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Cloning, Isolation, and Properties of a New Homologous Exoarabinase from the Penicillium canescens Fungus

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Abstract

A novel exo-arabinase (GH93, exo-ABN) enzyme produced by the ascomycete Penicillium canescens has been studied. Cloning of the abn1 gene coding for exo-ABN into the recipient P. canescens strain RN3-11-7 yielded recombinant producing strains characterized by a high yield of extracellular exo- ABN production (20–30% of the total amount of extracellular protein). Chromatographic purification yielded a homogenous exo-ABN with a molecular weight of 47 kDa, as shown by SDS-PAGE. The enzyme showed high specific activity towards linear arabinan (117 U/mg) and low specific activity towards branched arabinan and arabinoxylan (4–5 U/mg) and para-nitrophenyl-α-L-arabinofuranoside (0.3 U/mg), whereas arabinogalactan and para-nitrophenyl-α-L-arabinopyranoside, the substrates that contained the pyranose form of arabinose, were not hydrolyzed. Arabinohexaose was the major product of linear arabinan hydrolysis. Exo-ABN had a pH optimum at 5.0 and a temperature optimum at 60°C. The enzyme was stable in a broad pH range (4.0–7.0) and upon heating to 50°C during 180 min. Extensive hydrolysis of linear and branched arabinans by exo- and endo-arabinase mixtures, arabinofuranosidase, and arabinofuran-arabinoxylan hydrolase has been performed. The degree of substrate conversion amounted to 67 and 83% of the maximal possible value, respectively.

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Authors and Affiliations

  1. Fundamentals of Biotechnology Federal Research Center, Russian Academy of Sciences, Moscow, 119071, Russia

    M. V. Semenova, P. V. Volkov, A. M. Rozhkova, I. N. Zorov & A. P. Sinitsyn

  2. Faculty of Chemistry, Moscow State University, Moscow, 119991, Russia

    I. N. Zorov & A. P. Sinitsyn

Authors
  1. M. V. Semenova
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  2. P. V. Volkov
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  3. A. M. Rozhkova
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  4. I. N. Zorov
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  5. A. P. Sinitsyn
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Correspondence to M. V. Semenova.

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Original Russian Text © M.V. Semenova, P.V. Volkov, A.M. Rozhkova, I.N. Zorov, A.P. Sinitsyn, 2018, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2018, Vol. 54, No. 4.

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Semenova, M.V., Volkov, P.V., Rozhkova, A.M. et al. Cloning, Isolation, and Properties of a New Homologous Exoarabinase from the Penicillium canescens Fungus. Appl Biochem Microbiol 54, 387–395 (2018). https://doi.org/10.1134/S0003683818040130

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  • DOI: https://doi.org/10.1134/S0003683818040130

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