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Introduction of Two Halo-Alkali-thermo-stable Biocatalysts: Purification and Characterization

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Abstract

Halophilic bacterium, producing two extracellular proteases, was isolated from Badab-Sourt, Iran. 16S rDNA analysis shown high similarity with the genera Idiomarina. Maximum secretion of enzymes was observed during the early-stationary phase of bacterial growth. Both extracellular proteases were purified by three purification steps; ammonium sulfate precipitation, Q-Sepharose, and Sephadex G-200 chromatography columns. The molecular weight of purified enzymes was determined by SDS-PAGE gel electrophoresis with approximate masses of 48 and 51 kDa (45.57 and 80.30 U/mg specific activity). Proteases synthesized by strain S-18 were affected by medium compositions. Optimum concentration of substrate, pH, and temperature for both enzymes activity were 0.5% casein, 9.0, and 50 °C. However, purified proteases showed different activity at various salt concentrations, which their maximum activity was determined in the presence of 7.5 and 5% NaCl. The activity of enzymes increased in the presence of metal ions such as Mn2+ and Cu2+ and decreased by the presence of Hg2+ and Fe2+. Both proteases were strongly inhibited by SDS, while DDT, EDTA, and 2-Mercaptoethanol could stimulate their activity. The results of present research might be interesting issue for industrial applications and biotechnological processes.

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Acknowledgements

Authors are grateful Pasture Institute (Amol Branch) for molecular analysis and also thankful Dr. Farzad Mokhtari, Dr. Arastoo Badoei-Dalfard, Mr. Jaber Abbaszadeh, Mr. Shayan Abbasi, Mr. Mohammad Mehdi Choolaei, and Mr. Nima Allah-Yari for their valuable advice. Also, authors wish to thank Neuro-organic Lab. (Institute of biochemistry and biophysics, Tehran University, Iran.)

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Authors and Affiliations

  1. Microbiology Lab., Department of Molecular and Cell Biology, Faculty of Basic Science, University of Mazandaran, P.O Box 57416-13535, Babolsar, Mazandaran, Iran

    Leila Satari Faghihi & Salman Ahmady-Asbchin

  2. Biochemistry Lab., Department of Molecular and Cell Biology, Faculty of Basic Science, University of Mazandaran, P.O Box 57416-13535, Babolsar, Mazandaran, Iran

    Bagher Seyedalipour

  3. Neuro-Organic Lab., Biochemistry Department, Institute of Biochemistry and Biophysics (IBB), University of Tehran, Tehran, Iran

    Gholamhossein Riazi

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  1. Leila Satari Faghihi
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  2. Bagher Seyedalipour
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  3. Gholamhossein Riazi
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  4. Salman Ahmady-Asbchin
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Correspondence to Bagher Seyedalipour or Salman Ahmady-Asbchin.

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Satari Faghihi, L., Seyedalipour, B., Riazi, G. et al. Introduction of Two Halo-Alkali-thermo-stable Biocatalysts: Purification and Characterization. Catal Lett 148, 831–842 (2018). https://doi.org/10.1007/s10562-018-2295-6

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  • DOI: https://doi.org/10.1007/s10562-018-2295-6

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