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文章摘要
Ni2+螯合的琼脂糖凝胶载体用于重组酶SUMO-Hep I-His的固定化
Immobilization of an Sumo-Heparinase I-His Fusion Enzyme on Ni2+ Chelating Agarose Gel Carrier
  
DOI:10.3969/j.issn.1673-1689.2018.08.007
中文关键词: 肝素酶I  GLK-Gel Ni  固定化  酶活
英文关键词: Hep I,GLK-Gel Ni,immobilization,enzyme activity
基金项目:
作者单位
张轩月 江南大学 药学院江苏 无锡 214122 
程咏梅 无锡贝迪生物工程股份有限公司江苏 无锡 214092 
宋志新 常州生化药业(江苏)有限公司江苏 常州213000 
张华 常州生化药业(江苏)有限公司江苏 常州213000 
陈敬华 江南大学 药学院江苏 无锡 214122 
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中文摘要:
      以Ni2+螯合的琼脂糖凝胶(GLK-Gel Ni) 作为固定化载体,对融合肝素酶Sumo-Hep I-His的固定化进行了研究,实现了融合酶的一步纯化和固定化。通过对固定化条件的优化,得到较优的固定化体系:载体与粗酶质量比为30∶1,固定化pH为7.0,吸附时间为6 h,该条件下获得的固定化酶酶活10.07±0.35 IU/mL载体。酶学性质研究结果表明:固定化酶在30 ℃的热稳定性相对于游离酶显著提高,固定化酶半衰期是游离酶的8倍,最适反应温度提高了3 ℃,反应pH的耐受性也有了明显的提高。此外,与游离酶相比,固定化酶的储藏稳定性和可重复利用性良好,在4 ℃下放置60 d能保持76%的初始活性;固定化酶重复使用8次后,酶活仍剩余75.8%;而且GLK-Gel Ni载体本身具有良好的重复利用性,重复固定5次Sumo-Hep I-His后,载体对酶的活性吸附仍能达到88.9%。整体而言,固定化Sumo-Hep I-His显现了较好的工业应用潜能。
英文摘要:
      In this study,immobilization of Sumo-Hep I-His was investigated using Ni2+ chelating agarose gel as carrier. Under optimal conditions,i.e.,weight ratio of barrier to rude enzyme 30∶1,pH 7.0 and immobilized time 6 h,Sumo-Hep I-His was successfully immobilized with an apparent enzymatic activity of 10.07±0.35 IU/mL carrier. Compared with free enzyme,immobilized Sumo-Hep I-His showed great improvement in thermo stability (the half-live was 8 times of free Hep I at 30 ℃ as well as pH stability. The storage and operation stability of the immobilized enzyme was also significantly improved,as 76% relative enzymatic activity was remained after storing for 60 days at 4 ℃ and 75.8% relative enzymatic activity was remained after 8 reaction cycles. GLK-Gel Ni gel also showed good recycling stability that 88.9% adsorptive capacity was remained after 5 cycles immobilization. Conclusively,immobilized Hep I showed great potential in industrial application.
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