Study on catalytic properties of porcine pancreatic lipase modified by chiral proline ionic liquids

XU Chao; XUE Yu; CHEN Hongyue; HU Yi

doi:10.11949/0438-1157.20190074

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Study on catalytic properties of porcine pancreatic lipase modified by chiral proline ionic liquids

Biochemical engineering and technology | 更新时间：2026-07-24

- Study on catalytic properties of porcine pancreatic lipase modified by chiral proline ionic liquids
- CIESC Journal Vol. 70, Issue 6, Pages: 2221-2228(2019)
- 作者机构：
  
  南京工业大学材料化学工程国家重点实验室，南京工业大学药学院，江苏南京 211816
- 作者简介：
- 基金信息：
- DOI：10.11949/0438-1157.20190074
  CLC： Q 814
- Received：25 January 2019，
  
  Revised：2019-03-22，
  
  Online First：15 June 2020，
  
  Published：05 June 2019
- 稿件说明：
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徐超, 薛誉, 陈虹月, 胡燚. 手性脯氨酸类离子液体化学修饰猪胰脂肪酶催化性能研究[J]. 化工学报, 2019, 70(6): 2221-2228

XU Chao, XUE Yu, CHEN Hongyue, HU Yi. Study on catalytic properties of porcine pancreatic lipase modified by chiral proline ionic liquids[J]. CIESC Journal, 2019, 70(6): 2221-2228
徐超, 薛誉, 陈虹月, 胡燚. 手性脯氨酸类离子液体化学修饰猪胰脂肪酶催化性能研究[J]. 化工学报, 2019, 70(6): 2221-2228 DOI： 10.11949/0438-1157.20190074.

XU Chao, XUE Yu, CHEN Hongyue, HU Yi. Study on catalytic properties of porcine pancreatic lipase modified by chiral proline ionic liquids[J]. CIESC Journal, 2019, 70(6): 2221-2228 DOI： 10.11949/0438-1157.20190074.

摘要

通过化学修饰同时提高脂肪酶包括选择性在内的酶学性能具有重要意义。以

-乙酰-脯氨酸作为手性源制备的四种离子液体对猪胰脂肪酶（PPL）进行化学修饰，通过三硝基苯磺酸法（TNBS）测定了修饰度，采用水解4-硝基苯酚棕榈酸酯的反应测定酶活。正如预期的一样，所有修饰酶的酶学性能均得到显著改善。其中修饰酶的热稳定性、有机溶剂耐受性、不同温度和pH下的耐受性甚至对映体选择性均得到同时提升。结果也表明，离子液体的组成及构型对修饰酶的酶学性能有较大程度的影响，经过[BMIM

]

[N-AC-L-pro

]

修饰后的PPL各项催化性能明显优于[BMIM

]

[N-AC-D-pro

]

，而经过[N-AC-L-pro

]

[Cl

]

和[N-AC-D-pro

]

[Cl

]

修饰的PPL催化性能差异较小。经过[N-AC-L-pro

]

[Cl

]

修饰后PPL展现出最高的酶活，达到原酶的2.5倍。经过[BMIM

]

[N-AC-L-pro

]

修饰后的PPL展现出最佳的酶学性能，在酶活提高了1.0倍的同时，热稳定性提高了2.6倍（50℃，2.5 h），对映体选择性提高了1.5倍，在强极性质子溶剂甲醇中的稳定性也提升了1.3倍，且在不同温度、pH下也展现出了较好的稳定性与耐受性。通过荧光光谱和圆二色谱表征证明，手性离子液体成功嫁接到PPL的表面，引起脂肪酶构象的改变，从而促使酶的催化性能发生改善。通过该研究，手性离子液体被证实为是一种可用于酶分子改造并提升选择性在内各项催化性能的新型且有效的修饰剂。

Abstract

It is of great value to improve the enzymatic properties including selectivity by chemical modification at the same time. Four kinds of ionic liquids

prepared from

-acetyl-proline as chiral source

were used to modify porcine

pancreatic lipase (PPL). Modification degree was determined by 2

6-trinitrobenzene sulfonic acid (TNBS) method

and enzyme activity was determined by the hydrolysis of 4-nitrophenyl palmitate (pNPP). As expected

the catalytic properties of PPL were changed obviously after modification. The thermal stability

organic solvent tolerance

adaptability to temperature and pH changes

even enantio selectivity of all the modified enzymes was improved simultaneously. Research results also showed that the composition and configuration of ionic liquids (Ils)had an important influence on the enzymatic properties of modified enzymes. The PPL modified by [BMIM

]

[N-AC-L-pro

]

showed obvious better catalytic performance than that modified by [BMIM

]

[N-AC-D-pro

]

while the catalytic properties of PPL modified by [N-AC-L-pro

]

[Cl

]

and [N-AC-D-pro

]

[Cl

]

only showed slight difference. PPL modified by [N-AC-L-pro

]

[Cl

]

showed the highest activity

increasing by 2.5 folds as much as the original enzyme. Overall

PPL modified by [BMIM

]

[N-AC-L-pro

]

demonstrated the best enzymatic performance: the hydrolysis activity was increased by 1.0 time

the optimum temperature was raised to 55℃

the thermostability was improved by 2.6 times（50℃，2.5 h）

the enantio selectivity was increased by 1.5 times

the stability in the strong polar protonic solvent of methanol was enhanced by 1.3 times

and also showed better tolerance of temperature and pH. The fluorescence spectroscopy and circular dichroism spectroscopy (CD) characterizations confirmed that chiral proline ionic liquids were grafted onto the surface of PPL successfully

resulting in the conformation change of PPLs

which led to the improvements of the catalytic performance of modified PPLs. Through this study

chiral ionic liquids have proven to be a new and effective modifier for various catalytic properties of enzyme molecular modification and selectivity enhancement.

关键词

Keywords

references

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