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 Shi

Research output: Contribution to journalJournal articleResearchpeer-review


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 CaCl 2 , 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.
Original languageEnglish
JournalBioprocess and Biosystems Engineering
Issue number7
Pages (from-to)1185-1194
Publication statusPublished - 2019


  • Phospholipase D
  • System engineering
  • Heterologous expression
  • Phosphatidylserine

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