马铃薯细胞壁中果胶酶的提取及特性研究
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(1.贵州大学酿酒与食品工程学院 贵阳 550025;2.贵州医科大学公共卫生学院 贵阳 550025;3.贵州省农业科学院食品加工研究所 贵阳 550025;4.贵州省农业科学院生物技术重点实验室 贵阳 550025)

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贵州省基础研究计划项目(黔科合基础-ZK【2021】重点039);国家自然科学基金项目(32060545,32260602)


Extraction and Characterization of Pectinase from Potato Cell Wall
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(1.School of Liquor & Food Engineering, Guizhou University, Guiyang 550025;2.School of College of Public Health, Guizhou Medical University, Guiyang 550025;3.Institute of Food Processing Technology, Guizhou Academy of Agricultural Sciences, Guiyang 550025;4.Guizhou Provincial Key Laboratory of Agricultural Biotechnology, Guizhou Academy of Agricultural Sciences,Guiyang 550025)

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    摘要:

    通过硫酸铵梯度沉淀结合三相分离法来分离纯化马铃薯细胞壁果胶酶,探究环境条件对马铃薯细胞壁果胶酶活性的影响及其酶学特性。结果表明,当硫酸铵饱和度为60%时,果胶酶达到最大沉淀点。经两次三相分离纯化后果胶酶比活力达到2 012.40 U/mg,纯化倍数5.72,回收率82%。其最适温度20 ℃,最适pH 4.8。Ca2+、Cu2+可显著促进果胶酶酶活力,其中Ca2+对果胶酶促进作用最明显(251%),而Zn2+、Na+对果胶酶活性没有影响,K+、Mg2+对果胶酶有显著抑制作用。在最适温度和最适pH值下反应40 min时酶活力最大(4 667.10 U/g),果胶酶的动力学常数中Km=0.13 mg/mL,Vmax=277.80 U/g。比较果胶酶对胶凝化的马铃薯果胶和原果胶的酶解效果,结果胶凝化果胶对马铃薯细胞壁果胶酶有抗性。

    Abstract:

    The potato cell wall pectinase was isolated and purified by ammonium sulfate gradient precipitation with a combination of three phase partitioning method, and the effects of environmental conditions on the activity and enzymatic features of cell wall pectinase in potato were studied. The results showed that when the saturation of ammonium sulfate was 60%, pectinase reached the maximum precipitation point. Through a process of two three phase partitioning and purification, the specific activity of pectinase, the purification fold and the recovery rate reached 2 012.40 U/mg, 5.72 and 82%. The optimal temperature was 20 ℃ and the optimal pH was 4.8. Ca2+ and Cu2+ could significantly amp up pectinase activity, and Ca2+ possessed the strongest promoting effect on pectinase(251%), while Zn2+ and Na+ had no effect on pectinase activity, Whereas K+ and Mg2+ had a significant inhibiting effect on pectinase activity. The enzymatic activity reached to the climax (4 667.10 U/g) when the reaction was conducted at the optimal temperature and pH for 40 min. The kinetic constants of pectinase were Km=0.13 mg/mL and Vmax=277.80 U/g. Simultaneousl enzymolysis effect the gelled potato pectin and the original pectin were compared. It turned out to be that gelled pectin was resistant to potato cell wall pectinase.

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周盼,雷丹丹,李昊昕,秦礼康,董楠,刘嘉.马铃薯细胞壁中果胶酶的提取及特性研究[J].中国食品学报,2024,24(12):215-223

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  • 收稿日期:2023-12-07
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  • 在线发布日期: 2025-01-23
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