一种高耐热乳糖酶的异源表达、固定化及酶学性质研究

乳糖酶在乳制品生产中具有重要作用,但是目前商用的乳糖酶耐热性低,可重复利用率差。为了解决以上问题,该实验用枯草芽孢杆菌WB800异源表达了激烈热球菌(Pyrococcus furiosus)来源的β-半乳糖苷酶(β-galactosidase,BgaS)。该酶最适温度为90℃,最适pH值为7,具有良好的应用潜力。为了提高该乳糖酶的重复利用率,该实验用壳聚糖微球为载体,采用交联法实现了BgaS的固定化并对交联条件进行优化,确定最适交联条件为:戊二醛26.50 g/L,交联温度35℃,交联时间2.5 h,酶添加量5.94 U/mL,优化后酶活力回收率达55.11%。固定化酶相较游离酶,最适pH与最适温度相同,且在非最适pH条件下,固定化酶的相对酶活力较游离酶普遍提升了18.23%以上,并且固定化酶在95℃条件下孵育90 min仍保有80%以上的相对活性,较游离酶提升了11.31%。并且固定化酶回收利用率高,操作稳定性好,最适条件下处理3 h即可使牛乳中乳糖含量降低至0.5%以下,达到无乳糖水平,为工业化应用提供了技术支撑。

Removal of ammonium-N from ammonium-rich sewage using an immobilized Bacillus subtilis AYC bioreactor system

A self-design bioreactor system employing a fixed bed operation process with immobilized Bacillus subtilis AYC beads Ibr NH4^-N removal from slightly polluted water was proposed. Polyvinyl alcohol and Na-alginate were used as a gel matrix to entrap Bacillus subtilis AYC to form the immobilized beads. The NH4＋-N removal process was studied in a intermittent operation mode to examine the start-up and steady state behaviors of the immobilized AYC in the reactor. The results indicated that the reactor was in the start-up state during the first week. NH4＋-N began to be steadily removal since the second week, and the nitrogen removal rate was between 84.61% and 96.19% when the hydraulic retention time （HRT） was 30 rain. To apply Bacillus subtilis AYC to develop a practical nitrogen removal system and further understand its nitrogen removal ability, the bioreactor was continuously operated under different experimental perameters. The results showed that under the optimum conditions of an HRT of 20 min and DO of 3.77-5.80 mg/L, the NH4＋-N removal rate reached 99.55%. The NH4＋-N removal rate increased as the C/N ratio increased. However, a high C]N may cause a high residual carbon level in the effluent, therefore, the most suitable C/N ratio was 10. In addition, the results showed that the bioreactor system could remove many types of nitrogen such as NH4＋-N, NO3--N and organic-N, and had a good performance for inorganic nitrogen removal from sewage.

壳聚糖固定化枯草芽孢杆菌水溶液中铜离子的吸附(英文)

研究了壳聚糖固定化枯草芽孢杆菌吸附铜离子的性能,分析了pH、吸附剂投加量、温度、铜离子初始浓度和时间对铜离子吸附的影响。结果表明,pH对铜离子的吸附有较大的影响。此外,固定化枯草芽孢杆菌吸附剂比空白吸附剂具有更强的吸附性能。整个吸附过程符合朗缪尔吸附模型(R2=0.994),最大吸附量为100.70mg/L。动力学模型拟合结果表明,实验符合准二级动力学模型,线性相关指数大于0.999。吸附剂能在0.1mol/LNaOH溶液中被成功解吸。

壳聚糖固定化AS1．398中性蛋白酶稳定性的研究

对以壳聚糖为载体，由戊二醛作交联剂。制备的固定化ＡＳ１．３９８中性蛋白酶的稳定性进行了研究。实验结果表明，固定化ＡＳ１．３９８中性蛋白酶对热、乙醇、尿素以及ｐＨ值的稳定性均有明显的提高。