Interplay between Residual Protease Activity in Commercial Lactases and the Subsequent Digestibility of β-Casein in a Model System
Abstract
:1. Introduction
2. Results and Discussion
2.1. Proteolysis of β-casein (β-CN) during Storage
2.2. Formation of Bitter and N-Terminal Glycated Peptide
2.3. Change in Digestibility
3. Materials and Methods
3.1. Materials
3.2. Storage Experiment
3.3. In Vitro Digestion
3.4. SDS-PAGE
3.5. DH
3.6. LC-ESI-MS/MS Ion Trap
3.7. Statistics Analysis
4. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Sample Availability: Samples of the compounds β-CN and glucose are available from the authors. |
Sample | Representative Cleavage Sites |
---|---|
β-CN + Lactase A (day 10) | F (190), I (207), I (208) |
β-CN + Lactase A (day 30) | A (177), P (206), K (176), I (207), F (190), I (208), L (191), F (205), Y (193) |
β-CN + Lactase A (day 60) | A (177), F (205), P (206), K (176), I (207), F (190), I (207), I (208), L (191), E (195), |
β-CN + Lactase B (day 10) | F (190), V (170), P (206), F (205), Y (193), A (189), Q (194), L (191), L (192) |
β-CN + Lactase B (day 30) | F (190), V (170), P (206), F (205), Y (193), A (189), Q (194), L (192), A (177), P (204) |
β-CN + Lactase B (day 10) | I (207), F (190), P (206), I (208), Y (193), A (189), Q (194), L (192), A (177), S (164), I (207) |
No. | Peptides | Identified Mass (Da) | Q Value | Origin |
---|---|---|---|---|
1 | G(203)-V(209) | 642.37 | 2305 | β-CN + Lactase B |
2 | G(199)-V(209) | 1150.69 | 1869 | β-CN + Lactase B |
3 | E(195)-I(208) | 1489.87 | 1868 | β-CN + Lactase B |
4 | E(195)-I(209) | 1588.93 | 1856 | β-CN + Lactase B |
5 | L(171)-I(187) | 1948.06 | 1840 | β-CN + Lactase A |
6 | Y(193)-I(208) | 1780.99 | 1808 | β-CN + Lactase B |
7 | Y(193)-V(209) | 1880.06 | 1801 | β-CN + Lactase A or B |
8 | F(190)-P(206) | 1928.06 | 1792 | β-CN + Lactase A |
9 | V(178)-I(187) | 1214.61 | 1786 | β-CN + Lactase A |
10 | G(199)-I(207) | 938.53 | 1767 | β-CN + Lactase B |
11 | V(170)-Q(182) | 1434.82 | 1759 | β-CN + Lactase B |
12 | F(190)-F(205) | 1831.00 | 1741 | β-CN + Lactase B |
13 | L(191)-P(206) | 1780.99 | 1739 | β-CN + Lactase A or B |
14 | V(178)-R(183) | 758.41 | 1738 | β-CN + Lactase B |
15 | A(177)-V(209) | 3720.03 | 1737 | β-CN + Lactase B |
16 | Y(193)-I(207) | 1667.90 | 1734 | β-CN + Lactase B |
17 | Q(194)-V(209) | 1716.99 | 1730 | β-CN + Lactase B |
18 | D(184)-I(207) | 2696.44 | 1729 | β-CN + Lactase B |
19 | V(170)-I(187) | 2047.13 | 1724 | β-CN + Lactase A |
20 | L(171)-P(186) | 1834.98 | 1704 | β-CN + Lactase A |
No. | N-Terminally Glycated Peptides | Identified Mass (Da) | Theoretical Mass (Da) | Origin |
---|---|---|---|---|
1 | Q(194)EPVLGPVRGPFPI | 1667.90 | 1668.15 | β-CN + Lactase B |
2 | Q(188)AFLLYQEPVLGPVRGP | 2046.09 | 2046.37 | β-CN + Lactase A |
3 | Q(194)EPVLGPVRG | 1212.59 | 1212.64 | β-CN + Lactase B |
4 | Q(194)EPVLGPVRGP | 1309.83 | 1309.69 | β-CN + Lactase B |
5 | F(190)LLYQEPVLGP | 1536.60 | 1535.81 | β-CN + Lactase A or B |
6 | Y(193)QEPVLGPVRGPFP | 1716.35 | 1716.87 | β-CN + Lactase B |
Sample (Day 30) | DH (%) | ||||
---|---|---|---|---|---|
G0 | G15 | G120 | GI15 | GI120 | |
β-CN (control) | 0.36 ± 0.06a | 4.37 ± 0.37a | 9.97 ± 0.87a | 44.28 ± 3.63a | 58.91 ± 2.81a |
β-CN+Lactase A | 2.71 ± 0.42b | 10.72 ± 1.83b | 13.74 ± 2.10b | 60.82 ± 4.17b | 69.19 ± 4.37b |
β-CN+Lactase B | 4.15±0.04c | 16.57 ± 2.19c | 22.41 ± 3.04c | 67.53 ± 5.82b | 74.85 ± 5.80c |
Samples | Peptides after G Digestion | Peptides after G+I Digestion | ||||
---|---|---|---|---|---|---|
Total | Missed | New | Total | Missed | New | |
β-CN (control) | 63 | _ | _ | 53 | _ | _ |
β-CN + Lactase A | 61 | 16 | 14 | 55 | 6 | 8 |
β-CN + Lactase B | 80 | 15 | 32 | 62 | 10 | 19 |
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Zhao, D.; Le, T.T.; Larsen, L.B.; Nian, Y.; Wang, C.; Li, C.; Zhou, G. Interplay between Residual Protease Activity in Commercial Lactases and the Subsequent Digestibility of β-Casein in a Model System. Molecules 2019, 24, 2876. https://doi.org/10.3390/molecules24162876
Zhao D, Le TT, Larsen LB, Nian Y, Wang C, Li C, Zhou G. Interplay between Residual Protease Activity in Commercial Lactases and the Subsequent Digestibility of β-Casein in a Model System. Molecules. 2019; 24(16):2876. https://doi.org/10.3390/molecules24162876
Chicago/Turabian StyleZhao, Di, Thao T. Le, Lotte Bach Larsen, Yingqun Nian, Cong Wang, Chunbao Li, and Guanghong Zhou. 2019. "Interplay between Residual Protease Activity in Commercial Lactases and the Subsequent Digestibility of β-Casein in a Model System" Molecules 24, no. 16: 2876. https://doi.org/10.3390/molecules24162876