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Induction, purification and characterization of α-N-acetylgalactosaminidase from Aspergillus Niger

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Abstract

A set of filamentous fungi (42 strains) was screened for α-N-acetylgalactosaminidase activity, and a series of inducers and different cultivation conditions were tested. Enzyme production by the best producer Aspergillus niger CCIM K2 was optimized and scaled up. α-N-Acetylgalactosaminidase was purified to apparent homogeneity by cation exchange chromatography, gel filtration, and chromatofocusing, and basic biochemical data of the enzyme were determined: The native molecular weight was estimated by gel filtration to be approximately 440 kDa, the molecular weight of the subunit was determined to be 76 kDa and the pI = 4.8. The K M was 0.73 mmol/l for o-nitrophenyl 2-acetamido-2-deoxy-α-D-galactopyranoside (o-NP-α-GalNAc), and optimum enzyme activity was achieved at pH 1.8 and 55 °C. This α-N-acetylgalactosaminidase is a retaining-type glycosidase, and it was N-deglycosylated without any loss of activity.

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Acknowledgment

This project was supported in part by the Ministry of Education, Czech Republic, project LC 06010; by the Grant Agency of the Czech Republic, grant no. 203/05/0172; and by institutional research concept (Institute of Microbiology) AV0Z50200510. Thanks are due to Dr. M. Kuzma (Institute of Microbiology, Prague) for the NMR measurements.

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Correspondence to V. Křen.

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Weignerová, L., Filipi, T., Manglová, D. et al. Induction, purification and characterization of α-N-acetylgalactosaminidase from Aspergillus Niger . Appl Microbiol Biotechnol 79, 769–774 (2008). https://doi.org/10.1007/s00253-008-1485-3

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Keywords

  • α-N-Acetylgalactosaminidase
  • α-GalNAc
  • Aspergillus niger
  • Glycosidase library