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Thermoanaerobacterium saccharolyticum JW/SL-YS485来源的GH11家族木聚糖酶的克隆表达及其应用 被引量:2
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作者 裴建军 李杰 +1 位作者 李琦 赵林果 《化工进展》 EI CAS CSCD 北大核心 2018年第4期1544-1551,共8页
对Thermoanaerobacterium saccharolyticum JW/SL-YS485来源的GH11家族木聚糖酶基因xyn11进行了克隆表达研究。xyn11基因全长636bp,编码211氨基酸。通过构建重组质粒,使其在大肠杆菌中实现了活性表达,酶活达到13.8U/mL。重组酶通过热处... 对Thermoanaerobacterium saccharolyticum JW/SL-YS485来源的GH11家族木聚糖酶基因xyn11进行了克隆表达研究。xyn11基因全长636bp,编码211氨基酸。通过构建重组质粒,使其在大肠杆菌中实现了活性表达,酶活达到13.8U/mL。重组酶通过热处理和Ni亲和层析达到电泳纯,SDS-PAGE显示其分子量为20000,与理论分子量吻合。重组酶的最适反应温度为65℃,最适反应pH为4.5,在pH 3.5~6.0范围内保持较高的稳定性,60℃保温1h,酶活还残存60%,Cu^(2+)对该酶有激活作用。重组酶底物特异性强,且不能降解木二糖和木三糖。重组酶以榉木木聚糖为底物,其V_(max)和K_m分别为9809U/mg和5.9mg/mL。榉木木聚糖质量浓度为25g/L,重组木聚糖酶用量为20U/g,在50℃、pH 4.5条件下水解12h低聚木糖的得率为27.8%,水解产物主要由木二糖和木三糖组成,且不产生任何木糖。结果表明,该木聚糖酶催化特性优异,可用于低聚木糖的制备。 展开更多
关键词 木聚 嗜热糖杆菌 水解 低聚木
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Isolation and Production of Novel β-galactosidase from a Newly Isolated, Moderate Thermophile, Bacillus sp. Strain B1.1
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作者 Phimchanok Jaturapiree Suganya Phuengjayaeam +2 位作者 Pomtip Seangsawang Witsanu Srila Chirakam Muangnapoh 《Journal of Food Science and Engineering》 2012年第7期395-402,共8页
The enzyme β-galactosidase (lactase; EC 3.2.1.23) is a commercially important enzyme due to its various applications in dairy and food industries, which are based on the β-galactosidase-catalysed hydrolysis of lac... The enzyme β-galactosidase (lactase; EC 3.2.1.23) is a commercially important enzyme due to its various applications in dairy and food industries, which are based on the β-galactosidase-catalysed hydrolysis of lactose into glucose and galactose. The objectives of this work were to identify novel and attractive sources of this industrially relevant enzyme, and to study the effect of selected growth parameters (carbon source, lactose concentration, nitrogen source, peptone concentration, initial pH and temperature) on the formation of β-galactosidase. Based on a screening of isolates from Tha Pai hot spring, Mae Hong Son Province, Thailand, strain BI.1 was selected for further studies. Strain BI.1 is a Gram-positive, rod-shaped, catalase-positive bacterium that forms endospores. Based on the sequence of the 16S rDNA determined, this isolate is most closely related to Anoxybacillus sp. and Bacillus sp., and hence the strain is designated as Bacillus sp. B 1. I.β-Galactosidase was produced by this strain with lactose and peptone as carbon and nitrogen sources, respectively. Optimal enzyme production occurred at an initial culture pH of 8.5 and at 45 ℃. Under these optimum culture conditions, maximal volumetric and specific β-galactosidase activity of 0.478 U mL^-1 and 0.338 U mg^-1 protein, respectively, were obtained after 13 h of cultivation in a medium contain 2.5% lactose, 2.0% peptone, 0.3% K2HPO4, 0.1% KH2PO4 and 0.05% MgSOa·7H2O. 展开更多
关键词 Β-GALACTOSIDASE ISOLATION PRODUCTION THERMOPHILE prebiotic.
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