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Functional properties of a manganese-activated exo-polygalacturonase produced by a thermotolerant fungus Aspergillus niveus

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Abstract

A thermotolerant fungus identified as Aspergillus niveus was isolated from decomposing materials and it has produced excellent levels of hydrolytic enzymes that degrade plant cell walls. A. niveus germinated faster at 40 °C, presenting protein levels almost twofold higher than at 25 °C. The crude extract of the A. niveus culture was purified by diethylaminoethyl (DEAE)-cellulose, followed by Biogel P-100 column. Polygalacturonase (PG) is a glycoprotein with 37.7 % carbohydrate, molecular mass of 102.6 kDa, and isoelectric point of 5.4. The optimum temperature and pH were 50 °C and 4.0–6.5, respectively. The enzyme was stable at pH 3.0 to 9.0 for 24 h. The DEAE-cellulose derivative was about sixfold more stable at 60 °C than the free enzyme. Moreover, the monoaminoethyl-N-aminoethyl-agarose derivative was tenfold more stable than the free enzyme. PG was 232 % activated by Mn2+. The hydrolysis product of sodium polypectate corresponded at monogalacturonic acid, which classifies the enzyme as an exo-PG. The K m, V max, K cat, and K cat/K m values were 6.7 mg/ml, 230 U/mg, 393.3/s, and 58.7 mg/ml/s, respectively. The N-terminal amino acid sequence presented 80 % identity with PglB1, PglA2, and PglA3 putative exo-PG of Aspergillus fumigatus and an exo-PG Neosartorya fischeri.

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Acknowledgments

This work was supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and Conselho de Desenvolvimento Científico e Tecnológico (CNPq). J.A.J. and M.L.T.M.P. are research fellows of CNPq. A.M., A.R.L.D., and T.M.S. were recipients of a FAPESP Fellowship. We thank Dr. Dalton Amorim and Diego Fachin for the image support and Ricardo Alarcon and Mauricio de Oliveira for the technical assistance.

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Authors and Affiliations

  1. Departamento de Bioquímica e Imunologia, Faculdade de Medicina e Imunologia de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil

    Alexandre Maller & André Ricardo de Lima Damásio

  2. Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Av. Bandeirantes, 3900, 14040-901, Ribeirão Preto, SP, Brazil

    Tony Marcio da Silva, João Atílio Jorge, Hector Francisco Terenzi & Maria de Lourdes Teixeira de Moraes Polizeli

  3. Departamento de Biofísica, Universidade Federal de São Paulo, Ribeirão Preto, Brazil

    Izaura Yoshico Hirata

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  1. Alexandre Maller
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  2. Tony Marcio da Silva
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  3. André Ricardo de Lima Damásio
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  4. Izaura Yoshico Hirata
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  6. Hector Francisco Terenzi
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  7. Maria de Lourdes Teixeira de Moraes Polizeli
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Correspondence to Maria de Lourdes Teixeira de Moraes Polizeli.

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Maller, A., da Silva, T.M., Damásio, A.R.d.L. et al. Functional properties of a manganese-activated exo-polygalacturonase produced by a thermotolerant fungus Aspergillus niveus . Folia Microbiol 58, 615–621 (2013). https://doi.org/10.1007/s12223-013-0249-3

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