食品科学 ›› 2019, Vol. 40 ›› Issue (10): 84-91.doi: 10.7506/spkx1002-6630-20180614-264

• 生物工程 • 上一篇    下一篇

氨基化Fe3O4-SiO2固定化磷脂酶A1

李进红,操丽丽,庞 敏,潘丽军,侯志刚,水龙龙,鲍 赛,姜绍通*   

  1. 合肥工业大学食品与生物工程学院,安徽省农产品精深加工重点实验室,安徽 合肥 230009
  • 出版日期:2019-05-25 发布日期:2019-05-31
  • 基金资助:
    “十三五”国家重点研发计划重点专项(2016YFD0401401-2)

Immobilization of Phospholipase A1 on Amino-Modified Fe3O4-SiO2

LI Jinhong, CAO Lili, PANG Min, PAN Lijun, HOU Zhigang, SHUI Longlong, BAO Sai, JIANG Shaotong*   

  1. Key Laboratory for Agriculture Products Processing of Anhui Province, School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, China
  • Online:2019-05-25 Published:2019-05-31

摘要: 以磁性Fe3O4-SiO2纳米颗粒为载体,研究固定化条件对磷脂酶活力的影响,通过响应面试验得到最优固定化条件为:固定化pH 6.7、固定化温度30 ℃、固定化时间7.9 h、戊二醛质量分数8.3%、加酶量8.2 mL/50 mg,在此条件下酶活力回收率能达到63.6%,蛋白固载率68%。并对制备的固定化磷脂酶的化学组分、形态结构和粒径进行分析,结果表明磷脂酶固定化效果较好,粒径均一,载体平均粒径为200 nm左右。固定化酶热稳定性、pH值稳定性和贮藏稳定性增强,最适反应温度为50 ℃,最适pH 6.0,重复操作10 次后保留60%以上的初始酶活力。

关键词: 磷脂酶A1, 磁性纳米粒子, 固定化

Abstract: In this investigation, Fe3O4-SiO2 magnetic nanoparticles were used as a carrier to study the effects of different operating conditions on the immobilization of phospholipase A1. The optimal conditions determined using response surface methodology were as follows: pH 6.7, temperature 30 ℃, immobilization time 7.9 h, glutaraldehyde concentration 8.3%, and amount of enzyme 8.2 mL/50 mg. Under these conditions, the recovery of enzymatic activity was as high as 63.6% and the percentage of protein immobilization was 68%. The chemical composition, morphological structure and particle size of the immobilized phospholipase were characterized. The results showed that the efficiency of phospholipase immobilization was good; uniform particle size distribution was observed with a mean particle size of about 200 nm. The thermal, pH and storage stability of the immobilized enzyme were enhanced relative to the free one. The optimum reaction temperature for the immobilized enzyme was 50 ℃ and optimum pH 6.0. Over 60% of the enzymatic activity remained after tenth repeated use.

Key words: phospholipase A1, magnetic nanoparticles, immobilization

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