Yu Ping, Zhang Yishu, Gu Donglu
a College of Food Science and Biotechnology, Zhejiang Gongshang University , Hangzhou , Zhejiang Province , People's Republic of China.
Bioengineered. 2017 Sep 3;8(5):613-623. doi: 10.1080/21655979.2017.1292188. Epub 2017 Feb 16.
Alkaline pectinase has important applications in the pretreatment of waste water from food processing and in both the fabric and paper industries. In this study, a 2-level factorial design was used to screen significant factors that affect the activity of alkaline pectinase, and the response surface methodology (RSM) with a Box-Behnken design (BBD) was used to optimize their concentrations. Starch, peptone, KHPO and KHPO·3HO were found to significantly affect the activity of alkaline pectinase. Their optimal concentrations were as follows: 4.68% starch, 1.6% peptone, 0.26% KHPO and 0.68% KHPO·3HO. Under the above conditions, the activity of alkaline pectinase was significantly improved to 734.11 U/mL. Alkaline pectinase was purified to homogeneity with a recovery rate of 9.6% and a specific activity of 52372.52 U/mg. Its optimal temperature and pH were 50°C and 8.6, respectively. The purified enzyme showed strong thermo-stability and good alkali resistance. In addition, the activity of alkaline pectinase was improved in the presence of Mg. Cu, Mn, and Co significantly inhibited its activity. This study provides an important basis for the future development and use of a heat-tolerant alkaline pectinase from B. subtilis ZGL14.
碱性果胶酶在食品加工废水预处理以及纺织和造纸工业中都有重要应用。在本研究中，采用二水平析因设计筛选影响碱性果胶酶活性的显著因素，并使用Box-Behnken设计（BBD）的响应面法（RSM）优化其浓度。发现淀粉、蛋白胨、KH₂PO₄和KH₂PO₄·3H₂O对碱性果胶酶的活性有显著影响。它们的最佳浓度如下：4.68%淀粉、1.6%蛋白胨、0.26% KH₂PO₄和0.68% KH₂PO₄·3H₂O。在上述条件下，碱性果胶酶的活性显著提高到734.11 U/mL。碱性果胶酶被纯化至同质，回收率为9.6%，比活性为52372.52 U/mg。其最佳温度和pH分别为50°C和8.6。纯化后的酶表现出较强的热稳定性和良好的耐碱性。此外，在Mg²⁺存在下碱性果胶酶的活性提高。Cu²⁺、Mn²⁺和Co²⁺显著抑制其活性。本研究为未来开发和利用枯草芽孢杆菌ZGL14的耐热碱性果胶酶提供了重要依据。
