Isolation and characterization of proline-rich peptides from Indian indigenous cow colostrum
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Date
2022
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ICAR-NDRI, KARNAL
Abstract
The present investigation was aimed to isolate and characterize a group of peptides known
as Proline rich peptides (PRP) or Colostrinin from Indian Indigenous cow colostrum. PRP was
isolated from colostrum of three indigenous cow breeds viz. Gir, Sahiwal and Tharparkar and
a cross breed Karan Fries, at different milking intervals (1, 12, 24, and 36 h postpartum). The
compositional variation of colostrum with milking interval showed that colostrum of Indigenous
breeds except for Fat, had significantly (P<0.05) higher Protein, Lactose, Salt and Total solids
content than crossbreed. Except for lactose all the other parameters were maximum in the
first milking hour, while they decreased significantly (P<0.05) as milking intervals proceeded
postpartum. Protein (% w/w) was highest in Gir colostrum (16.1±0.02% w/w) and lowest in
Karan Fries (15.10±0.06% w/w) at 1 h postpartum. PRP isolation was optimized using
membrane filtration and two different PRP fractions viz. 10 kDa and 3 kDa peptide fractions
obtained were analysed for variation among breeds with respect to milking intervals. Peptide
content was highest at 1 h postpartum and decreased significantly (P<0.05) upto 36 h
postpartum. Peptide content was highest in Gir (0.310±0.002 mg/ml; 0.192±0.002 mg/ml) and
lowest in Karan Fries (0.265±0.002 mg/ml; 0.161±0.003 mg/ml) in 10 kDa and 3 kDa peptide
fractions, respectively at 1 h postpartum. SDS-PAGE analysis of 10 kDa and 3 kDa peptide
fractions at 1h and 12 h postpartum, showed the presence of PRP’s. Functional properties of
PRP revealed that samples obtained after 1 h postpartum had higher activity than those
obtained at 12 h postpartum. ACE inhibitory activity was highest in Gir (84.2±0.796;
80.7±1.04) and lowest in Karan Fries (69.1±1.59; 58.3±2.39) in 10 kDa and 3 kDa fractions,
respectively. IC50 value (ABTS) was lowest for Gir (141.11 μg/ml; 219.58 μg/ml) followed by
Tharparkar (142.55 μg/ml; 261.44 μg/ml), Sahiwal (172.24 μg/ml; 311 μg/ml) and Karan Fries
(268.26 μg/ml; 457.29 μg/ml) in 3 kDa and 10 kDa fractions, respectively at 1 h postpartum.
Qualitative analysis of phagocytic activity in PRP showed the peritoneal macrophages actively
showing phagocytic activity in the presence of PRP’s. Lymphocyte proliferation index of PRP
showed that Karan Fries (1.59±0.083) had pro-inflammatory activity, while Gir (1.21±0.024),
Sahiwal (1.16±0.045) and Tharparkar (1.17±0.032) had moderate pro- and better antiinflammatory
activity at 1 h postpartum. From the present investigation, it was concluded that
PRP from Indian indigenous cow colostrum has superior functional attributes viz. ACE
inhibitory activity, antioxidant activity and immunomodulatory properties compared to the PRP
obtained Karan Fries cow.