24 kDa Trypsin: A predominant protease purified from the viscera of hybrid catfish (Clarias macrocephalus × Clarias gariepinus)

Trypsin was purified to homogeneity from the viscera of hybrid catfish (Clarias macrocephalus × Clarias gariepinus) through ammonium sulphate fractionation and a series of chromatographies including Sephacryl S-200, Sephadex G-50 and DEAE-cellulose. It was purified to 47.6-fold with a yield of 12.7%. Based on native-PAGE, the purified trypsin showed a single band. The molecular weight of purified trypsin was estimated as 24 kDa by size exclusion chromatography and SDS–PAGE. The optimum pH and temperature for N^α-p-tosyl-l-arginine methyl ester hydrochloride (TAME) hydrolysis were 8.0 and 60 °C, respectively. Trypsin was stable to heat treatment up to 50 °C, and over a pH range of 6.0–11.0. Trypsin was stabilized by calcium ion. The trypsin activity was strongly inhibited by soybean trypsin inhibitor and N-p-tosyl-l-lysine chloromethyl ketone and partially inhibited by ethylenediaminetetraacetic acid. Activity decreased continuously as NaCl concentration (0–30%) increased. Apparent K_m value of trypsin was 0.3 mM and K_cat value was 92.1 S⁻¹ for TAME. The N-terminal amino acid sequence of 20 residues of trypsin was IVGGYECQAHSQPPTVSLNA, which is highly homologous with trypsins from other species of fish.     

Functional properties and antioxidative activity of protein hydrolysates from toothed ponyfish muscle treated with viscera extract from hybrid catfish

Functional properties and antioxidative activity of a protein hydrolysate prepared from toothed ponyfish (Gazza minuta) muscle, using viscera extract from hybrid catfish (Clarias macrocephalus × Clarias gariepinus), with a degree of hydrolysis (DH) of 70%, were investigated. The protein hydrolysate had a good solubility. It was soluble over a wide pH range (3–9), in which more than 77% solubility was obtained. The emulsifying activity index of the protein hydrolysate decreased with increasing concentration (P < 0.05). Conversely, the foaming abilities increased as the hydrolysate concentrations increased (P < 0.05). Protein hydrolysate exhibited the increases in 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH), 2,2‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulphonic acid) (ABTS) radical scavenging activities, ferric reducing power (FRAP) and metal chelating activity as hydrolysate concentration increased (P < 0.05). ABTS radical scavenging activity of protein hydrolysate was stable when heated at 100 °C for 180 min and subjected to a wide pH range (1–11). Therefore, protein hydrolysate from the muscle of toothed ponyfish produced by viscera extract from hybrid catfish can be used as a promising source of functional peptides with antioxidant properties.     

Use of pyloric caeca extract from bigeye snapper (Priacanthus macracanthus) for the production of gelatin hydrolysate with antioxidative activity

Proteolytic activity of pyloric caeca extract (PCE) from bigeye snapper (Priacanthus macracanthus) was studied. The highest activity was observed at 55 °C and pH 8.0 when casein, N^α-Benzoyl-dl-arginine-p-nitroanilide (BAPNA) and N^α-p-Tosyl-l-arginine methyl ester hydrochloride (TAME) were used as the substrates. The activity was inhibited markedly by 1 mg/ml soybean trypsin inhibitor, whereas E-64, pepstatin A and EDTA exhibited a negligible effect on activity. The results suggested that a trypsin-like enzyme was most likely the major proteinase in PCE. As determined by activity staining, two proteolytic activity bands with apparent molecular weights of 55 and 24 kDa were found. Gelatin hydrolysate from bigeye snapper skin prepared using PCE exhibited the increases in 2,2-Diphenyl-1-picrylhydrazyl (DPPH), 2,2-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities and ferric reducing antioxidative power (FRAP) as degrees of hydrolysis (DHs) increased (P < 0.05). Hydrolysates derived from gelatin using Alcalase combined with PCE showed the highest ABTS radical scavenging activity (P < 0.05). Gelatin hydrolysate prepared using Alcalase in combination with Neutrase or PCE at 500 and 1000 ppm, respectively, retarded the oxidation in both linoleic acid oxidation and lecithin liposome oxidation systems. The antioxidative peptide of gelatin hydrolysate had a molecular weight of 1.7 kDa.     

The effect of enzymatic hydrolysis time and temperature on the properties of protein hydrolysates from Persian sturgeon (Acipenser persicus) viscera

Protein hydrolysate was prepared from the viscera of Persian sturgeon (Acipenser persicus), a major sturgeon species in the Caspian Sea. Hydrolysis was performed at three different temperatures (35, 45 and 55 °C), pH 8.5, using commercially available Alcalase^® and an enzyme to substrate ratio of 0.1 AU/g viscera protein over a 205 min incubation period. Protein and lipid content of the hydrolysate were 65.82%, and 0.18%, respectively. Protein recovery and degree of hydrolysis ranged from 34.97% to 61.96% and 13.32% to 46.13%, respectively. The highest degree of hydrolysis was observed at 55 °C after 205 min (p < 0.05). The amino acid score of the hydrolysates was similar to that of the FAO/WHO reference protein. It is revealed that Persian sturgeon visceral protein hydrolysate amino acid fulfils adult human requirements. Based on National Research Council guidelines, phenylalanine is the first limiting amino acid in the hydrolysate. However, the hydrolysate has the potential for application as an ingredient in formulated diets.     

Antioxidant and biochemical properties of protein hydrolysates prepared from Silver carp (Hypophthalmichthys molitrix)

The antioxidant and biochemical properties of enzymatically hydrolyzed silver carp (Hypophthalmichthys molitrix) protein were studied. The molecular weight of the main peaks of the hydrolysates by both Alcalase and Flavourzyme was lower than 5000 Da. The hydrolysates treated by Alcalase for ?1.5 h (hydrolysis time) showed that the relative proportion of <1000 Da fraction was more than 60%. For the biochemical properties, hydrolysis by both enzymes increased protein solubility to above 75% over a wide pH range; and when the hydrolysis time was prolonged (>3 h), the colour of the hydrolysates turned yellowish. The protein hydrolysates exhibited significant hydroxyl radical-scavenging activity and inhibited linoleic acid peroxidation. For Alcalase treatment, the hydroxyl radical-scavenging activity and the inhibition of linoleic acid peroxidation of hydrolysates appeared to reach a maximum level for 1.5, 2.0 h of hydrolysis, respectively; and their antioxidant activity was close to that of α-tocopherol in a linoleic acid emulsion system, and carnosine in the 2-deoxyribose oxidation system. The hydrolysate with lower molecular weight distribution possessed stronger Fe²⁺ chelation ability at a sample concentration of 5.0 mg/mL. The results suggested that the antioxidant activity of silver carp protein hydrolysates were related to its degree of hydrolysis (DH), hydrolysis time and molecular weight.     

Purification and characterization of trypsin from the viscera of sardine (Sardina pilchardus)

Trypsin from the viscera of Sardina pilchardus was purified by fractionation with ammonium sulphate, heat treatment and Sephadex G-100 gel filtration with a ninefold increase in specific activity and 9% recovery. The molecular weight of the enzyme was estimated to be 25,000 Da on SDS–PAGE. This enzyme showed esterase-specific activity on Nα-benzoyl-l-arginine ethyl ester (BAEE). The purified enzyme was inhibited by benzamidine, a synthetic trypsin inhibitor, and phenylmethylsulphonyl fluoride (PMSF) a serine-protease inhibitor, but was not inhibited by the β-mercaptoethanol. The optimum pH and temperature for the enzyme activity were pH 8.0 and 60 °C, respectively. The relative activity at pH 9.0 was 95.5% and the enzyme showed pH stability between 6.0 and 9.0. The N-terminal amino acid sequence of the first 12 amino acids of the purified trypsin was IVGGYECQKYSQ. S. pilchardus trypsin, which showed high homology to other fish trypsins, had a charged Lys residue at position 9, where Pro or Ala are common in fish trypsins. The enzyme was strongly inhibited by Zn²⁺ and Cu²⁺.     

Manipulating the taste-related composition of strawberry fruits (Fragaria × ananassa) from different cultivars using deficit irrigation

Demand and, consequently, production of strawberry fruits has increased over the past few years and, as a result, the water abstractions for cultivation of this fruit have risen considerably. To limit the amounts of water used for several horticultural crops, water deficit irrigation (DI) has been seen as a potential alternative for new cultivation systems. DI in strawberry fruits is generally associated with reduction in berry size and yield; however, a recent study demonstrated that DI on strawberry can increase the concentration of some taste- and health-related compounds in fruits from cv. Elsanta. Hence, the aim of the present study was to further corroborate such findings and to assess the response (and variability) among different strawberry cultivars (namely Christine, Elsanta, Florence, Sonata and Symphony) to imposed water-DI conditions. Water-DI affected both fruit physiology and biochemistry. Nevertheless, the response to drought stress was different for each of the cultivars tested. Plants from cvs. Elsanta, Sonata and Symphony showed a greater reduction in berry size, accompanied by a significant increase in dry matter content for fruit harvested from DI-treated plants. Concomitant to this, and where dry matter was increased, the concentrations of sugars and some acids were generally higher in DI-derived fruit. In contrast, cvs. Florence and Christine did not show significant variations in berry weight or any of the target analytes measured when grown under the conditions imposed in this study. The results presented herein suggest that reducing water irrigation between flower initiation and fruit harvest may be a viable technique for increasing the concentrations of taste-related compounds in cvs. Elsanta, Sonata and Symphony and it may not have a negative impact on overall fruit size of cvs. Christine and Florence.     

Fishmeal-based diet decreases the redness of sunshine bass (Morone chrysops × Morone saxatilis) fillets

The objectives of the present study were to determine the effects of feeding a fishmeal-based diet on color attributes and lipid oxidation in sunshine bass (Morone chrysops × Morone saxatilis) fillets during retail display. A balanced diet containing 30 percent fishmeal (FM) or a diet containing poultry byproduct meal as a complete replacement of fishmeal (PB) was fed to sunshine bass for fifteen months. Harvested fish were filleted, overwrapped with polyvinyl chloride film and stored at 2 °C (REF) or over ice (ICE), under an illuminated retail display. Samples (n = 6) were analyzed after 0, 3, 6, or 9 d storage for color attributes (CIE L^∗, a^∗, b^∗, hue angle and chroma), thiobarbituric acid-reactive substances (TBARS), and pH. TBARS and pH increased (P < 0.05) during storage, indicating progress in lipid oxidation and protein changes. FM fillets demonstrated lower (P < 0.05) a^∗ (redness) value and greater (P < 0.05) hue angle than PB fillets. Since consumer acceptance of sunshine bass is dependant upon its white flesh, fishmeal supplementation could be used as a dietary strategy to improve fish marketability.     

The effects of pretreatments on antioxidative activities of protein hydrolysate from the muscle of brownstripe red snapper (Lutjanus vitta)

Different pretreatments of mince from brownstripe red snapper (Lutjanus vitta) including 1) washing; 2) membrane separation; 3) washing followed by membrane separation and 4) membrane separation followed by washing were conducted prior to hydrolysis. Among the resulting minces, that subjected to membrane separation with subsequent washing (MS/W) contained the lowest remaining myoglobin content, phospholipid content, heme iron and non-heme iron contents (p < 0.05) and showed the lowest TBARS values throughout 9 days of storage at 4 °C in the presence and absence of 0.15 mol L⁻¹ cupric acetate (p < 0.05). When hydrolysates from 1) mince, 2) MS/W and 3) protein isolate from MS/W (PI) with different degree of hydrolysis (DH) (20, 30 and 40%) were prepared using proteases from pyloric caeca of brownstripe red snapper, antioxidative activities determined by DPPH, ABTS radical scavenging activities, ferric reducing antioxidant power and metal chelating activity varied with hydrolysates and DH. Antioxidative activities increased with increasing DH up to 40% (p < 0.05). At all DH tested, hydrolysate prepared from MS/W exhibited the highest antioxidative activities determined by all assays, compared to those from mince and PI (p < 0.05). Hydrolysate from MS/W with 40% DH had the molecular weight lower than 6.5 kDa as determined by SDS-PAGE. In liposome oxidation system, the addition of hydrolysate from MS/W resulted in the lower TBARS, compared with the control throughout the incubation period of 48 h at room temperature (25-28 °C). Therefore, fish mince with membrane separation followed by washing was the most appropriate source for production of hydrolysate possessing antioxidative activity with the lowered amount of lipids and pro-oxidants.     

Lipid oxidation and fishy odour development in protein hydrolysate from Nile tilapia (Oreochromis niloticus) muscle as affected by freshness and antioxidants

Lipid oxidation and fishy odour development in protein hydrolysate from fresh and ice-stored Nile tilapia (Oreochromis niloticus) were investigated. During iced storage of 18 days, heme iron content decreased with a concomitant increase in non-heme iron content (P < 0.05). Peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) values increased. Phospholipid content decreased with a corresponding increase in free fatty acid content. The results suggested that lipid hydrolysis and oxidation took place during storage. When protein hydrolysates were produced from fresh and 18 days ice-stored Nile tilapia muscle, higher lipid oxidation and fishy odour/flavour along with higher amount volatile compounds were obtained in hydrolysate for unfresh sample (P < 0.05). However, the addition of mixed antioxidants during hydrolysis process markedly lowered lipid oxidation, b^∗, ΔC^∗, ΔE^∗ values, fishy odour/flavour as well as the formation of volatile compounds in the resulting hydrolysates prepared from both fresh and unfresh samples. Therefore, hydrolysate from Nile tilapia muscle with reduced fishy odour and lighter colour could be prepared by using fresh fish and incorporation of mixed antioxidants during hydrolysis.     

Crude palm oil from interspecific hybrid Elaeis oleifera × Elaeis guineensis: Fatty acid regiodistribution and molecular species of glycerides

The composition and structure of triacylglycerols (TAGs) and partial glycerides of crude palm oil obtained from interspecific hybrid Elaeis oleifera × Elaeis guineensis, grown in Colombia, were fully characterised and compared to data obtained by analysing crude African palm oil. Hybridisation appears to substantially modify the biosynthesis of fatty acids (FAs) rather than their assembly in TAGs. In fact, total FAs analysis showed significant differences between these two types of oil, with hybrid palm oil having a higher percentage of oleic acid (54.6 ± 1.0 vs 41.4 ± 0.3), together with a lower saturated fatty acid content (33.5 ± 0.5 vs 47.3 ± 0.1), while the percentage of essential fatty acid, linoleic acid, does not undergo significant changes. Furthermore, 34 TAG types were identified, with no qualitative differences between African and E. guineensis × E. oleifera hybrid palm oil samples. Short and medium chain FAs (8:0, 10:0, 12:0, 14:0) were utilised, together, to build a restricted number of TAG molecular species. Oil samples from the E. guineensis × E. oleifera hybrid showed higher contents of monosaturated TAGs (47.5–51.0% vs 36.7–37.1%) and triunsaturated TAGs (15.5–15.6% vs 5.2–5.4%). The sn-2 position of TAGs in hybrid palm oil was shown to be predominantly esterified with oleic acid (64.7–66.0 mol% vs 55.1–58.2 mol% in African palm oil) with only 10–15% of total palmitic acid and 6–20% of stearic acid acylated in the secondary position. The total amount of diacylglycerols (DAGs) was in agreement with the values of free acidity; DAG types found were in agreement with the representativeness of different TAG species.     

Flavour differences of cooked longissimus muscle from Chinese indigenous pig breeds and hybrid pig breed (Duroc × Landrace × Large White)

The objectives of this study were to investigate the flavour quality and volatile aroma compounds of cooked longissimus muscle from five typical Chinese indigenous pig breeds: Lantang (LT), Dahuabai (HB), Laiwu (LW), Rongchang (RC), Tongcheng (TC) and typical hybrid pig breed Duroc × Landrace × Large White (DLW). The chemical compositions of the main meat flavour precursors of the longissimus muscle from all six breeds were also examined. Distinct differences in amino acid composition and fatty acid composition of longissimus muscle, between the breeds, were observed. Among all six breeds, LW and RC had the highest intramuscular fat content and the lowest crude protein content; DLW had the lowest longissimus muscle fat content and the highest crude protein content. One-way analysis of variance showed that 23 volatile compounds were significantly affected by breed. Sensory analysis indicated that cooked longissimus muscle from DLW had the lowest pork flavour intensity and flavour-liking, compared with the Chinese indigenous breeds. LW and HB showed the highest pork flavour intensity and flavour-liking in cooked longissimus muscle.     

Expression of genes related to quality of Longissimus dorsi muscle meat in Nellore (Bos indicus) and Canchim (5/8 Bos taurus × 3/8 Bos indicus) cattle

This study was performed to compare CAPN1, CAPN2, CAST, TG, DGAT1 and LEP gene expressions and correlate them with meat quality traits in two genetic groups (Nellore and Canchim) in order to assess their expression profile and use their expression profile as genetic markers. We analyzed 30 young bulls (1 year old), 15 of each genetic group. Samples of the Longissimus dorsi muscle were collected for analysis of: total lipids (TL) and meat tenderness measured as Warner-Bratzler shear force (SF) and myofibrillar fragmentation (MFI) at day of slaughter and 7 days of aging. Gene expression profiles were obtained via RT-qPCR. TL and MFI showed differences between breeds, higher MFI in Canchim and higher TL in Nellore. Calpains showed no differential expression between groups, as did DGAT1, TG, and LEP. CAST was expressed more in the Nellore cattle. The only significant within-breed correlation (0.79) between gene expression and meat traits was found for DGAT1 and MFI in Canchim breed. Although the number of animals used in this study was small, the results indicate that the increased expression of CAST in Nellore may reflect tougher meat, but the lack of correlations with the meat traits indicates it is not a promising genetic marker.     

Characterisation of trypsin purified from the viscera of Tunisian barbel (Barbus callensis) and its application for recovery of carotenoproteins from shrimp wastes

Trypsin was purified from the viscera of barbel by precipitation using ammonium sulphate (0-80%), Sephadex G-100, and Mono Q-Sepharose ion exchange chromatography. The trypsin was purified 27-fold, with 79 U/mg specific activity and 31% recovery. The enzyme had a molecular weight of 24 kDa; purified trypsin appeared as a single band on native-PAGE. The optimum pH and temperature for enzyme activity were pH 10.0 and 55 °C with BAPNA used as a substrate. The N-terminal amino acid sequence of the first 12 amino acids of the purified trypsin was IVGGYECTPYSQ. The Michaelis-Menten constant (K_m) and catalytic constant (k_cat) values of the enzyme were 0.018 mM and 1.21 s⁻¹, respectively. The study also investigated the effects of purified trypsin on the recovery of carotenoproteins from shrimp (Parapenaeus longirostris) shells through hydrolysis using 1.0 U barbel trypsin/g shrimp shells for 1 h at 30 °C. The freeze-dried carotenoproteins recovered contained 71.09% protein, 16.47% lipid, 7.78% ash, and 1.79% chitin.     

Effect of chitosan coating combined with postharvest calcium treatment on strawberry (Fragaria × ananassa) quality during refrigerated storage

Strawberries (Fragaria × ananassa Duch.) were coated with either 1% or 1.5% chitosan (CS) or chitosan combined with calcium gluconate (CaGlu). Following treatment, strawberries were stored at 10 °C and 70 ± 5% RH for one week. The effectiveness of the treatments in extending fruit shelf-life was evaluated by determining fungal decay, respiration rate, quality attributes and overall visual appearance. No sign of fungal decay was observed during the storage period for fruit coated with 1.5% CS (with or without the addition of CaGlu) or 1% CS + 0.5% CaGlu. By contrast, 12.5% of the strawberries coated with 1% CS lacking calcium salt were infected after five days of storage. The chitosan coating reduced respiration activity, thus delaying ripening and the progress of fruit decay due to senescence. Chitosan coatings delayed changes in weight loss, firmness and external colour compared to untreated samples. Strawberries coated with 1.5% chitosan exhibited less weight loss and reduced darkening than did those treated with 1% chitosan, independently of the presence or absence of CaGlu. However, addition of calcium to the 1% chitosan solution increased the firmness of the fruit. Coated samples had greater visual acceptability than had untreated fruits. The addition of calcium gluconate to the chitosan coating formulation increased the nutritional value by incrementing the calcium content of the fruit.     

Changes in heme proteins and lipids associated with off-odour of seabass (Lates calcarifer) and red tilapia (Oreochromis mossambicus × O. niloticus) during iced storage

Changes in heme proteins and lipids associated with off-odour development in seabass (Lates calcarifer) and red tilapia (Oreochromis mossambicus × O. niloticus) muscles during 15 days of iced storage were studied. Fresh seabass contained the higher contents of myoglobin and heme iron, compared with red tilapia (P < 0.05). An increase in metmyoglobin proportion was observed during storage. After 3 days of storage, a decreased heme iron content and a concomitant increase in non-heme iron content were noticeable in both fish (p < 0.05). Oxidation of myoglobin and released non-heme iron were associated with lipid oxidation. The increases in oxidation products and free fatty acids were observed as the storage time progressed. Fishy and rancid odours were detected at day 6 of storage for both fish and a higher intensity was found in seabass muscle. Thus, the off-odour in fish muscle was mostly governed by lipid oxidation and species specific.     

Structural identification of the main ellagitannins of a boysenberry (Rubus loganbaccus × baileyanus Britt.) extract by LC–ESI-MS/MS,MALDI-TOF-MS and NMR spectroscopy

Four ellagitannins from boysenberry, a cross between Rubus loganbaccus and Rubus baileyanus Britt., were isolated by preparative HPLC and the exact structures determined by a combination of LC–ESI-MS/MS, MALDI-TOF-MS and NMR spectroscopy. The two most abundant ellagitannins were identified as sanguiin H-6, which is known to be abundant in Rubus species, and the other was identified as an isomer of sanguiin H-10, which has not previously been reported in Rubus. The two less abundant ellagitannins were identified as sanguiin H-2 and [galloyl–bis-HHDP–glucose]₂-gallate. Sanguiin H-2 has been previously reported in Rubus, whereas both sanguiin H-2 and [galloyl–bis-HHDP–glucose]₂-gallate have been previously reported as hot-water degradation products of lambertianin C. Even though lambertianin C is reported to be a major ellagitannin in other Rubus species, it was not found in any of the fractions, suggesting that both sanguiin H-2 and [galloyl–bis-HHDP–glucose]₂-gallate are present naturally in boysenberry.     

Antioxidative activity and functional properties of protein hydrolysate of yellow stripe trevally (Selaroides leptolepis) as influenced by the degree of hydrolysis and enzyme type

Antioxidative activity and functional properties of protein hydrolysates from yellow stripe trevally (Selaroides leptolepis) meat, hydrolyzed by Alcalase 2.4L (HA) and Flavourzyme 500L (HF) with different degrees of hydrolysis (DH) were investigated. As the DH increased, DPPH radical-scavenging activity and reducing power of HA decreased (p < 0.05) but no differences were observed for HF (p > 0.05). Metal chelating activity of both HA and HF increased with increasing DH (p < 0.05). HF generally had a higher (p < 0.05) chelating activity than had HA at the same DH tested. At low DH (5%), HA exhibited a better DPPH radical-scavenging activity while, at high DH (25%), HF had a higher (p < 0.05) reducing power. For the functional properties, hydrolysis by both enzymes increased protein solubility to above 85% over a wide pH range (2–12). When the DH increased, the interfacial activities (emulsion activity index, emulsion stability index, foaming capacity, foam stability) of hydrolysates decreased (p < 0.05), possibly caused by the shorter peptide chain length. At the same DH, the functionalities of protein hydrolysate depended on the enzyme used. The results reveal that antioxidative activity and functionalities of protein hydrolysates from yellow stripe trevally meat were determined by the DH and by the enzyme type employed.     

The formation of 2,5-dimethyl-4-hydroxy-2H-furan-3-one by cell-free extracts of Methylobacterium extorquens and strawberry (Fragaria × ananassa cv. Elsanta)

The formation of 2,5-dimethyl-4-hydroxy-2H-furan-3-one (DMHF), using the bacterial cells of Methylobacterium extorquens (strain with CABI registration number: IMI 369321) and strawberry cells (Fragaria × ananassacv. Elsanta), was studied. A combined mixture of the bacterial cell-free extract plus the strawberry extract was used as the enzymatic source. The pH of all the experimental aliquots was pH 6.0. The studied substrates were short-chain alcohols, carbohydrates and one amino acid. DMHF was analyzed by HPLC with UV detection at 280 nm. When 1,2-propanediol, 1-propanol, sucrose and fructose were used as substrates, the best production rates of DMHF were achieved, having a combined mixture of the bacterium and strawberry (minus the achenes) as the enzymic source. On the other hand, glucose, 1,2-propanediol, 2-propanol and fructose were the best substrates for the production of DMHF when a combined mixture of the bacterium and achenes served as the enzymic source. All results were obtained by comparing the production of DMHF from the strawberry extract alone and the bacterium plus the strawberry, without the achenes (in the first case) and the production of DMHF from the achene extract alone and the bacterium plus the achenes (in the second case). Moreover, the dehydrogenation activities of these three enzyme sources were determined by measuring the absorption of NADH at 340 nm. The substrate which was used for this series of the experiments was 1,2-propanediol. These results indicate the undoubted cooperation between the plant and the bacterial cells. This paper reports, for the first time, the formation of DMHF by bacterial and plant cell-free extracts.     

The application of high-concentration short-time chlorine dioxide treatment for selected specialty crops including Roma tomatoes (Lycopersicon esculentum), cantaloupes (Cucumis melo ssp. melo var. cantaloupensis) and strawberries (Fragaria × ananassa)

The effects of high-concentration short-time chlorine dioxide (ClO₂) gas treatment on food-borne pathogens inoculated onto the surface of tomatoes, cantaloupes, and strawberries were studied. Produce were spot-inoculated with a mixture of Salmonella enterica (serotypes Montevideo, Javiana and Baildon), Escherichia coli O157:H7 (serotypes 204 P, EDL 933 and C792) or Listeria monocytogenes (serotypes Scott A, F 5069 and LCDC 81-861), and treated with ClO₂ gas at 10 mg/l for 180 s. After ClO₂ gas treatment, surviving populations were determined and shelf-life studies were conducted (microbial spoilage population, change in color and overall appearance). Significant microbial reduction (p < 0.05) was observed for all treated samples. Nearly a 5 Log CFU/cm²Salmonella reduction was found on tomatoes, cantaloupe and strawberries, while a ∼3 Log CFU/cm² reduction was observed for E. coli and Listeria on all produce surfaces. E. coli and Listeria appeared to be more resistant to ClO₂ gas as compared to Salmonella spp. Treatments significantly (p < 0.05) reduced initial microflora population, while produce color surface was not significantly influenced, as compared to the control (p > 0.05). Results obtained suggest the potential use of high-concentration short-time ClO₂ gas treatment as an effective online pathogen inactivation technology for specialty crops in large-scale produce packing operations.