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Purification, immobilization and characterization of linoleic acid isomerase on modified palygorskite

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Abstract

Linoleic acid isomerase from Lactobacillus delbrueckii subsp. bulgaricus 1.1480 was purified by DEAE ion-exchange chromatography and gel filtration chromatography. An overall 5.1% yield and purification of 93-fold were obtained. The molecular weight of the purified protein was ~41 kDa which was analyzed by SDS-PAGE. The purified enzyme was immobilized on palygorskite modified with 3-aminopropyltriethoxysilane. The immobilized enzyme showed an activity of 82 U/g. The optimal temperature and pH for the activity of the free enzyme were 30 °C and pH 6.5, respectively; whereas those for the immobilized enzyme were 35 °C and pH 7.0, respectively. The immobilized enzyme was more stable than the free enzyme at 30–60 °C, and the operational stability result showed that more than 85% of its initial activity was retained after incubation for 3 h. The K m and V max values of the immobilized enzyme were found to be 0.0619 mmol l−1 and 0.147 mmol h−1 mg−1, respectively. The immobilized enzyme had high operational stability and retained high enzymatic activity after seven cycles of reuse at 37 °C.

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Acknowledgments

This work was supported by the National Basic Research Program of China (973) (No. 2007CB714304), the National High Technology Research and Development Program of China (No. 2009AA033004), Jiangsu Key Technology Research and Development Program (No. BE2009363), and Jiangsu Postdoctoral Sustentation Foundation (No. 0901011C).

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Authors and Affiliations

  1. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Food Science and Light Industry, Nanjing University of Technology, No. 5 Xinmofan Road, Gulou District, Nanjing, 210009, People’s Republic of China

    Qinghong You, Hong Xu & Li Sang

  2. Huaiyin Institute of Technology, School of Life Science and Chemical Engineering, Jiangsu, 223001, People’s Republic of China

    Qinghong You, Xiulian Yin & Xu Gu

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  1. Qinghong You
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  2. Xiulian Yin
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  3. Xu Gu
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  4. Hong Xu
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Correspondence to Hong Xu.

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You, Q., Yin, X., Gu, X. et al. Purification, immobilization and characterization of linoleic acid isomerase on modified palygorskite. Bioprocess Biosyst Eng 34, 757–765 (2011). https://doi.org/10.1007/s00449-011-0525-z

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