灰绿青霉阿魏酸酯酶的分离纯化和酶学性质研究Study on Separation,Purification and Enzymatic Properties of Ferulic Acid Esterase from Penicillium glaucum
徐大伟;高兆建;张铁柱;杨杰;王东星;杜永凯;
摘要(Abstract)：
[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。
关键词(KeyWords)： 灰绿青霉;阿魏酸酯酶;纯化;酶学性质
基金项目(Foundation): 大学生创新创业基金项目(201311998053Y);; 徐州市科技计划项目(XF13C011,XF13C027)
作者(Author): 徐大伟;高兆建;张铁柱;杨杰;王东星;杜永凯;
Email:
DOI: 10.13989/j.cnki.0517-6611.2016.15.008
参考文献(References)：
- [1]YANG S,TANG L,YAN Q,et al.Biochemical characteristics and gene cloning of a novel thermostable feruloyl esterase from Chaetomium sp.[J].Journal of molecular catalysis B:Enzymatic,2013,97:328-336.
- [2]方园,欧仕益,张宁.阿魏酸酯酶的制备及其酶学性质[J].暨南大学学报(自然科学与医学版),2012(5):481-485.
- [3]GOPALAN N,RODRGUEZ-DURAN LV,SAUCEDO-CASTANEDA G,et al.Review on technological and scientific aspects of feruloyl esterases:A versatile enzyme for biorefining of biomass[J].Bioresource technology,2015,193:534-544.
- [4]LIU Q,ZHOU D Y,CHEN L,et al.Effects of feruloyl esterase,non-starch polysaccharide degrading enzymes,phytase,and their combinations on in vitro degradation of rice bran and nutrient digestibility of rice bran based diets in adult cockerels[J].Livestock science,2015,178:255-262.
- [5]ZENG W,CHEN H Z.Air pressure pulsation solid state fermentation of feruloyl esterase by Aspergillus niger[J].Bioresource technology,2009,100(3):1371-1375.
- [6]徐腾洋,方若思,董亚晨,等.黑曲霉XTY液体深层发酵产阿魏酸酯酶的培养基优化研究[J].中国食品学报,2013(6):111-116.
- [7]FAZARY A E,ISMADJI S,JU Y.Stability and solubility studies of native and activated Aspergillus awamori feruloyl esterase[J].Journal of molecular catalysis B:Enzymatic,2009,59(1/2/3):190-196.
- [8]ZHANG S,ZHAI H,WANG L,et al.Expression,purification and characterization of a feruloyl esterase A from Aspergillus flavus[J].Protein expression and purification,2013,92(1):36-40.
- [9]DEBEIRE P,KHOUNE P,JELTSCH J,et al.Product patterns of a feruloyl esterase from Aspergillus nidulans on large feruloyl-arabino-xylo-oligosaccharides from wheat bran[J].Bioresource technology,2012,119:425-428.
- [10]龚燕燕,殷欣,邬敏辰,等.宇佐美曲霉阿魏酸酯酶A基因的克隆表达及酶学性质研究[J].食品与生物技术学报,2013(7):706-712.
- [11]李夏兰,范韵敏,方柏山.来自桔青霉的阿魏酸酯酶的分离纯化、理化性质[J].微生物学报,2010(8):1058-1064.
- [12]陈锦,胡茂丰,吴章华,等.高产阿魏酸酯酶菌株的筛选及其产酶条件优化[J].食品工业科技,2015(8):218-221.
- [13]LAEMMLI U K.Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J].Nature,1970,227:68012-68015.