CAJ | 학술논문

目的:获得库拉索芦荟过氧化氢酶纯品并对其性质进行研究.方法:采用硫酸铵分级沉淀,DEAE-Sepharose阴离子交换层析,CM-Sepharose阳离子交换层析和Sephacryl S-200凝胶层析对酶蛋白进行分离纯化,采用SDS-PAGE鉴酶的纯度、分子质量.结果:从芦荟中分离纯化获得电泳纯的过氧化氧酶,纯化倍数为228.05倍,酶活力回收率为14.10%,比活力达17427.30U/mg.该酶的分子质量为239.90kD,亚基分子质量为60.60kD.推测该酶由4个相同亚基构成.该酶最适温度为45℃,最适pH值为7.5,以过氧化氢为底物测定该酶的表观K_m为34mmol/L.结论:成功分离纯化获得过氧化氢酶,该酶具有良好的热稳定性和酸碱耐受性.
목적:획득고랍색호회과양화경매순품병대기성질진행연구.방법:채용류산안분급침정,DEAE-Sepharose음리자교환층석,CM-Sepharose양리자교환층석화Sephacryl S-200응효층석대매단백진행분리순화,채용SDS-PAGE감매적순도、분자질량.결과:종호회중분리순화획득전영순적과양화양매,순화배수위228.05배,매활력회수솔위14.10%,비활력체17427.30U/mg.해매적분자질량위239.90kD,아기분자질량위60.60kD.추측해매유4개상동아기구성.해매최괄온도위45℃,최괄pH치위7.5,이과양화경위저물측정해매적표관K_m위34mmol/L.결론:성공분리순화획득과양화경매,해매구유량호적열은정성화산감내수성.
Objective: To obtain catalase from aloe and explore its enzymaological properties. Methods: Catalase was isolated and purified through fractional ammonium sulfate precipitation, ion-exchange chromatography on DEAE-Sepharose, ion-exchange chromatography on CM-Sepharose and gel filtration on Sephacryl S-200. SDS-PAGE was used to identify the purity and relative molecular mass of the catalase. Results: Aloe catalase revealed a 228.05-fold purification and a recovery rate of activity of 14.10%. The specific activity of the purified aloe catalase was measured to be 17427.30 U/mg. Aloe catalase was characteristic of 239.90 kD relative molecular mass and 60.60 kD molecular mass of subunit. This demonstrates that the enzyme consists of four identical subunits. Moreover, the optimal reaction temperature and pH of the enzyme were 45 ℃ and 7.5, and its apparent K_m towards hydrogen peroxide as the substrate was 34 mmol/L. Conclusion: Aloe catalase has been successfully isolated and purified, which exhibits excellent thermostability and acid-alkali tolerance.