固定化磷脂酰胆碱水相酶催化合成磷脂酰丝氨酸

朱帅; 黄澳; 刘夏忠; 刘云

doi:10.13386/j.issn1002-0306.2022060070

固定化磷脂酰胆碱水相酶催化合成磷脂酰丝氨酸

doi: 10.13386/j.issn1002-0306.2022060070

朱帅^1,,
黄澳¹,
刘夏忠²,
刘云^1, ,

1.
北京化工大学生命科学与技术学院，北京 100029
2.
河北美业斯维生物技术有限公司，河北保定 072650

基金项目: 企业委托项目（H2019167）。

详细信息

作者简介:
朱帅（1997−），男，硕士研究生，研究方向：功能性油脂研发，E-mail：1144289755@qq.com

通讯作者:
刘云（1972−），男，博士，教授，研究方向：生物催化与食品科学，E-mail：liuyun@mail.buct.edu.cn

中图分类号: TS253
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- 文章访问数: 51
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- PDF下载量: 6
- 被引次数: 0
出版历程
- 收稿日期: 2022-06-10
- 网络出版日期: 2022-12-16
- 刊出日期: 2023-01-17

Enzyme Catalytic Synthesis of Phosphatidylserine in Aqueous Phase through Immobilization of Phosphatidylcholine

ZHU Shuai^1
,,
HUANG Ao¹,
LIU Xiazhong²,
LIU Yun^{1
, ,}

1.
College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China
2.
Hebei Meiyesiwei Biotechnology Ltd. Co., Baoding 072650, China

摘要

摘要: 本文构建了一种固定化底物磷脂酰胆碱（phosphatidylcholine，PC）水相催化体系，磷脂酶D转化PC合成磷脂酰丝氨酸（phosphatidylserine，PS）新技术。探讨了沉淀剂种类、载体类型和底物初始添加量等主要因素，对固定化底物PC过程中PC吸附率的影响，获得了最佳工艺参数。在此基础上，进一步研究了固定化底物PC酶法合成PS的时间反应动力学和酶重复利用特性。结果表明，催化反应24 h后，PS得率为98.74%，磷脂酶D重复使用10次，PS得率仍然大于60%。文章还比较了固定化底物水相催化体系和双液相催化体系酶法合成PS效果，结果发现，固定化底物水相催化体系中PS得率和酶循环利用特性均优于双液相催化体系。本文研究结果为今后酶法改性PC合成PS提供了一种新的催化反应体系。
- 磷脂酰胆碱（PC） /
- 磷脂酰丝氨酸（PS） /
- 酶法改性 /
- 固定化底物 /
- 双液相催化体系
Abstract: This work reported a novel technology on bioconversion of phosphatidylcholine (PC) into phosphatidylserine (PS) by phospholipase D (PLD) in an aqueous phase catalytic system through immobilization of substrate phosphatidylcholine (PC). The major factors affecting PC adsorption efficiency were investigated to obtain the optimal parameters, such as the kinds of precipitant reagents, carrier types, and substrate loading concentration. Furthermore, the time kinetics of PS catalytic synthesis by PLD through immobilization of substrate PC and the recycling utilization of enzyme PLD were also examined. Results showed that the yield of PS achieved 98.74% after 24 h reaction, and PLD had an excellent recycling utilization. When it was reused for 10 batches, PLD could still yield more than 60% of PS. In the end, the efficiency of PS catalytic synthesis by PLD was also compared between through immobilization of substrate PC and in two-liquid phase system. It was found that higher yield of PS and more excellent stability of PLD were achieved through immobilization of substrate PC than in two-liquid phase system. These findings would provide a novel catalytic system for PC bioconversion into PS in the future.
- phosphatidylcholine (PC) /
- phosphatidylserine (PS) /
- enzymatic modification /
- immobilization of substrate /
- two-liquid phase system

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图 1 PC、PS混合标样的HPLC色谱图及其标准曲线

Figure 1. HPLC spectrum of PC and PS mixture standards and their standard curves

注：a：PC、PS混合标样的HPLC色谱图；b：PC标曲；c：PS标曲。

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图 2 不同沉淀剂（a）、底物初始加入量（b）和载体（c）对PC吸附率的影响

Figure 2. Effect of precipitants (a), carriers (b) and substrate loading (c) on PC adsorption rate

注：图a中柱状上标注不同字母表示显著性差异（P<0.05）。

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图 3 不同载体吸附PC前后的SEM形态

Figure 3. SEM images of carriers before and after adsorption of PC

注：a、b硅胶；c、d二氧化硅；e、f大孔树脂；g、h微晶纤维素；i、j氧化铝；k、l活性炭。

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图 4 时间反应动力学（a）和酶重复利用（b）

Figure 4. Time reaction kinetics (a) and enzyme recycling (b)

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图 5 两种不同催化体系的PS得率比较（a）和双液相体系中磷脂酶D的循环利用（b）

Figure 5. PS yield comparison for two kinds of catalytic system (a) and recycling utilization of enzyme for a two-liquid phase system (b)

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