中压液相色谱快速分离纯化Kluyveromyces fragilis β-D-半乳糖苷酶  

Purification of Kluyveromyces fragilis β-Galactosidase by Medium-Pressure Liquid Chromotography

作　　者：刘建福[1] 李素芬[2] 陈占洲[2] 陈庆森[1] 王璋[3] 

机构地区：[1]天津商学院天津食品生物技术重点实验室,天津300134 [2]河北工程大学农学院,河北邯郸056001 [3]江南大学食品学院,江苏无锡214036

出　　处：《食品与生物技术学报》2007年第1期116-119,共4页Journal of Food Science and Biotechnology

摘　　要：应用AKTA explorer 100型中压液相色谱快速纯化工艺,系统分离纯化了脆壁克鲁维酵母(Kluyveromyces fragilis)β-D-半乳糖苷酶。K.fragilis发酵生产的菌体细胞经过破碎、离心后的上清液中含有K.fragilis β-D-半乳糖苷酶。粗酶经过AKTA explorer 100中压液相色谱的HiprepR16/10 Source 30Q强阴离子交换柱和Hioad26/60 Superdex凝胶过滤色谱连续两步纯化,酶的回收率为27%,纯化倍数为42。纯化的K.fragilis β-D-半乳糖苷酶在垂直板十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)上,呈现一条染色谱带,表明纯化的酶具有较高的纯度。K.fragilisLFS-8611 β-D-半乳糖苷酶相对分子质量约为60000。The β-D-galactosidase from Kluyveromyces fragilis was purified by medium-pressure liquid chromatography. The enzyme was purified about 42 fold with a yield of 270% total activity by successive Hiprep^R 16/10 Source30Q and Hioad26/60 Superdex medium-pressure liquid chromatography. When the purified enzyme samples were subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis （SDS-PAGE）, single bands were observed for the enzyme samples, this suggested that the enzyme samples were almost pure. The molecular weight estimated for K. fragilis LFS-8611 β-D-galactosidase by sodium SDS-PAGE was 60 000.

关 键 词：中压液相色谱 脆壁克鲁维酵母 Β-D-半乳糖苷酶 分离纯化 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]