Impact of Extraction Method on the Detection of Quality Biomarkers in Normal vs. DFD Meat
The objective of this work was to demonstrate how the extraction method affects the reliability of biomarker detection and how this detection depends on the biomarker location within the cell compartment. Different extraction methods were used to study the sarcoplasmic and myofibrillar fractions of...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-05-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/10/5/1097 |
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| author | Laura González-Blanco Yolanda Diñeiro Andrea Díaz-Luis Ana Coto-Montes Mamen Oliván Verónica Sierra |
| author_facet | Laura González-Blanco Yolanda Diñeiro Andrea Díaz-Luis Ana Coto-Montes Mamen Oliván Verónica Sierra |
| author_sort | Laura González-Blanco |
| collection | DOAJ |
| description | The objective of this work was to demonstrate how the extraction method affects the reliability of biomarker detection and how this detection depends on the biomarker location within the cell compartment. Different extraction methods were used to study the sarcoplasmic and myofibrillar fractions of the <i>Longissimus thoracis et lumborum</i> muscle of young bulls of the Asturiana de los Valles breed in two quality grades, standard (Control) or dark, firm, and dry (DFD) meat. Protein extractability and the expression of some of the main meat quality biomarkers—oxidative status (lipoperoxidation (LPO) and catalase activity (CAT)), proteome (SDS-PAGE electrophoretic pattern), and cell stress protein (Hsp70)—were analyzed. In the sarcoplasmic fraction, buffers containing Triton X-100 showed significantly higher protein extractability, LPO, and higher intensity of high-molecular-weight protein bands, whereas the TES buffer was more sensitive to distinguishing differences in the protein pattern between the Control and DFD meat. In the myofibrillar fraction, samples extracted with the lysis buffer showed significantly higher protein extractability, whereas samples extracted with the non-denaturing buffer showed higher results for LPO, CAT, and Hsp70, and higher-intensity bands in the electrophoretic pattern. These findings highlight the need for the careful selection of the extraction method used to analyze the different biomarkers considering their cellular location to adapt the extractive process. |
| first_indexed | 2024-03-10T11:23:26Z |
| format | Article |
| id | doaj.art-efdefbd4162d4c3fbf08256f240a0ca5 |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-10T11:23:26Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-efdefbd4162d4c3fbf08256f240a0ca52023-11-21T19:52:06ZengMDPI AGFoods2304-81582021-05-01105109710.3390/foods10051097Impact of Extraction Method on the Detection of Quality Biomarkers in Normal vs. DFD MeatLaura González-Blanco0Yolanda Diñeiro1Andrea Díaz-Luis2Ana Coto-Montes3Mamen Oliván4Verónica Sierra5Área de Sistemas de Producción Animal, Servicio Regional de Investigación y Desarrollo Agroalimentario (SERIDA), Ctra. AS-267, PK 19, 33300 Villaviciosa, Asturias, SpainÁrea de Sistemas de Producción Animal, Servicio Regional de Investigación y Desarrollo Agroalimentario (SERIDA), Ctra. AS-267, PK 19, 33300 Villaviciosa, Asturias, SpainDepartment of Molecular Biology, Faculty of Medicine, University of Cantabria, Av. Herrera Oria, s/n, 39011 Santander, Cantabria, SpainInstituto de Investigación Sanitaria del Principado de Asturias (ISPA), Av. del Hospital Universitario, s/n, 33011 Oviedo, Asturias, SpainÁrea de Sistemas de Producción Animal, Servicio Regional de Investigación y Desarrollo Agroalimentario (SERIDA), Ctra. AS-267, PK 19, 33300 Villaviciosa, Asturias, SpainÁrea de Sistemas de Producción Animal, Servicio Regional de Investigación y Desarrollo Agroalimentario (SERIDA), Ctra. AS-267, PK 19, 33300 Villaviciosa, Asturias, SpainThe objective of this work was to demonstrate how the extraction method affects the reliability of biomarker detection and how this detection depends on the biomarker location within the cell compartment. Different extraction methods were used to study the sarcoplasmic and myofibrillar fractions of the <i>Longissimus thoracis et lumborum</i> muscle of young bulls of the Asturiana de los Valles breed in two quality grades, standard (Control) or dark, firm, and dry (DFD) meat. Protein extractability and the expression of some of the main meat quality biomarkers—oxidative status (lipoperoxidation (LPO) and catalase activity (CAT)), proteome (SDS-PAGE electrophoretic pattern), and cell stress protein (Hsp70)—were analyzed. In the sarcoplasmic fraction, buffers containing Triton X-100 showed significantly higher protein extractability, LPO, and higher intensity of high-molecular-weight protein bands, whereas the TES buffer was more sensitive to distinguishing differences in the protein pattern between the Control and DFD meat. In the myofibrillar fraction, samples extracted with the lysis buffer showed significantly higher protein extractability, whereas samples extracted with the non-denaturing buffer showed higher results for LPO, CAT, and Hsp70, and higher-intensity bands in the electrophoretic pattern. These findings highlight the need for the careful selection of the extraction method used to analyze the different biomarkers considering their cellular location to adapt the extractive process.https://www.mdpi.com/2304-8158/10/5/1097cellular compartmentsprotein extractabilitysarcoplasmic proteinsmyofibrillar proteinsmeat quality biomarkersDFD meat |
| spellingShingle | Laura González-Blanco Yolanda Diñeiro Andrea Díaz-Luis Ana Coto-Montes Mamen Oliván Verónica Sierra Impact of Extraction Method on the Detection of Quality Biomarkers in Normal vs. DFD Meat Foods cellular compartments protein extractability sarcoplasmic proteins myofibrillar proteins meat quality biomarkers DFD meat |
| title | Impact of Extraction Method on the Detection of Quality Biomarkers in Normal vs. DFD Meat |
| title_full | Impact of Extraction Method on the Detection of Quality Biomarkers in Normal vs. DFD Meat |
| title_fullStr | Impact of Extraction Method on the Detection of Quality Biomarkers in Normal vs. DFD Meat |
| title_full_unstemmed | Impact of Extraction Method on the Detection of Quality Biomarkers in Normal vs. DFD Meat |
| title_short | Impact of Extraction Method on the Detection of Quality Biomarkers in Normal vs. DFD Meat |
| title_sort | impact of extraction method on the detection of quality biomarkers in normal vs dfd meat |
| topic | cellular compartments protein extractability sarcoplasmic proteins myofibrillar proteins meat quality biomarkers DFD meat |
| url | https://www.mdpi.com/2304-8158/10/5/1097 |
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