红曲霉发酵龙虾壳产酸性蛋白酶分离纯化及酶学性质研究  

Purification and Study Enzymatic Properties of Acid Protease from Fermentation of Crayfish Shell with Monascus

作　　者：李莹莹 刘枣 李娇 蒋潇 吴滢滢 王伟平 LI Yingying;LIU Zao;LI Jiao;JIANG Xiao;WU Yingying;WANG Weiping(School of Biological Engineering and Food,Hubei Univ.of Tech.,Wuhan 430068,China)

机构地区：[1]湖北工业大学生物工程与食品学院,湖北武汉430068

出　　处：《湖北工业大学学报》2024年第5期55-59,107,共6页Journal of Hubei University of Technology

基　　金：国家自然科学基金(31101349)。

摘　　要：小龙虾虾壳废弃物排放造成环境污染和资源浪费,而红曲霉可以转化利用虾壳蛋白质。以小龙虾虾壳为唯一氮源,烟灰色红曲霉BQ2液态发酵诱导其产胞外酸性蛋白酶。该酸性蛋白酶经硫酸铵盐析沉淀和DEAE-Sepharose CL-6B阴离子交换柱层析分离纯化,酶活为29.10 U/mL,纯化倍数为11.42,相对分子质量为65000。对其酶学性质进行研究,结果表明该酸性蛋白酶的最适反应pH为4.0,最适反应温度为40℃。该酶属于天冬氨酸蛋白酶,Pepstatin A能强烈抑制其活性,Cu^(2+)和Ca^(2+)能抑制该蛋白酶活性,而Mg^(+)、Zn^(2+)和Fe^(3+)对该蛋白酶的活性具有促进作用。Crayfish shell waste discharge causes environmental pollution and waste of resources.Monascus can convert and utilize shrimp shell proteins.In this paper,crayfish shells were used as the only nitrogen source,and Monascus fuliginosus BQ2 liquid fermentation induced the production of extracellular acid protease.The acid protease was purified by ammonium sulfate and DEAE-Sepharose CL 6B anion exchange column.The enzyme activity was 29.10U/mL,purification ratio was 11.42,and molecular weight was 65 KDa.Its enzymatic properties were investigated.The results showed that the optimal pH value for the enzyme activity was 4.0 and the optimal reaction temperature was 40℃.Pepstatin A strongly inhibited the enzyme activity and the enzyme belongs to the aspartic proteases.Cu ^(2+)and Ca ^(2+)inhibited the activity of the protease.Mg^(+),Zn ^(2+)and Fe ^(3+)promoted the activity of the protease.

关 键 词：红曲霉 小龙虾壳 酸性蛋白酶 酶学性质 

分 类 号：Q556[生物学—生物化学]