The insertion of bioactive peptides at the C-terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activity
Background: The 11S globulin from amaranth is the most abundant storage protein in mature seeds and is well recognized for its nutritional value. We used this globulin to engineer a new protein by adding a four valine-tyrosine antihypertensive peptide at its C-terminal end to improve its functionali...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2019-01-01
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| Series: | Electronic Journal of Biotechnology |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0717345818300447 |
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| author | Edgar Espinosa-Hernández Jocksan Ismael Morales-Camacho D. Alejandro Fernández-Velasco Claudia G. Benítez-Cardoza Flor de Fátima Rosas-Cárdenas Silvia Luna-Suárez |
| author_facet | Edgar Espinosa-Hernández Jocksan Ismael Morales-Camacho D. Alejandro Fernández-Velasco Claudia G. Benítez-Cardoza Flor de Fátima Rosas-Cárdenas Silvia Luna-Suárez |
| author_sort | Edgar Espinosa-Hernández |
| collection | DOAJ |
| description | Background: The 11S globulin from amaranth is the most abundant storage protein in mature seeds and is well recognized for its nutritional value. We used this globulin to engineer a new protein by adding a four valine-tyrosine antihypertensive peptide at its C-terminal end to improve its functionality. The new protein was named AMR5 and expressed in the Escherichia coli BL21-CodonPlus(DE3)-RIL strain using a custom medium (F8PW) designed for this work. Results: The alternative medium allowed for the production of 652 mg/L expressed protein at the flask level, mostly in an insoluble form, and this protein was subjected to in vitro refolding. The spectrometric analysis suggests that the protein adopts a β/α structure with a small increment of α-helix conformation relative to the native amaranth 11S globulin. Thermal and urea denaturation experiments determined apparent Tm and C1/2 values of 50.4°C and 3.04 M, respectively, thus indicating that the antihypertensive peptide insertion destabilized the modified protein relative to the native one. AMR5 hydrolyzed by trypsin and chymotrypsin showed 14- and 1.3-fold stronger inhibitory activity against angiotensin I-converting enzyme (IC50 of 0.034 mg/mL) than the unmodified protein and the previously reported amaranth acidic subunit modified with antihypertensive peptides, respectively. Conclusion: The inserted peptide decreases the structural stability of amaranth 11S globulin and improves its antihypertensive activity.How to cite: Espinosa-Hernández E, Morales-Camacho JI, Fernández-Velasco DA, et al. The insertion of bioactive peptides at the C terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activity. Electron J Biotechnol 2019;37. https://doi.org/10.1016/j.ejbt.2018.11.001. Keywords: Amaranth, Antihypertensive peptides, Bioactive, Culture media design, Globulin, Mature seeds, Potato waste, Protein engineering, Protein expression, Protein stability, Storage protein, Thermal stability |
| first_indexed | 2024-12-12T17:31:51Z |
| format | Article |
| id | doaj.art-97021b6b914b4794835cae83d2f1b07e |
| institution | Directory Open Access Journal |
| issn | 0717-3458 |
| language | English |
| last_indexed | 2024-12-12T17:31:51Z |
| publishDate | 2019-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Electronic Journal of Biotechnology |
| spelling | doaj.art-97021b6b914b4794835cae83d2f1b07e2022-12-22T00:17:20ZengElsevierElectronic Journal of Biotechnology0717-34582019-01-01371824The insertion of bioactive peptides at the C-terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activityEdgar Espinosa-Hernández0Jocksan Ismael Morales-Camacho1D. Alejandro Fernández-Velasco2Claudia G. Benítez-Cardoza3Flor de Fátima Rosas-Cárdenas4Silvia Luna-Suárez5Centro de Investigación en Biotecnología Aplicada, Instituto Politécnico Nacional, CIBA-IPN, Ex-Hacienda San Juan Molino, carretera estatal Tecuexcomac-Tepetitla Km 1.5, 90700 Tepetitla, Tlaxcala, MéxicoDepartamento de Ingeniería Química, Alimentos y Ambiental, Universidad de las Américas Puebla, 72830 Cholula, Puebla, MéxicoLaboratorio de Fisicoquímica e Ingeniería de Proteínas, Departamento de Bioquímica, Facultad de Medicina, UNAM, 04510 Ciudad de México, MéxicoLaboratorio de Investigación Bioquímica, Instituto Politécnico Nacional, ENMyH-IPN, 07320 Ciudad de México, MéxicoCentro de Investigación en Biotecnología Aplicada, Instituto Politécnico Nacional, CIBA-IPN, Ex-Hacienda San Juan Molino, carretera estatal Tecuexcomac-Tepetitla Km 1.5, 90700 Tepetitla, Tlaxcala, MéxicoCentro de Investigación en Biotecnología Aplicada, Instituto Politécnico Nacional, CIBA-IPN, Ex-Hacienda San Juan Molino, carretera estatal Tecuexcomac-Tepetitla Km 1.5, 90700 Tepetitla, Tlaxcala, México; Corresponding author.Background: The 11S globulin from amaranth is the most abundant storage protein in mature seeds and is well recognized for its nutritional value. We used this globulin to engineer a new protein by adding a four valine-tyrosine antihypertensive peptide at its C-terminal end to improve its functionality. The new protein was named AMR5 and expressed in the Escherichia coli BL21-CodonPlus(DE3)-RIL strain using a custom medium (F8PW) designed for this work. Results: The alternative medium allowed for the production of 652 mg/L expressed protein at the flask level, mostly in an insoluble form, and this protein was subjected to in vitro refolding. The spectrometric analysis suggests that the protein adopts a β/α structure with a small increment of α-helix conformation relative to the native amaranth 11S globulin. Thermal and urea denaturation experiments determined apparent Tm and C1/2 values of 50.4°C and 3.04 M, respectively, thus indicating that the antihypertensive peptide insertion destabilized the modified protein relative to the native one. AMR5 hydrolyzed by trypsin and chymotrypsin showed 14- and 1.3-fold stronger inhibitory activity against angiotensin I-converting enzyme (IC50 of 0.034 mg/mL) than the unmodified protein and the previously reported amaranth acidic subunit modified with antihypertensive peptides, respectively. Conclusion: The inserted peptide decreases the structural stability of amaranth 11S globulin and improves its antihypertensive activity.How to cite: Espinosa-Hernández E, Morales-Camacho JI, Fernández-Velasco DA, et al. The insertion of bioactive peptides at the C terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activity. Electron J Biotechnol 2019;37. https://doi.org/10.1016/j.ejbt.2018.11.001. Keywords: Amaranth, Antihypertensive peptides, Bioactive, Culture media design, Globulin, Mature seeds, Potato waste, Protein engineering, Protein expression, Protein stability, Storage protein, Thermal stabilityhttp://www.sciencedirect.com/science/article/pii/S0717345818300447 |
| spellingShingle | Edgar Espinosa-Hernández Jocksan Ismael Morales-Camacho D. Alejandro Fernández-Velasco Claudia G. Benítez-Cardoza Flor de Fátima Rosas-Cárdenas Silvia Luna-Suárez The insertion of bioactive peptides at the C-terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activity Electronic Journal of Biotechnology |
| title | The insertion of bioactive peptides at the C-terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activity |
| title_full | The insertion of bioactive peptides at the C-terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activity |
| title_fullStr | The insertion of bioactive peptides at the C-terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activity |
| title_full_unstemmed | The insertion of bioactive peptides at the C-terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activity |
| title_short | The insertion of bioactive peptides at the C-terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activity |
| title_sort | insertion of bioactive peptides at the c terminal end of an 11s globulin changes the structural stability and improves the antihypertensive activity |
| url | http://www.sciencedirect.com/science/article/pii/S0717345818300447 |
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