文章摘要
Production, Purification and Characterization of Alkaline β-Glucosidase Isolated from Agrobacterium sp. PJD-1-1
来源于土壤农杆菌PJD-1-1的碱性β-葡萄糖苷酶的生产、纯化及酶学性质研究
  
DOI：doi:10.3969/j.issn.1005-7021.2017.05.006
中文关键词: alkalinity  β-glucosidase  ion exchange chromatography  metal ions；Agrobacterium sp. PJD-1-1
英文关键词: 碱性  β-葡萄糖苷酶  离子交换层析  金属离子  土壤农杆菌PJD-1-1
基金项目:Fund for Agro scientific Research in the Public Interest (201203072)； Natural Science Foundation of Jiangxi Province (20142BAB214010)； Research and Development Projects of Jiangxi Province (20161BBF60138) 公益性行业(农业) 科研专项(201203072); 江西省青年科学基金资助项目(20142BAB214010); 江西省重点研发计划项目(20161BBF60138)
作者单位
Yao Jian Inst. of Agric. Appl. Microbiol., Jiangxi Acad. of Agric. Sci., Nanchang 330200 
CHEN Qing-long Inst. of Agric. Appl. Microbiol., Jiangxi Acad. of Agric. Sci., Nanchang 330200 
ZHONG Guo-xiang Inst. of Agric. Appl. Microbiol., Jiangxi Acad. of Agric. Sci., Nanchang 330200 
GUI Lun Inst. of Agric. Appl. Microbiol., Jiangxi Acad. of Agric. Sci., Nanchang 330200 
BI Wen-hui Dept. of Food Sci. & Engin., Shangdong Agric. & Engin. Uni., Jinan 250100 
摘要点击次数: 2021
全文下载次数: 405
中文摘要:
      In order to screen novel β-glucosidase producing strains from environment, one targeted novel strain PJD-1-1 producing β-glucosidase were isolated from putrefied sugarcane leaves with screening and spreading plate. 16S rDNA analysis revealed it was a novel Agrobacterium sp. When the strain was incubated at initial pH 7.0, 20 ℃ with lactose as carbon and NaNO3 as nitrogen sources, the maximum enzyme activity was 3.92 U/mg. β-glucosidase from this strain was purified using (NH4)2SO4 precipitation followed by dextran gel filtration chromatography and ion exchange chromatography. A purifying fold of 4.85 with gaining rate of 8.0% was obtained. SDA-PAGE analysis of the purified enzyme showed that it was a clear and pure band with molecular mass of ca. 40 kDa. The most optimum activity of the enzyme was at 50 ℃ and pH at 8.0. The enzyme could maintain stability under the conditions below 50 ℃. Hg2+ and Ag+ heavily inhibited the enzyme activity suggesting that the active catalytic sites of the enzymes might possess thiol radical. Ba2+, Ca2+, Pb2+, Co2+, Zn2+, Mn2+, Na+, K+, EDTA, and urea had no obvious effects on the enzyme activity. It is concluded that the novel strain Agrobacterium sp. PJD-1-1 producing β-glucosidase was successfully screened from putrefied sugar cane leaves. The produced enzyme had thermal stability, alkaline feature and metal ions tolerance made it useful in the food and broad potential applications in other fields.
英文摘要:
      为从环境中筛选新的β-葡萄糖苷酶生产菌株，利用筛选平板涂布法从腐败的甘蔗叶中筛选目的菌株，从中筛选到1株能产β-葡糖糖苷酶的菌株PJD-1-1，16S rDNA鉴定该菌株为新型的土壤农杆菌。该菌株在20 ℃，初始pH 7.0，以乳糖为碳源，NaNO3为氮源的条件下，其酶活性最高为3.92 U/mg。经硫酸铵沉淀、葡聚糖凝胶过滤层析和离子交换层析纯化后（纯化倍数4.85，得率8.0%），SDA-PAGE分析表明得到一条分子量大小为40 kDa条带；该酶的最适反应温度和pH分别为50 ℃和 8.0；在低于50 ℃条件下保有较好的稳定性；Hg2+和 Ag+对酶活性有明显的抑制作用，表明该酶的催化活性中心可能含有硫醇基，Ba2+、Ca2+、Pb2+、Co2+、Zn2+、Mn2+、Na+、 K+、EDTA和尿素对酶活性没有明显的影响。该酶的温度稳定性、碱性特征以及对金属离子的耐受性等使其在食品及其他领域具有广泛的应用潜力。
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