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中国精品科技期刊2020
李明,余华顺,喻晨,等. 桔青霉产核酸酶P1酶分离纯化及其酶学性质[J]. 食品工业科技,2021,42(7):89−94. doi: 10.13386/ j.issn1002-0306.2020050212.
引用本文: 李明,余华顺,喻晨,等. 桔青霉产核酸酶P1酶分离纯化及其酶学性质[J]. 食品工业科技,2021,42(7):89−94. doi: 10.13386/ j.issn1002-0306.2020050212.
LI Ming, YU Huashun, YU Chen, et al. Isolation, Purification and Enzymatic Properties of Nuclease P1 Fermented by Penicillium citrinum [J]. Science and Technology of Food Industry, 2021, 42(7): 89−94. (in Chinese with English abstract). doi: 10.13386/j.issn 1002-0306.2020050212.
Citation: LI Ming, YU Huashun, YU Chen, et al. Isolation, Purification and Enzymatic Properties of Nuclease P1 Fermented by Penicillium citrinum [J]. Science and Technology of Food Industry, 2021, 42(7): 89−94. (in Chinese with English abstract). doi: 10.13386/j.issn 1002-0306.2020050212.

桔青霉产核酸酶P1酶分离纯化及其酶学性质

Isolation, Purification and Enzymatic Properties of Nuclease P1 Fermented by Penicillium citrinum

  • 摘要: 桔青霉(Penicillium citrinum)产核酸酶P1浓缩液采用活性炭脱色、硫酸铵分级沉淀、脱盐和凝胶层析等分离技术,得到核酸酶P1纯组分,并研究了该酶的酶学性质。该酶纯化后比酶活达到33967 U/mg,纯化倍数为8.48倍;该酶的米氏常数Km、最大反应速度Vm和催化常数Kcat分别为2.50 mmol/L、0.0864 mmol/(mL·min)和252.43 s−1。该酶最适温度为75 ℃,热稳定范围60~75 ℃;最适pH为5.5,pH稳定范围为4.0~6.0;Zn2+在1 mmol/L条件下对核酸酶P1有很好的激活作用,Cu2+和Co2+对该酶的抑制作用明显,而Ni2+、Fe2+、Mn2+等离子具有不同程度的抑制作用。本研究对于该酶的广泛应用奠定了科学基础。

     

    Abstract: The nuclease P1 was purified to obtain pure component by activated carbon decolorization, (NH4)2SO4 precipitation, desalination and gel chromatography and its enzymatic properties was investigated. This purified enzyme had a specific activity of 33967 U/mg protein after 8.48-fold purification. The Michaelis constant (Km), the maximum reaction rates (Vm) and the catalytic constant (Kcat) of the purified enzyme were 2.50 mmol/L, 0.0864 mmol/(mL·min) and 252.43 s−1, respectively. The optimization pH and temperature for the nuclease P1 were at pH5.5 and 75 ℃. The enzyme was stable in the temperature range from 60 to 75 ℃ and in the pH range from 4.0 to 6.0. Zn2+ had a positive effect on the enzyme activity, while Cu2+ was a strong inhibitor of nuclease P1, Ni2+、Fe2+、Mn2+ had the different inhibition. This research laid a scientific foundation for the extensive application of the enzyme.

     

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