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Streptomyces violaceoruber Phospholipase A2: Expression in Pichia pastoris, Properties, and Application in Oil Degumming

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Abstract

The phospholipase A2 (PLA2) from Streptomyces violaceoruber was successfully expressed in the methylotrophic yeast Pichia pastoris GS115 under the control of AOX1 promoter for the first time. The maximum activity of the recombinant PLA2 (rPLA2) reached 34.7 ± 0.2 U/mL, and specific activity was 170 ± 4 U/mg after purification. On the sodium dodecyl sulfate polyacrylamide gel electrophoresis of the culture supernatants, three bands of 21, 18, and 14.3 kDa were observed. The peptide mass fingerprinting analysis showed that all of these three bands were rPLA2 from S. violaceoruber. By the treatment with Endo H and PNGase F, it indicated that the rPLA2 occurred N-glycosylation. The enzymatic properties of this enzyme were determined. The rPLA2 exhibited a lower optimum pH (pH = 6.0) compared to the wild-type enzyme, which was a desirable property in the application of oil degumming. In the enzymatic degumming process, the phosphorus content decreased from 261.77 ± 3.51 mg/kg to 20.74 ± 0.23 mg/kg, which is very promising for the industrial application.

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Acknowledgments

Financial support from the National High Technology Research and Development Program of China (863 Program) (No. 2012AA022207), the National Key Basic Research and Development Program of China (973 Program) (No. 2011CB710800), the High-end Foreign Experts Recruitment Program (GDW20123200113), and the 111 Project (111-2-06) are greatly appreciated.

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Authors and Affiliations

  1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, China

    Aixia Liu, Xiao-Wei Yu, Chong Sha & Yan Xu

  2. State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, Jiangsu, China

    Aixia Liu, Xiao-Wei Yu, Chong Sha & Yan Xu

Authors
  1. Aixia Liu
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  2. Xiao-Wei Yu
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  3. Chong Sha
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  4. Yan Xu
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Correspondence to Xiao-Wei Yu or Yan Xu.

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Liu, A., Yu, XW., Sha, C. et al. Streptomyces violaceoruber Phospholipase A2: Expression in Pichia pastoris, Properties, and Application in Oil Degumming. Appl Biochem Biotechnol 175, 3195–3206 (2015). https://doi.org/10.1007/s12010-015-1492-7

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  • DOI: https://doi.org/10.1007/s12010-015-1492-7

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