Oscar Martínez-Alvarez

Biochemical and antioxidant properties of peptidic fraction of carotenoproteins generated from shrimp by-products by enzymatic hydrolysis

The composition, functional properties and in vitro antioxidative activity of the peptidic fraction of carotenoproteins from shrimp (Parapenaeus longirostris) by-products generated by enzymatic treatment with Alcalase was evaluated. The peptidic fraction of carotenoproteins (PFCP) contained 80.8 ± 0.21% protein, 2.74 ± 0.3% lipid, 14.4 ± 0.14% ash, 1.13 ± 0.08% chitin and 1.08 ± 0.02 μg total carotenoid/g of sample. The amino acid profile of PFCP showed a high percentage of essential amino acids, such as arginine, lysine, histidine and leucine. Therefore, PFCP had a high nutritional value and could be used as a supplement to poorly balanced dietary proteins. PFCP showed an excellent solubility and possessed interfacial properties, which were governed by their concentrations. The antioxidant activities of PFCP at different concentrations were evaluated using various in vitro antioxidant assays, including the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical method, reducing power, chelating effects assay and β-carotene bleaching. The antioxidant activity of PFCP, based on their protection of supercoiled DNA strand from scission by peroxyl and hydroxyl radicals into the nicked circular form was also investigated. Results from this study suggest that the peptidic fraction of carotenoproteins is a good source of natural antioxidants and peptides with interesting functionalities.

Recovery, viscoelastic and functional properties of Barbel skin gelatine: investigation of anti-DPP-IV and anti-prolyl endopeptidase activities of generated gelatine polypeptides.

The characteristics and functional properties of gelatine from freshwater fish skin (Barbus callensis) were investigated. The gelatine extraction efficiency was improved by an acid-swelling process in the presence of barbel crude acid protease extract. Barbel skin gelatine (BSG) contained 92.15% protein, 0.31% lipid and 0.72% ash. The amino acid profile of BSG showed a high percentage of imino acids. The electrophoretic profile showed that BSG is mainly composed of α- and β-components. BSG showed an excellent solubility and possessed interfacial properties, which were governed by the protein concentration. Biological activities of the hydrolysates obtained after digestion of BSG with several commercial proteases were evaluated. The results suggested that these hydrolysates are a good source of natural inhibitors of dipeptidyl peptidase-IV and prolyl endopeptidase and could potentially be used as dietary ingredients in the management of type 2-diabetes and/or neuropathological disorders.

Evidence of an active laccase-like enzyme in deepwater pink shrimp (Parapenaeus longirostris)

This paper demonstrates the presence of an active laccase-like enzyme from deepwater pink shrimp (Parapenaeus longirostris) using polyacrylamide gel electrophoresis. This enzyme was found in all anatomical parts of the deepwater pink shrimp, but particularly in the cephalothorax, and became active during the course of storage. Gel staining with laccase-specific substrates such as ADA, DMP and DAB was used to characterize a protein of around 44kDa as containing laccase activity. The enzyme was inhibited by a specific inhibitor, CTAB. 4-Hexylresorcinol, a specific inhibitor of polyphenoloxidase (PPO), did not inhibit the laccase-like enzyme. Low concentrations of antioxidants ascorbic acid or sodium metabisulphite were sufficient to inhibit the laccase-like enzyme. ABTS and DMP were subsequently used to characterize the enzyme. Given the evidence of this enzyme in deepwater pink shrimp, new melanosis-inhibiting compounds that are suitable for consumption need to be found to complement specific inhibitors of PPO activity.

Enhanced recovery of alkaline protease from fish viscera by phase partitioning and its application

BackgroundToo many different protein and enzyme purification techniques have been reported, especially, chromatographic techniques. Apart from low recovery, these multi-step methods are complicated, time consuming, high operating cost. So, alternative beneficially methods are still required. Since, the outstanding advantages of aqueous two phase system (ATPS) such as simple, low cost, high recovery and scalable, ATPS have been used to purify various enzymes. To improve purification efficiency, parameters affected to enzyme recovery or purity was investigated. The objectives of the present study were to optimize of alkaline protease recovery from giant catfish fish viscera by using ATPS and to study of hydrolytic patterns against gelatin.ResultsUsing 70% (w/w) crude enzyme extract (CE) in system (15% PEG2000-15% sodium citrate) provided the highest recovery, PF and KE. At unmodified pH (8.5) gave the best recovery and PF with compare to other pHs of the system. The addition of 1% (w/w) NaCl showed the recovery (64.18%), 3.33-fold and 15.09 of KE compared to the system without NaCl. After addition of 10% (w/w) sodium citrate in the second ATPS cycle, the highest protease recovery (365.53%) and PF (11.60-fold) were obtained. Thus, the top phase from the system was subjected to further studied. The protein bands with molecular weights (MWs) of 20, 24, 27, 36, 94 and 130 kDa appeared on the protein stained gel and also exhibited clear zone on casein-substrate gel electrophoresis. The β, α1, α2 of skin gelatin extensively degraded into small molecules when treated with 10 units of the extracted alkaline protease compared to those of the level of 0.21 units of Flavourzyme.ConclusionsRepetitive ATPS is the alternative strategy to increase both recovery and purity of the alkaline protease from farmed giant catfish viscera. Extracted alkaline protease exposed very high effectiveness in gelatin hydrolysis. It is suggested that the alkaline protease from this fish viscera can further be used in protein hydrolysate production.

Gelatin hydrolysates from farmed Giant catfish skin using alkaline proteases and its antioxidative function of simulated gastro-intestinal digestion

This work aims to evaluate the ability of different alkaline proteases to prepare active gelatin hydrolysates. Fish skin gelatin was hydrolysed by visceral alkaline-proteases from Giant catfish, commercial trypsin, and Izyme AL®. All antioxidant activity indices of the hydrolysates increased with increasing degree of hydrolysis (P<0.05). The hydrolysates obtained with Izyme AL® and visceral alkaline-proteases showed the highest and lowest radical scavenging capacity, while prepared with commercial trypsin was the most effective in reducing ferric ions and showed the best metal chelating properties. The hydrolysate obtained with Izyme AL® showed the lowest iron reducing ability, but provided the highest average molecular weight (⩾ 7 kDa), followed by commercial trypsin (2.2 kDa) and visceral alkaline-proteases (1.75 kDa). After in vitro gastrointestinal digestion, the hydrolysates showed significant higher radical scavenging, reducing ferric ions and chelating activities. Gelatin hydrolysates, from fish skin, could serve as a potential source of functional food ingredients for health promotion.

Fish skin gelatin hydrolysates produced by visceral peptidase and bovine trypsin: Bioactivity and stability

The peptidase from the viscera of farmed giant catfish was used for producing gelatin hydrolysates (HG) and compared with those produced from commercial bovine trypsin (HB). The degree of hydrolysis (DH) observed suggests that proteolytic cleavage rapidly occurred within the first 120min of incubation, and there was higher DH in HG than in HB. HG demonstrated the highest ACE-inhibitory activity, DPPH, ABTS radical scavenging activity, and FRAP. HB showed the highest FRAP activity. The DPPH radical scavenging activity of HG was quite stable over the pH range of 1-11, but it increased slightly when the heating duration time reached 240min at 100°C. The ACE-inhibitory activity of HG showed the highest stability at a pH of 7, and it remained very stable at 100°C for over 15-240min. The visceral peptidase from farmed giant catfish could be an alternative protease for generating protein hydrolysates with desirable bioactivities. The resulting hydrolysates showed good stability, making them potential functional ingredients for food formulations.

Angiotensin-I-Converting Enzyme Inhibitory and Antioxidant Activities of Protein Hydrolysate from Muscle of Barbel (Barbus callensis)

The present study investigated angiotensin-I-converting enzyme (ACE) inhibitory and antioxidant activities of barbel muscle protein hydrolysate prepared with Alcalase. The barbel muscle protein hydrolysate displayed a high ACE inhibitory activity ( mg/mL). The antioxidant activities of protein hydrolysate at different concentrations were evaluated using various in vitro antioxidant assays, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical method and reducing power assay. The barbel muscle protein hydrolysate exhibited an important radical scavenging effect and reducing power. These results obtained by in vitro systems obviously established the antioxidant potency of barbel hydrolysate to donate electron or hydrogen atom to reduce the free radical. Furthermore, these bioactive substances can be exploited into functional foods or used as source of nutraceuticals.

Thermoseparating Aqueous Two-Phase System for the Separation of Alkaline Proteases from Fish Viscera

The objective of this study was to isolate alkaline proteases from farmed giant catfish viscera by using a thermoseparating aqueous two-phase system (T-ATPS). Different phase partitioning parameters—type of salts, NaCl addition, and temperature—were optimized. The optimum system contained 40% EOPO3900-10% MgSO4 with 17%NaCl, induced phase separation at 55°C, and provided the highest recovery (77.98 %) and purity (PF: 21.50-fold) with a partition coefficient (KE) of 11.90 and a volume ratio (VR) of 0.19. In the recycling step, the mixture of the bottom salt-rich phase and bottom EOPO-rich phase in the ratio of 0.5:1.5 (w/w) provided the highest recovery and purity. Hence, the total recovery of 91.62% was obtained from the separation system. Three major clear zones (24, 36, and 130 kDa) can be distinctively observed on casein-substrate gel electrophoresis for protease activity staining. Major protein components of perilla seed hydrolysates were completely hydrolyzed by the alkaline proteases at 70 units while those of red kidney bean isolate were more resistant to protein hydrolysis. Fifty units of Flavourzyme® hydrolyzed completely both plant proteins into small peptides or amino acids. As a result, T-ATPS could be used as an alternative method for the separation of enzymes from various sources with acceptable recovery. In addition, the obtained alkaline proteases can be further used in preparation of protein hydrolysates.

Glycosaminoglycans from grey triggerfish and smooth hound skins: Rheological, Anti-inflammatory and wound healing properties

The present study aims to evaluate for the first time the wound healing and in vivo anti-inflammatory effects of glycosaminoglycans from skins of smooth hound (SHSG) and grey triggerfish (GTSG). Thermal analysis of GTSG and SHSG was evaluated using differential scanning calorimetry (DSC). The rheologie properties and water absorption capacity of two gels prepared from SHSG and GTSG were also studied. The application of GTSG and SHSG based gels on dermal full-thickness excision wounds in a mouse model, enhanced significantly wound healing activity and a total closure was achieved after eleven days of wound induction for SHSG. Further, histological examination of biopsies showed advanced tissue regeneration, characterized by the presence of well-organized stratum of both derma and epidermis. The anti-inflammatory evaluation of GTSG and SHSG in mice showed a significant inhibition of edema paw, after 5 h of carrageenan injection. The edema inhibition was 91.6% and 90% for SHSG and GTSG, respectively at the dose of 50 mg/kg. Furthermore, the histological evaluation and the superoxide dismutase, catalase and malondialdehyde level in muscle tissue were investigated. In summary, this work demonstrates that both GTSG and SHSG could be promising drugs with good wound healing and anti-inflammatory effects in animal model.

Fermented Seafood Products and Health

Abstract Fermented seafood products are traditional foods in many countries. These products are sources of healthy molecules, although occasionally contain undesirable compounds, such as high-salt content, biogenic amines, or pathogens. Natural antioxidants such as free amino acids (FAAs) and desirable Maillard reaction products have been found in some fermented products, in addition to peptides exhibiting angiotensin I-converting enzyme-inhibiting activity and hence potential antihypertensive effects. The presence of other healthy molecules (eg, insulin-secretion stimulating peptides, anticoagulants, immune modulators, antidepressors, bacteriocins, and others) have also been reported in fermented seafood products. This chapter summarizes the most recent advances in the search for bioactive molecules in fermented seafood products with an interest in human health. This chapter also discusses different technological approaches to eliminating undesirable compounds that are associated with health risks and to improving product quality.