Anti-Oxidative and Anti-Aging Activities of Porcine By-Product Collagen Hydrolysates Produced by Commercial Proteases: Effect of Hydrolysis and Ultrafiltration

Abstract

To investigate methods for improving the processing of porcine waste, porcine skin was hydrolyzed using different commercially available proteases (Alcalase, Flavorzyme, Neutrase, Bromeline, Protamex, and Papain) under several optimal conditions. Following enzymatic hydrolysis, the collagen hydrolysates (CHs) were fractionated by molecular weight (3 kDa) via membrane ultrafiltration. The CHs were analyzed for physical properties (pH, protein recovery, free amino group content, molecular weight distribution, and amino composition) as well as for functional properties (antioxidant activities and anti-aging activities). Among the CHs, CHs hydrolyzed by Alcalase (CH-Alcalase) exhibited the highest degree of hydrolysis compared to other CHs. Both "CH-Alcalase" and "CH-Alcalase < 3 kDa" fractions showed a considerably high antioxidant activity and collagenase inhibition activity. Therefore, resulting bioactives have potential for development as antioxidants and anti-aging ingredients in the food, cosmetics, and pharmaceuticals, from animal by-products.

Keywords: anti-aging; antioxidant; collagen hydrolysate; commercial proteases; hydrolysis.

Figure 1

Change in pH of collagen hydrolysates depending on incubation time.

Figure 2

Change in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) patterns of collagen hydrolysates hydrolyzed by different proteases: (A) Alcalase, (B) Flavorzyme, (C) Neutrase, (D) Bromeline, (E) Protamex, and (F) Papain.

Figure 3

Change in protein recovery (A) and free amino groups (B) of collagen hydrolysates (CH) depending on incubation time. Data expressed as mean ± standard deviation (SD). Data denoted by different letters (a–d) show statistically significant differences depending on incubation time (p < 0.05).

Figure 4

Change in molecular weight distribution of collagen hydrolysates (CH).

Figure 5

Antioxidant activities of collagen hydrolysates (CH) in 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging (A) and reducing power (B) assays. Data denoted by different letters (a–c) show statistically significant differences depending on different treatments (p < 0.05). Data denoted by different letters (A–D) show statistically significant differences depending on concentration (p < 0.05).