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磷脂酶D的制备及催化磷脂酰胆碱合成磷脂酰丝氨酸
Preparation of phospholipase D and catalytic synthesis of phosphatidylserine from phosphatidylcholine
  
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中文关键词:  穗色链霉菌  磷脂酶D  磷脂酰丝氨酸  两相催化
英文关键词:Streptomyces racemochromogenes  phospholipase D  phosphatidylserine  biphasic catalysis
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Author NameAffiliation
QIAN Juan, PANG Yang, WANG Xin  
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中文摘要:
      选用穗色链霉菌（Streptomyces racemochromogenes）作为磷脂酶 D 的生产菌株进行了酶的制备及条件优化，并在两相体系中进行了酶催化磷脂酰胆碱合成磷脂酰丝氨酸的研究。结果表明，最适的产酶条件为葡萄糖 10 g/L、酵母粉5 g/L、鱼粉蛋白胨 5 g/L、司班-60 10 g/L、氯化钙 3 g/L，接种量 1%、培养基初始 pH 7.0、培养温度 30℃，在最适产酶条件下酶活达到2 967 U/L，是优化前 的2.12 倍；最适的催化工艺条件为有机相二氯甲烷、反应温度35℃、pH 5.5、酶用量1.15 U/mL、钙离子浓度10 mmol/L，在该催化工艺条件下反应10 h后磷脂酰胆碱的生成率达94%。
英文摘要:
      Streptomyces racemochromogenes was used as the producing strain to prepare phospholipase D and the preparation conditions were optimized. The synthesis of phosphatidylserine from phosphatidylcholine catalyzed by phospholipase D in biphasic catalysis system was studied. The results showed that the optimal conditions for enzyme production were obtained as follows：glucose 10 g/L, yeast extract 5 g/L, fishmeal peptone 5 g/L, Span-60 10 g/L, calcium chlodride 3 g/L, inoculation amount 1%, initial pH of medium 7.0 and cultural temperature 30℃. Under these conditions, the enzyme activity reached 2 967 U/L, which was 2.12 times of that before optimization. The optimal catalytic process conditions were obtained as follows：dichloromethane as organic phase, reaction temperature 35℃, pH 5.5, enzyme dosage 1.15 U/mL, Ca2+concentration 10 mmol/L. Under these conditions, the yield of phosphatidylcholine reached 94%.
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