This study aimed to express recombinant phospholipase D (PLD) and explore its application in biocatalytic synthesis of phosphatidylserine (PS). The recombinant strain Escherichia coli BL21 (DE3)/pET-28a(+)-sspld was constructed, followed by optimizing its cultivation conditions to study its enzymatic properties. Besides, the preparation process of PS by the strain was also optimized. The results showed that a 60 kDa recombinant PLD was successfully expressed. The maximum activity of PLD produced reached 38.58 U/mL, which was 2.26-fold higher than that before optimization. Moreover, the optimum pH of PLD was 7.5 and its optimum reaction temperature was 60 ℃. Furthermore, the optimum reaction condition to prepare PS was as follows: reacted at 40 ℃ and dissolved 64 mg PC60 in 8 mL ethyl acetate and 160 mg L-ser in 4 mL PLD. Under this condition, the PS conversion rate was 28.0% and had the highest yield of 1.34 g/L. In conclusion, the recombinant strain exhibited favorable catalytic performance, which lays a foundation for enzymatic preparation of PS.
HOU Haijuan
,
GONG Jinsong
,
ZHAI Shen
,
XU Min
,
ZHOU Wenbin
,
CHEN Jiepeng
,
DUAN Lili
,
ZHANG Xiaomei
,
XU Zhenghong
,
SHI Jinsong
. Recombinant expression of phospholipase D and its application in synthesizing phosphatidylserine[J]. Food and Fermentation Industries, 2019
, 45(13)
: 9
-14
.
DOI: 10.13995/j.cnki.11-1802/ts.020355
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