Antioxidant properties of papain mediated protein hydrolysates from fresh water carps (Catla catla, Labeo rohita and Cirrhinus mrigala) and its application on inhibition of lipid oxidation in oil sardine mince during ice storage.

Fish protein hydrolysates (FPH) from fresh water carps Catla catla, Labeo rohita and Cirrhinus mrigala were prepared with 5, 10, 15 and 20% degree of hydrolysis (DH) using papain enzyme. FPH were evaluated for antioxidant properties using in vitro assays such as DPPH free radical scavenging activity (at 10 mg/ml), ferric reducing antioxidant power assay (at 20 mg/ml) and linoleic acid peroxidation inhibition activity (at 10 mg/ml). Antioxidant properties of FPH varied with species and DH. The DPPH radical scavenging activity, linoleic acid peroxidation inhibition activity and ferric reducing antioxidant power (as absorbance at 700 nm) of FPH from carps was in the range of 59-92%, 52-85% and 0.388-0.663 respectively. Based on the overall antioxidant activity, FPH from C. catla with 20% DH was added to oil sardine mince at different concentration (0.1, 0.2 and 0.4%) and found to inhibit effectively the formation of peroxides and malonaldehyde in dose dependent manner. FPH from C. catla with 20% DH was fractionated using size exclusion chromatography and had three different peptide fractions with the approximate molecular weight of 6561-2106 Da (fraction 1), 1942-994 Da (fraction 2) and 935-383 Da (fraction 3). The present study showed promising results that the fish protein hydrolysates from fresh water carps muscle proteins can be used as natural antioxidants in food system. Production of fish protein hydrolysates with nutraceutical properties could be the way forward for better utilization and value addition.

Exploiting of Secondary Raw Materials from Fish Processing Industry as a Source of Bioactive Peptide-Rich Protein Hydrolysates.

Developing peptide-based drugs are very promising to address many of the lifestyle mediated diseases which are prevalent in a major portion of the global population. As an alternative to synthetic peptide-based drugs, derived peptides from natural sources have gained a greater attention in the last two decades. Aquatic organisms including plants, fish and shellfish are known as a rich reservoir of parent protein molecules which can offer novel sequences of amino acids in peptides, having unique bio-functional properties upon hydrolyzing with proteases from different sources. However, rather than exploiting fish and shellfish stocks which are already under pressure due to overexploitation, the processing discards, regarded as secondary raw material, could be a potential choice for peptide based therapeutic development strategies. In this connection, we have attempted to review the scientific reports in this area of research that deal with some of the well-established bioactive properties, such as antihypertensive, anti-oxidative, anti-coagulative, antibacterial and anticarcinogenic properties, with reference to the type of enzymes, substrate used, degree of particular bio-functionality, mechanism, and wherever possible, the active amino acid sequences in peptides. Many of the studies have been conducted on hydrolysate (crude mixture of peptides) enriched with low molecular bioactive peptides. In vitro and in vivo experiments on the potency of bioactive peptides to modulate the human physiological functions beneficially have demonstrated that these peptides can be used in the prevention and treatment of non-communicable lifestyle mediated diseases. The information synthesized under this review could serve as a point of reference to drive further research on and development of functionally active therapeutic natural peptides. Availability of such scientific information is expected to open up new zones of investigation for adding value to underutilized secondary raw materials, which in turn paves the way for sustainability in fish processing. However, there are significant challenges ahead in exploring the fish waste as a source of bioactive peptides, as it demands more studies on mechanisms and structure-function relationship understanding as well as clearance from regulatory and statutory bodies before reaching the end user in the form of supplement or therapeutics.

Antioxidant and emulsion properties of freshwater carps (Catla catla, Labeo rohita, Cirrhinus mrigala) protein hydrolysates prepared using flavorzyme.

Fish protein hydrolysates (FPHs) were prepared from freshwater carps (Catla catla, Labeo rohita, and Cirrhinus mrigala) using flavorzyme at different degrees of hydrolysis (DHs) ranging from 5 to 20%. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical-scavenging activity of the FPHs prepared from the three species were in the range of 50-82%; the ferric reducing power of the FPHs prepared from catla was the highest. The linoleic acid peroxidation inhibition activity of the prepared FPHs varied from 71 to 91%. The emulsion activity index of the FPHs prepared from catla and rohu decreased significantly with an increase in the DH (p < 0.05). The emulsion stability index of the FPHs prepared from the three species was the highest at 20% DH. FPHs prepared from freshwater carps possess good antioxidant and surface-active properties and are therefore suitable to be used as natural antioxidants in health-food formulation and as water-soluble antioxidants in the food-processing industry.

Bioactive and functional properties of protein hydrolysates from fish frame processing waste using plant proteases.

Enzymatic conversion of fish frame waste of threadfin breams (Nemipterus japonicus) to protein hydrolysate could be a solution for minimizing the pollution issues related to seafood processing operations and a way for the value addition to processing by-products. Protein hydrolysates from fish frame waste (FW) of thread fin breams (N. japonicus) were prepared and evaluated for bioactive properties such as angiotensin-I-converting enzyme (ACE) inhibitory activity and antioxidant and functional properties as a function of degree of hydrolysis (DH). Two different plant proteases, papain and bromelain, were used to prepare fish protein hydrolysates (FPH) and designated as HP (hydrolysates prepared using papain) and HB (hydrolysates prepared using bromelain). The ACE inhibitory activity of HP samples was higher at 5 and 10 % DH than that of the HB samples at DH 15 %, and there was no significant difference (p < 0.05). Antioxidant properties (2, 2 diphenyl-1-picrylhydrazyl [DPPH] radical scavenging activity, ferric reducing power and lipid peroxidation inhibition) of hydrolysates increased with increase in DH. The HB samples at DH 15 % had significantly higher antioxidant properties than HP samples (p < 0.05). The solubility of HP and HB samples was high in a wide range of pH and increased with DH. The functional properties of HP and HB samples decreased significantly with increase in DH (p < 0.05). The fractionation of the HB-DH 15 % sample yielded three peptide fractions with the approximate molecular weight of peptides in the range of 7562-812 Da. Relatively, bromelain enzyme is more effective in producing the FPH with desirable bioactive and functional properties.

Effect of oven drying and freeze drying on the antioxidant and functional properties of protein hydrolysates derived from freshwater fish (Cirrhinus mrigala) using papain enzyme.

Fish protein hydrolysate (FPH) was prepared from fresh water fish Cirrhinus mrigala using papain and dried in oven (OD-FPH) and freeze dryer (FD-FPH). The electron micrographs of FD-FPH samples showed porous structure. The browning intensity of OD-FPH samples was higher than the FD-FPH samples. The DPPH (2, 2 Diphenyl-1-picrylhydrazyl) free radical scavenging activity and linoleic acid peroxidation inhibition activity of FPH were not affected by oven drying process. The sequential digestion of FPH with pepsin and pancreatin reduced the antioxidant properties in both OD-FPH and FD-FPH samples. The solubility of proteins in OD-FPH was lower at pH 5 while for that of FD-FPH it was at pH 7 with water as solvent. The surface active properties of FD-FPH samples were higher than OD-FPH samples. The oven drying of fish protein hydrolysates may be advocated considering the properties and cost of production.