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Partial enrichment of phospholipids by enzymatic hydrolysis and membrane filtration of whey protein phospholipid concentrate
Whey protein phospholipid concentrate (WPPC) contains high amounts of phospholipids (PL; 4.5 ± 1%) but there is interest in further enriching the PL content for nutritional and functional applications. Chemical methods were unsuccessful in separating PL from proteins due to the presence of protein-f...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10285240/ https://www.ncbi.nlm.nih.gov/pubmed/37360124 http://dx.doi.org/10.3168/jdsc.2022-0322 |
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author | Swaminathan, A.V. Molitor, M.S. Burrington, K.J. Otter, D. Lucey, J.A. |
author_facet | Swaminathan, A.V. Molitor, M.S. Burrington, K.J. Otter, D. Lucey, J.A. |
author_sort | Swaminathan, A.V. |
collection | PubMed |
description | Whey protein phospholipid concentrate (WPPC) contains high amounts of phospholipids (PL; 4.5 ± 1%) but there is interest in further enriching the PL content for nutritional and functional applications. Chemical methods were unsuccessful in separating PL from proteins due to the presence of protein-fat aggregates. Instead, we explored hydrolysis of the proteins to peptides with the objective of removing peptides, thereby concentrating the PL fraction. We used microfiltration (MF) with a pore size of 0.1 µm to help reduce protein/peptide retention. Hydrolyzing proteins should facilitate passage of low molecular weight peptides through the MF membrane, while concentrating fat and PL in the MF retentate. Bench-top experiments were performed to select the proteolytic enzyme that resulted in the most extensive hydrolysis of proteins in WPPC from among 5 different commercial proteases. Sodium dodecyl sulfate-PAGE analysis was performed to measure the extent of protein hydrolysis over a period of 4 h. Alcalase enzyme was found to exhibit the highest proteolytic activity at conditions of pH 8 and temperature 55°C. The intensity of major protein bands (milkfat globule membrane proteins, caseins, β-lactoglobulin) in WPPC decreased in sodium dodecyl sulfate-PAGE profiles as hydrolysis progressed, along with the appearance of low molecular weight bands. Pilot-scale MF production, coupled with diafiltration (DF), of the hydrolyzed sample aided in the removal of peptides that caused an ~18% reduction in protein content with the final retentate having a total PL content of 9.3% dry basis (db) with protein and fat contents at approximately 43.8 ± 0.4% (db) and 48.9 ± 1.2% (db), respectively. The MF permeate had minimal fat content, indicating that there was no transmission of lipids or PL through the membrane during the MF/DF process. Confocal laser scanning microscopy and particle size analysis of enzyme hydrolyzed solution revealed that protein aggregates were still present after 1 h of hydrolysis. Complete removal of proteins and peptides was not achieved by this process, suggesting that a combination of enzymes would be needed for further hydrolysis of protein aggregates in WPPC solution to further enrich the PL content. |
format | Online Article Text |
id | pubmed-10285240 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-102852402023-06-23 Partial enrichment of phospholipids by enzymatic hydrolysis and membrane filtration of whey protein phospholipid concentrate Swaminathan, A.V. Molitor, M.S. Burrington, K.J. Otter, D. Lucey, J.A. JDS Commun Dairy Foods Whey protein phospholipid concentrate (WPPC) contains high amounts of phospholipids (PL; 4.5 ± 1%) but there is interest in further enriching the PL content for nutritional and functional applications. Chemical methods were unsuccessful in separating PL from proteins due to the presence of protein-fat aggregates. Instead, we explored hydrolysis of the proteins to peptides with the objective of removing peptides, thereby concentrating the PL fraction. We used microfiltration (MF) with a pore size of 0.1 µm to help reduce protein/peptide retention. Hydrolyzing proteins should facilitate passage of low molecular weight peptides through the MF membrane, while concentrating fat and PL in the MF retentate. Bench-top experiments were performed to select the proteolytic enzyme that resulted in the most extensive hydrolysis of proteins in WPPC from among 5 different commercial proteases. Sodium dodecyl sulfate-PAGE analysis was performed to measure the extent of protein hydrolysis over a period of 4 h. Alcalase enzyme was found to exhibit the highest proteolytic activity at conditions of pH 8 and temperature 55°C. The intensity of major protein bands (milkfat globule membrane proteins, caseins, β-lactoglobulin) in WPPC decreased in sodium dodecyl sulfate-PAGE profiles as hydrolysis progressed, along with the appearance of low molecular weight bands. Pilot-scale MF production, coupled with diafiltration (DF), of the hydrolyzed sample aided in the removal of peptides that caused an ~18% reduction in protein content with the final retentate having a total PL content of 9.3% dry basis (db) with protein and fat contents at approximately 43.8 ± 0.4% (db) and 48.9 ± 1.2% (db), respectively. The MF permeate had minimal fat content, indicating that there was no transmission of lipids or PL through the membrane during the MF/DF process. Confocal laser scanning microscopy and particle size analysis of enzyme hydrolyzed solution revealed that protein aggregates were still present after 1 h of hydrolysis. Complete removal of proteins and peptides was not achieved by this process, suggesting that a combination of enzymes would be needed for further hydrolysis of protein aggregates in WPPC solution to further enrich the PL content. Elsevier 2023-02-24 /pmc/articles/PMC10285240/ /pubmed/37360124 http://dx.doi.org/10.3168/jdsc.2022-0322 Text en © 2023. https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Dairy Foods Swaminathan, A.V. Molitor, M.S. Burrington, K.J. Otter, D. Lucey, J.A. Partial enrichment of phospholipids by enzymatic hydrolysis and membrane filtration of whey protein phospholipid concentrate |
title | Partial enrichment of phospholipids by enzymatic hydrolysis and membrane filtration of whey protein phospholipid concentrate |
title_full | Partial enrichment of phospholipids by enzymatic hydrolysis and membrane filtration of whey protein phospholipid concentrate |
title_fullStr | Partial enrichment of phospholipids by enzymatic hydrolysis and membrane filtration of whey protein phospholipid concentrate |
title_full_unstemmed | Partial enrichment of phospholipids by enzymatic hydrolysis and membrane filtration of whey protein phospholipid concentrate |
title_short | Partial enrichment of phospholipids by enzymatic hydrolysis and membrane filtration of whey protein phospholipid concentrate |
title_sort | partial enrichment of phospholipids by enzymatic hydrolysis and membrane filtration of whey protein phospholipid concentrate |
topic | Dairy Foods |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10285240/ https://www.ncbi.nlm.nih.gov/pubmed/37360124 http://dx.doi.org/10.3168/jdsc.2022-0322 |
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