Enzymatic Production of Biologically Active 3-Methoxycinnamoylated Lysophosphatidylcholine via Regioselctive Lipase-Catalyzed Acidolysis
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents
2.2. Acidolysis Reaction of Egg-Yolk Phosphatidylcholine with 3-Methoxycinnamic Acid
2.3. Design of Experiment
2.4. Acidolysis of Egg-Yolk Phosphatidylcholine with 3-methoxycinnamic Acid Catalyzed by Novozym 435 in an Increased Scale
2.5. Methods of Analysis
2.5.1. Thin-Layer Chromatography (TLC)
2.5.2. Gas Chromatography (GC)
2.5.3. High-Performance Liquid Chromatography (HPLC)
3. Results and Discussion
3.1. Screening of Commercial Immobilized Lipases
3.2. Effect of Organic Solvents on Enzyme Activity
3.3. Model Fitting
3.4. Response Surface Metodology Analysis and Interpretation
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Run | Substrate Molar Ratio PC/3-OMe-CA | Enzyme Loading [%] | Reaction Time [Days] | Incorporation of 3-OMe-CA into PC/LPC [mol%] a (Experimental) | Incorporation of 3-OMe-CA to PC/LPC [mol%] (Predicted) |
---|---|---|---|---|---|
1 | 5 | 10 | 3 | 11 ± 0.4 | 13 |
2 | 15 | 10 | 3 | 19 ± 0.6 | 20 |
3 | 5 | 30 | 3 | 34 ± 1.2 | 33 |
4 | 15 | 30 | 3 | 49 ± 2.3 | 47 |
5 | 5 | 20 | 2 | 10 ± 0.6 | 8 |
6 | 15 | 20 | 2 | 16 ± 0.9 | 14 |
7 | 5 | 20 | 4 | 15 ± 0.7 | 17 |
8 | 15 | 20 | 4 | 31 ± 1.1 | 33 |
9 | 10 | 10 | 2 | 10 ± 0.5 | 10 |
10 | 10 | 30 | 2 | 23 ± 0.9 | 26 |
11 | 10 | 10 | 4 | 20 ± 0.7 | 17 |
12 | 10 | 30 | 4 | 48 ± 1.8 | 48 |
13 | 10 | 20 | 3 | 19 ± 0.7 | 19 |
14 | 10 | 20 | 3 | 19 ± 0.6 | 19 |
15 | 10 | 20 | 3 | 19 ± 0.2 | 19 |
Evaluated Factors | Sum of Squares | Degrees of Freedom | Medium Square | F-Value | p-Value |
---|---|---|---|---|---|
(1) Substrate molar ratio (L) | 253.1250 | 1 | 253.125 | 27.3649 | 0.003379 |
Substrate molar ratio (Q) | 3.692 | 1 | 3.692 | 0.3992 | 0.555275 |
(2) Enzyme loading(L) | 1104.500 | 1 | 1104.500 | 119.4054 | 0.000112 |
Enzyme loading (Q) | 251.308 | 1 | 251.308 | 27.1684 | 0.003432 |
(3) Time of reaction (L) | 378.125 | 1 | 378.125 | 40.8784 | 0.001387 |
Time of reaction (Q) | 14.769 | 1 | 14.769 | 1.5967 | 0.262086 |
1 by 2 | 12.250 | 1 | 12.250 | 1.3243 | 0.301861 |
1 by 3 | 25.000 | 1 | 25.000 | 2.7027 | 0.161099 |
2 by 3 | 56.250 | 1 | 56.250 | 6.0811 | 0.056810 |
Error | 46.250 | 5 | 9.250 | ||
Total error | 2153.733 | 14 | |||
R2 = 0.98771 |
Fatty and 3-OMe-CA Acids | Native PC | Modified PC |
C16:0 (PA) | 36 | 10 |
C16:1 (OPA) | 3 | 1 |
C18:0 (SA) | 16 | 3 |
C18:1 (OA) | 23 | 13 |
C18:2 (LA) | 17 | 12 |
C20:4 (AA) | 5 | 3 |
3-OMe-CA | - | 58 |
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Okulus, M.; Rychlicka, M.; Gliszczyńska, A. Enzymatic Production of Biologically Active 3-Methoxycinnamoylated Lysophosphatidylcholine via Regioselctive Lipase-Catalyzed Acidolysis. Foods 2022, 11, 7. https://doi.org/10.3390/foods11010007
Okulus M, Rychlicka M, Gliszczyńska A. Enzymatic Production of Biologically Active 3-Methoxycinnamoylated Lysophosphatidylcholine via Regioselctive Lipase-Catalyzed Acidolysis. Foods. 2022; 11(1):7. https://doi.org/10.3390/foods11010007
Chicago/Turabian StyleOkulus, Marta, Magdalena Rychlicka, and Anna Gliszczyńska. 2022. "Enzymatic Production of Biologically Active 3-Methoxycinnamoylated Lysophosphatidylcholine via Regioselctive Lipase-Catalyzed Acidolysis" Foods 11, no. 1: 7. https://doi.org/10.3390/foods11010007