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.
出处
《食品与生物技术学报》
CAS
CSCD
北大核心
2007年第1期116-119,共4页
Journal of Food Science and Biotechnology
