Improving protein hydrolysis and digestibility in Arthrospira platensis biomass through recombinant peptidases (EC 3.4): Opportunities for monogastric animal diets
This study investigates the use of recombinant peptidases (EC 3.4) to improve protein hydrolysis and digestibility in Arthrospira platensis, with a focus on addressing the challenge of reduced protein bioavailability for monogastric animals due to resistant protein-pigment formations, such as phycoc...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-01-01
|
| Series: | Heliyon |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844024174919 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841533323650793472 |
|---|---|
| author | Maria P. Spínola Mónica M. Costa Rita S. Simões Vânia O. Fernandes Vânia Cardoso Virgínia M.R. Pires Cláudia Afonso Carlos Cardoso Narcisa M. Bandarra Carlos M.G.A. Fontes José A.M. Prates |
| author_facet | Maria P. Spínola Mónica M. Costa Rita S. Simões Vânia O. Fernandes Vânia Cardoso Virgínia M.R. Pires Cláudia Afonso Carlos Cardoso Narcisa M. Bandarra Carlos M.G.A. Fontes José A.M. Prates |
| author_sort | Maria P. Spínola |
| collection | DOAJ |
| description | This study investigates the use of recombinant peptidases (EC 3.4) to improve protein hydrolysis and digestibility in Arthrospira platensis, with a focus on addressing the challenge of reduced protein bioavailability for monogastric animals due to resistant protein-pigment formations, such as phycocyanin, and increased digesta viscosity caused by jellification properties. A library of 192 peptidases was generated, from which 142 soluble peptidases were expressed in Escherichia coli and subsequently screened for activity against an A. platensis suspension in vitro. Among these peptidases, six promising candidates were identified for protein and peptide extraction from the microalga. These enzymes were tested individually, and in a mix (MIX6), and compared to commercial trypsin and pancreatin. Protein content was determined using the Bradford method and potential peptide formation was measured via an o-phthaldialdehyde (OPA) assay. The protein solubility and hydrolysis, specifically of two main protein fractions (18–26 kDa and 40–48 kDa) along with minor fractions, were analysed via 14 % sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Results indicated that the enzyme ID 138, a serine-peptidase, significantly increased total peptide formation in the A. platensis supernatant, although it did not outperform other peptidases or enzyme mixtures. Notably, enzymes ID 152, derived from a marine bacterium, and ID 153, another serine-peptidase, exhibited significant improvements in the extraction and hydrolysis of one protein fraction (18–26 kDa), possibly corresponding to a phycocyanin fraction. While no synergistic effects were observed among peptidases, further investigations are warranted to understand the enzyme composition of MIX6, particularly enzymes ID 138, ID 152 and ID 153, and their potential to enhance the bioavailability of A. platensis proteins for monogastric animals when incorporated into dietary formulations. |
| format | Article |
| id | doaj-art-9bab60b39dff4c18b4f31c584ecd0da7 |
| institution | Kabale University |
| issn | 2405-8440 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj-art-9bab60b39dff4c18b4f31c584ecd0da72025-01-17T04:51:20ZengElsevierHeliyon2405-84402025-01-01111e41460Improving protein hydrolysis and digestibility in Arthrospira platensis biomass through recombinant peptidases (EC 3.4): Opportunities for monogastric animal dietsMaria P. Spínola0Mónica M. Costa1Rita S. Simões2Vânia O. Fernandes3Vânia Cardoso4Virgínia M.R. Pires5Cláudia Afonso6Carlos Cardoso7Narcisa M. Bandarra8Carlos M.G.A. Fontes9José A.M. Prates10CIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade Técnica, 1300-477, Lisboa, Portugal; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. Da Universidade Técnica, 1300-477, Lisboa, PortugalCIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade Técnica, 1300-477, Lisboa, Portugal; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. Da Universidade Técnica, 1300-477, Lisboa, PortugalCIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade Técnica, 1300-477, Lisboa, Portugal; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. Da Universidade Técnica, 1300-477, Lisboa, Portugal; NZYTech - Genes and Enzymes, Estrada do Paço do Lumiar, Campus do Lumiar, Edifício E, 1649-038, Lisboa, PortugalCIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade Técnica, 1300-477, Lisboa, Portugal; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. Da Universidade Técnica, 1300-477, Lisboa, Portugal; NZYTech - Genes and Enzymes, Estrada do Paço do Lumiar, Campus do Lumiar, Edifício E, 1649-038, Lisboa, PortugalCIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade Técnica, 1300-477, Lisboa, Portugal; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. Da Universidade Técnica, 1300-477, Lisboa, Portugal; NZYTech - Genes and Enzymes, Estrada do Paço do Lumiar, Campus do Lumiar, Edifício E, 1649-038, Lisboa, PortugalCIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade Técnica, 1300-477, Lisboa, Portugal; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. Da Universidade Técnica, 1300-477, Lisboa, Portugal; NZYTech - Genes and Enzymes, Estrada do Paço do Lumiar, Campus do Lumiar, Edifício E, 1649-038, Lisboa, PortugalDivAV - Division of Aquaculture and Upgrading, Portuguese Institute for the Sea and Atmosphere, Rua Alfredo Magalhães Ramalho, 6, 1495-006, Lisbon, Portugal; CIIMAR, Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua dos Bragas 289, 4050-123, Porto, PortugalDivAV - Division of Aquaculture and Upgrading, Portuguese Institute for the Sea and Atmosphere, Rua Alfredo Magalhães Ramalho, 6, 1495-006, Lisbon, Portugal; CIIMAR, Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua dos Bragas 289, 4050-123, Porto, PortugalDivAV - Division of Aquaculture and Upgrading, Portuguese Institute for the Sea and Atmosphere, Rua Alfredo Magalhães Ramalho, 6, 1495-006, Lisbon, Portugal; CIIMAR, Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Rua dos Bragas 289, 4050-123, Porto, PortugalCIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade Técnica, 1300-477, Lisboa, Portugal; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. Da Universidade Técnica, 1300-477, Lisboa, Portugal; NZYTech - Genes and Enzymes, Estrada do Paço do Lumiar, Campus do Lumiar, Edifício E, 1649-038, Lisboa, PortugalCIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. da Universidade Técnica, 1300-477, Lisboa, Portugal; Associate Laboratory for Animal and Veterinary Sciences (AL4AnimalS), Faculdade de Medicina Veterinária, Universidade de Lisboa, Av. Da Universidade Técnica, 1300-477, Lisboa, Portugal; Corresponding author. Faculdade de Medicina Veterinária, Av. da Universidade Técnica, Pólo Universitário do Alto da Ajuda, 1300-477, Lisboa, Portugal.This study investigates the use of recombinant peptidases (EC 3.4) to improve protein hydrolysis and digestibility in Arthrospira platensis, with a focus on addressing the challenge of reduced protein bioavailability for monogastric animals due to resistant protein-pigment formations, such as phycocyanin, and increased digesta viscosity caused by jellification properties. A library of 192 peptidases was generated, from which 142 soluble peptidases were expressed in Escherichia coli and subsequently screened for activity against an A. platensis suspension in vitro. Among these peptidases, six promising candidates were identified for protein and peptide extraction from the microalga. These enzymes were tested individually, and in a mix (MIX6), and compared to commercial trypsin and pancreatin. Protein content was determined using the Bradford method and potential peptide formation was measured via an o-phthaldialdehyde (OPA) assay. The protein solubility and hydrolysis, specifically of two main protein fractions (18–26 kDa and 40–48 kDa) along with minor fractions, were analysed via 14 % sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Results indicated that the enzyme ID 138, a serine-peptidase, significantly increased total peptide formation in the A. platensis supernatant, although it did not outperform other peptidases or enzyme mixtures. Notably, enzymes ID 152, derived from a marine bacterium, and ID 153, another serine-peptidase, exhibited significant improvements in the extraction and hydrolysis of one protein fraction (18–26 kDa), possibly corresponding to a phycocyanin fraction. While no synergistic effects were observed among peptidases, further investigations are warranted to understand the enzyme composition of MIX6, particularly enzymes ID 138, ID 152 and ID 153, and their potential to enhance the bioavailability of A. platensis proteins for monogastric animals when incorporated into dietary formulations.http://www.sciencedirect.com/science/article/pii/S2405844024174919Arthrospira platensisMicroalgaProtein hydrolysisProtein digestibilityRecombinant peptidase |
| spellingShingle | Maria P. Spínola Mónica M. Costa Rita S. Simões Vânia O. Fernandes Vânia Cardoso Virgínia M.R. Pires Cláudia Afonso Carlos Cardoso Narcisa M. Bandarra Carlos M.G.A. Fontes José A.M. Prates Improving protein hydrolysis and digestibility in Arthrospira platensis biomass through recombinant peptidases (EC 3.4): Opportunities for monogastric animal diets Heliyon Arthrospira platensis Microalga Protein hydrolysis Protein digestibility Recombinant peptidase |
| title | Improving protein hydrolysis and digestibility in Arthrospira platensis biomass through recombinant peptidases (EC 3.4): Opportunities for monogastric animal diets |
| title_full | Improving protein hydrolysis and digestibility in Arthrospira platensis biomass through recombinant peptidases (EC 3.4): Opportunities for monogastric animal diets |
| title_fullStr | Improving protein hydrolysis and digestibility in Arthrospira platensis biomass through recombinant peptidases (EC 3.4): Opportunities for monogastric animal diets |
| title_full_unstemmed | Improving protein hydrolysis and digestibility in Arthrospira platensis biomass through recombinant peptidases (EC 3.4): Opportunities for monogastric animal diets |
| title_short | Improving protein hydrolysis and digestibility in Arthrospira platensis biomass through recombinant peptidases (EC 3.4): Opportunities for monogastric animal diets |
| title_sort | improving protein hydrolysis and digestibility in arthrospira platensis biomass through recombinant peptidases ec 3 4 opportunities for monogastric animal diets |
| topic | Arthrospira platensis Microalga Protein hydrolysis Protein digestibility Recombinant peptidase |
| url | http://www.sciencedirect.com/science/article/pii/S2405844024174919 |
| work_keys_str_mv | AT mariapspinola improvingproteinhydrolysisanddigestibilityinarthrospiraplatensisbiomassthroughrecombinantpeptidasesec34opportunitiesformonogastricanimaldiets AT monicamcosta improvingproteinhydrolysisanddigestibilityinarthrospiraplatensisbiomassthroughrecombinantpeptidasesec34opportunitiesformonogastricanimaldiets AT ritassimoes improvingproteinhydrolysisanddigestibilityinarthrospiraplatensisbiomassthroughrecombinantpeptidasesec34opportunitiesformonogastricanimaldiets AT vaniaofernandes improvingproteinhydrolysisanddigestibilityinarthrospiraplatensisbiomassthroughrecombinantpeptidasesec34opportunitiesformonogastricanimaldiets AT vaniacardoso improvingproteinhydrolysisanddigestibilityinarthrospiraplatensisbiomassthroughrecombinantpeptidasesec34opportunitiesformonogastricanimaldiets AT virginiamrpires improvingproteinhydrolysisanddigestibilityinarthrospiraplatensisbiomassthroughrecombinantpeptidasesec34opportunitiesformonogastricanimaldiets AT claudiaafonso improvingproteinhydrolysisanddigestibilityinarthrospiraplatensisbiomassthroughrecombinantpeptidasesec34opportunitiesformonogastricanimaldiets AT carloscardoso improvingproteinhydrolysisanddigestibilityinarthrospiraplatensisbiomassthroughrecombinantpeptidasesec34opportunitiesformonogastricanimaldiets AT narcisambandarra improvingproteinhydrolysisanddigestibilityinarthrospiraplatensisbiomassthroughrecombinantpeptidasesec34opportunitiesformonogastricanimaldiets AT carlosmgafontes improvingproteinhydrolysisanddigestibilityinarthrospiraplatensisbiomassthroughrecombinantpeptidasesec34opportunitiesformonogastricanimaldiets AT joseamprates improvingproteinhydrolysisanddigestibilityinarthrospiraplatensisbiomassthroughrecombinantpeptidasesec34opportunitiesformonogastricanimaldiets |