肌酐水解酶菌株S-09发酵条件优化及酶的分离纯化  

Optimization of fermentation conditions of creatininase-producting Exiguobacterium sp. S-09 and separation and purification of creatininase

作　　者：石群[1,2] 张庆芳[1,2] 杨丽娜[1,2] 任楠楠[1,2] 乔慧[1,2] 迟乃玉[1,2] SHI Qun ZHANG Qingfang YANG Lina REN Nannan QIAO Hui CHI Naiyu(School of Life Science and Biotectmology, Dalian University, Dalian 116622, China Liaoning Marine Microbial Engineering and Technology Center, Dalian 116622, China)

机构地区：[1]大连大学生命科学与技术学院,辽宁大连116622 [2]辽宁省海洋微生物工程技术研究中心,辽宁大连116622

出　　处：《中国酿造》2017年第4期20-25,共6页China Brewing

基　　金：国家高技术研究发展计划‘863计划’(2007AA021306)

摘　　要：该实验以海洋来源的微小杆菌(Exiguobacterium sp.)S-09为出发菌株,对其发酵培养基和产酶条件进行优化。并将粗酶液经硫酸铵盐析、透析、超滤离心和Sephadex G-100凝胶过滤层析进行分离纯化。结果表明,其最佳发酵培养基为0.5%的肌酐作为碳源、0.3%胰蛋白胨和0.2%玉米浆作为复合氮源,诱导物肌酸含量为0.3%;其最佳发酵条件为作用pH值7.5、温度25℃、装液量50 mL/250 mL、接种量3%。Fe^(2+)和Mn^(2+)能显著促进产酶。十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)凝胶电泳结果显示,纯化后的肌酐水解酶分子质量为24.3 ku。Using Exiguobacterium sp. S-09 from marine as original strain, the fermentation medium and creatininase-producing condition were opti- mized. The crude enzyme was separated and purified by salting out, dialysis, ultrafiltration and Sephadex G-100 gel filtration chromatography. The results showed that the optimum fermentation conditions were as follows： creatinine 0.5%, tryptone 0.3%, corn steep liquor 0.2%, creatine 0.3%, initial pH 7.5, culture temperature 25℃, liquid volume 100 ml/250 ml, inoculum 3%. Fe〉 and Mn^2＋ can strongly activate enzyme production. SDS-PAGE electrophoresis showed that molecular mass of creatininase purified was about 24.3 ku.

关 键 词：肌酐水解酶 培养基 发酵条件 优化 分离纯化 

分 类 号：Q93[生物学—微生物学]