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A Fungal Alpha-Galactosidase from Pseudobalsamia microspora Capable of Degrading Raffinose Family Oligosaccharides

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Abstract

An alpha-galactosidase was purified from Pseudobalsamia microspora (PMG) to 1224.1-fold with a specific activity of 11,274.5 units/mg by ion-exchange chromatography and gel filtration. PMG is a monomeric protein with a molecular mass of 62 kDa as determined by SDS-PAGE and by gel filtration. Chemical modification using N-bromosuccinimide (NBS) resulted in a complete abrogation of the activity of PMG, suggesting that Trp is an amino acid essential to its activity. The activity was strongly inhibited by Hg2+, Cd2+, Cu2+, and Fe3+ ions. Three inner peptide sequences for PMG were obtained by liquid chromatography–tandem mass spectrometry (LC–MS–MS) analysis. When 4-nitrophenyl α-d-glucopyranoside (pNPGal) was used as substrate, the optimum pH and temperature of PMG were 5.0 and 55 °C, respectively. The Michaelis constant (K m) value of the alpha-galactosidase on pNPGal was 0.29 mM, and the maximal velocity (V max) was 0.97 μmol ml−1 min−1. Investigation by thin-layer chromatography (TLC) demonstrated its ability to hydrolyze raffinose and stachyose. Hence, it can be exploited in degradation of non-digestible oligosaccharides from food and feed industries.

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Acknowledgments

This work was financially supported by National Grants of China (Biomass dissociation and low-molecular fragment green monomerization and transformation, 2010CB732202).

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Authors and Affiliations

  1. State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing, 100193, People’s Republic of China

    Dongxue Yang, Fang Du & Hexiang Wang

  2. Institute of Biotechnology and Germplasmic Resource, Yunnan Academy of Agricultural Science, Kunming, 650223, People’s Republic of China

    Guoting Tian & Yongchang Zhao

  3. College of Food Science and Technology, Nanjing Agricultural University, Weigang, Nanjing, 210095, People’s Republic of China

    Liyan Zhao

  4. School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, People’s Republic of China

    Tzi Bun Ng

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  1. Dongxue Yang
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  2. Guoting Tian
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  3. Fang Du
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  4. Yongchang Zhao
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  5. Liyan Zhao
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  6. Hexiang Wang
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Correspondence to Hexiang Wang or Tzi Bun Ng.

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Yang, D., Tian, G., Du, F. et al. A Fungal Alpha-Galactosidase from Pseudobalsamia microspora Capable of Degrading Raffinose Family Oligosaccharides. Appl Biochem Biotechnol 176, 2157–2169 (2015). https://doi.org/10.1007/s12010-015-1705-0

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  • DOI: https://doi.org/10.1007/s12010-015-1705-0

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