Extraction, Characterization, and Application of Agricultural and Food Processing By-Products

By-products originating from agricultural and food processing are considerable disposal problem for the industry because these waste streams emerge in huge quantities and often have direct impact on the economy and environmental pollution. However, at the same time, it constitute a rich but yet underutilized source of valuable components, which may find application as ingredients in the food and non-food industries. As a result, numerous projects are currently directed toward the utilization of agricultural and food processing by-products such as animal-based (skin, bone, flesh, and internal organs) and plant-based biomaterials (fruit peels and seeds, rice bran, and etc.). In the present work, we would like to focus on the potentialities and the current research of the compounds and extracts deriving from agro-industrial disposable wastes in the food-related utilization. The potential of selected by-products as a source of bioactive/functional compounds in terms of emerging and conventional techniques for extraction, characterization, biological activity monitoring and application of the extracts or isolated compounds as functional food ingredients or bio-based materials for food packaging are highlighted. Considering environmental effect and economic loss, agricultural and food processing by-products should be utilized in various innovative processes in the cause of beneficial product derivation

Three-phase partitioning and proteins hydrolysis patterns of alkaline proteases derived from fish viscera

In this study, the recovery alkaline proteases from farmed giant catfish viscera were isolated by using three-phase partitioning (TPP). Factors affecting partitioning efficiency such as salts, solvent types, pH, and incubation temperatures were studied. Furthermore, the application of extracted alkaline proteases on proteins hydrolysis was also determined. The system consisted of crude enzyme extract:t-butanol 1:0.5 (w/v), 50% sodium citrate, pH 8.0 with incubation temperature of 25 °C provided the highest enzyme recovery (220%). The smear protein bands with molecular weight of 20, 24, and 215 kDa of TPP fractions appeared on the protein stained gel. Two major clear zones (24 and 130 kDa) in the interphase were observed on casein-substrate gel electrophoresis. Extracted alkaline proteases showed relatively high effective in protein hydrolysis. As a result, TPP provided high enzyme recovery and could be applied to other enzymes. The obtained alkaline proteases can be further applied in preparation of protein hydrolysates. © 2014 Elsevier B.V. All rights reserved.Peer Reviewe

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

Abstract Background Too 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. Results Using 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. Conclusions Repetitive 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.Financial support from Mae Fah Luang University and the Thailand Research Fund through the Royal Golden Jubilee Ph.D. Program (Grant No. PHD 0113/2554) to Ms. Sunantha Ketnawa are acknowledged.Peer Reviewe

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 120 min 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 240 min 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 15–240 min. 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.This work was supported by the Thailand Research Fund (TRF). Miss Sunantha Ketnawa was sponsored through the Royal Golden Jubilee Ph.D. Program (Grant No. PHD 0113/2554). Mae Fah Luang University and the TRF Distinguished Research Professor Grant also provided some financial support.Peer Reviewe

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

© 2015 Elsevier Ltd. Abstract 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.This study was financed by the Spanish Ministry of Economy and Competitiveness under project AGL2011-2760. Ms. Sunantha Ketnawa was funded by the Royal Golden Jubilee Ph.D. Program (Grant No. PHD 0113/2554) from the Thailand Research FundPeer Reviewe

Extraction and biochemical characterization of peptidases from giant catfish viscera by aqueous two-phase system

Peptidases were extracted from the viscera of farmed giant catfish (Pangasianodon gigas) in an aqueous two-phase system (ATPS) of 15% (w/w) polyethylene glycol (PEG-2000) and 15% (w/w) sodium citrate. The recovery of the enzymes was 273% with 12-fold purification. Protein pattern, activity and inhibitory staining confirmed that the proteins with molecular weights of 12 and 31kDa were a mixture of proteolytic enzyme. The optimum pH and temperature of the enzyme were 8.0 and 70C, respectively. Besides, it retained more than 50% of activity at the highest salt concentration (30% w/v). The enzyme was strongly inhibited by serine protease inhibitors (>80% inhibition), while low inhibition with cysteine-, aspartic- and metallo-protease inhibitors (+, Cu2+ and Fe2+. The peptidases from the giant catfish viscera have potential applications in food processing where high temperatures (60-70C) and/or high salt content are used, or even in detergent formulation. Practical Applications: Extraction and characterization of a proteolytic enzyme from viscera, fishery processing by-product, could add value to it. According to its properties, the isolated peptidases could be potentially applied to food industry, especially in fish sauce production, protein hydrolysate production, or in neutraceutical and in detergent industry.This research was supported by a grant from Mae Fah Luang University and Thailand Research Fund through the Royal Golden Jubilee PhD Program (Grant No. PHD4030113/ 2554) to Ms. Sunantha Ketnawa.Peer Reviewe

Physical, chemical, and microbiological properties of fish tofu containing shrimp hydrolysate

Fish tofu was formulated by adding shrimp hydrolysate at levels of 0.05, 1.00, and 2.00 % (w/w) of ingredient formulation. The physicochemical, microbiological, and sensorial characteristics were evaluated for 14 days of storage at 4 °C. Greater incorporation of hydrolysate resulted in a tofu which was more resistant to lipid oxidation, with lower thiobarbituric acid-reactive substances (TBARS) values during storage, according to the rancidity score. However, the overall acceptability score was lower than in fish tofu without the hydrolysate. The counts of microbiological indicators, including total viable counts, yeast and molds were significantly (P < 0.05) affected by the addition of 2.00 % hydrolysate to the fish tofu. The surface of fish tofu with hydrolysate was lighter (L) and yellower (b), whilst the inside was darker (L), and there were higher whiteness values from storage day 0 to 14. The fish tofu with hydrolysate had a softer texture when compared with the control sample, which was evidenced by the instrumental analysis. Therefore, shrimp hydrolysate could be used as a natural substance in meat products to delay lipid oxidation as well as decrease microbial spoilage, therefore improving nutritional quality as a low energetic value food, and increasing the shelf-life of food products.Peer Reviewe

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 55ºC 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 recycling step, the mixture of 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, TATPS could be used as an alternative method for separation of enzymes from various sources with acceptable recovery. Besides, the obtained alkaline proteases can be further used in preparation of protein hydrolysates.Peer Reviewe

Whole Wheat Bread Enriched with Cricket Powder as an Alternative Protein

The current market trends in modern sedentary lifestyles drive the development of new functional products able to fulfill consumers’ demand for a healthy diet. Whole wheat bread contains more protein and fiber than white bread; however, it could be improved in terms of protein content and quality. Cricket powder, which contains high protein (55.11, wt%), could be used as an alternative source to tackle those deficiencies in such bread. Hence, the study aimed to apply cricket powder in the whole wheat bread formula to enrich protein content, indispensable amino acids and determine their physico-chemical properties, consumers’ acceptance, and shelf-life storage. The results showed that all enriched bread presented high protein (18.97–25.94, wt%), fat (10.91–15.07, wt%), and ash (2.09–2.33, wt%) with the increment of the cricket powder than those of the control bread. Enriched breads’ crust colors were not significantly different (p > 0.05), while crumb colors were darker (L* = 55.64–64.48) compared to the control (L* = 69.98). Enriched bread had a hard texture and required a lot of chewing force compared to the control. Furthermore, all samples yielded a shelf-life of 5 days when monitoring the mold growth. From the results, the bread enriched with 20% cricket powder yielded the best consumers’ acceptance score of 77%. It was predominantly high in indispensable amino acids such as leucine, phenylalanine, lysine, and arginine. Therefore, cricket powder could be a novel alternative protein source and successfully utilized in whole wheat bread to enhance its protein content and indispensable amino acids with consumers’ acceptance responding to the current market trend