Hydrolytic Stability at Intermediate pHs of the Common Purine Nucleotides in Food, Inosine-5'-monophosphate, Guanosine-5'-monophosphate and Adenosine-5'-monophosphate

When the purine nucleotides, inosine-5′-monophosphate (IMP), guanosine-5′-monophosphate (GMP) and adenosine-5′-monophosphate (AMP) were subjected to canning temperatures (121°C) at pHs between 3 and 8, extensive phosphate bond hydrolysis to the corresponding nucleoside occurred. The half life for hydrolysis at pH 5 was 63, 41, and 51 min for IMP, GMP, and AMP, respectively. Rates of hydrolysis were even greater at pH 3 because both hydrolysis of the phosphate bond and the glycosidic bond occurred simultaneously. An Arrhenius plot of data collected at different temperatures illustrated that the nucleotides were very stable at room temperature. Half lives for IMP, GMP, and AMP were estimated to be 36, 19, and 40 yr, respectively, at 23°C and a pH of 5.     

Quality changes and storage life of common carp (Cyprinus carpio) at various storage temperatures

The shelf life and freshness changes in pond-grown common carp (Cyprinus carpio L) during storage at 0–2°C, 5–6°C and room temperature (26–29°C) were investigated by sensory, microbiological, physical and chemical analyses. The effect of gutting on the shelf life during storage at 0–2°C was examined. Iodine/starch and potassium sorbate were examined for their effects on shelf life of whole fish stored at 0–2°C and 5–6°C. Sensory results indicated that the whole fish had a maximum shelf life of 24 to 25 days at 0– 2°C. The life of the fish to the point beyond which it would be unsuitable for sale (commercial shelf life) was 17 days at 0–2°C. Storage at 5–6°C shortened shelf life 2- to 2.5-fold. At room temperature (26–29°C), spoilage was evident after 13 h. Gutting the carp shortened its storage potential at 0–2°C. Iodine treatment of this species stored at 0–2°C and at 5–6°C did not extend shelf life. The maximum shelf life of sorbate-treated fish at 0–2°C and 5–6°C was extended by 1–2 days, commercial shelf life by 3–4 days. Total volatile basic nitrogen, pH and penetrometer analyses were not reliable indicators of changes in freshness during shelf life. Thiobarbituric acid values were not useful as rancid odours or flavours were not detected during storage.     

Effect of lightly salt and sucrose on rigor mortis changes in silver carp (Hypophthalmichthys molitrix) stored at 4°C

To evaluate the effect of salt and sucrose on rigor mortis changes in silver carp during 72 h at 4 °C, physical (texture parameters, water holding capacity (WHC), cooking loss, L* value) and chemical adenosine triphosphate (ATP‐related compounds, K value, pH value) characteristics were studied. Silver carp were subjected to following treatments: left untreated (CK), treated with 1.8% salt (S) and 1.8% salt incorporating 1.8% sucrose (S+S). Curing treatments predominantly increased IMP content, WHC and texture parameters, retarded cooking loss and discoloration compared with untreated samples. The highest IMP concentration measured in S and S+S all occurred at 4 h, which was prior to that of CK (48 h). K value of S+S did not follow a linear relationship with time. The results indicated that the best eating times of CK, S and S+S were 2, 4 and 4 h at 4 °C, respectively, and curing treatments improved the quality of silver carp during postmortem process.     

Sensory Changes in Umami Taste of Inosine 5'-Monophosphate Solution after Heating

Discrimination in umami taste of inosine 5′-monophosphate (IMP) solution caused by thermal degradation was investigated by sensory evaluation. The difference threshold of umami taste of 0.005% IMP solution in the presence of 0.05% monosodium glutamate (MSG) was 0.002%. The difference threshold of a 0.005% IMP solution decreased by about one half when heated at 95°C for 15 h. Inosine, one of the main products of the thermal degradation of IMP, had a bitter taste. The detection threshold of inosine varied widely among panelists. Heating a 0.005% IMP solution at 95°C for 15 h formed inosine at about one tenth of its lowest detection threshold.     

Short Thermal Treatment Effect on Carp Myofibril and Sarcoplasmic Reticulum: Possible Mechanisms in Rigor Mortis Acceleration by "Arai" Treatment

Inorganic γ-phosphate liberation from adenosine 5′-triphosphate (ATP) by carp myofibrillar ATPase was measured at 0-60°C to elucidate mechanisms in rigor mortis acceleration of sliced carp muscle during washing at a moderately high temperature. ATP splitting within 20 sec in the presence of 5 mM MgCl₂ plus 0.25 mM CaCl₂ was maximal at 45°C, which agreed well with the commercially adopted condition for preparing carp “arai” muscle. In addition, the maximum Ca²⁺ uptake by sarcoplasmic reticulum was observed at 30°C and decreased at higher temperatures. The acceleration of carp muscle rigor mortis at around 45°C was suggested to be partly due to enhancement of myofibrillar Mg²⁺-ATPase activity by increase of intracellular Ca²⁺ concentration.     

Effect of Adenosine-Nucleotides and Their Derivatives on the Denaturation of Myofibrillar Proteins in vitro during Frozen Storage at −20°C

To investigate the effect of adenosine-nucleotides and their derivatives on the denaturation of myofibrillar proteins, 235 nmoles/mL adenosine-5′-diphosphate (ADP), adenosine-5′-monophosphate (AMP), inosine-5′-monophosphate (IMP), inosine (HxR), or hypoxantine (Hx), was added to 3 mg/mL actomyosin (AM) solution suspended in 0.10M KC1 solution and stored at ?20°C for 12 wk. The AM was extracted from milktish dorsal muscle. Protein denaturation was evaluated by measuring solubility, Ca-ATPase and Mg(EGTA)-ATPase activity of AM, by analyzing changes in electrophoretic profiles and transmission electron microscopy. Inosine and hypoxanthine accelerated protein denaturation compared to control samples. Infrared spectrum analyses indicated that negatively charged groups of these nucleotides interacted with amino or imino groups on AM after addition. ADP, AMP, and IMP had a protective effect on denaturation of AM during frozen storage at ?20°C.     

Hypoxanthine Ratio Determination in Fish Extract Using Capillary Electrophoresis and Immobilized Enzymes

Fish freshness was assessed using capillary electrophoresis and an immobilized enzyme procedure to monitor degradation of inosine-5′-monophosphate (IMP), inosine (HxR) and hypoxanthine (Hx). The enzymatic method used an amperometric probe at + 0.7 V (platinum vs silver/silver chloride) with immobilized xanthine oxidase, catalase, nucleoside phosphorylase, and nucleotidase for converting Hx, HxR or IMP to uric acid. Capillary electrophoresis resolved IMP, inosine and Hx by migration rates resulting from an applied electric field (416 V/cm, 50 μA). Components were detected at 250 nm. The H ratio of Hx/[IMP + HxR + Hx] and simplified K value of [HxR + Hx]/ [IMP + HxR + Hx] were determined in cod, salmon and trout stored on ice (0-4°C) and at 20°C. The two procedures agreed and for all species H ratio and K values increased with storage time.     

Thermal Degradation of Flavor Enhancers, Inosine 5'-monophosphate, and Guanosine 5'-monophosphate in Aqueous Solution

The mechanism of thermal degradation of inosine 5′-monophosphate (IMP) and guanosine (GMP) was investigated kinetically in aqueous solution as a function of pH and temperature. The degradation of IMP and GMP followed first order kinetics. The rate constants were considerably affected by pH and temperature. The half-life times at 100°C were: IMP, 8.7 hr (pH 4.0), 13.1 hr (pH 7.0), 46.2 hr (pH 9.0); GMP, 6.4 hr (pH 4.0), 8.2 hr (pH 7.0), 38.5 hr (pH 9.0). These times were shortened to about one-third by raising the temperature 10°C. The predominant degradation products were nucleosides and phosphoric acid, indicating that the main reaction of the thermal degradation was the hydrolysis of the phosphoric ester bond in the nucleotides.     

Lipid Peroxidation by Microsomal Fractions Isolated from Light and Dark Muscles of Herring (Clupea harengus)

Enzymic lipid peroxidation by light and dark muscle microsomes of herring (Clupea harengus) required ATP or ADP, NADH and Fe. NADPH could not effectively replace NADH. Inhibition was observed at high concentrations of ADP and NADH but not Fe. The optimal pH for the reaction of both types of microsomes was between 6 and 7. The average peroxidation rate was 362 and 1143 nmoles MDA per mg protein per hr at 6°C for the light and dark muscle microsomes, respectively. The energy of activation for the light and dark muscle microsomes was similar. The light muscle microsomes lost activity faster than the dark muscle microsomes when exposed to 35°C. Ferrous ion stimulated enzymic lipid peroxidation of light and dark muscle microsomes over that observed with ferric ion.     

Concentration of Umami Compounds in Pork Meat and Cooking Juice with Different Cooking Times and Temperatures

This study examined the concentrations of umami compounds in pork loins cooked at 3 different temperatures and 3 different lengths of cooking times. The pork loins were cooked with the sous vide technique. The free amino acids (FAAs), glutamic acid and aspartic acid; the 5′‐nucleotides, inosine‐5′‐monophosphate (IMP) and adenosine‐5′‐monophosphate (AMP); and corresponding nucleoside inosine of the cooked meat and its released juice were determined by high‐performance liquid chromatography. Under the experimental conditions used, the cooking temperature played a more important role than the cooking time in the concentration of the analyzed compounds. The amino acid concentrations in the meat did not remain constant under these experimental conditions. The most notable effect observed was that of the cooking temperature and the higher amino acid concentrations in the released juice of meat cooked at 80 °C compared with 60 and 70 °C. This is most likely due to the heat induced hydrolysis of proteins and peptides releasing water soluble FAAs from the meat into the cooking juice. In this experiment, the cooking time and temperature had no influence on the IMP concentrations observed. However, the AMP concentrations increased with the increasing temperature and time. This suggests that the choice of time and temperature in sous vide cooking affects the nucleotide concentration of pork meat. The Sous vide technique proved to be a good technique to preserve the cooking juice and the results presented here show that cooking juice is rich in umami compounds, which can be used to provide a savory or brothy taste.     

Inhibition of Listeria monocytogenes and Salmonella by Combinations of Oriental Mustard,Malic Acid,and EDTA

The antimicrobial activities of oriental mustard extract alone or combined with malic acid and EDTA were investigated against Salmonella spp. or Listeria monocytogenes at different temperatures. Five strain Salmonella or L. monocytogenes cocktails were separately inoculated in Brain Heart Infusion broth containing 0.5% (w/v) aqueous oriental mustard extract and incubated at 4 °C to 21 °C for 21 d. For inhibitor combination tests, Salmonella Typhimurium 02:8423 and L. monocytogenes 2–243 were individually inoculated in Mueller Hinton broth containing the mustard extract with either or both 0.2% (w/v) malic acid and 0.2% (w/v) EDTA and incubated at 10 °C or 21 °C for 10 to 14 d. Mustard extract inhibited growth of the L. monocytogenes cocktail at 4 °C up to 21 d (2.3 log₁₀ CFU/mL inhibition) or at 10 °C for 7 d (2.4 log₁₀ CFU/mL inhibition). Salmonella spp. viability was slightly, but significantly reduced by mustard extract at 4 °C by 21 d. Although hydrolysis of sinigrin in mustard extract by both pathogens was 2 to 6 times higher at 21 °C than at 4 °C to 10 °C, mustard was not inhibitory at 21 °C, perhaps because of the instability of its hydrolysis product (allyl isothiocyanate). At 21 °C, additive inhibitory effects of mustard extract with EDTA or malic acid led to undetectable levels of S. Typhimurium and L. monocytogenes by 7 d and 10 d, respectively. At 10 °C, S. Typhimurium was similarly susceptible, but combinations of antimicrobials were not more inhibitory to L. monocytogenes than the individual agents.     

Inhibition of Shigella sonnei and Shigella flexneri in Hummus Using Citric Acid and Garlic Extract

Hummus (chickpea dip) is a ready‐to‐eat product that may pose a significant risk to human if pathogens are present. Several organisms including Shigella spp. have been isolated from hummus. However, studies on the survival and inhibition of Shigella spp. in food are scarce. This study investigated the growth pattern of Sh. sonnei and Sh. flexneri in hummus at different temperatures (4, 10, and 24 °C). Additionally, the inhibitory activity of different concentrations of citric acid (CA) (0.5%, 1.0%, and 2.0%) and garlic extract (GE) (1.0%, 2.0%, and 3.0%) against Sh. sonnei and Sh. flexneri inoculated into hummus and stored at 4 and 10 °C was investigated. Both Shigella spp. survived well at 4 °C, while both grew to >7.0 log₁₀ after 4 d at 10 °C or 1 d at 24 °C. At 4 °C, CA at 0.5% and 1.0% resulted in a slight reduction in the count (approximately 1.0 log₁₀); a complete elimination of Sh. sonnei was attained by using 2.0% CA. However, approximately 3.0 log₁₀ reduction in Sh. sonnei was obtained at 10 °C. For Sh. flexneri, CA at 0.5% and 1.0% resulted in a bacteriostatic inhibition. GE at 1.0% and 2.0% resulted in approximately 1.0 to 2.0 log₁₀ reduction in Sh. sonnei count at 4 °C, while at 3.0% GE, approximately 4.0 and 3.0 log₁₀ reductions were obtained at 4 and 10 °C, respectively. In comparison, the 2.0% and 3.0% GE resulted in a bacteriostatic effect against Sh. flexneri at 4 and 10 °C.     

Nonvolatile taste compounds in cooked Chinese Nanjing duck meat following postproduction heat treatment

Abstract: Taste‐active compounds, including free amino acids, succinic acid and 5′‐nucleotides, and chemical components including moisture, pH, protein, crude fat, and sodium chloride were evaluated in cooked and packaged Chinese Nanjing ducks following heat treatments including control, 99 °C for 40 min, 108 °C for 20 min, 92 °C microwave followed by water heating, 95 °C for 30 min, 121 °C for 25 min. Heat treatment decreased (P < 0.05) the content of Alamine and moisture and reduced the pH value in muscle, but increased (P < 0.05) the protein and 5′‐nucleotides content. The 99 °C group had a significantly lower crude fat content compared with other groups (P < 0.05). The succinic acid content in the control group was significantly higher than in the 121 °C group (P < 0.05). Groups treated at higher temperatures (108 °C, 121 °C, and microwave) had similar equivalent umami concentrations and 5′‐nucleotides, free amino acids content, as well as the derived bitter and sweet taste components compared with the groups treated at lower temperatures. It can be speculated that these differences account for the enhanced flavor of the meat in the 99 °C, 108 °C, 121 °C, and microwave groups compared with the untreated control. Therefore, heat treatment at lower temperature after packaging may prolong product shelf life without any detrimental effects on taste. The results of this study indicate that it is important to use lower temperatures in this type of food processing. However, it may be possible to modify the primary processing steps to improve the content of umami‐like taste compounds such as 5′‐nucleotides. Practical Application: Heat treatment of packaged products is an effective method for eradication of microbes, therefore increasing the shelf‐life. However, such treatment can result in major changes in the sensory perception of meat products, particularly the formation of off‐flavors. This study investigated changes in taste‐active compounds in duck meat following a number of types of heat treatment.     

Optimization of Fermentation Parameters for the Production of Concentrated Starters from Nonbitter Streptococcus lacti INIA 12

Streptococcus lactis INIA 12, a selected nonbitter strain, reached its maximum counts and its highest lactic acid production in a culture medium containing 125 g/L skim milk powder, fortified with 5 g/L yeast extract and digested with 10 ppm papain for 20 min at 65°C. Lactose added to the medium did not enhance growth rate or biomass production. A growth temperature of 32°C and the maintenance of pH at 6.80, with 10N NaOH as the neutralizer, were the optimum fermentation parameters in batch cultures. In ten 40-L fermentations carried out at 32°C and pH 6.80, with a 5% inoculum, a 0.2 kg/cm² nitrogen head space pressure and a stirring rate of 80 rpm, maximum counts of S. lactis (10¹⁰ CFU/mL) were reached after incubation for 6 hr at 32°C and pH 6.80.     

Rheological and physico‐chemical properties of gelatin extracted from the skin of a few species of freshwater carp

The physico‐chemical and rheological properties of gelatin from the skins of three different freshwater carp species, namely Catla catla, (catla) Cirrhinus mrigala (mrigal) and Labeo rohita (rohu), have been assessed and compared with that of gelatin from porcine skin. The average solids yield from the three species of carp varied in the range of 11.8–14.1%. The amino acid profile showed that the porcine gelatin had a higher proportion of imino acids and glycine than carp skins gelatin. The average molecular weight of carp skins gelatin as determined using a gel filtration technique was 233 kDa, while that of porcine skin gelatin was 282 kDa. The gelling temperature of carp skins gelatin was in the range of 6–15.7 °C, and the melting temperature was 17.9–23.7 °C as determined using a controlled stress rheometer. A higher gelling and melting temperature was observed for porcine skin gelatin.     

Myofibrillar protein degradation of carp (Labeo rohita (Hamilton)) muscle after post‐mortem unfrozen and frozen storage

The degradation of myofibrillar proteins of rohu carp (Labeo rohita (Hamilton)) muscle was analysed after post‐mortem storage. Muscle fillets were kept either unfrozen at 2 °C for up to 15 days or frozen at ?8 °C or ?20 °C for up to 6 months. A co‐ordinated histochemical, biochemical and electrophoretic study showed a differential response of the carp muscle, revealing clear degenerative/degradative changes specific to the post‐mortem storage temperatures. The myofibrillar protein fractions, namely myosin light chains and α‐actinin, showed degradative changes during the above storage conditions, whereas other protein fractions in the high‐molecular‐weight range fragmented to give lower‐molecular‐weight proteins. The importance of the post‐mortem storage temperature for controlling the degradation of the myofibrillar proteins was emphasised. This is the first report on this popular fish species, known for its culinary importance, showing that specific protein fractions of the myofibrils degrade during post‐mortem storage. © 2001 Society of Chemical Industry     

Formation of selected biogenic amines in carp meat

Changes in biogenic amine content in carp (Cyprinus carpio) muscle were studied. Fish halves and minced fish meat were stored at 3 and 15 °C. Both the temperature of storage and the type of meat processing had statistically important effects on the amine content. In another set of experiments, temperature and the preservative effects of Purac at various concentrations were tested. Purac can extend the shelf‐life of fish halves stored at 3 °C by about 5 days. Putrescine concentration is proposed as a chemical indicator of carp meat quality. Decomposition is apparent when the putrescine content in the meat exceeds 20 mg kg^?1. © 2002 Society of Chemical Industry     

Enzymatic Production of Ribonucleotides from Autolysates of Kluyveromyces marxianus Grown on Whey

After 20h fermentation of medium containing 5% (w/v) dehydrated whey, at 30°C, pH 4.5, yeast cells were harvested, diluted in 0.1M KH₂PO₄, and autolyzed at different pHs (6.5–7.5) and temperatures (45–55°C). Phosphodiesterase (0.2–1.0% w/v, 65°C, pH 6.5, 6h) and adenyl deaminase (0.5-1.0% w/v, 60°C, pH 5.5, 4h) were added to the autolysates. After heat treatment (100°C, 15 min), samples were analyzed by RP-HPLC and LC/MS. Production of 5′-ribonucleotides was maximized at 50°C, pH 6.5. Yields of 5′-AMP (800 μg/g of biomass) and 5′-GMP (2000 μg/g) increased considerably after addition of 1.0% phosphodiesterase. 5′-IMP increased only after addition of 1.0% adenyl deaminase.     

Determination of Inosine-5-Monophosphate in Fish Tissue with an Enzyme Sensor

An enzyme sensor for IMP consisted of immobilized enzymes and an oxygen electrode. A nucleotidase (E.C. 3.1.3.5.), nucleoside phosphorylase (E.C. 2.4.2.1.) and xanthine oxidase (E.C. 1.2.3.2.) were immobilized on a membrane prepared from cellulose triacetate, 1,8-diamino-4-aminomethyloctane and glutaraldehyde. Optimum conditions for IMP determination were pH 7.8, 30°C, a flow rate of 1 ml min⁻¹ and sample volume of 20 μl. A calibration curve for IMP was linear up to 15 mM. This sensor could be applied to the determination of IMP in meats of sea bass, mackerel, yellowfish and saurel. Good correlative results were observed between the values obtained by the sensor and the conventional method. The sensor was stable for more than 15 days at 5°C and 100 assays.     

Effects of Hydrolysates from Silver Carp (Hypophthalmichthys molitrix) Scales on Rancidity Stability and Gel Properties of Fish Products

This study aimed to evaluate the effectiveness of hydrolysates, which were obtained from the scales of silver carp (Hypophthalmichthys molitrix) by papain, flavourzyme, and Alcalase 2.4 L, as natural antioxidants in silver carp mince and surimi gels during storage at 4 °C. The hydrolysates that possess greater in vitro antioxidant activities (DPPH radical-scavenging activity, Fe²⁺-chelating activity, and reducing power), including hydrolysates catalyzed by papain at 10 min (HP), flavourzyme at 5 min (HF), and Alcalase 2.4 L at 5 min (HA), were chosen as additives. Color, cooking loss, conjugated dienes (CDs), thiobarbituric acid reactive substances (TBARS), fatty acids, and sensory scores of mince were measured on days 0, 2, 4, 6, and 8 during 4 °C storage; additionally, whiteness, breaking force, deformation, gel strength, and sensory score of surimi gels were measured on days 1, 3, 5, 7, 9, and 11 during 4 °C storage. The results indicate that HA was conducive to lowering the cooking loss of mince and that HF significantly (P?<?0.05) reduced the CDs value of mince. For surimi gels, HF improved whiteness, deformation, and gel strength. Hence, HF could serve as a natural antioxidant during early oxidation and improve gel formation of silver carp products.