Gel Forming Ability and Other Properties of Eleven Underutilized Tropical Marine Fish Species

Abstract

The gel forming ability and other characteristics of the mince of 11 underutilized marine fish were studied. They were Bombay duck, silverbelly, sea catfish, silver jewfish, jewelled shad, queenfish, Spanish mackerel, hardtail, Indian tuna, tripletail and false conger eel. Mince was prepared from fillet and a portion of the mince was washed two times with cold water (5°C) containing 0.1% NaCl. Both washed and unwashed mince were ground with 3% NaCl. Ground paste was then stuffed into plastic tube and heated for one- and two-step heating. In the one-step heating, the tubes were subjected to 25°, 30°, 35°, 40°, 50°, 60°, 70° and 80°C for 60, 120 and 180 min. In the two-step heating, the tubes were pre-heated at 25°, 30°, 35°, 40°, 50°, 60°, 70° and 80°C for 60, 120 and 180 min. After the pre-heating, the tubes were immediately subjected to 85°C for 30 min. The gel was subjected to puncture, folding, expressible moisture and sensory tests.

Two-step heating distinctly improved the gel strength compared to the one-step heating. The improvement due to two-step heating was more at low preheating temperatures from 25-35°C. Washing improved the texture and color of all of the gels except Bombay duck and decreased the extent of gel-disintegration in silverbelly, queenfish, sea catfish and hardtail. The gels were set optimally at 35°-40°C for most species. Species variation in the disintegration of the gels was observed. Bombay duck mince produced very weak gel. Neither two-step heating nor washing could improve the gel quality of Bombay duck mince. Our data suggested that jewelled shad, queenfish, silver jewfish, sea catfish, tripletail and false conger eel could be suitable as the material for surimi.     

Gel Forming Ability of Tropical Tilapia Surimi as Compared with Alaska Pollock and Pacific Whiting Surimi

Abstract

Setting and thermal treatment effects on texture and color of tropical tilapia surimi gels were compared to Alaska pollock and Pacific whiting gels. Heat treatments that most favored intrinsic gelling factors of a fish species exhibited strong gel formation. Whiteness values increased as total thermal inputs increased, which reflect the increasing opacity of the gels. Pollock gels were generally the strongest and whitest. Tilapia gel quality was generally second to pollock gels, however, in heat treatments using setting temperatures ≥ 40 °C, tilapia gels were comparable (60 °C setting) or superior (40 °C setting) to pollock gels. The optimum heat treatment for tilapia surimi appeared to be a 40° C setting for 1 hr followed by a 90 °C cook for 15 min. SDS-PAGE patterns of gels prepared with 60°C setting followed by 90 °C cooking elucidated the various degree of protein degradation depending upon the species in a descending order of whiting, tilapia, and pollock.     

Thermal gelation properties of white croaker, walleye pollack and deepsea bonefish surimi after suwari treatment at various temperatures

The effects of setting (suwari) at around 40 °C on the breaking strength and breaking strain rate of thermal gels treated at 85 °C for 20 min during the following processing step were examined in association with the polymerization and degradation of myosin heavy chains (MHCs) for surimi prepared from white croaker, walleye pollack and deepsea bonefish. In the case of white croaker and walleye pollack, maximum values of breaking strength and breaking strain rate were obtained after suwari at 30–40 °C for both 30 and 60 min, at which temperature MHCs were polymerized. In comparison, these textual properties of the thermal gels decreased in surimi prepared from deepsea bonefish after suwari at around 38 °C for 30 min and at around 32 °C for 60 min, with concomitant degradation products of MHC. The textual properties of deepsea bonefish after suwari at temperatures >45 °C tended to be almost the same as those after suwari at temperatures of <30 °C, where neither polymerization nor degradation of MHC was observed.     

Comparative Properties of Bluefish (Pomatomus saltatrix) Gels Formulated by High Hydrostatic Pressure and Heat

High pressure was applied at levels of 300 to 3,742 atm for 30 min to formulate gels from bluefish meat paste, and the properties of the resulting gels were compared with those of heat-induced gels formulated at 90°C for 20 min, or at 60°C for 60 min. The moisture content of the pressure-induced gels was similar to that of the heat-induced gels, while protein contents and pH values of pressurized gels were slightly lower than those formulated by heat. Gels formed by pressure were more translucent as compared with those formulated by heat. Texture measurement indicated that there were no significant differences between the elasticities of the gels obtained under various pressures, although gel firmness increased with pressure. Overall, the heat-induced gels formulated at 90°C for 20 min were firmer but had similar elasticities to pressure-induced gels, while gels formulated at 60°C for 60 min were comparatively softer and had lower elasticities. The salt-extractable protein and protein digestibility studies indicated that pressure treatment formed gels with less protein denaturation and which were more digestible than the fish gels formulated by heat. The results from proteolytic activity studies showed that the pressure range used in this study was less effective in inactivating the endogenous proteases in the fish flesh than heat.     

Digestive protease characterization, localization and adaptation in blacklip abalone (Haliotis rubra Leach)

Digestive protease in blacklip abalone (Haliotis rubra Leach) from whole gut extracts was found to have dual pH optima at pH 3 and 10 and an optimum temperature of 45 °C. Over the biologically relevant range of pH (5–8), protease activity dropped to a minimum at pH 5 (53% of the maximal rate at pH 3) rising gradually and continuously up to (and beyond) pH 8. Over the biologically relevant range of temperatures (9–24 °C), protease activity increased continuously with activity at 24 °C being 75% higher than activity at 9 °C. Protease digestion was relatively uniform in gut extracts from sections containing gut contents. Digestion thus appears to be significantly extracellular (in the lumen of the gut). Analysis of gut sections washed free of food matter suggests the anatomical origin of protease from activity of gut regions in the order: digestive gland » salivary gland ≈ stomach > crop > intestine > upper oesophagus. Whole gut protease levels did not alter in response to a high protein artificial diet.     

In vitro assay of plant protease inhibitors from four different sources on digestive proteases of rohu, Labeo rohita (Hamilton), fingerlings

The in vitro inhibitory effect of protease inhibitors from four seed extracts (soybean, grasspea, black gram and horse gram) on digestive proteases of rohu was assessed by enzyme inhibition assay and substrate sodium dodecyl sulphate‐polyacrylamide gel electrophoresis. High proteolytic activity was detected in the intestinal extract of rohu (Labeo rohita) fingerlings at two different pH ranges (8–8.5 and 10–11). That protein digestion occurs mainly in the alkaline condition in this fish without a stomach is evident from very high trypsin activity (0.95±0.04 benzoyl‐dl ‐arginine‐p‐nitroanilide U mg protein⁻¹) in the intestine. In case of grass pea seed, more than 50% inhibition of alkaline protease activity was recorded when the ratio of inhibitor to enzyme was 9.41 μg U⁻¹. More than 40% inhibition of protease activity was recorded in case of horse gram seed when the ratio of inhibitor to enzyme was 5.51 μg U⁻¹. Black gram at 11.0 μg U⁻¹ and soybean seed proteins at 62.75 μg U⁻¹ resulted in 50% and more than 30% inhibition of digestive protease activity in rohu fingerlings respectively. A plot of the inhibition values obtained by changing the relative concentrations of enzyme/inhibitor resulted in different dose–response curves for different protein sources. The use of substrate gel electrophoresis allowed the visualization of the aforementioned differences in inhibition. Each seed extract produced a characteristic profile of protease inhibition. It is concluded that protease inhibitors present in plant protein sources adversely affect the digestive proteases in fish and hence there is a need to eliminate/reduce the amount of such inhibitors through proper processing before incorporation into aquafeeds.     

Autolysis and Characterization of Sarcoplasmic and Myofibril Associated Proteinases of Oxeye Scad (Selar boops) Muscle

ABSTRACT

The autolytic profile of oxeye scad mince was characterized. Mince showed higher proteolytic activity than washed mince. The highest autolysis was observed at 65 and 60°C for mince and washed mince, respectively. Both mince and washed mince showed the optimum pH for autolysis at pH 9.0, and their activities decreased with increasing NaCl concentration (0–3.5%). Autolysis of washed mince was markedly inhibited by soybean trypsin inhibitor (SBTI), suggesting that myofibril-associated proteinase was serine proteinase. Sarcoplasmic proteinase was characterized to be heat-activated alkaline proteinase having the optimal pH and temperature of 9.0 and 60°C, respectively. The activities were stable at pH range of 8.0–11.0 at 20–40°C. The crude proteinase was inhibited by N-p-tosyl-L-lysine chloromethyl ketone, SBTI, and phenylmethanesulfonyl fluoride, suggesting the predominance of serine proteinases, especially trypsin. NaCl suppressed the activity while β-mercaptoethanol, dithiothreitol, and CaCl₂ activated the activity. Therefore, trypsin-like proteinase is a major endogenous proteinase responsible for autolysis in oxeye scad muscle. The present results can be used as scientific guidelines to predict the gel strength of surimi made from oxeye scad muscle.     

Effect of Protease Treatment on Oil Extractability of Carotenoids from Shrimp Waste

Crustacean waste is an important source of natural carotenoids. Carotenoids in shrimp waste occur as a complex with proteins. Investigations were carried out to assess the effect of hydrolyzing shrimp waste with different proteases on the subsequent recovery of carotenoids in vegetable oil. Hydrolyzing the shrimp waste with the bacterial protease alcalase gave higher extraction yields in oil (28.6 μg/g waste) compared to hydrolyzing with the plant protease papain (24.8 μg/g) or the animal protease trypsin (25.3 μg/g). Optimized conditions for alcalase treatment and extraction of carotenoids was determined to be hydrolysis of shrimp waste with 0.75% of alcalase at 37°C for 150 min, adding sunflower oil to hydrolyzed waste in a ratio of 2:1, and heating at 70°C for 90 min. Prior hydrolysis of waste considerably reduced the heating time required for maximum extraction of pigments, which may help in preventing the carotenoid degradation during prolonged heating in oil.     

Improvement of Cold and Thermally Induced Gelation of Giant Squid (Dosidicus gigas) Surimi

Depending on the season of capture, giant squid (Dosidicus gigas) surimi processed by isoelectric precipitation presents low gel strength. Addition of microbial transglutaminase (MTGase) and application of high isostatic pressure (300 MPa) to improve physicochemical properties were assayed for purposes of making “suwari” gels and heated gels for use in restructured products which have a raw or cooked appearance. The physicochemical properties of both pressurized and unpressurized gels induced by application of 30°C/1 h improved when MTGase was added. In contrast, addition of MTGase was less effective in gels subsequently heated at 90°C/30 min after 30°C/1 h. High pressure treatment for 30 min at 300 MPa and 15°C helped to produce gels with better mechanical and water binding properties, whether treated for 30°C/1 h only or for 30°C/1 h plus 30-min heat treatment at 90°C. High pressure treatment also reduced lightness.     

Optimum formulation of feed for rohu, Labeo rohita (Hamilton), with biofloc as a component

The present study was conducted to design an optimum feed mix for the growth of rohu (Labeo rohita) in light-limited indoor culture with biofloc as a component along with fish feed. Eighteen 700-L tanks were utilized for producing biofloc using aquaculture effluent from a nearby carp culture pond. Fifteen different feed mixes were prepared using fish feed and biofloc in dry (4 % moisture content) and wet (90 % moisture content) forms at different proportions and used in feeding trials (three replications) conducted in forty-five 50-L glass aquarium stocked with three rohu fingerlings with average individual weight of 20 ± 1.5 g for a period of 90 days. Fish survival was 100 % in all the treatments. Mixture design was used to obtain a solution of best combination of feed source to obtain the optimum growth parameters of rohu. Optimum growth parameters (net yield, specific growth rate, protein efficiency ratio and feed conversion ratio) of rohu were obtained at feed mix containing 50 % fish feed and 50 % wet floc. The nutritional quality of biofloc was found to be quite suitable for rohu. The images of 3-week-old biofloc captured in scanning electron microscope (SEM) indicated the presence of different types of bacteria, algae, protozoa, rotifers, etc. in different sizes ranging from 10 to 100 μm.     

Purification and characterization of stomach protease from the turbot (Scophthalmus maximus L.)

Proteolytic activity in the different parts of the digestive tract of the turbot (Scophthalmus maximus L.) were studied in this work. One pure protease was isolated from turbot stomach and its behavior was studied. Results showed the optimum pH for proteases in the different parts of the digestive tract of the turbot were pH 2.0 for the stomach, pH 8.0 for the pylorus cecum, pH 8.0 for the foregut, pH 8.5 for the midgut, and pH 8.0 for the hindgut. The activity of proteases in the different parts of the digestive tract were in the sequence pylorus cecum protease > stomach protease > foregut protease > midgut protease > hindgut protease. The stomach protease was purified by ammonium sulfate precipitation and column chromatography on DEAE-Sepharose F.F. and Sephadex G-100. The purified enzyme gave a single band in SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Its molecular weight was found to be approximately 42,000 Da. The enzyme is stable at pH 1.0–9.0 and at temperatures below 40°C. Its activity was maximum at pH 2.0 and 40°C. When reaction time was prolonged the optimum temperature of the enzyme tended to decline. The enzyme was activated by Mn²⁺ and Cu²⁺ and inactivated by Fe³⁺. It was fully inhibited by pepstatin and partially inhibited by PMSF, TPCK, PCMB, and NBS. These results imply the enzyme is a pepsin.     

Measurement of Morphometric Dimensions and Mechanical Properties of Rohu Fish for Design of Processing Machines

Indian major carps (IMC) viz. rohu, catla, and mrigal are the most important fresh water species. They are marketed locally without post-harvest processing and value addition because of the lack of suitable processing machines. The present study was undertaken to measure the morphometric and mechanical properties of rohu fish for the development of fish processing machinery, and the same can be adjusted for the other IMC. Twenty-five samples of rohu fish (Labeo rohita) were caught, with weight ranging from 290 to 2,100 g (harvestable size). The measured morphometric dimensions were the body depth (10.14 ± 1.63 cm), thickness (5.9 ± 0.85 cm), head length (9.35 ± 1.04 cm), standard length (37.54 ± 4.17 cm), total length (45.89 ± 5.68 cm), and fish backbone thickness (11.2 ± 4.91 mm). Mechanical properties including the angle of repose over steel surface (20°–22°) and plastic surface (18°–20°) were recorded. The maximum power consumption for slicing the fish lengthwise was recorded as 460 W. The measured morphometric and mechanical properties are useful in limiting the adjustments, design of conveyors, capacity, and power requirement of fish processing machines for rohu fish.     

Effect of setting conditions on mechanical properties of acid-induced Kamaboko gel from squid <Emphasis Type="Italic">Todarodes pacificus</Emphasis> mantle muscle meat

Squid Todarodes pacificus suwari gels, set at various temperatures and times, and acid-induced kamaboko gel, which was prepared by soaking suwari gel in 5% acetic acid for 20 h, were studied to evaluate the mechanical properties that are affected by setting conditions. Unset squid meat paste did not form a gel when soaked in acetic acid. The breaking strength of both suwari gel and acid-induced kamaboko gel showed a tendency to increase with setting temperature and time. SDS-PAGE analysis of suwari gel and acid-induced kamaboko gel, which were set at various temperatures and times, showed that myosin heavy chain (MHC) was observed at 30°C only for the first hour. The intensity of the MHC band at 30°C gradually decreased with setting time, while the intensity of the polymer band gradually increased with setting time. These results suggest that the protein-protein bonds in suwari gel affect the final texture of acid-induced kamaboko gel. Based on the analysis of the mechanical properties, and in consideration of the fact that the purpose of this experiment was to reduce energy usage, the best setting condition was determined to be 40°C for 3 h.     

Digestive proteases of three carps Catla catla, Labeo rohita and Hypophthalmichthys molitrix: partial characterization and protein hydrolysis efficiency

Characteristics and functional efficacy of digestive proteases of Catla catla, catla, Labeo rohita, rohu and Hypophthalmichthys molitrix, silver carp were studied. Total protease activity was significantly (P < 0.05) higher in rohu (1.219 ± 0.059 U mg protein⁻¹ min⁻¹) followed by silver carp (1.084 ± 0.061 U mg  protein⁻¹ min⁻¹), and catla (0.193 ± 0.006 U mg  protein⁻¹ min⁻¹). Trypsin activity of silver carp and rohu was 89–91% higher than catla. Chymotrypsin activity was significantly (P < 0.05) higher in silver carp compared with rohu and catla. The protease activity of rohu and silver carp displayed bell‐shaped curves with maximum activity at pH 9; whereas in catla, maximum activity was found between pH 8 and 11. Inhibition of protease activity with soybean trypsin inhibitor (SBTI), phenylmethylsulfonyl fluoride (PMSF) revealed the presence of serine proteases and inhibition of activity with N‐α‐p‐tosyl‐L‐lysine‐chloromethyl ketone (TLCK) and N‐tosyl‐L‐phenylalanychloromethane (TPCK) indicated the presence of trypsin‐like and chymotrypsin‐like enzymes in all these three carps. SDS‐PAGE showed the presence of several protein bands ranging from 15.3 to 121.9 kDa in enzyme extracts of catla, rohu and silver carp. The substrate SDS‐PAGE evidenced the presence of various protease activity bands ranging from 21.6–93.7, 21.6–63.8 and 26.7–98.5 kDa for catla, rohu and silver carp respectively. In pH‐stat hydrolysis of Chilean fishmeal showed significantly (P < 0.05) higher degree of hydrolysis compared with soybean meal, silver cup (a commercial fish feed of Mexico) and wheat flour, with enzyme preparations of three fishes. The rate of hydrolysis was significantly (P < 0.05) higher in silver carp compared with others.     

Stress mitigating and growth enhancing effect of dietary tryptophan in rohu (Labeo rohita,Hamilton, 1822) fingerlings

An experiment was conducted to study the stress mitigation and growth enhancing role of dietary l-tryptophan (TRP) under thermal stress in rohu, Labeo rohita fingerlings for 45 days. Seven hundred and twenty fishes were distributed in three major groups that are ambient temperature (26 °C), 34 and 38 °C in triplicate following a complete randomized design. Acclimation of fishes to 34 and 38 °C over average ambient temperatures were carried out at 1 °C/day. Each group was fed with a diet supplemented with 0, 0.36, 0.72 or 1.42 % l-TRP. Results showed that blood glucose and serum cortisol level were found to be significantly higher (p < 0.05) in the higher temperature groups than the ambient temperature group. Similarly, aminotransferase, lactate dehydrogenase, malate dehydrogenase, CAT, superoxide dismutase activities were found to be significantly higher (p < 0.05) in the control groups (0 % l-TRP) and decreasing activities of these enzymes were observed with the increasing level of dietary l-TRP. In different temperature groups, l-TRP-supplemented groups were found to have higher (p < 0.05) growth, RGR and PER. The results obtained in the present study indicate that dietary l-TRP mitigates thermal stress and enhances growth. From the present study, we can conclude that dietary supplementation of l-TRP at the 0.72 % level in the diet is found to be optimum to reduce thermal stress even up to 38 °C in rohu, L. rohita. The baseline data obtained here could be useful for the farmers to formulate feeds to culture the fish in different agro-climatic zones.     

Antioxidant Properties of Scomber japonicus Hydrolysates Prepared by Enzymatic Hydrolysis

The antioxidant properties of the Pacific chub mackerel (Scomber japonicus) muscle protein hydrolysates prepared by enzymatic hydrolysis were investigated. After enzyme hydrolysis at 50°C for 60 min, more than 80% of the S. japonicus muscle protein was hydrolyzed. The highest 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity (71.69%) occurred in whole muscle protein hydrolysates treated at 50°C for 30 min with Protamex, and the highest 2,2?-azino-bis(3-ethylbenzothiazolin-6-sulfonic acid) (ABTS) radical-scavenging activity (95.39%) was observed in white muscle protein hydrolysates treated at 50°C for 30 min with Neutrase. The highest superoxide dismutase (SOD)-like activity (32.84%) was recorded in white muscle protein hydrolysates treated at 50°C for 120 min with Protamex. Changes in the molecular weight distribution of S. japonicus muscle proteins after enzymatic hydrolysis were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). A robust and a convenient enzyme hydrolysis technique for obtaining S. japonicus muscle protein hydrolysates with useful biological activities, within a short time (<2 h) is proposed.     

Characterization of a Heat Stable Protease from Arrowtooth Flounder;

Sodium dodecyl sulfate gel electrophoresis revealed rapid proteolytic degradation of myosin heavy chain in heated arrowtooth flounder muscle. A proteolytic enzyme of approximately 32,000 molecular weight was extracted from the muscle and purified 125 fold. Activity of the semi-purified enzyme at 55°C was optimal against casein at pH 6.0-7.0. Incubation with chemical reagents indicated the involvement of sulfhydryl groups in enzyme activity.     

Purification and properties of a histidine decarboxylase from Staphylococcus epidermidis TYH1 isolated from Japanese fish-miso

Histidine decarboxylase (HDC) from Staphylococcus epidermidis TYH1, a halotolerant histamine-producing bacterium isolated from Japanese fermented fish-miso, was purified to homogeneity for the first time. The enzyme was purified 182-fold from cell-free extracts by ammonium sulfate precipitation, anion exchange chromatography and gel filtration chromatography. The N-terminal amino acid sequences of two polypeptide chains of 27–30 and 7–9 kDa were highly homologous with those of α- and β-chains of other staphylococcal HDCs. The optimum pH and temperature for the enzyme were 6.0 and 60 °C, respectively. This enzyme did not decarboxylate lysine, arginine, tyrosine, tryptophan or ornithine. The enzyme activity decreased with the addition of NaCl. At pH 4.8, the V _max and K _m values were 45.5 μmol histamine min^?1 mg^?1 and 1.10 mmol/L, respectively. Moreover, this enzyme was resistant to heat treatment (80 °C for 15 min) and was stable upon freezing at ?30 °C for 7 days. The very similar physiological properties of this enzyme and the almost identical N-terminal amino acid sequence to that of the HDC from S. capitis indicated that this enzyme may be evolutionally highly conserved in the genus Staphylococcus. The biophysical properties of staphylococcal HDC were elucidated using native purified enzyme.     

Triploidization in rohu × mrigal hybrid and comparison of growth performance of triploid hybrid

The present study was aimed at the identification of treatment optima to induce triploidy in ‘Labeo rohita (rohu) × Cirrhinus cirrhosus (mrigal)’ hybrid using heat shock treatment. The eggs were exposed at four different temperature regimes viz., 38, 39, 40 and 41°C for 1–3 min, applied 3–5 min after fertilization. After 4 min of fertilization, heat shock treatments for 1 and 1.5 min durations were found the best inducing triploidy up to 100% and 96% respectively. Survival rates upto yolk sac absorption were found to be 73% and 71% in rohu and mrigal, 68% and 67% in the reciprocal diploid hybrids and 61% and 60% in the reciprocal triploid hybrids (RTH). Triploidy was confirmed by chromosome counting that revealed the diploid chromosome number of rohu and mrigal at 2n = 50 and in their triploid hybrid chromosome number was found to be 3n = 75. Growth rate of the RTH showed a significant difference (P < 0.05) from the single species and the diploid hybrids. Triploids also showed higher survival rate over the diploids.     

Degradation of myofibrils in cultured yellowtail <Emphasis Type="Italic">Seriola quinqueradiata</Emphasis> burnt meat: effects of a myofibril-bound EDTA-sensitive protease

“Burnt meat” is a term used to describe the white (pale, grainy, exudative) muscle of yellowtail or tuna. It (with lightness parameter, L* ≥ 55) was observed after 2 h storage in the suffocate in air (SA) 29°C group and after 4 h storage in the spinal cord destruction (SCD) 29°C group. In the SA 17°C group, burnt meat was also observed after 4 h storage. In contrast, the meat in the SCD 17°C group was normal until after 12 h storage. The myosin heavy chain (MHC) was more degraded than the other myofibrillar proteins, and some protein bands increased in the burnt meat. The protease that leads to the degradation of MHC was investigated using myofibrils from the meat. EDTA completely suppressed the degradation, indicating that a myofibril-bound EDTA-sensitive protease (MBESP) may exist in yellowtail muscle and this caused the degradation of MHC. The optimum pH and temperature of MBESP in yellowtail were 5.0 and 50–60°C, respectively.