胞外单宁酶纯化及酶学性质的研究.PDF

※生物工程 食品科学 2011, Vol. 32, No. 13 239 1,2 1, 2, 1,2 3 2,3 2, 3 王维维 ，邱树毅 * ，谢晓莉 ，吴鑫颖 ，李广神 ，何顺荣 (1.贵州大学生命科学学院，贵州 贵阳 550025 ；2.贵州省发酵工程与生物制药重点实验室，贵州 贵阳 550003 ； 3.贵州大学化学与化工学院，贵州 贵阳 550003) 采用黑曲霉 B0201 固态生料发酵生产胞外单宁酶，经过硫酸铵分级沉淀、DEAE- 纤维素层析、葡聚糖 G-150 凝胶柱层析纯化，得到电泳纯的单宁酶，纯化倍数达到 18.44。对该酶的性质研究表明：单宁酶的相对分 子质量大小约为 58800 ，该酶的最适反应温度是 40 ℃，最适反应pH 值是 5.0 ；以PG 为底物，米氏常数K m 为 1.08mmol/L，Vmax 为 104.1 μmol/(L ·min) 。 单宁酶；生料发酵；纯化 Purification and Enzymatic Properties of Extracellular Tannase from Aspergillus niger 1,2 1,2, 1,2 3 2,3 2,3 WANG Wei-wei ，QIU Shu-yi *，XIE Xiao-li ，WU Xin-ying ，LI Guang-shen ，HE Shun-rong (1. College of Life Science, Guizhou University, Guiyang 550025, China ； 2. Guizhou Province Key Laboratory of Fermentation and Biopharmacy, Guiyang 550003, China ； 3. School of Chemistry and Chemical Engineering, Guizhou University, Guiyang 550003, China) Abstract ：Extraction with sodium citrate-citric acid buffer, (NH4)2 SO4 fractional precipitation, DEAE-cellulose column separation and Sephadex G-150 column purification were sequentially carried out to obtain an electrophoretically pure extracel- lular tannase from solid state cultures of Aspergillus niger B0201, with a purification fold of 18.44. The enzyme had a molecular weight of roughly 58800 and the optimum reaction temperature and pH were 40 ℃ and 5.0, respectively. When the substrate was propyl gallate (PG), the Km was 1.08 mmol/L and the Vmax 104.1 μmol/(L ·min). Key words ：tannase ；fermentation without cooking ；purification 中图分类号：