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| 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 |
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| 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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| 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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