Biochemistry Laboratory, Department of Life Science, Faculty of Science, University of Sfax, PB 802, 3018 Sfax, Tunisia.
Int J Biol Macromol. 2013 Sep;60:128-33. doi: 10.1016/j.ijbiomac.2013.05.002. Epub 2013 Jun 1.
A newly isolated bacterial strain from Tunisian thermal source was selected for its ability to produce extracellular levanase when grown on levan substrate. The optimization of carbon source, nitrogen source, temperature and initial pH of the growth medium in submerged liquid cultures were investigated. In fact, levan was found to be a good inducer of levanase enzymes. The optimal temperature and pH of the levanase activity were 40 °C and 6.4, respectively. This enzyme exhibited a remarkable stability and retained 75% of its original activity at 55 °C for more than 1 h at pH 6.4. Crude enzyme of the strain rich in levanase was established for the hydrolysis of levan in order to produce fructooligosaccharides with variable degrees of polymerization which could be used in important fields such medicine, food-processing industry and cosmetic. The extracellular levanase of the strain was then, partially purified as determined by SDS-PAGE. The purification was achieved by ammonium sulfate precipitation, gel filtration and DEAE cellulose chromatographies.
从突尼斯热泉中分离到的一株新的细菌菌株,因其在蔗聚糖底物上生长时能够产生胞外蔗聚糖酶而被选中。研究了在液体深层培养中优化碳源、氮源、温度和初始 pH 值对生长培养基的影响。事实上,蔗聚糖被发现是一种很好的蔗聚糖酶诱导剂。该酶的最适温度和 pH 值分别为 40°C 和 6.4。该酶在 pH 值为 6.4 时表现出显著的稳定性,在 55°C 下超过 1 小时仍保留其原始活性的 75%。为了水解蔗聚糖以生产具有不同聚合度的果寡糖,该菌株富含蔗聚糖酶的粗酶制剂被建立,这些果寡糖可用于医学、食品加工和化妆品等重要领域。然后,通过 SDS-PAGE 确定对该菌株的胞外蔗聚糖酶进行了部分纯化。通过硫酸铵沉淀、凝胶过滤和 DEAE 纤维素层析进行了纯化。