该研究采用溶胶-凝胶法包埋丙酮酸氧化酶,首先利用单因素实验确定了溶胶-凝胶法中每个步骤的最优条件,然后利用正交实验得到最优条件为采用5.0 mmol/L二辛基琥珀酸钠条件下振荡10 min,加入0.672 mmol/L四甲氧基硅烷吸附5 h,包埋24 h。结果表明丙酮酸氧化酶的剩余活力比对照组提高了14%,且丙酮酸氧化酶热稳定性在20~30℃时比对照组提高了2~10倍。该结果为多亚基酶的固定化提供了参考,也为丙酮酸氧化酶的应用提供了技术支持。
Pyruvate oxidase, having multi subunits, plays an important role in food and fermentation industry. In recent years, genetic technology help to greatly increase the yield of pyruvate oxidase. However, the stability of pyruvate oxidase needs to be further improved. In this study, pyruvate oxidase was encapsulated by sol-gel technology. Firstly, single factor experiments were carried out to obtain the best value of each factor. Then an orthogonal experiment was used to optimize the conditions for pyruvate oxidase encapsulation by sol-gel technology. In this study, the optimized condition is: shaking in 5.0 mmol/L dioctyl sulfosuccinate sodium solution for 10 min; adding 0.672 mmol/L tetramethoxysilane for 5 h adsorption and then 24 h for encapsulation. The results showed that the final encapsulated pyruvate oxidase residual activity increased by 14% compared with control sample. And two to ten times increased in thermal stability of the encapsulated pyruvate oxidase than control sample at 20-30 ℃. These results provided a reliable methodology for pyruvate oxidase immobilization and application.
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