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Isolation of mitogenic glycophosphopeptides from cheese whey protein concentrate
S S Yun1, Y Sugita-Konishi, S Kumagai
1College of Agriculture and Veterinary Medicine, Nihon University, Tokyo, Japan.
Insights
Researchers found that a cheese whey protein concentrate (CWPC) fraction exhibits strong immune-stimulating properties. This glycophosphopeptide (GPP) component, rich in carbohydrates, activates immune cells, suggesting potential applications in immunology.
Area of Science:
- Immunology
- Biochemistry
- Food Science
Background:
- Cheese whey protein concentrate (CWPC) is a dairy by-product.
- The immunological functions of CWPC components are not fully understood.
Purpose of the Study:
- To investigate the immunological function of CWPC.
- To identify and characterize the active component responsible for mitogenic activity.
Main Methods:
- Gel filtration and anion exchange chromatography were used to isolate fractions from CWPC.
- Mitogenic activity was assessed using murine splenocytes.
- SDS-PAGE, Pronase digestion, and affinity chromatography were employed for characterization.
Main Results:
- A CWPC fraction displayed high mitogenic activity, comparable to lipopolysaccharide (LPS).
- The active component was identified as a glycophosphopeptide (GPP), containing calcium, phosphorus, and carbohydrates.
- Pronase digestion did not abolish activity, suggesting carbohydrates are crucial.
- The GPP separated into two components on an anti-beta-caseinophosphopeptide (beta-CPP) column, both retaining mitogenic activity and N-linked oligosaccharide chains.
Conclusions:
- Cheese whey contains a glycophosphopeptide (GPP) with significant mitogenic activity.
- The carbohydrate moiety of the GPP appears to be critical for its immune-stimulating effects.
- These findings highlight the potential immunomodulatory properties of dairy by-products.
Abstract:
We investigated the immunological function of cheese whey protein concentrate (CWPC), which is a by-product of cheese production, using mitogenic activity in murine splenocytes as an index. A fraction isolated by gel filtration and anion exchange chromatography of CWPC showed high mitogenic activity, comparable to the activity of lipopolysaccharide (LPS). The fraction was detected as a single band on SDS-PAGE. It contained calcium, inorganic phosphorus, and carbohydrate, indicating the active component to be a glycophosphopeptide (GPP). Since Pronase digestion of GPP did not reduce its mitogenic activity, carbohydrate rather than peptide may be important in the activity. When applied on an anti-beta-caseinophosphopeptide (beta-CPP) antibody affinity column, the GPP was separated into two components, one with affinity to beta-CPP and the other without such affinity. Both the components contained N-linked oligosaccharide chains and had the mitogenic activity. These results demonstrate that cheese whey contains a GPP having strong mitogenic activity.