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Isolation of a Ribonuclease with Antiproliferative and HIV-1 Reverse Transcriptase Inhibitory Activities from Japanese Large Brown Buckwheat Seeds

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Abstract

A ribonuclease, with a molecular mass of 22.5 kDa and an N-terminal sequence exhibiting resemblance to previously isolated buckwheat storage proteins and allergens, was isolated from Japanese large brown buckwheat seeds. The ribonuclease was purified using a simple protocol that comprised ion exchange chromatography on Q-Sepharose and DEAE-cellulose and gel filtration on Superdex 75. The ribonuclease exhibited low activity toward poly U, lower activity toward poly C, and very low activity toward poly A and poly G. The enzyme was activated upon exposure to 10 mM of Fe2+ and Zn2+ ions but was inhibited by Ca2+, Mg2+, and Mn2+ ions at the same concentration. The optimum pH and optimum temperature for the enzyme were pH 9 and 60 °C, respectively. It inhibited proliferation of HepG2 hepatoma and MCF 7 breast cancer cells, with an IC50 value of 79.2 and 63.8 μM, respectively. It potently inhibited HIV-1 reverse transcriptase activity with an IC50 of 48 μM. However, there were no antifungal and mitogenic activities.

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Authors and Affiliations

  1. Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou, Fujian, 350002, China

    Susu Yuan, Juan Yan & Xiujuan Ye

  2. School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China

    Tzibun Ng

  3. Key Laboratory of Plant Virology of Fujian Province, Institute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou, Fujian, 350002, China

    Susu Yuan, Juan Yan, Xiujuan Ye & Zujian Wu

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  1. Susu Yuan
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  2. Juan Yan
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  3. Xiujuan Ye
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  4. Zujian Wu
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Correspondence to Xiujuan Ye, Zujian Wu or Tzibun Ng.

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Yuan, S., Yan, J., Ye, X. et al. Isolation of a Ribonuclease with Antiproliferative and HIV-1 Reverse Transcriptase Inhibitory Activities from Japanese Large Brown Buckwheat Seeds. Appl Biochem Biotechnol 175, 2456–2467 (2015). https://doi.org/10.1007/s12010-014-1438-5

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