Kishore Devesh, Kayastha Arvind M
School of Biotechnology, Faculty of Science, Banaras Hindu University, Varanasi 221005, India.
Food Chem. 2012 Oct 1;134(3):1650-7. doi: 10.1016/j.foodchem.2012.03.055. Epub 2012 Mar 21.
Response surface methodology was advantageously used to optimally immobilise a β-galactosidase from chick pea onto alkylamine glass using Box-Behnken experimental design, resulting in an overall 91% immobilisation efficiency. Analysis of variance was performed to determine the adequacy and significance of the quadratic model. Immobilised enzyme showed a shift in the optimum pH; however, optimum temperature remained unaffected. Thermal denaturation kinetics demonstrated significant improvement in thermal stability of the enzyme after immobilisation. Galactose competitively inhibits the enzyme in both soluble and immobilised conditions. Lactose in milk whey was hydrolysed at comparatively higher rate than that of milk. Immobilised enzyme showed excellent reusability with retention of more than 82% enzymatic activity after 15 uses. The immobilised enzyme was found to be fairly stable in both dry and wet conditions for three months with retention of more than 80% residual activity.
采用响应面法，利用Box-Behnken实验设计将鹰嘴豆中的β-半乳糖苷酶最佳地固定在烷基胺玻璃上，固定化效率总体达到91%。进行方差分析以确定二次模型的充分性和显著性。固定化酶的最适pH发生了变化；然而，最适温度未受影响。热变性动力学表明，固定化后酶的热稳定性有显著提高。在可溶性和固定化条件下，半乳糖对该酶均有竞争性抑制作用。乳清中的乳糖水解速率相对高于牛奶。固定化酶具有出色的可重复使用性，15次使用后仍保留超过82%的酶活性。发现固定化酶在干燥和潮湿条件下三个月内相当稳定，残留活性保留率超过80%。
