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Purification and Characterization of a Cold Alkaline Protease from a Psychrophilic Pseudomonas aeruginosa HY1215

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Abstract

A novel alkaline protease was purified from Pseudomonas aeruginosa HY1215 using ammonium sulfate, DEAE-Sepharose and Sephacryl S-200 gel filtration chromatographic techniques. The protease had a relative molecular weight of 32.8 KDa by SDS-PAGE, and the optimal temperature and pH for excellent stability and activity were determined as 25 °C and 10.0, respectively. Within the pH range of 7.0–11.0, the protease had a good stability, which could retain more than 80 % of its original activity; in the temperature range of 15–35 °C, the protease had a higher activity, and its activity at 20 °C could amount to 85 % of the maximum activity at 25 °C. Besides, the enzyme activity showed a valuable stability towards several commercially available surfactants (Tween-80, Tween-40, and Triton X-100) and bleaches (H2O2) even when their concentrations ranged up to 2.0 and 1.6 %.

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Acknowledgments

This research work has been supported by National Natural Science Foundation of China (No. 41376175) and Qingdao Municipal Science and Technology Plan Project (14-2-4-11-jch).

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Authors and Affiliations

  1. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, 266071, People’s Republic of China

    Jian-Hua Hao & Mi Sun

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  1. Jian-Hua Hao
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  2. Mi Sun
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Correspondence to Mi Sun.

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Hao, JH., Sun, M. Purification and Characterization of a Cold Alkaline Protease from a Psychrophilic Pseudomonas aeruginosa HY1215. Appl Biochem Biotechnol 175, 715–722 (2015). https://doi.org/10.1007/s12010-014-1315-2

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  • DOI: https://doi.org/10.1007/s12010-014-1315-2

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