Wang Yanhua, Zhu Kai, Liu Hui, Han Pingfang, Wei Ping
Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China.
Sheng Wu Gong Cheng Xue Bao. 2009 Dec;25(12):2036-41.
We immobilized Candida sp. lipase onto seven kinds of industrial adsorption and ion exchange resins. By determining the activity of each immobilized enzyme, the weakly basic anionic exchange resin of D301 showed the best results for the immobilization of Candida sp. lipase. Comparing the scanning electron micrographs of D301 with Novozym 435 (immobilized Candida antarctica lipase B from Novo Nordisk Corp.), we selected D301 as a carrier for the immobilization of Candida sp. lipase. And we pretreated the resin D301 with the bifunctional agent glutaraldehyde and crosslinked it with Candida sp. lipase. The optimal conditions for the immobilization of Candida sp. lipase were as follows: 8 mL of the amount of 5% glutaraldehyde solution, five hours of the time pretreated D301 with glutaraldehyde, 1.0 g/L the concentration of Candida sp. lipase used, pH of the phosphate buffered, 6.0 and 10 hours of time for immobilization, respectively. The activity of immobilized enzyme was over 35 U/mg and the efficiency of immobilization was around 3.5 Ul(mg x h).
我们将假丝酵母脂肪酶固定在七种工业吸附和离子交换树脂上。通过测定每种固定化酶的活性,D301弱碱性阴离子交换树脂在固定假丝酵母脂肪酶方面表现出最佳效果。将D301的扫描电子显微镜图像与诺维信435(诺和诺德公司固定化的南极假丝酵母脂肪酶B)进行比较后,我们选择D301作为固定假丝酵母脂肪酶的载体。然后我们用双功能试剂戊二醛对树脂D301进行预处理,并使其与假丝酵母脂肪酶交联。固定假丝酵母脂肪酶的最佳条件如下:5%戊二醛溶液用量8 mL、用戊二醛预处理D301的时间为5小时、所用假丝酵母脂肪酶的浓度为1.0 g/L、磷酸盐缓冲液的pH值为6.0、固定化时间分别为10小时。固定化酶的活性超过35 U/mg,固定化效率约为3.5 U/(mg·h)。