A novel fluorescence sensor for milk clotting enzyme chymosin using peptide as substrate and covalent organic framework nanosheet as fluorescence quencher
Chymosin is one of the critical enzymes in cheese making. Herein, we proposed a novel fluorometric assay for chymosin determination. Firstly, covalent organic frameworks (COF) were synthesized and exfoliated to 2-dimensional COF nanosheets (COF NS) by ultrasound treatment. Gold nanoparticles (Au NPs...
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Tsinghua University Press
2024-11-01
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| Series: | Food Science and Human Wellness |
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| Online Access: | https://www.sciopen.com/article/10.26599/FSHW.2023.9250042 |
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| author | Xiaokang Lu Tianle Qi Linjiang Guo Lin Xiao Hanbin Xu Guobao Ning Hui Zhao Canpeng Li |
| author_facet | Xiaokang Lu Tianle Qi Linjiang Guo Lin Xiao Hanbin Xu Guobao Ning Hui Zhao Canpeng Li |
| author_sort | Xiaokang Lu |
| collection | DOAJ |
| description | Chymosin is one of the critical enzymes in cheese making. Herein, we proposed a novel fluorometric assay for chymosin determination. Firstly, covalent organic frameworks (COF) were synthesized and exfoliated to 2-dimensional COF nanosheets (COF NS) by ultrasound treatment. Gold nanoparticles (Au NPs) were loaded with COF NS to prepare AuNPs/COF NS (Au@COF NS). Secondly, rhodamine B (RhB) modified substrate peptide (Pep) for chymosin was linked with Au@COF NS to construct a Pep-Au@COF NS nanocomposite. For the sensing principle, fluorescence of RhB was quenched by Au@COF NS and the fluorescence intensity was weak due to the fluorescence resonance energy transfer between COF NS and RhB of Pep. However, in the presence of chymosin, the RhB was released by specific cleavage of the substrate peptide by chymosin and resulted in the recovery of fluorescence. The increased fluorescence intensity was proportional to the increase of chymosin concentration and thus a "turn on" fluorescent sensor for chymosin was constructed. The sensor showed a linear range in the concentration of 0.05-60.00 μg/mL for the detection of chymosin with a detection limit of 20 ng/mL. The sensor was used to quantify chymosin in rennet product with good selectivity, which has the potential applications in cheese manufacturing. |
| format | Article |
| id | doaj-art-8dcc2efc57ec43c089abf0103dcce9fb |
| institution | Kabale University |
| issn | 2097-0765 2213-4530 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Tsinghua University Press |
| record_format | Article |
| series | Food Science and Human Wellness |
| spelling | doaj-art-8dcc2efc57ec43c089abf0103dcce9fb2025-01-10T06:57:02ZengTsinghua University PressFood Science and Human Wellness2097-07652213-45302024-11-011363606361310.26599/FSHW.2023.9250042A novel fluorescence sensor for milk clotting enzyme chymosin using peptide as substrate and covalent organic framework nanosheet as fluorescence quencherXiaokang Lu0Tianle Qi1Linjiang Guo2Lin Xiao3Hanbin Xu4Guobao Ning5Hui Zhao6Canpeng Li7School of Chemical Science and Technology, Yunnan University, Kunming 650091, ChinaSchool of Chemical Science and Technology, Yunnan University, Kunming 650091, ChinaSchool of Chemical Science and Technology, Yunnan University, Kunming 650091, ChinaSchool of Chemical Science and Technology, Yunnan University, Kunming 650091, ChinaSchool of Chemical Science and Technology, Yunnan University, Kunming 650091, ChinaSchool of Chemical Science and Technology, Yunnan University, Kunming 650091, ChinaLaboratory for Conservation and Utilization of Bio-Resource, Yunnan University, Kunming 650091, ChinaSchool of Chemical Science and Technology, Yunnan University, Kunming 650091, ChinaChymosin is one of the critical enzymes in cheese making. Herein, we proposed a novel fluorometric assay for chymosin determination. Firstly, covalent organic frameworks (COF) were synthesized and exfoliated to 2-dimensional COF nanosheets (COF NS) by ultrasound treatment. Gold nanoparticles (Au NPs) were loaded with COF NS to prepare AuNPs/COF NS (Au@COF NS). Secondly, rhodamine B (RhB) modified substrate peptide (Pep) for chymosin was linked with Au@COF NS to construct a Pep-Au@COF NS nanocomposite. For the sensing principle, fluorescence of RhB was quenched by Au@COF NS and the fluorescence intensity was weak due to the fluorescence resonance energy transfer between COF NS and RhB of Pep. However, in the presence of chymosin, the RhB was released by specific cleavage of the substrate peptide by chymosin and resulted in the recovery of fluorescence. The increased fluorescence intensity was proportional to the increase of chymosin concentration and thus a "turn on" fluorescent sensor for chymosin was constructed. The sensor showed a linear range in the concentration of 0.05-60.00 μg/mL for the detection of chymosin with a detection limit of 20 ng/mL. The sensor was used to quantify chymosin in rennet product with good selectivity, which has the potential applications in cheese manufacturing.https://www.sciopen.com/article/10.26599/FSHW.2023.9250042covalent organic frameworkfluorescence sensorchymosinfluorescence resonance energy transfer |
| spellingShingle | Xiaokang Lu Tianle Qi Linjiang Guo Lin Xiao Hanbin Xu Guobao Ning Hui Zhao Canpeng Li A novel fluorescence sensor for milk clotting enzyme chymosin using peptide as substrate and covalent organic framework nanosheet as fluorescence quencher Food Science and Human Wellness covalent organic framework fluorescence sensor chymosin fluorescence resonance energy transfer |
| title | A novel fluorescence sensor for milk clotting enzyme chymosin using peptide as substrate and covalent organic framework nanosheet as fluorescence quencher |
| title_full | A novel fluorescence sensor for milk clotting enzyme chymosin using peptide as substrate and covalent organic framework nanosheet as fluorescence quencher |
| title_fullStr | A novel fluorescence sensor for milk clotting enzyme chymosin using peptide as substrate and covalent organic framework nanosheet as fluorescence quencher |
| title_full_unstemmed | A novel fluorescence sensor for milk clotting enzyme chymosin using peptide as substrate and covalent organic framework nanosheet as fluorescence quencher |
| title_short | A novel fluorescence sensor for milk clotting enzyme chymosin using peptide as substrate and covalent organic framework nanosheet as fluorescence quencher |
| title_sort | novel fluorescence sensor for milk clotting enzyme chymosin using peptide as substrate and covalent organic framework nanosheet as fluorescence quencher |
| topic | covalent organic framework fluorescence sensor chymosin fluorescence resonance energy transfer |
| url | https://www.sciopen.com/article/10.26599/FSHW.2023.9250042 |
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