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. 2021 Nov 24:4:829-839.
doi: 10.1016/j.crfs.2021.11.007. eCollection 2021.

Copper chelating protein hydrolysate from Salvia hispanica L. by pepsin-pancreatin treatment

Latha B V  1 , Likhitha R  2 , Chethan Kumar M  1
Affiliations

Copper chelating protein hydrolysate from Salvia hispanica L. by pepsin-pancreatin treatment

Latha B V et al. Curr Res Food Sci. .

Abstract

Salvia hispanica L. (Chia) seeds are good source of proteins with diverse health benefits. The seed protein was extracted through alkaline solubilisation followed by acid precipitation to separate fibres and are digested sequentially by pepsin and pancreatin. Enzyme-substrate ratio, temperature and contact time had high impact on degree of hydrolysis affecting their chelating ability. Maximum degree of hydrolysis (14.06%) and maximum copper chelation (74.98%) was obtained at 4% w/w enzyme-substrate ratio at 37 °C for 4 h. Copper chelating enzymatic hydrolysate was isolated by HiTrap chelating column and purified further by rpHPLC. Out of nine fractions obtained by rpHPLC the sixth fraction with 93.09 ± 0.16% of copper chelating activity and 82.91 ± 0.52% of antioxidant activity was further characterized as Copper chelating Chia Protein Hydrolysate (CCPH). Ultraviolet spectroscopy and fluorescence spectroscopic studies revealed the interaction of the major chelating sites of the CCPH with the copper divalent ion. The purified CCPH was subjected to LC-MS/ESI-TOF analysis from which six major intense peaks obtained with m/z value ranging from 0.4 kDa to 2.5 kDa were identified and sequenced using Mascot database. The functional behaviour and the binding capacity of these peptides were analysed by their amino acid composition. The CCPH was stable in a simulated gastric condition and its chelating ability remained unaltered. These results explored an informative bioactive peptides with varied activity and one valuable among is the copper chelating with antioxidant property. Furthermore, these Chia seed protein hydrolysates can be useful as dietary supplements to enhance mineral bioavailability.

Keywords: Chia seed; Copper chelating peptides; Protein hydrolysate.

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Conflict of interest statement

The authors declare that they have no competing financial interests and personal relationship that could have appeared to influence the work reported in this paper.

Figures

Image 1
Graphical abstract
Fig. 1
Fig. 1
Degree of hydrolysis (DH) and percentage copper chelation (PCC) of chia seed protein at different temperature and enzyme concentrations.
Fig. 2
Fig. 2
Analytical rpHPLC elution profile of (A) pepsin-pancreatin hydrolysed chia seed protein (CPH) and (B) purified copper chelating chia protein hydrolysate (CCPH).
Fig. 3
Fig. 3
Percentage antioxidant activity and chelation of copper ions by chia seed protein and its hydrolysates.
Fig. 4
Fig. 4
Structural analysis of purified copper chelating CCPH using (A) Fluorescence quenching at wavelength 310 nm (B) UV spectra with different CuSO4 concentrations over the wavelength range from 190 to 400 nm.
Fig. 5
Fig. 5
Q-TOF mass spectrometer analysis of Hi-Trap and rpHPLC purified CCPH fraction showing six major ions with m/z value of 0.562 kDa, 0.508 kDa, 0.433 kDa, 0.482 kDa, 2.522 kDa and 2.332 kDa respectively.
Fig. 5
Fig. 5
Q-TOF mass spectrometer analysis of Hi-Trap and rpHPLC purified CCPH fraction showing six major ions with m/z value of 0.562 kDa, 0.508 kDa, 0.433 kDa, 0.482 kDa, 2.522 kDa and 2.332 kDa respectively.
Fig. 5
Fig. 5
Q-TOF mass spectrometer analysis of Hi-Trap and rpHPLC purified CCPH fraction showing six major ions with m/z value of 0.562 kDa, 0.508 kDa, 0.433 kDa, 0.482 kDa, 2.522 kDa and 2.332 kDa respectively.
See this image and copyright information in PMC

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