食品科学 ›› 2019, Vol. 40 ›› Issue (2): 25-31.doi: 10.7506/spkx1002-6630-20180514-194

• 食品化学 • 上一篇    下一篇

环氧基修饰磁性微球固定化磷脂酶A1

鲍赛,操丽丽,庞敏,潘丽军,侯志刚,水龙龙,李进红,姜绍通*   

  1. (合肥工业大学食品科学与工程学院,安徽省农产品精深加工重点实验室,安徽?合肥 230009)
  • 出版日期:2019-01-25 发布日期:2019-01-22
  • 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 into Epoxy-Modified Magnetic Microspheres

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

  1. (Key Laboratory for Agriculture Products Processing of Anhui Province, School of Food and Engineering, Hefei University of Technology, Hefei 230009, China)
  • Online:2019-01-25 Published:2019-01-22

摘要: 以自制的环氧基修饰的Fe3O4磁性微球为载体固定化磷脂酶A1,采用响应面试验优化固定化条件,并研究固定化酶的酶学性质。结果表明:最佳固定化条件为缓冲液pH?4.0、固定化时间1.9?h、酶液添加量3.6?mL/g,得到的酶活力为3?675?U/g,固定化率为61.1%。固定化酶与游离酶比较,最适pH值向碱性方向偏移1.0,最适温度提高5?℃;贮藏稳定性有一定程度的提高;在重复菜籽油脱胶8?个批次后,仍保留81.1%的酶活力。此外,通过X射线衍射、衰减全反射傅里叶变换红外光谱、扫描电子显微镜和透射电子显微镜等现代分析手段对载体进行表征,表明成功制备纳米尺寸的微球,且微球表面成功修饰上环氧基团。

关键词: 环氧基, Fe3O4磁性微球, 磷脂酶A1, 固定化

Abstract: The epoxy-modified Fe3O4 magnetic microspheres prepared in our laboratory were used as the carrier to immobilize phospholipase A1 (PLA1). Response surface methodology was used to optimize the immobilization conditions. Furthermore, the properties of the immobilized enzyme were studied. The results showed that the optimal immobilization conditions were determined as follows: buffer pH 4.0, reaction time 1.9 h and enzyme dosage 3.6 mL/g. The activity of the immobilized enzyme obtained under these conditions was 3 675 U/g with an immobilization efficiency of 61.1%. Compared with the free enzyme, the optimal reaction pH was shifted to alkaline pH of 1.0 and the optimal reaction temperature of the immobilized enzyme was increased by 5?℃. The storage stability of the immobilized enzyme was increased as well. And the immobilized enzyme retained about 81.1% of the initial activity after the 8th use for rapeseed oil degumming. Moreover, modern analytical methods such as X-ray diffraction (XRD), attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR), scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to characterize the carrier. The results showed that the nano-sized microspheres were successfully prepared with epoxy-modified surfaces.

Key words: epoxy, Fe3O4 magnetic microspheres, phospholipase A1 (PLA1), immobilization

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