基于自诱导仿生矿化固定化蛋白酶的研究-生物转化与生物催化专业论文.docxVIP

万方数据 万方数据 Research of Immobilization of Proteases Based on Biomimetic mineralization By Autoinduction Abstract Biomimetic mineralization process is used for protease immobilized at room temperature, ambient pressure and neutral pH, which not only provides a new avenue for enzyme immobilization technology, but also further expands the application of biomimetic mineralization. This paper puts biomimetic mineralization processes on the immobilization of bromelain and papain by auto-induction, the study details and the results were recapitulated as follows: Bromelain was immobilized via biomimetic zirconium process by auto- induction. Based on studying the influences of different factors on immobilization effect, we obtained the optimal conditions of immobilized bromelain were: 0.2 mol·L-1 K2ZrF6, 6 mg·mL-1 bromelain, volume ratio of K2ZrF6 and bromelain was 1:2, phosphate buffer was pH 7.5 and the concentration was 0.1 mol·L-1, under the optimum immobilization conditions, activity of immobilized enzyme was about 4 U·mg-1, recovery and entrapped efficiency were about 60 % and 63 %, respectively. It was found that the optimum pH of free and immobilized bromelain were 7.0 and 7.5, respectively, while the optimum temperature were 50 ℃ and 55 ℃ , and under the optimum pH and temperature, activity of immobilized bromelain was about 18 U·mg-1. Furthermore, the immobilized bromelain demonatrated excellent pH, temperature, storage and recycling stability. Kinetic analyses showed the affinity of immobilized bromelain weaken due to the steric hindrance. We studied the biomimetic zirconium proess by auto-induced of bromelain, and proposed the mechanism of biomimetin mineralization. Papain was immobilized through biomimetic zirconium process by auto- induction By incubating papain in a zirconia precursor solution, the papain-zirconia composite formed rapidly, and papain wan encapsulated at the same time. Under optimal immobi- lization conditions, the acti