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Molecules
2018 Mar 06;233:. doi: 10.3390/molecules23030590.
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Enzyme-Assisted Extraction Optimization, Characterization and Antioxidant Activity of Polysaccharides from Sea Cucumber Phyllophorus proteus.
Qin Y
,
Yuan Q
,
Zhang Y
,
Li J
,
Zhu X
,
Zhao L
,
Wen J
,
Liu J
,
Zhao L
,
Zhao J
.
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Enzyme-assisted extraction optimization, characterization and in vitro antioxidant activity of polysaccharides from sea cucumber Phyllophorus proteus (PPP) were investigated in the present study. The optimal extraction conditions with a yield of 6.44 ± 0.06% for PPP were determined as follows: Extraction time of 2.89 h, ratio of extraction solvent to raw material of 16.26 mL/g, extraction pH of 6.83, exraction temperature of 50 °C and papain concentration of 0.15%. Three purified fractions, PPP-1a, PPP-1b and PPP-2 with molecular weights of 369.60, 41.73 and 57.76 kDa, respectively, were obtained from PPP by chromatography of FPA98Cl and Sepharose CL-6B columns. Analysis of monosaccharide compositions showed that PPP-1a consisted of N-acetyl-galactosamine (GalNAc), galactose (Gal) and fucose (Fuc), PPP-1b of Fuc as the only monosaccharide and PPP-2 of glucuronic acid, GalNAc and Fuc. Sulfate contents of PPP, PPP-1a, PPP-1b and PPP-2 were determined to be 21.9%, 20.6%, 25.2% and 28.0% (w/w), respectively. PPP and PPP-1a had higher molecular weight and intrinsic viscosity than those of the PPP-1b and PPP-2. PPP, PPP-1a, PPP-1b and PPP-2 exhibited obvious activities of scavenging 1,1-diphenyl-2-picrylhydrazyl radical, hydroxyl radical, superoxide radical and ABTS radical in different extent, which suggested that the polysaccharides from Phyllophorus proteus may be novel agents having potential value for antioxidation.
Figure 1. Effects of different extraction parameters such as enzyme concentration (A), temperature (B), ratio of solvent to raw material (C), pH (D), and time (E) on the yield of polysaccharides from sea cucumber Phyllophorus proteus. ★ indicates the optimal reaction condition for each single-factor test.
Figure 2. Response surface plots (A, C and E) and contour plots (B, D and F) showing the effects of variables (X1, ratio of extraction solvent to raw material; X2, extraction time; X3, extraction pH) and their mutual effects on the extraction yield of PPP.
Figure 3. HPLC profiles of PPP (A), PPP-1a (B), PPP-2 (C) and PPP-1b (D) (a), chromatograms of PMP derivatives of mixed monosaccharide standards (A), PPP-1a (B), PPP-1b (C), PPP-2 (D) and PPP (E) (b), and conductimetric titration curves of PPP-1a, PPP-1b and PPP-2 (c).
Figure 4. FI-IR (A) and 1H-NMR (B) spectra of PPP-1a, PPP-1b and PPP-2.
Figure 5. Scavenging effects of PPP, PPP-1a, PPP-1b and PPP-2 on DPPH radical (A), hydroxyl radical (B), superoxide radical (C) and ABTS radical (D), respectively.
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