黑曲霉脂肪酶的分离纯化及酶学性质的研究
Purification and properties of the lipase from Aspergillus niger
作者：师慧;孟尧;范翔;李刚锐;孟延发;
Author：
收稿日期： 年卷（期）页码：2014,29(01):-47-50
期刊名称：华西药学杂志
Journal Name：WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字：黑曲霉;脂肪酶;脂肪酶性质;分离纯化
Key words：
基金项目：“十一五”国家科技支撑计划重点项目(2008BA163B07)
中文摘要
目的以黑曲霉发酵液为原料,分离纯化脂肪酶,并研究其酶学性质。方法以硫酸铵沉淀、Q-Sepharose Fast Flow和Sephacryl S-200凝胶层析等方法分离纯化脂肪酶,利用SDS-PAGE、PAGE、IEF-PAGE等方法进行鉴定。结果所得脂肪酶为单一条带,等电点为3.2,天然分子质量为43.7 kDa。以p-NPP为底物,在pH6、50℃条件下,K m=15.67 mmol·L-1;脂肪酶对多种油脂均有水解作用;此酶在65℃以下,pH3~12稳定,在50℃、pH4表现出相对高的酶活性;Ca2+、Co2+、Mn2+、Ni2+、卵磷脂、胆酸钠对脂肪酶活性有激活作用,Hg2+有抑制作用;PMSF对此脂肪酶有特异性修饰作用。结论确定了黑曲霉脂肪酶的部分酶学性质,为脂肪酶的应用提供了理论依据。
参考文献
[1]Shu CH,Xu CJ,Lin GC.Purification and partial characterization of a lipase from Antrodia cinnamomea[J].Process Biochem,2006,41:734-738.
[2]lker S,Karaoglu SA.Purification and characterization of an extracellular lipase from Mucor hiemalis f.corticola isolated from soil[J].J Biosci Bioeng,2012,114:385-390.
[3]López C,Guerra NP,Rúa ML.Purification and characterization of two isoforms from Candida rugosa lipase B[J].Biotechnol Lett,2000,22:1291-1294.
[4]Mahadik ND,Puntambekar US,Bastawde KB,et al.Production of acidic lipase by Aspergillus niger in solid state fermentation[J].Process Biochem,2002,38:715-721.
[5]汪家政,范明.蛋白质技术手册[M].北京:科学出版社,2002:77-100.
[6]Yadav RP,Saxena RK,Gupta R,et al.Purification and characterization of a regiospecific lipase from Aspergillus terreus[J].Biotechnol Appl Biochem,1998,28:243-249.
[7]Mhetras NC,Bastawde KB,Gokhale DV.Purification and characterization of acidic lipase from Aspergillus niger NCIM 1207[J].Bioresource Technol,2009,100:1486-1490.
[8]Liu ZQ,Zheng XB,Zhang SP,et al.Cloning,expression and characterization of a lipase gene from the Candida antarctica ZJB09193 and its application in biosynthesis of vitamin A esters[J].Microbiol Res,2012,167:452-460.
[9]Sarkar P,Yamasaki S,Basakc S,et al.Purication and characterization of a new alkali-thermostable lipase from Staphylococcus aureus isolated from Arachis hypogaea rhizosphere[J].Process Biochem,2012,47:858-866.
[10]Nthangeni MB,Patterton HG,Tonder Avan,et al.Overexpression and properties of a purified recombinant Bacillus licheniformis lipase:A comparative report on Bacillus lipases[J].Enzyme and Microb Tech,2001,28:705-712.
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