Isolation, Purification and Enzymatic Properties of Nuclease P1 Fermented by <i>Penicillium citrinum</i>

Ming LI; Huashun YU; Chen YU; Yao WU; Dachun GONG

doi:10.13386/j.issn1002-0306.2020050212

Volume 42 Issue 7

Mar. 2021

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Science and Technology of Food Industry > 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.

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

Ming LI^{1, 2, 3,},
Huashun YU^{2, 3},
Chen YU^{2, 3},
Yao WU^{2, 3},
Dachun GONG^{1, 2, 3, ,}

1.
Three Gorges University, Yichang 443002, China
2.
China Key Laboratory of Light Industry Functional Yeast, China Three Gorges University, Yichang 443002, China
3.
Angel Yeast Co.,Ltd, Special Enzyme Division, Yichang 443003, China

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Received Date: May 19, 2020
Available Online: January 27, 2021

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Abstract

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 (K_m), the maximum reaction rates (V_m) and the catalytic constant (K_ca_t) 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.
- nuclease P1,
- purification,
- kinetic parameter,
- enzymatic property

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References (27)

References

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