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Extraction of microbial transglutaminase from Amycolatopsis sp. fermentation broth using aqueous two-phase system

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Abstract

Partitioning of microbial transglutaminase (MTG) from Amycolatopsis sp. in the polyethylene glycol (PEG)/salt-based ATPS was investigated for the first time. The key parameters such as the molecular weight of PEG (PEG 600-6000), the type and concentration of phase-forming salt (ammonium sulfate or phosphates), the pH of system (pH 5.0-8.5), and the concentration of neutral salt (0-6% NaCl, w/w) were determined. The partition coefficient of the enzyme was not linear with PEG molecular weight; PEG1000 gave better yield than others. The concentration of PEG1000, ammonium sulfate and NaCl, and the system pH showed effects with different extents on specific activity (SA) and yield of the enzyme. In the ATPS of 26% w/w PEG 1000 and 19% w/w ammonium sulfate in the presence of 5% w/w NaCl and at pH 6.0, MTG was partitioned into the PEG-rich phase with a maximum yield of 86.51% and SA was increased to 0.83. The results of SDS-PAGE showed the MTG produced by the test strain differed from the enzymes reported before. Thus, this study proves that ATPS can be used as a preliminary step for partial purification of MTG from Amycolatopsis sp. fermentation broth.

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Authors and Affiliations

  1. College of Food Science and Technology, Engineering Technology Center for Agricultural Products Processing of Hebei Province, Agricultural University of Hebei, Baoding, 071001, China

    Nan Shi, Hongwei Xu, Kaiyuan Guo, Chunyu Kang, Wei Zhang, Yingying Zhang & Jianxin Tan

  2. Key Laboratory of Microbial Diversity Research and Application of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China

    Nan Shi & Liping Zhang

Authors
  1. Nan Shi
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  2. Hongwei Xu
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  3. Kaiyuan Guo
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  4. Chunyu Kang
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  5. Wei Zhang
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  6. Yingying Zhang
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  7. Liping Zhang
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  8. Jianxin Tan
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Correspondence to Liping Zhang or Jianxin Tan.

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Shi, N., Xu, H., Guo, K. et al. Extraction of microbial transglutaminase from Amycolatopsis sp. fermentation broth using aqueous two-phase system. Korean J. Chem. Eng. 34, 2248–2254 (2017). https://doi.org/10.1007/s11814-017-0105-2

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  • DOI: https://doi.org/10.1007/s11814-017-0105-2

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