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Molecules
2020 Jun 08;2511:. doi: 10.3390/molecules25112663.
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Response Factorial Design Analysis on Papain-Generated Hydrolysates from Actinopyga lecanora for Determination of Antioxidant and Antityrosinase Activities.
Bahari AN
,
Saari N
,
Salim N
,
Ashari SE
.
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Actinopyga lecanora (A. lecanora) is classified among the edible species of sea cucumber, known to be rich in protein. Its hydrolysates were reported to contain relatively high antioxidant activity. Antioxidants are one of the essential properties in cosmeceutical products especially to alleviate skin aging. In the present study, pH, reaction temperature, reaction time and enzyme/substrate ratio (E/S) have been identified as the parameters in the papain enzymatic hydrolysis of A. lecanora. The degree of hydrolysis (DH) with antioxidant activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric-reducing antioxidant power (FRAP) assays were used as the responses in the optimization. Analysis of variance (ANOVA), normal plot of residuals and 3D contour plots were evaluated to study the effects and interactions between parameters. The best conditions selected from the optimization were at pH 5.00, 70 °C of reaction temperature, 9 h of hydrolysis time and 1.00% enzyme/substrate (E/S) ratio, with the hydrolysates having 51.90% of DH, 42.70% of DPPH activity and 109.90 Fe2+μg/mL of FRAP activity. The A. lecanora hydrolysates (ALH) showed a high amount of hydrophobic amino acids (286.40 mg/g sample) that might be responsible for antioxidant and antityrosinase activities. Scanning electron microscopy (SEM) image of ALH shows smooth structures with pores. Antityrosinase activity of ALH exhibited inhibition of 31.50% for L-tyrosine substrate and 25.40% for L-DOPA substrate. This condition suggests that the optimized ALH acquired has the potential to be used as a bioactive ingredient for cosmeceutical applications.
Figure 1. Normal plot of residual for (a) degree of hydrolysis (DH), (b) 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and (c) ferric-reducing antioxidant power (FRAP) activity.
Figure 2. Three-dimensional (3D) contour plots showing the influence of variable parameters on the degree of hydrolysis (DH) response. (a) pH vs. reaction temperature, (b) reaction time vs. E/S ratio and (c) pH vs. reaction time.
Figure 3. Cube plots showing the interaction of three significant effects on (a) the degree of hydrolysis (DH) response, (b) 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging response and (c) ferric-reducing antioxidant power (FRAP) response.
Figure 4. 3D contour plots showing the influence of variable parameters on the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging response. (a) pH vs. reaction time, (b) pH vs. reaction temperature and (c) reaction temperature vs. E/S ratio.
Figure 5. 3D contour plots showing the influence of variable parameters on the ferric-reducing antioxidant power (FRAP) response. (a) pH vs. E/S ratio, (b) reaction temperature vs. E/S ratio and (c) reaction time vs. E/S ratio.
Figure 6. Scanning electron microscopy (SEM) image of papain-generated hydrolysates from Actinopyga lecanora.
Figure 7. Tyrosinase inhibition activity for an optimized A. lecanora hydrolysates sample.
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