Proteinase inhibitory activity of sarcoplasmic proteins from threadfin bream (Nemipterus spp.)

BACKGROUND: Thailand is the second largest surimi producer in the world and 50% of surimi is produced from threadfin bream. During surimi processing, sarcoplasmic proteins are removed through water washing and discarded in the waste stream. This study was aimed at investigating the proteinase inhibitory activity of sarcoplasmic proteins. RESULTS: Sarcoplasmic proteins from threadfin bream (TBSP) exhibited inhibitory activity toward trypsin but did not inhibit papain and chymotrypsin. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis under non‐reducing condition stained by trypsin inhibitory activity revealed three protein bands of molecular mass of 95, 41 and 37 kDa. Inhibitory activity of TBSP reached a maximum when subjected to 45 °C and completely disappeared at 60 °C. The breaking force and deformation of lizardfish surimi gel with added TBSP and pre‐incubated at 37° for 20 min increased with additional levels of TBSP (P < 0.05). Trichloroacetic acid–oligopeptide content of lizardfish surimi gel with added TBSP decreased with the addition of 4 g kg^?1 TBSP (P < 0.05). Retention of myosin heavy chain (MHC) increased when TBSP concentration was increased. TBSP effectively protected MHC from proteolysis at 37 °C to a similar extent as egg white powder, but efficacy of TBSP was not observed at 65 °C. CONCLUSION: TBSP could be applied to reduce proteolytic degradation of lizardfish surimi or other surimi associated with trypsin‐like proteinase, rendering an improvement in surimi gelation set at 37–40 °C. Copyright © 2009 Society of Chemical Industry     

Structure and functional properties of modified threadfin bream sarcoplasmic protein

Surimi wash-water contains up to 30% of protein from fish muscle that is currently underutilized. This paper describes the effect of acetylation, succinylation, trypsin hydrolysis or pre-heating at 55 °C on the emulsification and foaming properties of a threadfin bream sarcoplasmic protein (TBSP) model for wash-water protein. Multiple regression analysis showed that emulsification and foaming characteristics were differentially affected by TBSP surface hydrophobicity (S₀), solubility in water (S_W) and free amino group (fNH₂) concentration. Emulsification activity index (EAI) for TBSP was most enhanced by succinylation, whereas the foaming capacity (FC) was more effectively extended by trypsin hydrolysis. Structure–function relationships for emulsification were different from those associated with foaming or for ensuring the stability of these food dispersions. This study suggests that surimi wash-water protein functionality can be improved by protein modification. Further strategies may be needed to stabilize fish protein stabilized emulsions and foams.     

Processing optimization and characterization of gelatin from lizardfish (Saurida spp.) scales

Response surface methodology (RSM) with a 4-factor, 5-level central composite design (CCD) was conducted to ascertain the optimum gelatin extraction conditions from lizardfish scales. The effects of concentration of NaOH (%, X₁), treatment time (h, X₂), extraction temperature (°C, X₃) and extraction time (h, X₄) were determined. The responses included extraction yield (%), gel strength (g) at 9-10 °C and viscosity (cP) at 25 °C. The results showed the optimum conditions for the highest values of the three responses when a concentration of NaOH at 0.51%, a treatment time at 3.10 h, an extraction temperature at 78.5 °C and an extraction time at 3.02 h. The predicted responses were 10.7% extraction yield, 240 g gel strength and 5.61 cP viscosity. The experimental values were 10.6 ± 0.82% extraction yield, 252 ± 1.21 g gel strength and 7.50 ± 0.28 cP viscosity. The physicochemical properties of the lizardfish scales gelatin were characterized and the results indicated high protein and low ash content. Texture profile analysis (TPA) with compression was carried out at 30% deformation. The lizardfish scales gelatin was found to contain 20.4% imino acids (proline and hydroxyproline). Furthermore, slightly loose strands of the gel microstructure were observed using scanning electron microscopy (SEM).     

Effect of chymotrypsin digestion followed by polysaccharide conjugation or transglutaminase treatment on functional properties of millet proteins

Millet protein was solubilized by chymotrypsin; the soluble protein was conjugated to galactomannan under controlled conditions (60 °C, 76% RH) or polymerised by transglutaminase (TGase). SDS–PAGE patterns showed that the conjugated and polymerised proteins had higher molecular mass bands above the stacking gel. SDS–PAGE patterns also indicated that the digest was conjugated to galactomannan and polymerised by TGase. The free amino groups (OD₃₄₀) of the conjugated and polymerised digest were greatly reduced. Although the chymotrypsin digest was considerably insoluble between pH 2.0 and 5.0, galactomannan conjugate was completely soluble at all levels of pH. TGase polymer was slightly insoluble at pH 4.0. Galactomannan conjugate resisted heat-induced aggregation, even after heating at 90 °C for 20 min, while TGase polymer resisted heat-induced aggregation up to 70 °C, after which its solubility started to decline. The emulsifying properties of the conjugate and the polymerized proteins were greatly improved, compared to the native and chymotrypsin digests.     

Whey protein concentrate: Autolysis inhibition and effects on the gel properties of surimi prepared from tropical fish

Effects of whey protein concentrate (WPC) on autolysis inhibition and gel properties of surimi produced from bigeye snapper (Priacanthus tayenus), goatfish (Mulloidichthys vanicolensis), threadfin bream (Nemipterus bleekeri) and lizardfish (Saurida tumbil) were investigated. WPC (0–3%) showed inhibitory activity against autolysis in all surimi at both 60 and 65 °C in a concentration-dependent manner. Myosin heavy chain (MHC) of surimi was more retained in the presence of WPC. Breaking force and deformation of kamaboko gels of all surimi increased as added levels of WPC increased (P < 0.05). This was associated with lower levels of protein degradation, as evidenced by the decrease in trichloroacetic acid-soluble peptide content (P < 0.05). WPC at 3% (w/w) significantly decreased the whiteness of gels. However, water-holding capacity of kamaboko gels was improved with increasing concentration of WPC. The microstructure of surimi gels generally became denser with the addition of WPC.     

Gel‐enhancing effect and protein cross‐ linking ability of tilapia sarcoplasmic proteins

BACKGROUND: Tilapia (Oreochromis niloticus) sarcoplasmic proteins contain substantial transglutaminase (TGase) activity. The enzyme catalyzes the protein cross‐linking reaction, resulting in a more elastic gel. The objective was to investigate the gel‐enhancing effect of sarcoplasmic proteins from tilapia as related to TGase activity. RESULTS: Total TGase activity of sarcoplasmic proteins concentrate (SpC) increased about 3.6‐fold after ultrafiltration using 30 kDa membrane, but specific activity remained unchanged, indicating minimal TGase purification by ultrafiltration. Addition of 1 mg mL^?1 SpC containing 40 units TGase activity induced cross‐linking of tilapia actomyosin, and the extent of cross‐linking increased with added level of SpC. Myosin heavy chain (MHC) and troponin were preferably cross‐linked by tilapia SpC, while actin and tropomyosin were not affected. Higher retention of MHC was observed concomitantly with greater content of cross‐linked protein when SpC was added to lizardfish surimi. Lizardfish surimi with 10 g kg^?1 SpC added and pre‐incubated at 37 °C for 1 h exhibited 91.6% and 26.7% increase in breaking force and deformation, respectively, when compared to the control. CONCLUSIONS: Residual TGase activity in SpC played an important role in catalyzing the protein cross‐linking and enhancing actomyosin gelation. SpC could be a potential ingredient for improving textural properties of fish protein gel. Copyright © 2007 Society of Chemical Industry     

Effect of porcine plasma protein and setting on gel properties of surimi produced from fish caught in Thailand

Effects of porcine plasma protein (PPP) and high temperature setting on gel properties of surimi from bigeye snapper, bigeye croaker, threadfin bream and barracuda were investigated. PPP was effective in increasing breaking force and deformation of kamaboko gels set at 40°C for 30 min and heated at 90°C for 20 min. The optimum levels of PPP were 0.5, 0.5, 1.5 and 1.5 g/100 g and the optimum setting times were 2, 1.5, 1.5 and 2 h for bigeye snapper, bigeye croaker, threadfin bream and barracuda surimi, respectively. However, the addition of PPP significantly decreased whiteness (P<0.05). An increase in gel-forming ability of surimi with PPP coincided with a decrease in solubility in mixture of SDS, urea and β-mercaptoethanol, indicating the formation of nondisulfide covalent bond induced by both endogenous and plasma transglutaminase. The results supported that PPP improve the gelation of surimi in combination with setting.     

Effect of several cryoprotectants on the physicochemical and rheological properties of suwari gels from frozen squid surimi made by two methods

Functionality of squid surimi (Dosidicus gigas) made by two methods (isoelectric precipitation A, and acid washing, B) and stored for 6 months at −15 °C, was analysed as a function of several cryoprotectants. The cryoprotectant effect was studied in terms of the ability to form suwari gels (SA and SB) from the two kinds of surimi. Chemical analyses to detect protein aggregation, dynamic oscillatory tests at constant temperature (10 °C) and temperature sweep tests from 10 to 90 °C were performed.     

Effect of formaldehyde on protein cross-linking and gel forming ability of surimi from lizardfish induced by microbial transglutaminase

Impact of formaldehyde (FA) at various levels (0–9 μmol/g surimi) on gel properties of surimi from lizardfish added with microbial transglutaminase (MTGase) was studied. During iced storage of 10 days, total and free FA in lizardfish flesh increased continuously (P < 0.05). In the presence of FA, breaking force of gels slightly increased, whilst the deformation decreased (P < 0.05). The addition of MTGase (0.4 units/g surimi) was able to increase gel strength and water holding capacity of resulting gel. Nevertheless, gel strengthening effect of MTGase was lowered when FA at higher level was present. Myosin heavy chain (MHC) dominantly underwent polymerisation to a higher extent when either MTGase or FA was added. The higher reduction in ε-amino group content was observed in natural actomyosin (NAM) when FA at higher levels (0–30 μmol/g protein) was incorporated. Acyl transfer reaction mediated by MTGase was impeded in NAM containing FA, especially at higher levels. Generally, FA had an adverse effect on cross-linking ability towards surimi proteins induced by MTGase. Therefore, cross-linking and gel-forming ability of lizardfish surimi could be maximised by MTGase when surimi contained no FA.     

Segregative interactions and competitive binding of Ca in gelling mixtures of whey protein isolate with Naκ-carrageenan

Gelling mixtures of Na⁺κ-carrageenan with whey protein isolate (WPI) at pH 7.0 have been studied rheologically and by differential scanning calorimetry (DSC), with comparative measurements for the individual constituents of the mixtures. The concentration of WPI was held fixed at 10.0 wt% and carrageenan concentration was varied in the range 0.05–3.0 wt%. Ca²⁺ cations, which have been shown previously to be particularly effective in inducing gelation of κ-carrageenan, were introduced as CaCl₂. The concentration of CaCl₂ used in most of the experiments was 8 mM, but other concentrations were also studied. Mixtures were prepared in the solution state at 45 °C, and showed no evidence of either phase separation or complex formation. Rheological changes were monitored by low-amplitude oscillatory measurements of storage modulus, G′, during (i) cooling (1 °C/min) and holding at 5 °C, to induce gelation of the carrageenan in the presence of non-gelled WPI; (ii) heating and holding at 80 °C to dissociate the carrageenan network and induce gelation of WPI; (iii) cooling and holding again at 5 °C, to give composite networks with both components gelled; and (iv) re-heating to 80 °C to dissociate the carrageenan network. Gel structure was characterised further by creep–recovery measurements at the end of each holding period, and by torsion measurements at 5 °C, before and after thermal gelation of WPI.     

Effect of salts on the alkaline degradation of β-lactoglobulin gels and aggregates: Existence of a dissolution threshold

The effect of NaCl and CaCl₂ on the alkaline degradation of β-lactoglobulin gels and aggregates, and particularly on the onset of dissolution, is studied. For gels, measurements of solubility in 0.063–0.5 M NaOH at 20 °C show the existence of a practical dissolution threshold in NaCl concentration, lying between 0.24 and 0.47 M. For aggregates, destruction of soluble β-lactoglobulin in alkali, followed by size exclusion chromatography, yields similar results. Furthermore, during dissolution of a gel in alkali at high NaCl concentrations, the protein aggregates released are very large (e.g. ∼40% are larger than 200 kDa). CaCl₂ is found to cause similar inhibition of dissolution to NaCl, but at concentrations about 30× lower (∼10 mM). The threshold is hypothesised to arise from a combination of physical entanglements caused by the high protein concentration under conditions where little swelling occurs, and hydrophobic/electrostatic interactions between aggregates favoured by the high concentration of salts.     

Diffusing-wave spectroscopy investigation of heated reconstituted skim milks containing calcium chloride

The effect of heat treatment on reconstituted 10 wt% skim milks containing up to 20 mM added CaCl₂ at different pH values (pH 6.0–7.2), was investigated both in situ and after cooling of the heat treated milks. Measurements of pH in situ showed that pH decreased at high temperature, that the decrease in pH increases with the increase in the initial pH and that the magnitude of the decrease in pH was greater for milks with added CaCl₂. Marked increases in the viscosity at 25 °C of heated milks indicated that milks without added CaCl₂ with initial pH ≤6.2 and milks with 10 mM added CaCl₂ with initial pH ≤6.4, heated at 90 °C were not heat stable. At a given heating temperature, it was possible to superimpose the measured viscosity of samples with or without added CaCl₂ on the same curve when these were plotted as a function of the pH at that temperature instead of the initial milk temperature. To further demonstrate this finding a DWS experimental set-up was built and in situ measurements were performed on the milk samples heated at 75 °C for 10 min. The DWS measurements showed that ηa, the product of the viscosity with particle size, can also be superimposed for all the measured samples, when plotted as a function of the pH at the heating temperatures. Both the viscosity and DWS data demonstrate the importance of the pH at the heating-temperatures in influencing heat-induced changes in milk.     

Is intraspecific variability of growth and mycotoxin production dependent on environmental conditions? A study with Aspergillus carbonarius isolates

The aim of this study was to assess the impact of suboptimal environmental conditions on the intraspecific variability of A. carbonarius growth and OTA production using thirty isolates of A. carbonarius.Three a_w/temperature conditions were tested, one optimal (0.98a_w/25 °C) and two suboptimal: 0.90a_w/25 °C and 0.98a_w/37 °C as suboptimal water activity and temperature, respectively, which might take place through over ripening and dehydration of grapes. For each condition, 12 Petri dishes were inoculated, and colony growth and OTA production were measured over time.ANOVA revealed significant differences among μ and λ within the 30 assayed isolates. Coefficients of variation (CV%) revealed a wider dispersion of growth rates at 0.90a_w/25 °C compared to 0.98a_w/25 °C, and a more than 4-fold higher CV at 0.98a_w/37 °C compared to 0.98a_w/25 °C. However, dispersion of lag phases was similar at 0.98a_w/25 °C and 0.90a_w/25 °C and wider at 0.98a_w/37 °C.There were significant differences (p < 0.05) among OTA levels (ng/mm²) for the different conditions, values being lower under marginal conditions, and particularly at 0.98a_w/37 °C. Coefficients of variation (CV%) revealed a wider dispersion of OTA production at 0.90a_w/25 °C compared to 0.98a_w/25 °C, while CV at 0.98a_w/37 °C was similar to that at 0.98a_w/25 °C.In order to address the strain variability in growth initiation and prove the well-established notion of reducing OTA in foods by preventing fungal growth, a greater number of strains should be included when developing models for conditions that are suboptimal both for a_w for OTA production and temperature levels for growth.     

Trypsin Inhibitory Activity and Gel‐Enhancing Effect of Sarcoplasmic Proteins from Common Carp

Abstract: Proteinase inhibitory activity of sarcoplasmic protein (SP) extracted from common carp (Cyprinus carpio) muscle and its gel‐improving ability were investigated. SPs displayed 89% and 54% inhibitory activity toward trypsin at 40 and 65 °C, respectively. Protein bands with molecular mass of 69, 50, 44, 41, and 35 kDa appeared on trypsin inhibitory activity staining under nonreducing condition when incubated at 40 °C, while 2 protein bands at 54 and 35 kDa were observed at 65 °C. Addition of SP at 0.18 g protein/100 g increased textural properties of threadfin bream surimi gel. However, when SP was added in combination with various CaCl₂ concentrations (0.1% to 0.5%) it did not further improve textural properties as compared to the addition of SP alone. Retention of myosin heavy chain of threadfin bream surimi was greater with the addition of SP. These results indicated that the gel‐enhancing effect of common carp SP was due to the inhibitory activity toward endogenous trypsin‐like proteinases in threadfin bream surimi. Practical Application: Sarcoplasmic protein from common carp muscle could be used as a functional protein ingredient that minimizes muscle proteolysis and improves textural properties of surimi containing trypsin‐like endogenous proteinases.     

Interaction between calcium chloride and some carbohydrates as seen by mutual diffusion at 25 °C and 37 °C

Ternary mutual diffusion coefficients measured by Taylor dispersion method (D₁₁, D₂₂, D₁₂ and D₂₁) are reported for aqueous solutions of calcium chloride (CaCl₂) and some carbohydrates (glucose, fructose and sucrose) at 25 °C and 37 °C at carrier concentrations from 0.000 to 0.100 M, for each solute, respectively.     

Impact of zinc salts on heat-induced aggregation of natural actomyosin from yellow stripe trevally

Impact of zinc sulphate (ZnSO₄) and zinc chloride (ZnCl₂) on heat-induced aggregation of natural actomyosin (NAM) extracted from yellow stripe trevally (Selaroides leptolepis) was investigated. In the presence of ZnSO₄ or ZnCl₂, the transition temperature (T_max) of myosin shifted from 47.83 ± 0.30 °C to 46.05 ± 0.36 and 46.49 ± 0.49 °C, with the coincidental decreases in ΔH from 1.07 ± 0.03 J/g to 0.63 ± 0.02 and 0.67 ± 0.04 J/g, respectively (P < 0.05). Additionally, Ca²⁺–ATPase activity of NAM decreased with increasing the concentrations of ZnSO₄ or ZnCl₂ during heating up to 40 °C. During heating from 20 to 75 °C, higher turbidity, surface hydrophobicity and disulphide bond formation were obtained in NAM added with ZnSO₄ or ZnCl₂ at temperatures ranging from 40 to 75 °C, compared with the control. Nevertheless, a higher aggregation was found in NAM added with ZnSO_4, compared with ZnCl_2. Zeta potential (ζ) analysis suggested that the surface of NAM added with ZnSO₄ became less negatively charged, compared with that of ZnCl₂ counterpart. Transmission electron microscopy showed that the structure of NAM was highly interconnected, finer and denser when zinc salts, especially ZnSO₄ were incorporated. Therefore, ZnSO₄ could be used to induce aggregation of fish muscle proteins, thereby improving gelling property of fish mince or surimi.     

The effects of different cooking regimes on the cook yield and tenderness of non-injected and injection enhanced forequarter beef muscles

The aim was to investigate the effect of different cooking regimes on the cook yield and tenderness of non-injected and brine injected (0.5% residual NaCl) bovine M. triceps brachii caput longum (TB), M. supraspinatus (SP) and M. pectoralis profundus (PP). Injected and non-injected TB, SP and PP muscle sections (400 g) were (a) conventionally oven cooked to 72 °C or cooked slowly (using a Δ10 programme) to 72 °C or (b) cooked in a water bath to 72 °C or cooked in a water bath to 55 °C and held at this temperature for 2 h before heating to 72 °C. In addition, injected PP muscle sections were oven cooked to 69 °C and held at this temperature for up to 12 h. Slow cooking using a Δ10 programme had no significant (P < 0.05) effect on WBSF values of injected or non-injected SP and TB muscles but significantly (P < 0.05) decreased the WBSF values of injected and non-injected PP muscles when compared to conventional cooking. Slow cooking significantly (P < 0.05) increased % cook yield of injected PP, SP and TB muscles. Staged cooking significantly (P < 0.05) decreased the WBSF values and had no significant effect on sensory tenderness ratings of non-injected TB, SP and PP muscles and injected PP muscles. Staged cooked injected or non-injected PP, SP and TB muscles had lower % cook yield values than those cooked straight to 72 °C. Increasing the cooking time of injected PP muscles at 69 °C to 8 and 12 h decreased % cook yield, decreased WBSF values and increased sensory tenderness ratings. It also alleviated the problem of residual chewiness which was evident in PP muscles cooked using the other regimes.     

Colour and sarcoplasmic protein evaluation of pork following water bath and ohmic cooking

The objective of this study was to investigate the effects of ohmic (OH) and waterbath (WB) cooking on colour attributes and sarcoplasmic changes of porcine longissimus dorsi muscle at the same endpoint temperatures (EPTs; range 10 °C–80 °C). The OH treatment was carried out at 10 V cm^− 1, and the WB temperature at 85 °C. The colour parameters, deoxymyoglobin% (DeoMb) and metmyoglobin% (MetMb) of the OH-cooked meat were significantly lower (P < 0.05) than those obtained by WB-cooking at the same EPTs (range 60 °C–80 °C). SDS-PAGE analysis showed that the meat treated with WB-cooking had a lower sarcoplasmic protein solubility (5.97 mg/g vs.14.89 mg/g, P < 0.05) and fainter protein bands than that of OH-cooking thus, indicating paler colour, and lower water-holding capacity especially in WB-cooked meat at EPTs above 40 °C. Strong correlations among lightness, browness, metmyoglobin% and soluble proteins were observed in meat following OH-cooking.     

Thermal stability of soy protein isolate and hydrolysate ingredients

The objective of this study was to characterize the effects of pH, protein concentration and calcium supplementation on thermal stability, at 140 °C, of soy protein isolate (SPI) and soy protein hydrolysate (SPH) ingredients. Increasing pH between 6.4 and 7.5 led to significantly (p < 0.05) higher mean heat coagulation times (HCTs) at 140 °C, for all soy protein ingredients at 1.8, and 3.6% (w/v) protein. Increasing protein concentration from 1.8 to 7.2% (w/v) led to shorter HCTs for protein dispersions. Calcium supplementation up to 850 mg/L, except in the case of supplementation of SPI 1 with calcium citrate (CaCit), decreased HCT for soy protein ingredient dispersions, at pH 6.4 – 7.5. No significant differences (p < 0.05) were found in mean HCT for dispersions supplemented with calcium chloride (CaCl₂) and those supplemented with CaCit at 450, 650 and 850 mg/L Ca²⁺, in the pH range 6.4–7.5.