产碱性木聚糖酶菌株的筛选及酶学性质.pdf

HEREDITAS (Beijing) 2012 3 , 34(3): 356―365 ISSN 0253-9772 研究报告 DOI: 10.3724/SP.J.1005.2012.00356 产碱性木聚糖酶菌株的筛选及酶学性质 单志琼, 周峻岗, 周宇飞, 袁汉英, 吕红 , , 200433 从青海盐碱湖土壤中筛选到 25 株产碱性木聚糖酶的菌株, 其中编号为 QH14 的菌株产酶量达 648.79 U/mL, 纯化后比活可达 1148.56 U/mg。16 SrDNA 鉴定表明菌株 QH14 属于短小芽孢杆菌, 命名为 Bacillus sp. QH14 。从该菌株的基因组中克隆获得了碱性木聚糖酶编码基因XynQH14 , 并在大肠杆菌 E.coliBL21(DE3) 中获 得重组表达。通过 Ni-NTA 亲和层析分离纯化后的重组 QH14 木聚糖酶比活达 700.47 U/mg 。该碱性木聚糖酶 的酶促反应最适温度为 60 ℃, 最适 pH 为 9.2; 55 ℃处理 1h 仍保持 50% 的活力; 在 pH7.0~11 条件下 37 ℃处理酶 液 24 h 后均保持 80% 以上的活力, 且在 pH11 缓冲溶液中 50 ℃处理 24 h 仍保持 31.02% 的酶活, 显示了该碱性 木聚糖酶较好的热稳定性和碱稳定, 提示该碱性木聚糖酶在制浆造纸、纺织等行业的应用潜力。 木聚糖酶; 短小芽孢杆菌; 耐热性; 耐碱性 Isolation and characterization of an alkaline xylanasefrom a newly isolated Bacillus sp. QH14 SHAN Zhi-Qiong, ZHOU Jun-Gang, ZHOU Yu-Fei,YUAN Han-Ying, LV Hong State Key Laboratory of Genetic Engineering, Institute of Genetics , School of Life Sciences, Fudan University, Shanghai 200433, China Abstract: Twenty-fivealkaline xylanase producing strains were isolated from Qinghai Lake side soil samples. Among these strains, QH14 produced 648.79 U/mLxylanase, and the enzymatic specific activity was 1148.56 U/mg after purifica- tion. This alkaline xylanase producing strain belongs to genus Bacillus based on16S rDNA sequencing analysis and then was designated as Bacillus sp. QH14. The alkalinexylanaseencoding gene, XynQH14 , was cloned from Bacillus sp. QH14 and expressed in Escherichiacoli BL21 (DE3). The specific activity of the recombinant xylanase XynQH14 was 700.47 Umg−1 after purification by Ni-NTA affinity chromatography. The opt