Lipoxygenase in Sweet Corn Germ: Isolation and Physicochemical Properties

Off-flavor and off-aroma development, which may be catalyzed by lipoxygenase (LPO), are common in frozen stored sweet corn. Lipoxygenase activity in the germ fraction of sweet com (Zea nruys L. cv. Jubilee) was determined and compared with that in the degermed fraction. Lipoxygenase activity/g germ was about three times greater than that of the degermed fraction, Optimized procedures for isolation of lipoxygenase from the germ fraction were developed. Lipoxygenase was isolated by preparation as an acetone powder, extraction with 0.2M TrisHCI, pH 8.0 (4°C), fractionation with 40–60% saturated ammonium sulfate and dialysis. Optimum pH was 6–7 and temperature 50°C for activity of partially purified lipoxygenase. The enzyme appeared stable at pH 5–8 and ～90% of original activity was inactivated after heating in pH 7 buffer at 70°C for 3 min.     

Characterization of endogenous enzymes of milled rice and its application to rice cooking

The relationship between the activities of endogenous enzymes in milled rice and accumulation of chemical components in the rice grains during cooking was investigated. Maltose and soluble starch were optimally hydrolyzed around 60 °C by both crude extracts and purified α-glucosidases of milled rice. Gelatinization onset temperature of rice flour was determined to be 63 °C with DSC analysis under usual cooking condition. Hydrolytic activities on carbohydrates and proteolytic activities of milled rice were enhanced at pH 5 compared to pH 7 at the range of 4–80 °C. When rice was cooked at pH 5, glucose and amino acids highly accumulated with soaking for 16 h before heating. We propose a new method of cooking with prolonged soaking at acidic pH, which is effective for increasing the amounts of chemical components in cooked rice.     

Characterization of Seed Lipoxygenase of Phaseolus vulgaris cv, Haricot

Phaseolus vulgaris cv, haricot seed lipoxygenase was extracted and partially purified. The precipitation of an active lipoxygenase fraction with solid ammonium sulfate (at 20–50% of saturation) increased its activity by a factor of 3. The pH for optimum activity was 7.3. The addition of 40 mM potassium cyanide resulted in a doubling of the enzyme activity. Enzyme activity was completely lost on storage at 4°C for 48 hr. The activity of haricot lipoxygenase extract was considerably greater on linoleic acid than on its esters, and decreased in the order: mono- > di > trilinolein. The haricot lipoxygenase shares many of the characteristics of the corresponding enzyme found in several seed sources.     

Control of Enzymatic Browning in Processed Mushrooms (Agaricus bisporus)

Control of polyphenol oxidase (E.C. 1.14.18) activity by the use of citric acid was investigated. The enzyme was inactivated at pH 4.0 and was stable to 10 min exposures at 25°C in the pH range 4.0–8.0. At pH 6.5 the enzyme was active at 45°C but not at 70°C and thermal inactivation followed pseudo first-order kientics. At pH 6.5 the activation energy (E_a) for enzyme inactivation was 41.1 Kcal/mole while at pH 3.5 two rate constants and hence two values for E_a were observed. Between 0–5 min E_a for inactivation of polyphenol oxidase was 8.7 Kcal/mole and >5 min Ea was 21.8 Kcal/mole.     

Heat Inactivation of Trypsin Inhibitors in Soymilk at Ultra-High Temperatures

Soymilk samples at pH 7.5, 6.5 and 2 were subjected to heat treatment at 93°C and indirect ultra-high temperature. When heated at 93°C, 121°C and 132°C, trypsin inhibitor activity (TIA) in soymilk was more heat-labile at high pH than at lower pH. However, the effect of pH on rate of thermal inactivation was less pronounced when the holding temperature was increased to 143°C and 154°C. The point on a curve relating holding temperature and holding time, indicating inactivation of 90% of the TIA in soymilk at pH 6.5 in the range 93–154°C, coincided with the thermal-death-time curve of the organism putrefactive anacrobe 3679 at about 125°C.     

Pasteurization of mixed mandarin and Hallabong tangor juice using pulsed electric field processing combined with heat

Effects of pulsed electric filed (PEF) processing combined with heating (H-PEF processing) on the inactivation of microorganisms and the physicochemical properties of mixed mandarin and Hallabong tangor (MH) juice were studied. Using a pilot-scale PEF system, MH juice, pre-heated at 55 °C, was PEF-treated at 19 kV/cm of electric field and 170 kJ/L of specific energy and the juice, pre-heated at 70 °C, was PEF-treated at 16 kV/cm and 100 kJ/L or 12 kV/cm and 150 kJ/L. H-PEF processing at 70 °C–16 kV/cm–100 kJ/L reduced the aerobe, yeast/mold, and coliform counts of MH juice by 3.9, 4.3, and 0.8 log CFU/mL, respectively, without affecting the ascorbic acid concentration and antioxidant capacity of juice. H-PEF processing changed juice color and browning degree (p < 0.05), but not total soluble solid content or pH. By controlling initial juice temperature and electric field strength, H-PEF processing can be an effective pasteurization method for mixed juice with minimal changes in quality.     

Pressure Induced Inactivation of Selected Food Enzymes

Pectinesterase, lipase, polyphenol oxidase, lipoxygenase, peroxidase, lactoperoxidase, phosphatase and catalase have been examined at distinct conditions within a pressure range of 0.1 to 900 MPa, temperatures from 25°C to 60°C, pH 3 to 7, and time of treatment of 2 min to 45 min. Results in model buffers made it possible to rank the enzymes according to their pressure induced inactivation in the following order: lipoxygenase, lactoperoxidase, pectinesterase, lipase, phosphatase, catalase, polyphenol oxidase, peroxidase. A combination of pressure with moderate temperature increased the degree of enzyme inactivation. Pressure treatment of real food systems showed a protective effect of food ingredients on the pressure inactivation of most enzymes evaluated. For example sucrose protected pectinesterase from inactivation by pressure while lactoperoxidase and lipoxigenase were as stable in milk as in buffer.     

Microwave heat treatment application to pasteurization of human milk

A prototype of microwave pasteurizer has been proposed as an alternative for holder pasteurization (HP) routinely used in Human Milk Bank (HMB), ensuring microbiological safety of human milk (HM). It was shown that the time of heat generation was about 15–16 min shorter by applying the microwave than in HP. Total inactivation of heat-sensitive bacteria Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis, suspended in milk, occurred in the temperature 62–72 °C in HP. In the case of heat-resistant enterococci the level of inactivation depended on the conditions of the process and the properties of the strains. The application of microwave heating allows to obtain lower D-value than those achieved during HP. The using of microwave heating at 62.5 or 66 °C for 5 or 3 min, respectively, allows to inactivation of HM microbiota. Appropriate microbiological quality of milk is critical for the effectiveness of the pasteurization process.Industrial relevanceLooking for new methods of donor human milk (HM) preservation is dictated by the necessity of providing microbiological safety and, at the same time, maintain its high nutritional and biological value. The holder pasteurization used in the Human Milk Banks (HMB) (heating at 62 °C for 30 min) leads to inactivation of all vegetative forms of microorganisms. Unfortunately, this method causes significant reduction of health benefitting properties of HM. The paper demonstrates the possibility of using the new microwave pasteurizer for preservation of HM, allowing for quick heating of milk to the appropriate temperature and maintaining it in these conditions for a required time. It was shown that the decimal reduction times (D) for strains inoculated to UHT or human milk are several times shorter by using microwave heating than in the commercial pasteurization method. The total inactivation of HM microbiota is obtained after heating at 62.5 and 66 °C for 5 and 3 min, respectively.     

Rheological and microstructural characterization of double cream cheese

Firmness at 5 °C, dynamic viscoelastic moduli (G' G) and capillary extrusion profiles at 20 °C, were obtained on double cream cheese after the different steps of processing (curd obtention, mixing at 70 °C and 500 r.p.m. with heat-denatured WPC, heating at 85 °C, homogenizing at 20 MPa (1st stage) and 5 MPa (2nd stage) and cooling to 20 °C and 13 °C) and storage 7–9 days at 5 °C. It was found that double cream cheese became firmer and more elastic after heating and homogenization, although it became softer and more viscous after mixing and cooling. Transmission Electron Microscopy (TEM) and cryo-Scanning Electron Microscopy (SEM) micrographs showed that rheology results could be related to aggregation (during heating and homogenization) and disruption (during cooling) of milk fat globule/casein complexes. Dispersion of homogenization clusters after cooling, and aggregation of milk fat globules during storage caused double cream cheese structural instability to appear. It was suggested that the heterogeneity of capillary extrusion profiles could be quantified through application of fractal concepts and Fourier analysis and related to structure and texture of double cream cheese.     

Thermal Inactivation of Clostridium perfringens Enterotoxin in Buffer and in Chicken Gravy

The time-temperature relationships for the inactivation of purified enterotoxin prepared from Clostridium perfringens were determined by Vero cell assay in two different heating menstrua. Enterotoxin diluted to an initial concentration of 12.5 μg/g in chicken gravy and in 0.1M phosphate buffer both at pH 6.1 was heated at temperatures of 59–65°C. Average inactivation times in gravy ranged from 72.8–1.5 min and in buffer from 149.4–2.4 min.     

Identification and characterization of lipoxygenase in fresh culinary herbs

The aim of this project was to study the biochemical characterization of lipoxygenase extracted from basil (Ocimum basilicum L.), rosemary (Rosmarinus officinalis L.), and sage (Salvia officinalis L.) leaves. The lipoxygenase extracted from these culinary herbs was frozen with liquid N₂ and ground into powder before adding ice-cold phosphate buffer (pH 7). The crude extracts from the three aromatic plants showed various specific activities and the highest specific activity and the lowest protein content were observed for the basil extracts. The optimum pH values for lipoxygenase extracted from basil and sage ranged between 5 and 6, while pH 6 was the optimum value for lipoxygenase activity for rosemary. The optimum temperature was 40°C for all of the crude extracts analyzed. Considering the biochemical characteristics evaluated for lipoxygenase from each culinary herb, suitable strategies for the inactivation of the enzyme can be proposed in order to reduce its detrimental effects on fresh aromatic herbs.     

Effect of pH-dependent,stationary phase acid resistance on the thermal tolerance of Escherichia coli O157:H7

The ability of pH-dependent, stationary phase acid resistance to cross-protect Escherichia coli O157:H7 against a subsequent lethal thermal stress was evaluated using microbiological media and three liquid foods. Three strains were grown for 18 h at 37°C in acidogenic (TSB+G, final pH 4·6–4·7) and non-acidogenic (TSB-G, final pH 7·0–7·2) media to provide stationary phase cells with and without induction of pH-dependent acid resistance. The cells were then heated in BHI broth (pH 6·0) at 58°C, using a submerged coil apparatus. The TSB+G grown strains had greatly increased heat resistance, with the heating time needed to achieve a five-log inactivation, being increased two- to four-fold. The z -values of TSB+G and TSB-G grown cells were 4·7°C and 4·3°C, respectively. Increases in heat resistance with TSB+G-grown E. coli O157:H7 were also observed using milk and chicken broth, but not with apple juice. However, cross-protection was restored if the pH of the apple juice was increased from 3·5 to 4·5. The data indicate that pH-dependent acid resistance provides E. coli O157:H7 with cross-protection against heat treatments, and that this factor must be considered to estimate this pathogen's thermal tolerance accurately.     

Use of a spouted bed to improve the storage stability of wheat germ followed in paper and polyethlyene packages

Stabilization of wheat germ by heating in a spouted bed for 180–540 s with air at 140–200 °C was studied. The lipase activity decreased by 6–65%. Wheat germ processed at 200 °C for 360 s was ranked highest in sensory evaluation, described as having ‘a golden color’ and ‘nutty flavor’, and its lipoxygenase activity had decreased by 91.2%. This product and raw wheat germ were stored in paper, polyethylene and vacuum‐packed polyethylene pouches at 5 °C, room temperature (18–26 °C) and 40 °C, and the moisture contents, water activities, free fatty acid contents and peroxide values were followed for 20 weeks. The increases were faster in paper pouches than in the polyethylene ones; vacuum packaging in polyethylene did not bring about significant improvement. The peroxide values of raw samples exceeded 10 meq O₂ kg^?1 oil after 3–23 days while those of the processed samples stored at room temperature or 5 °C were still less than 10 after 20 weeks. The free fatty acid content and peroxide value changes were expressed by zero order kinetics, resulting in similar activation energies for the raw and processed samples. Copyright © 2005 Society of Chemical Industry     

Myrosinase activity and total glucosinolate content of cruciferous vegetables,and some properties of cabbage myrosinase in Taiwan

The myrosinase activity and total glucosinolates of 10 different cruciferous vegetables and some properties of cabbage (Brassica oleracea L var capitata L) myrosinase were investigated. Radish, cabbage and broccoli showed higher myrosinase activity than other samples. No correlation (P > 0–05) was found between myrosinase activity and total glucosinolates of cruciferous vegetables tested. Myrosinase activity for white cabbage was higher than red cabbage. An optimal pH of myrosinase activity was observed at pH 8–0 for both white and red cabbages. The myrosinase extracted from cabbage was more stable in neutral or alkaline pH. The optimal temperature of myrosinase activity for both cabbages was about 60°C, but the myrosinase activity was destroyed after heating at 70°C for 30 min. The myrosinase activities of both white and red cabbages were activated by 10 mM and 5 mM ascorbic acid, respectively.     

Evaluation of Phosphatase Inactivation Kinetics in Milk Under Continuous Flow Microwave and Conventional Heating Conditions

Raw milk was subjected to conventional isothermal water bath heating, continuous flow microwave heating and continuous flow thermal holding in the pasteurization temperature range (60–75°C), and then immediately cooled in an ice-water bath. The associated alkaline phosphatase (ALP) residual activities were evaluated. Based on the gathered time-temperature profiles, the come-up time (CUT) and come-down time (CDT) contributions to enzyme inactivation were assessed and adjusted prior to first order rate kinetic data handling. The time-corrected D-values of ALP varied from 1250 s at 60°C to 1.7 s at 75°C with a z-value of 5.2°C under conventional batch heating conditions, 128 s at 65°C to 13.5 s at 70°C, with a z-value of 5.2°C under continuous-flow thermal holding condition, 17.6 s at 65°C to 1.7 s at 70°C with a z-value of 4.9°C under continuous-flow microwave heating condition. D values associated ALP inactivation under microwave heating were therefore an order of magnitude lower than under conventional thermal heating. The results thus emphasize that ALP inactivation occurred much faster under microwave heating condition than under conventional heating thereby confirming the existence of enhanced thermal effects from microwave. Because of the enhanced effects, MW pasteurization would reduce the severity of the treatment and hence potentially offer pasteurized milk of higher quality.     

Inactivation of pectin methylesterase and stabilization of opalescence in orange juice by dynamic high pressure

The effect of dynamic high pressure homogenization (DHP) alone or in combination with pre-warming on pectin methylesterase (PME) activity and opalescence stability of orange juice was studied. DHP without heating reduced PME activity by 20%. Warming the juice (50 °C, 10 min) prior to homogenization significantly increased the effectiveness of DHP. PME inactivation was further increased by adjusting the pH downward prior to treatment. Accelerated shelf-life study at 30 °C revealed that opalescence stability can be increased by several days by DHP treatment, even in the presence of active PME. These results suggest that the opalescence stability of orange juice treated by DHP does not depend entirely on PME activity but also on particle size reduction and structural changes to pectin resulting from the treatment. The freshness attributes of orange juice treated by warming was improved by DHP treatment.     

Inactivation and injury of Listeria monocytogenes as affected by heating and freezing

The effects of heating and freezing Listeria monocytogenes in tryptose phosphate broth an inactivation and injury were investigate. Four strains (Scott A, Brie-1, LCDC 81–861, and DA-3) were examined. Only slight decreases in the viable populations pf all four strains were detected after 75 min at 50°C. Populations of strains Scott A, LCDC 81–861, and DA-3 were reduced to non-detectable levels after 10 min at 60°C while viable cells of strain Brie-1 were recovered after heating at 60°C for 30 min. Viable populations of all four strains declined steadily at 52,54, and 56°C; similar inactivation rates were observed for strains Scott A, LcDC 81–861, and DA-3. Strain Brie-1 was consistently the most heat resistant. Substantial populations of viable cells of all strains heated at 52, 54, and 56°C were injured as evidenced by increased sensitivity to NaCl when plated on tryptose phosphate recover agar. Strain DA-3 was the most susceptible strain to sublethal heat injury. Viable populations of all four test strains were not appreciably reduced after 14 days at −18°C, although strain LCDC 81–861 lost viability at a more rapid rate at this temperature than did the other test strains. About 72, 82, 73, and 80% of Scott A, Brie-1, LCDC 81–861, and DA-3 cells, respectively, held at −18°C for 14 days were not receovered on tryptose phosphate agar (TPA) supplemented with 8% NaCl compared to populations recovered on TPA not supplemented with NaCl.     

Biologically Active Components Inactivation and Protein Insolubilization during Heat Processing of Soybeans

Inactivation of lipoxygenase, trypsin inhibitor, urease and retention of protein solubility during water blanching of dehulled soybeans at 90°, 95° and 100°C were investigated. Lipoxygenase was the most heat labile, followed by urease and then trypsin inhibitor. Processing time based on “acceptable inactivation time” (AI) was proposed. AI value was longest for trypsin inhibitor, followed by urease and then lipoxygenase. The combined effect of heat on protein solubility and biologically active components inactivation was expressed as “PDI at acceptable inactivation time” (PDIAI). PDIAI value for the processing “limiting factor,” trypsin inhibitor inactivation, at the three temperatures were 36.7%, 42.5%, and 34.8%, respectively.     

Phytate Reduction in Brown Beans (Phaseolus vulgaris L.)

Brown beans (Phaseolus vulgaris L.) were subjected to treatments to evaluate effects of pH, temperature, CaCl₂, tannase and fermentation on degradation of phytate. Soaking was performed at 21°C, 37°C and 55°C at pH 4.0, 6.0, 6.4, 7.0, and 8.0. Optimal conditions for phytate degradation were pH 7.0 and 55°C. After soaking 4, 8 or 17 hr at these conditions 79%, 87% and 98% of phytate was degraded, respectively. Addition of tannase enhanced reduction of phytate. Fermentation of presoaked whole beans resulted in reduction of 88% of phytate after 48 hr.     

Mechanical and thermal characteristics of amaranth starch isolated by acid wet-milling procedure

Effects of soaking conditions during acid wet-milling of amaranth grain on mechanical and thermal properties of amaranth starch were investigated. A factorial design based on temperature (40–60 °C) and SO₂ concentration (0.1–1.0 g/L) was used. Dynamic oscillatory tests involved heating and cooling cycles (25–90 °C) with tempering at 90 °C followed by a frequency sweep (0.1–20 Hz) at constant temperature. Thermal properties of starch suspensions were based on DSC tests. Viscoelastic modulus and thermal properties were affected by both factors, being significant the interaction effect. Maximum values of viscoelastic modulus at the end of heating and cooling steps were determined for samples corresponding to soaking conditions of 50 °C and 1.0 g/L SO₂. Based on Ross-Murphy index from frequency sweep analysis it was found that paste behavior occurs at 60 °C and 0.1 g/L, while at 47.4 °C and 0.72 g/L starch suspension gave a strong gel. Onset (61.7 °C) and peak (66.2 °C) temperatures showed a minimum at 50 °C and 1.0 g/L. As SO₂ concentration decreased, the end of gelatinization took place at lower temperature. Maximum gelatinization enthalpy (12.7 J/g) was found at 40 °C and 0.1 g/L SO₂.