Bacillus subtilis LC-9产β-1,3-1,4-葡聚糖酶的酶学性质与催化特性研究 |
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引用本文: | 刘娟娟,吴斌,何冰芳.Bacillus subtilis LC-9产β-1,3-1,4-葡聚糖酶的酶学性质与催化特性研究[J].安徽农业科学,2012(27):13243-13246. |
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作者姓名: | 刘娟娟 吴斌 何冰芳 |
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作者单位: | 南京工业大学,江苏南京,211816 |
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基金项目: | 中国国家基础研究计划(973计划)资助项目(2011CB707400);中国国家高新技术研究和发展计划(863计划)资助项目(2011AA02A202);江苏高校优势学科建设工程资助项目 |
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摘 要: | 目的]系统研究Bacillus subtilis LC-9所产β-1,3-1,4-葡聚糖酶的酶学性质,探讨其在催化应用上的可行性。方法]采用两步阴离子交换层析法对B.subtilis LC-9发酵液中的β-1,3-1,4-葡聚糖酶进行纯化,用SDS-PAGE检测其纯度,研究纯酶的酶学性质及该酶对β-葡聚糖和地衣多糖的降解特性,并采用TLC法检测其水解液中糖的成分。结果]从自行筛选的糖苷酶产生菌B.subtilis LC-9的发酵液中经两步纯化,得到电泳纯的β-1,3-1,4-葡聚糖酶,其比活力达3 502 U/mg,表观分子量为28 kDa;酶的最适反应pH和温度分别为6.5和45℃,且在45℃下稳定;该酶催化地衣多糖的Km值和Vmax分别为4.13 mg/ml和1 238μmol/min/mg,催化大麦β-葡聚糖的Km值和Vmax分别为3.84 mg/ml和1 427μmol/min/mg;该酶降解大麦β-葡聚糖产生寡聚糖,最终产物主要为三糖和四糖,而降解地衣多糖终产物主要为三糖。结论]从Bacillus subtilis LC-9发酵液中纯化获得了电泳纯的β-1,3-1,4-葡聚糖酶,性质研究表明该酶是专一性的β-1,3-1,4-葡聚糖酶,有望用于啤酒、饲料等领域。
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关 键 词: | B.subtilis LC- β- - -葡聚糖酶 纯化 酶学性质 |
Enzymatic and Catalytic Properties of β-1,3-1,4-glucanase from Bacillus subtilis LC-9 |
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Institution: | LIU Juan-juan et al(Nanjing University of Technology,Nanjing,Jiangsu 211816) |
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Abstract: | Objective] To systematically study the enzymatic properties of purified β-1,3-1,4-glucanase from B.subtilis LC-9,and to explore the possibility of its application in catalysis.Method] Purification of β-1,3-1,4-glucanase from B.subtilis LC-9 fermentation broth was carried out by a two-step ion exchange chromatography.The purity of the enzyme was identified by SDS-PAGE.Then,the enzymatic characterizations of the purified enzyme were studied.Finally,the products of the barley β-glucan and lichenan hydrolyzed by the enzyme were analyzed by TLC.Result] The enzyme was purified to homogeneity with a specific activity of 3 502 U/mg.The purified enzyme showed as a single protein band with an apparent molecular mass of 28 kDa.The optimum pH and temperature for the enzyme activity were 6.5 and 45 ℃,respectively.The purified enzyme was stable below 45 ℃.The kinetic parameters Km and Vmax were 4.13 mg/ml,1 238 μmol/min/mg for lichenan and 3.84 mg/ml,1 427 μmol/min/mg for barley β-glucan.The main products were cellubiosyltriose and cellutriosyltraose for barley β-glucan and cellubiosyltriose for lichenan.Conclusion] A β-1,3-,4-glucanase with high activity from B.subtilis LC-9 was purified.The enzyme exhibited strict specificity for β-1,3-1,4-D-glucans.The characterizations of this enzyme showed that the enzyme was suitable for application to the beer and feed industry. |
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Keywords: | B subtilis LC-9 β-1 3-1 4-glucanase Purification Enzymatic properties |
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