avicenna J. Med. Biotechnol. 2, 197–206.

    PubMed  CAS  Google Scholar 

  • Meza, R.A., Monroy, A.F., Mercado, M., Poutou, R.A., Rodríguez, P., and Pedroza, A.M. 2004. Study of the stability in real time of cryopreserved strain banks. Univ. Sci. 9, 35–42.

    Google Scholar 

  • Miech, C., Dierks, T., Selmer, T., von Figura, K., and Schmidt, B. 1998. Arysulfatase from Klebsiella pneumoniae carries a formylglycine generated from serine. J. Biol. Chem. 273, 4835–4837.

    Article  PubMed  CAS  Google Scholar 

  • Millat, G., Froissart, R., Maire, I. and Bozon, D. 1997. Characterization of iduronate sulfate sulfatase mutants affecting N-glycosilations sites and cysteine-84 residue. Biochem. J. 326, 243–247.

    PubMed  CAS  Google Scholar 

  • Muenzer, J., Gucsavas-Calikoglu, M., McCandless, S.E., Schuetz, T.J., and Kimura, A. 2007. A phase I/II clinical trial of enzyme replacement therapy in mucopolysaccharidosis II (Hunter syndrome). Mol. Genet. Metab. 90, 329–337.

    Article  PubMed  CAS  Google Scholar 

  • Muenzer, J., Wraith, J.E., Beck, M., Giugliani, R., Harmatz, P., Eng, C.M., Vellodi, A., Martin, R., Ramaswami, U., Gucsavas-Calikoglu, M., and et al. 2006. A phase II/III clinical study of enzyme replacement therapy with idursulfase in mucopolysaccharidosis II (Hunter syndrome). Gen. Med. 8, 599.

    Article  Google Scholar 

  • Nabavinia, M.S., Nasab, M.N., Meshkat, Z., Derakhshan, M., and Khaje-Karamadini, M. 2011. Construction and evaluation of an expression vector containing Mtb32C (Rv0125) of Mycobacterium tuberculosis. Avicenna J. Med. Biotechnol. 3, 207–210.

    PubMed  CAS  Google Scholar 

  • Nagai, K. and Thøgersen, H.C. 1984. Generation of beta-globin by sequence-specific proteolysis of a hybrid protein produced in Escherichia coli. Nature 309, 810–812.

    Article  PubMed  CAS  Google Scholar 

  • Nagai, K. and Thøgersen, H.C. 1987. Synthesis and sequence-specific proteolysis of hybrid proteins produced in Escherichia coli. Meth. Enzymol. 153, 461–481.

    Article  PubMed  CAS  Google Scholar 

  • Neufeld, E.F. and Muenzer, J. 2001. The mucopolysaccharidoses. In Scriver, C.R., Beaudet, A.L., Sly, W., and Valle, D. (eds.), The metabolic and molecular bases of inherited disease, pp. 3421–3452. McGraw-Hill, New York, N.Y., USA.

    Google Scholar 

  • Niederau, C., Vom Dahl, S., and Haussinger, D. 1998. First long-term results of imiglucerase therapy of type 1 Gaucher disease. Eur. J. Med. Res. 3, 25–30.

    PubMed  CAS  Google Scholar 

  • Nyhan, W.L. and Ozand, P.T. 1998. Hunter disease/mucopolysaccharidoses type II (MPSII) iduronate sulfatase deficiency, Atlas of Metabolic Diseases, pp. 455–461. Chapman and Hall Medical, New York, USA.

    Google Scholar 

  • Parenti, G., Meroni, G. and Ballabio, A. 1997. The sulfatase gene family. Curr. Opin. Gen. Devel. 7, 386–391.

    Article  CAS  Google Scholar 

  • Peña, O., Sosa, A., Echeverri, O., Sáenz, H., and Barrera, L.A. 2005. Producción de anticuerpos policlonales IgG contra la proteína Iduronato-2-sulfato sulfatasa y desarrollo de un sistema de detección para IDS humana recombinante. Biomédica. 25, 181–188.

    PubMed  Google Scholar 

  • Poutou, R.A. 2006. Expresión de sulfatasas humanas en Escherichia coli y Pichia pastoris, p. 162. Tesis Doctoral Thesis, Pontificia Universidad Javeriana, Bogotá, D.C., Columbia.

    Google Scholar 

  • Poutou, R.A., Córdoba, H., Quevedo, B.E., Landázuri, P., Echeverri, O.Y., Sáenz, H., Vanegas, A., Acero, J., Gónzalez, A., Herrera, J., and et al. 2005. Expresión de iduronato 2-sulfato sulfatasa humana recombinante (IDShr) en Pichia pastoris. Univ. Sci. 10, 75–96.

    CAS  Google Scholar 

  • Poutou-Piñales, R.A., Vanegas Ni~no, A., Landázuri, P., Sáenz, H., Lareo, L., Echeverri, O.Y., and Barrera Avellaneda, L.A. 2010. Human sulfatase transiently and functionally active expressed in E. coli K12. Elect. J. Biotechnol. 13, article 8.

  • Rodríguez, A., Espejo, Á.J., Hernández, A., Velásquez, O.L., Lizaraso, L.M., Cordoba, H.A., Alméciga-Díaz, C., Barrera, L.A., and Sanchez, O. 2010. Enzyme replacement therapy for Morquio A: an active recombinant N-acetylgalactosamine-6-sulfate sulfatase produced in Escherichia coli BL21. J. Ind. Microbiol. Biotechnol. 37, 1193–1201.

    Article  PubMed  Google Scholar 

  • Sáenz, H. 2005. Expresión, purificación parcial y estudios computacionales de la IDShr producida en Pichia pastoris, p. 180. Doctoral Thesis, Pontificia Universidad Javeriana, Bogotá, D.C., Colombia.

    Google Scholar 

  • Sáenz, H., Lareo, L., Poutou, R.A., Sosa, C., and Barrera, L.A. 2007. Predicción computacional de la estructura terciaria de la iduronato 2-sulfato sulfatasa humana. Biomédica. 27, 7–20.

    PubMed  Google Scholar 

  • Saluta, M. and Bell, P. 2005. Troubleshooting GST fusion protein expression in E. coli. Amersham Biosciences, Piscataway, NJ, USA.

    Google Scholar 

  • Sambrook, J. and Russell, D.W. 2001. Molecular cloning: A laboratory manual, p. 2100. Cold Spring Harbor Laboratory Press, New York, N.Y., USA.

    Google Scholar 

  • Scott, H.S., Blanch, L., Guo, X.-H., Freeman, C., Orsborn, A., Baker, C., Sutherland, G.R., Morris, C.P. and Hopwood, J.J. 1995. Cloning of the sulphamidase gene and identification of mutations in Sanfilippo A syndrome. Nat. Genet. 11, 465–467.

    Article  PubMed  CAS  Google Scholar 

  • Söding, J. 2005. Protein homology detection by HMM-HMM comparison. Bioinformatics 21, 951–960.

    Article  PubMed  Google Scholar 

  • Sosa, A.C., Espejo, A.J., Rodriguez, E.A., Lizaraso, L.M., Rojas, A., Guevara, J., Echeverri, O.Y., and Barrera, L.A. 2011. Development of a sandwich enzyme linked immunosorbent assay (ELISA) for the quantification of iduronate-2-sulfate sulfatase. J. Immunol. Methods 368, 64–70.

    Article  PubMed  CAS  Google Scholar 

  • Szameit, C., Miech, C., Balleinninger, M., Bernhard., S., von Figura, K., and Dierks, T. 1999. The iron sulfur protein AtsB is required for posttranslational formation of formylglycine in the Klebsiella sulfatase. J. Biol. Chem. 274, 15375–15381.

    Article  PubMed  CAS  Google Scholar 

  • Vervecken, W., Kaigorodov, V., Callewaert, N., Geysens, S., De Vusser, K., and Contreras, R. 2004. In vivo synthesis of mammalian-like, hybrid-type N-glycans in Pichia pastoris. Appl. Environ. Microbiol. 70, 2639–2646.

    Article  PubMed  CAS  Google Scholar 

  • Vincze, T., Posfai, J., and Roberts, R.J. 2003. NEBcutter: a program to cleave DNA with restriction enzymes. Nucleic Acid Res. 30, 3688–3691.

    Article  Google Scholar 

  • von Figura, K., Bemhard, S., Thorsten, S., and Dierks, T. 1998. A novel protein modification generating an aldehyde group in sulfatases: Its role in catalysis and disease. BioEssay 20, 505–510.

    Article  Google Scholar 

  • von Figura, K., Gieselman, V., and Jaeken, J. 2001. Metachromatic leukodystrophy. In Scriver, C.R., Beaudet, A., Sly, W., and Valle, D. (eds.), The metabolic and molecular bases of inherited disease, pp. 3695–3724. Mc Graw-Hill, New York, N.Y., USA.

    Google Scholar 

  • Voznyi, Y.V., Keulemans, L.M., and van Diggelen, O.P. 2001. A fluorogenic enzyme assay for the diagnosis of MPS II (Hunter disease). J. Inherit. Metabol. Dis. 24, 675–680.

    Article  CAS  Google Scholar 

  • Waldow, A., Schmidt, B., Dierks, T., von Bülow, R., and von Figura, K. 1999. Amino acids residues forming the active site of arylsulfatase A. J. Biol. Chem. 274, 12284–12288.

    Article  PubMed  CAS  Google Scholar 

  • Weaston, A. and Neufeld, E.F. 1982. Iduronate of human plasma. Meth. Enzymol. 83, 573–578.

    Article  Google Scholar 

  • Weinreb, N., Charrow, J., Anderson, H.C., Kaplan, P., Kolodny, E.H., Mistry, P., Pastores, G., Rosenbloom, B.E., Ronald, C., Wappner, R., and et al. 2002. Eficacia de la terapia de reemplazo enzimático en 1.028 pacientes con enfermedad de Gaucher tipo 1 después de 2 a 5 a.os de tratamiento: Comunicación del Gaucher registry. Amer. J. Med. 113, 1–8.

    Article  Google Scholar 

  • Zhao, Y., Liu, Z., Yu, S., Wen, S., Hammarstrom, L., and Rabbani, H. 2009. Construction of a high efficiency PCR products cloning T vector using pGEM-5zf (+). Avicenna J. Med. Biotechnol. 1, 37–39.

    PubMed  CAS  Google Scholar 

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  1. Grupo de Investigación en Enfermedades Cardiovasculares y Metabólicas (GECAVYME), Facultad de Ciencias de la Salud, Universidad del Quindío, Armenia-Quindío, Colombia

    Edwin David Morales-Álvarez, Claudia Marcela Rivera-Hoyos, Angélica María Baena-Moncada & Patricia Landázuri

  2. Departamento de Química, Grupo de Investigación en Genética, Biodiversidad y Manejo de Ecosistemas (GEBIOME), Facultad de Ciencias Exactas y Naturales, Universidad de Caldas, Manizales-Caldas, Colombia

    Edwin David Morales-Álvarez

  3. Laboratorio de Biotecnología Molecular. Grupo de Biotecnología Ambiental e Industrial (GBAI), Departamento de Microbiología, Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, D.C., Colombia

    Claudia Marcela Rivera-Hoyos & Raúl A. Poutou-Piñales

  4. Unidad de Biología Celular y Microscopía, Decanato de Ciencias de la Salud, Universidad Centroccidental Lisandro Alvarado, Barquisimeto, Venezuela

    Homero Sáenz-Suárez

  5. Instituto de Errores Innatos del Metabolismo (IEIM), Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, D.C., Colombia

    Luis A. Barrera & Olga Y. Echeverri-Peña

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  1. Edwin David Morales-Álvarez
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Morales-Álvarez, E.D., Rivera-Hoyos, C.M., Baena-Moncada, A.M. et al. Low-Scale expression and purification of an active putative iduronate 2-sulfate sulfatase-Like enzyme from Escherichia coli K12. J Microbiol. 51, 213–221 (2013). https://doi.org/10.1007/s12275-013-2416-2

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