磷脂酶D的纯化及催化制备磷脂酰丝氨酸研究
Purification of phospholipase D and catalytic preparation of phosphatidylserine
  
DOI:10.12166/j.zgyz.1003-7969/2020.01.021
中文关键词:  磷脂酶D  纯化  磷脂酰丝氨酸  酶法  丝氨酸  磷脂酰胆碱
英文关键词:phospholipase D  purification  phosphatidylserine  enzymatic method  serine  phosphatidylcholine
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Author NameAffiliation
XIA Yu ZHANG Tao JIANG Bo State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, Jiangsu, China 
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中文摘要:
      采用硫酸铵分级沉淀、离子交换层析和凝胶层析对广岛链霉菌SK43.001-11发酵液中的磷脂酶D进行纯化,并对双液相体系中酶法制备磷脂酰丝氨酸的工艺进行研究。结果表明,纯化后,磷脂酶D比酶活提高到49.48 U/mg,是粗酶液的54.37倍,回收率为32.31%。磷脂酶D双液相催化制备磷脂酰丝氨酸的最佳条件为:有机相为乙醚,pH 6.5,反应温度30 ℃,L-丝氨酸与磷脂酰胆碱质量比15∶ 1,有机相与水相体积比1∶ 1,反应时间2.5 h。在最佳条件下,磷脂酰丝氨酸生成率达到74.8%。
英文摘要:
      The phospholipase D(PLD) in the fermentation broth of Streptomyces hiroshimemsis SK43.001-11 was purified by ammonium sulfate fractionation, ion exchange chromatography and gel chromatography. The process of enzymatic preparation of phosphatidylserine catalyzed by PLD in two-liquid phase system was studied.The results showed that after purification, the specific enzyme activity of PLD increased to 49.48 U/mg,which was 54.37 times of the crude enzyme solution,and the recovery rate was 32.31%. The optimal conditions for the two-liquid phase catalytic reaction were obtained as follows: with diethyl ether as organic phase, pH 6.5, reaction temperature 30 ℃, mass ratio of L-serine to phosphatidylcholine 15∶ 1, volume ratio of organic phase to aqueous phase 1∶ 1 and reaction time 2.5 h. Under these conditions, the phosphatidylserine production rate reached 74.8%.
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