脂肪酶菌种的筛选、鉴定及酶学性质研究  

Isolated and Identification of Lipase Producing Strain and Studies on Its Properties

作　　者：胡长浩[1] 施文芳[1] 沈麟[1] 林琳[1] 

机构地区：[1]福建师范大学生命科学学院,福建福州350108

出　　处：《生物技术》2009年第2期53-57,共5页Biotechnology

基　　金：国家自然科学基金项目(No.30270033);福建省自然科学基金项目(C0410009);福建省科技创新平台建设计划项目(2006H0085)资助

摘　　要：目的:筛选耐热脂肪酶产生菌并研究其酶学特性。方法:以固体平板法筛选产酶菌株,单因素和分子进化树分析确定产酶条件、酶学性质及分类地位。结果:产酶条件初步确定为2%糊精、3%酵母膏、MgSO4·7ddH2O 0.1%、K2HPO40.1%、CaCl20.1%、三丁酸甘油酯乳化液0.1%,起始pH7.0,装量为50mL/250mL摇瓶,发酵温度及周期为35℃,22h。酶学性质表明:较适作用温度,pH分别为45℃,10.0,在pH5.0～11.0范围内稳定;该酶具有较好的热稳定性,60℃下温浴30min仍保留77%的活性;可水解长链油脂(C18∶0);0.01%的SDS及10mmol/L的Pb2+、Ba2+抑制该酶的活性,0.05%的EDTA没有影响,0.01%的Triton X-100、Tween80及10mmol/L的K+有促进作用;16S rDNA的分子进化树分析表明,该菌株与Staphylococcus aureus有最紧密的亲缘关系。结论:获得了一株耐热脂肪酶产生菌FL-12,并初步鉴定为Staphylococcus aureus。Objective：Isolated and identification of thermostable lipase producing strain and studies on its properties.Method： Isolated lipase producing strain,determine lipase producing conditions、lipase properties,taxonomic status by means of solid plate,single factor and molecular phylogenetic tree methods.Result： Bacterium FL-12 ,which can produce lipase , was isolated from hot spring and the optimum cultural and lipase secretion condition was optimized as follows： 2% dextrine,3% yeast extract, MgSO4·7ddH2O 0.1%, K2HPO4 0.1%, CaCl2 0.1%, tributyrylglycerols emulsion 0.1%, pH7.0 , 50mL/250mL flask and grow at 35℃ for 22h. 16S rDNA sequencing of the bacterium shows strong homology with Staphylococcus aureus. Lipase from FL-12 was purified and shows highest catalytic activity at pH 10.0 and 45℃. The enzyme stables at wide pH range of 5.0～11.0 and shows high thermostability which retain 77% activity when incubated at 60℃ for 30min. The enzyme catalyze C4 to C18 glyceride. The activity of the enzyme was improved by 0.01% TritonX-100 and K^＋ while inhibited by Pb^2＋, Ba^2＋, 0.01% SDS and 0.05% EDTA. Conclusion： A thermostable lipase producing strain was isolated and identification of Staphylococcus aureus.

关 键 词：葡萄球菌脂肪酶 热稳定性 筛选 酶学性质 16SrDNA 

分 类 号：Q939.115[生物学—微生物学]