摘要
[目的]研究青霉(Penicillium glaucum)XGY36胞外产阿魏酸酯酶的纯化和酶学性质。[方法]菌株胞外产酶发酵液经硫酸铵沉淀、DEAE-cellulose DE52离子交换柱层析、Sephadex-G75分子筛层析进行纯化,得到纯度较高的酶蛋白,并对其酶学性质进行研究。[结果]从菌株发酵液中分离纯化到电泳纯的阿魏酸酯酶,纯化倍数6.54,酶活性回收59.7%。阿魏酸酯酶经SDS-PAGE获得1个条带,其表观分子量为36.5 ku。催化底物阿魏酸乙酯的最适反应温度为50℃,最适p H为5.0。阿魏酸酯酶在60℃以下和p H 3.0~7.0范围内稳定。金属离子Zn^(2+)、Mg^(2+)、Ca^(2+)对酶有明显的激活作用,而Pb^(2+)、Hg^(2+)和Ag+对阿魏酸酯酶有强烈的抑制作用,Na+、K+、Mn^(2+)、Cu^(2+)和Fe^(2+)对酶活的影响不大。[结论]青霉阿魏酸酯酶具有潜在的应用价值,适合应用于食品、医药及生物等领域。
Objective The aim was to study purification and enzymatic properties of feruloyl esterase in the culture solution of Penicillium glau-cum XGY36.[Method] The feruloyl esterase was obtained in the culture solution of Penicillium glaucum XGY36 and was purified to be homoge-neous by the steps of ammonium sulfate precipitation,DEAE-cellulose DE52 chromatographies,and Sephadex G-75 chromatography.The enzyme protein with high purity was obtained,enzymatic properties were studied.[Result] Purification of about 6.54 fold was achieved with an overall yield of 59.7%.The molecular weight of purified feruloyl esterase was estimated to be about 36.5 ku by SDS-PAGE.The optimum temperature and pH of the enzyme activity were 50 ℃and 5.0,respectively in catalytic reaction of ethyl ferulate.The enzyme activity was stable under 60 ℃, and with pH range of 3.0-7.0.The activity of feruloyl esterase was highly enhanced by Zn2 ＋,Mg2 ＋and Ca2 ＋,whereas it was strongly inhibited by the metal ions Pb2 ＋,Hg2 ＋and Ag ＋,while Na ＋,K ＋,Mn2 ＋,Cu2 ＋and Fe2 ＋had no strong effect on feruloyl esterase activity.[Conclusion] Fer-uloyl esterase has potential applied value and fit to apply in the fields of food,medicine and biology etc..
出处
《安徽农业科学》
CAS
2016年第15期24-27,30,共5页
Journal of Anhui Agricultural Sciences
基金
大学生创新创业基金项目(201311998053Y)
徐州市科技计划项目(XF13C011
XF13C027)
关键词
灰绿青霉
阿魏酸酯酶
纯化
酶学性质
Penicillium glaucum
Ferulic acid esterase
Purification
Enzymatic properties
