Bacillus sublitis JH-1木聚糖酶的纯化及酶学性质  

Purification and characterization of xylanase from Bacillus sublitis JH-1

作　　者：邵婷婷[1] 王春明[1] 李蘅香 钟超[1] 雍晓雨[1] 贾红华[1] 周华[1] 韦萍[1] 

机构地区：[1]南京工业大学生物与制药工程学院,江苏南京211800

出　　处：《生物加工过程》2016年第5期51-55,共5页Chinese Journal of Bioprocess Engineering

基　　金：国家重点基础研究发展计划(973计划)(2013CB733500);国家科技支撑计划(2014BAC33B00)

摘　　要：对枯草芽孢杆菌Bacillus sublitis JH-1胞外木聚糖酶的纯化及酶学性质进行了研究。通过（NH4）2SO4分级沉淀法、透析除盐、DEAE-Sepharose FF弱阴离子交换层析等方法,从枯草芽孢杆菌Bacillus sublitis JH-1发酵液中分离纯化得到了电泳纯的木聚糖酶,其相对分子质量为3.45×10^4,比活力为75 899.68 U/mg。酶学性质研究结果表明：该酶的最适p H为6.0,在最适p H条件下保温2 h后仍能保持75%的活力,而p H越高,活力下降越快,表明为酸性木聚糖酶;最适温度为55℃,在50-60℃保温2 h后仍具有70%左右相对较高的活性,说明该酶具有较强的耐高温性;Fe^2＋、Mg^2＋、Ca^2＋、Zn^2＋、Ba^2＋和低浓度（5 mmol/L）的Fe^3＋对酶的活性有促进作用,而Mn2＋和高浓度（10mmol/L）的Fe^3＋对酶的活性有抑制作用。We purified and characterized xylanase from Bacillus sublitis JH-1. The crude enzyme was purified by ammonium sulfate fractionation,dialysis desalination and DEAE-Sepharose FF anion exchange chromatography.The specific activity was 75 899. 68 U / mg,and its subunit molecular weight was 3. 45 ×10^4 determined by SDS-PAGE.The xylanase showed optimal activity at p H 6. 0.It retained more than 75%of its maximum activity at p H 6. 0 for 2 h,and the higher the p H,the faster the activity lost. The optimal temperature was 55 ℃ and it retained more than 70% of its maximum activity between 50 and 60 ℃ for 2h. Metal ions Fe^2＋,Mg^2＋,Ca^2＋,Zn^2＋,Ba2＋and low concentration（ 5 mmol / L） of Fe^3＋have obviously promoting effect on xylanase activity,whereas Mn2＋and high concentration（ 10 mmol / L） of Fe^3＋inhibited the enzyme.

关 键 词：枯草芽孢杆菌 木聚糖酶 纯化 弱阴离子交换层析 酶学性质 

分 类 号：Q814[生物学—生物工程]