催化学报

催化学报 ›› 2016, Vol. 37 ›› Issue (11): 1891-1898.DOI: 10.1016/S1872-2067(16)62487-7

• 论文 • 上一篇    下一篇

磁性纳米颗粒固定化细菌纤维素酶的活性和稳定性

Kandasamy Selvam, Muthusamy Govarthanan, Duraisamy Senbagam, Seralathan Kamala-Kannan, Balakrishnan Senthilkumar, Thangasamy Selvankumar   

  1. a Muthayammal艺术与科学学院生物工程中心, 纳马卡尔 637408, 泰米尔纳德邦, 印度;
    b Mahendra艺术与科学学院生物工程系, 纳马卡尔 637501, 泰米尔纳德邦, 印度;
    c 室兰工业大学环境工程学院应用科学系, 北海道 620024, 日本;
    d 巴拉迪大学海洋生物工程系, 蒂鲁吉拉伯利 620024, 泰米尔纳德邦, 印度;
    e 全北国立大学环境和生物资源科学学院, 环境与生物科学高级研究所生物技术部, 益山 570752, 韩国;
    f Haramaya大学健康与医学科学学院医学微生物学系, 哈拉尔, 埃塞俄比亚
  • 收稿日期:2016-05-08 修回日期:2016-06-14 出版日期:2016-11-25 发布日期:2016-11-25
  • 通讯作者: Balakrishnan Senthilkumar,E-mail:nbsenthilkumar@gmail.com;Thangasamy Selvankumar,Tel:+91-9443286292, +91-9443470394; E-mail:t_selvankumar@yahoo.com

Activity and stability of bacterial cellulase immobilized on magnetic nanoparticles

Kandasamy Selvam, Muthusamy Govarthanan, Duraisamy Senbagam, Seralathan Kamala-Kannan, Balakrishnan Senthilkumar, Thangasamy Selvankumar   

  1. a Centre for Biotechnology, Muthayammal College of Arts and Science, Rasipuram, Namakkal 637408, Tamil Nadu, India;
    b PG and Research Department of Biotechnology, Mahendra Arts and Science College (Autonomous), Kalippatti, Namakkal 637501, Tamil Nadu, India;
    c Department of Applied Sciences, College of Environmental Technology, Muroran Institute of Technology, 27-1 Mizumoto, Muroran, Hokkaido 050-8585, Japan;
    d Department of Marine Biotechnology, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India;
    e Division of Biotechnology, Advanced Institute of Environment and Bioscience, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan 570752, South Korea;
    f Department of Medical Microbiology, College of Health and Medical Sciences, Haramaya University, P.O. Box 235, Harar, Ethiopia
  • Received:2016-05-08 Revised:2016-06-14 Online:2016-11-25 Published:2016-11-25
  • Contact: Balakrishnan Senthilkumar,E-mail:nbsenthilkumar@gmail.com;Thangasamy Selvankumar,Tel:+91-9443286292, +91-9443470394; E-mail:t_selvankumar@yahoo.com

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摘要：

在氨水溶液中进行Fe+2和Fe+3离子共沉淀并水热处理后制得磁性纳米颗粒Fe3O4，通过戊二醛活化将纤维素酶固定于其上.采用基于响应面法的Box-Behnken法（BBD）优化了制备条件，如磁性纳米颗粒浓度、戊二醛浓度、酶浓度和交联时间.BBD分析结果表明，用实验数据可合理调节二次模型.利用生成的基于统计数据的等高线评价了响应面的变化，以理解纳米颗粒和酶活性之间的关系.运用扫描电镜、X射线衍射和红外光谱表征了纳米颗粒上酶的尺寸、结构、形貌和结合情况.采用诸如pH值、温度、重复使用性和存储能力分析了固定化纤维素酶的活性和稳定性.发现固定后的纤维素酶表现出更好的稳定性和活性.

关键词: 磁性纳米颗粒, 固定化酶, 响应面法, 表征

Abstract:

Magnetic nanoparticles (Fe3O4) were synthesized by co-precipitating Fe2+ and Fe3+ ions in an am-monia solution and treating under hydrothermal conditions. Cellulase was immobilized onto Fe3O4 magnetic nanoparticles via glutaraldehyde activation. Using response surface methodology and Box-Behnken design, the variables such as magnetic nanoparticle concentration, glutaraldehyde concentration, enzyme concentration, and cross linking time were optimized. The Box-Behnken design analysis showed a reasonable adjustment of the quadratic model with the experimental data. Statistical contour plots were generated to evaluate the changes in the response surface and to understand the relationship between the nanoparticles and the enzyme activity. Scanning electron microscopy, X-ray diffraction analysis, and Fourier transform infrared spectroscopy were studied to characterize size, structure, morphology, and binding of enzyme onto the nanoparticles. The stabil-ity and activity of the bound cellulase was analyzed using various parameters including pH, tem-perature, reusability, and storage stability. The immobilized cellulase was compared with free cel-lulase and it shows enhanced stability and activity.

Key words: Magnetic nanoparticle, Immobilized enzyme, Response surface methodology, Characterization