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中国精品科技期刊2020
常祥祥,田永莉,颜娟,等. 木薯块根中β-葡萄糖苷酶的分离纯化及酶学性质[J]. 食品工业科技,2023,44(3):141−147. doi: 10.13386/j.issn1002-0306.2022050202.
引用本文: 常祥祥,田永莉,颜娟,等. 木薯块根中β-葡萄糖苷酶的分离纯化及酶学性质[J]. 食品工业科技,2023,44(3):141−147. doi: 10.13386/j.issn1002-0306.2022050202.
CHANG Xiangxiang, TIAN Yongli, YAN Juan, et al. Isolation, Purification and Characterization of β-Glucosidase from Cassava Roots[J]. Science and Technology of Food Industry, 2023, 44(3): 141−147. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022050202.
Citation: CHANG Xiangxiang, TIAN Yongli, YAN Juan, et al. Isolation, Purification and Characterization of β-Glucosidase from Cassava Roots[J]. Science and Technology of Food Industry, 2023, 44(3): 141−147. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2022050202.

木薯块根中β-葡萄糖苷酶的分离纯化及酶学性质

Isolation, Purification and Characterization of β-Glucosidase from Cassava Roots

  • 摘要: 研究了木薯块根中β-葡萄糖苷酶的分离纯化及酶学性质。以缓冲液从木薯块根中获得粗提酶液,粗酶酶活力为9.37 U/g木薯干重;再分别通过丙酮沉淀、离子交换层析和凝胶过滤层析进行纯化,β-葡萄糖苷酶酶活力为1.14 U/g木薯干重,经纯化β-葡萄糖苷酶纯度提高了14.62倍,总活力回收率为12.14%,电泳测得其分子量约70 kDa。该酶米氏常数Km为3.60 mmol/L,Vmax为12.36 µmol/(min·mg protein);其最适pH为7.0,pH在6.0~8.0之间有较好的稳定性;在40 ℃以内有良好稳定性,在4 ℃存放30 d酶活力剩余81.78%。Mn2+和K+对酶有一定的促进作用,Al3+、Cu2+、Mg2+、Zn2+、Ca2+、Ba2+、Na+、尿素和SDS对酶没有显著影响(P>0.05),而Fe3+、Fe2+、Ag+和EDTA均会不同程度地抑制酶活性,其中Ag+的抑制作用最强。研究结果可为今后木薯块根中的β-葡萄糖苷酶的应用提供理论依据。

     

    Abstract: The isolation, purification and enzymatic properties of β-glucosidase from cassava roots were studied. The crude enzyme extract was obtained from cassava roots with buffer solution, and the activity of crude enzyme was 9.37 U/g cassava dry weight. Purified by acetone precipitation, ion exchange chromatography and gel filtration chromatography, β-glucosidase activity was 1.14 U/g cassava dry weight, purified β-glucosidase purity increased by 14.62 times, the total activity recovery was 12.14%, the molecular weight of β-glucosidase was about 70 kDa. The Km and Vmax of the enzyme were 3.60 mmol/L and 12.36 µmol/(min·mg protein) respectively. The optimum pH was 7.0, and it was stable when the pH was between 6.0 and 8.0. It had good stability within 40 ℃, and 81.78% enzyme activity remained after 30 days at 4 ℃. Mn2+ and K+ promoted the enzyme to a certain extent. Al3+, Cu2+, Mg2+, Zn2+, Ca2+, Ba2+, Na+, urea and SDS had no significant effect on the enzyme activity (P>0.05). Fe3+, Fe2+, Ag+ and EDTA all inhibited the enzyme activity to varying degrees, among which Ag+ had the strongest inhibitory effect. The results can provide theoretical basis for the application of β-glucosidase in cassava roots in the future.

     

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