Phospholipase D engineering for improving the biocatalytic synthesis of phosphatidylserine

  • Hai-Juan Hou
  • Jin-Song Gong
  • Yu-Xiu Dong
  • Jiufu Qin
  • Heng Li
  • Hui Li
  • Zhen-Ming Lu
  • Xiao-Mei Zhang
  • Zheng-Hong Xu
  • Jin-Song ShiEmail author
Research Paper


Phosphatidylserine is widely used in food, health, chemical and pharmaceutical industries. The phospholipase D-mediated green synthesis of phosphatidylserine has attracted substantial attention in recent years. In this study, the phospholipase D was heterologously expressed in Bacillus subtilis, Pichia pastoris, and Corynebacterium glutamicum, respectively. The highest activity of phospholipase D was observed in C. glutamicum, which was 0.25 U/mL higher than these in B. subtilis (0.14 U/mL) and P. pastoris (0.22 U/mL). System engineering of three potential factors, including (1) signal peptides, (2) ribosome binding site, and (3) promoters, was attempted to improve the expression level of phospholipase D in C. glutamicum. The maximum phospholipase D activity reached 1.9 U/mL, which was 7.6-fold higher than that of the initial level. The enzyme displayed favorable transphosphatidylation activity and it could efficiently catalyze the substrates l-serine and soybean lecithin for synthesis of phosphatidylserine after optimizing the conversion reactions in detail. Under the optimum conditions (trichloromethane/enzyme solution 4:2, 8 mg/mL soybean lecithin, 40 mg/mL l-serine, and 15 mM CaCl2, with shaking under 40 °C for 10 h), the reaction process showed 48.6% of conversion rate and 1.94 g/L of accumulated phosphatidylserine concentration. The results highlight the use of heterologous expression, system engineering, and process optimization strategies to adapt a promising phospholipase D for efficient phosphatidylserine production in synthetic application.


Phospholipase D System engineering Heterologous expression Phosphatidylserine 



This work was financially supported by the National Natural Science Foundation of China (No. 21676121), the National first-class discipline program of Light Industry Technology and Engineering (No. LITE2018-18), the Six talent peaks project in Jiangsu Province (No. 2015-SWYY-006), and the Top-notch Academic Programs Project of Jiangsu Higher Education Institutions (No. PPZY2015B146).

Compliance with ethical standards

Conflict of interest

All the authors reviewed and agreed to submit this manuscript. The authors declare that they have no confict of interest.

Supplementary material

449_2019_2116_MOESM1_ESM.doc (1.5 mb)
Supplementary material 1 (DOC 1555 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Hai-Juan Hou
    • 1
  • Jin-Song Gong
    • 1
  • Yu-Xiu Dong
    • 1
  • Jiufu Qin
    • 3
  • Heng Li
    • 1
  • Hui Li
    • 1
  • Zhen-Ming Lu
    • 1
    • 2
  • Xiao-Mei Zhang
    • 1
  • Zheng-Hong Xu
    • 1
    • 2
  • Jin-Song Shi
    • 1
    Email author
  1. 1.Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Pharmaceutical SciencesJiangnan UniversityWuxiPeople’s Republic of China
  2. 2.National Engineering Laboratory for Cereal Fermentation Technology, School of BiotechnologyJiangnan UniversityWuxiPeople’s Republic of China
  3. 3.Novo Nordisk Foundation Center for BiosustainabilityTechnical University of DenmarkKongens LyngbyDenmark

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