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. 2016 Dec 27;15(1):3.
doi: 10.3390/md15010003.

Novel Peptide with Specific Calcium-Binding Capacity from Schizochytrium sp. Protein Hydrolysates and Calcium Bioavailability in Caco-2 Cells

Xixi Cai  1   2 , Jiaping Lin  3 , Shaoyun Wang  4
Affiliations

Novel Peptide with Specific Calcium-Binding Capacity from Schizochytrium sp. Protein Hydrolysates and Calcium Bioavailability in Caco-2 Cells

Xixi Cai et al. Mar Drugs. .

Abstract

Peptide-calcium can probably be a suitable supplement to improve calcium absorption in the human body. In this study, a specific peptide Phe-Tyr (FY) with calcium-binding capacity was purified from Schizochytrium sp. protein hydrolysates through gel filtration chromatography and reversed phase HPLC. The calcium-binding capacity of FY reached 128.77 ± 2.57 μg/mg. Results of ultraviolet spectroscopy, fluorescence spectroscopy, and infrared spectroscopy showed that carboxyl groups, amino groups, and amido groups were the major chelating sites. FY-Ca exhibited excellent thermal stability and solubility, which were beneficial to be absorbed and transported in the basic intestinal tract of the human body. Moreover, the calcium bioavailability in Caco-2 cells showed that FY-Ca could enhance calcium uptake efficiency by more than three times when compared with CaCl₂, and protect calcium ions against dietary inhibitors, such as tannic acid, oxalate, phytate, and Zn2+. Our findings further the progress of algae-based peptide-calcium, suggesting that FY-Ca has the potential to be developed as functionally nutraceutical additives.

Keywords: Schizochytrium sp.; bioavailability; calcium-binding peptide; protein hydrolysate; structure.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Elution profiles and calcium-binding capacities of calcium-binding peptides. (a) Sephadex G-25 gel filtration chromatography of SPH; (b) semi-preparative C18 RP-HPLC of fraction F3; and (c) analytic RP-HPLC of fraction 17 from semi-preparative HPLC.
Figure 2
Figure 2
Identification of the amino acid sequence of the calcium-binding peptide using LC-ESI-MS/MS.
Figure 3
Figure 3
UV spectra of FY with different CaCl2 concentrations over the wavelength range from 190 to 400 nm.
Figure 4
Figure 4
Fluorescence spectra of FY with different CaCl2 concentration over the wavelength range from 295 to 500 nm.
Figure 5
Figure 5
Fourier transform infrared (FTIR) spectra of FY and FY-Ca chelate in the regions from 4000 to 400 cm−1.
Figure 6
Figure 6
Typical TG-DSC thermograms of (a) FY and (b) FY-Ca chelate.
Figure 7
Figure 7
Calcium-releasing percentage of FY-Ca chelate and CaCl2 at different pH.
Figure 8
Figure 8
Cell uptake of FY-Ca chelate and CaCl2 in Caco-2 cell by Fluo-3-AM loading for fluorescence analysis.
Figure 9
Figure 9
Effect of FY-Ca chelate on calcium bioavailability under the action of dietary inhibition factors. The concentration of calcium was 10 mM and tannic acid/Ca, oxalate/Ca, phytate/Ca, or Zn/Ca = 20:1. * Statistical significance p < 0.05, compared with the CaCl2 control group. # Statistical significance p < 0.05, compared with the FY-Ca control group.
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