Stephanie Jung

Changes in lysosomal enzyme activities and shear values of high pressure treated meat during ageing

The effects of pressure treatment (520 MPa for 260 s at 10°C) on lysosomal enzyme activities, Warner-Bratzler (WB) shear values and physico-chemical parameters of post-rigor (day 2) beef (Biceps femoris and Longissimus dorsi muscles) were investigated immediately after treatment and during ageing. Activities of cathepsin D and acid phosphatase in pressurised meat samples were higher than in controls at 2 days post-mortem and throughout storage. This increase in activities was related to the breakdown of the lysosomal membrane and/or to enzyme activation. The WB peak forces were higher for pressurised samples of both muscles at 2 days than in the control and remained higher throughout ageing. The WB shear force deformation curves from pressurised samples showed that the increase in toughness was due to the increase in shear force of the myofibrillar component and not the collagen component. Pressurised muscle led to a significant decrease in sarcomere length and higher cooking loss. Post-rigor application of high pressure induced higher catheptic activity but no conclusive effect on the post-mortem rate of tenderization or tenderness of meat was observed.

Demulsification of oil-rich emulsion from enzyme-assisted aqueous extraction of extruded soybean flakes.

Extraction of soybean oil from flaked and extruded soybeans using enzyme-assisted aqueous extraction processing (EAEP) is a promising alternative to conventional hexane extraction. The efficiencies of four proteases releasing oil from extruded material were compared. Protex 51FP, Protex 6L and Protex 7L each extracted 90% of the total oil available while Protex 50FP gave similar extraction yield as the control (without enzyme treatment). During EAEP, however, a stable emulsion forms that must be broken in order to recover free soybean oil. The potential of various proteases and phospholipases to destabilize the emulsion was determined. Two enzymes, a phospholipase A2 (LysoMax) and a protease (Protex 51FP) were selected to determine the effect of enzyme concentration on demulsification. Although at a 2% concentration (w/w, enzyme/(cream+free oil)), each enzyme tested was effective in totally destabilizing the cream; the protease released significantly more free oil than did the phospholipase at concentrations less than 2%. At 0.2% concentration, 88 and 48% of free oil were obtained with the protease and phospholipase, respectively. Reducing the pH of the cream also destabilized the cream with maximum demulsification at the isoelectric point of soy proteins. These results provide destabilization strategies for the oil-rich emulsion formed during aqueous extraction processing of extruded flakes and significantly contribute to the development of this environmentally-friendly technology.

Factors affecting emulsion stability and quality of oil recovered from enzyme-assisted aqueous extraction of soybeans

The objectives of the present study were to assess how the stability of the emulsion recovered from aqueous extraction processing of soybeans was affected by characteristics of the starting material and extraction and demulsification conditions. Adding endopeptidase Protex 6L during enzyme-assisted aqueous extraction processing (EAEP) of extruded soybean flakes was vital to obtaining emulsions that were easily demulsified with enzymes. Adding salt (up to 1.5 mM NaCl or MgCl(2)) during extraction and storing extruded flakes before extraction at 4 and 30 degrees C for up to 3 months did not affect the stabilities of emulsions recovered from EAEP of soy flour, flakes and extruded flakes. After demulsification, highest free oil yield was obtained with EAEP of extruded flakes, followed by flour and then flakes. The same protease used for the extraction step was used to demulsify the EAEP cream emulsion from extruded full-fat soy flakes at concentrations ranging from 0.03% to 2.50% w/w, incubation times ranging from 2 to 90 min, and temperatures of 25, 50 or 65 degrees C. Highest free oil recoveries were achieved at high enzyme concentrations, mild temperatures, and short incubation times. Both the nature of enzyme (i.e., protease and phospholipase), added alone or as a cocktail, concentration of enzymes (0.5% vs. 2.5%) and incubation time (1 vs. 3 h), use during the extraction step, and nature of enzyme added for demulsifying affected free oil yield. The free oil recovered from EAEP of extruded flakes contained less phosphorus compared with conventional hexane-extracted oil. The present study identified conditions rendering the emulsion less stable, which is critical to increasing free oil yield recovered during EAEP of soybeans, an environmentally friendly alternative processing method to hexane extraction.

Efficiency of pretreatments for optimal enzymatic saccharification of soybean fiber

The effectiveness of several pretreatments [high-power ultrasound, sulfuric acid (H(2)SO(4)), sodium hydroxide (NaOH), and ammonium hydroxide (NH(3)OH)] to enhance glucose production from insoluble fractions recovered from enzyme-assisted aqueous extraction processing of extruded full-fat soybean flakes (FFSF) was investigated. Sonication of the insoluble fraction at 144 μm(pp (peak-to-peak)) for 30 and 60s did not improve the saccharification yield. The solid fractions recovered after pretreatment with H(2)SO(4) [1% (w/w), 90°C, 1.5h], NaOH [15% (w/w), 65°C, 17 h], and NH(3)OH [15% (w/w), 65°C, 17 h] showed significant lignin degradation, i.e., 81.9%, 71.2%, and 75.4%, respectively, when compared to the control (7.4%). NH(3)OH pretreatment resulted in the highest saccharification yield (63%) after 48 h of enzymatic saccharification. A treatment combining the extraction and saccharification steps and applied directly to the extruded FFSF, where oil extraction yield and saccharification yield reached 98% and 43%, respectively, was identified.

31P NMR Phospholipid Profiling of Soybean Emulsion Recovered from Aqueous Extraction

The quantity and composition of phospholipids in full-fat soybean flour, flakes, and extruded flakes and in the cream fraction recovered after aqueous extraction (AEP) and enzyme-assisted aqueous extraction (EAEP) of these substrates were studied with (31)P NMR. Extruded flakes had significantly more phosphatidic acid (PA) than flakes and flour prior to aqueous extraction. The PA content of the cream recovered after AEP and EAEP of extruded flakes was similar to that of the starting material, whereas the PA content of the creams from flour and flakes significantly increased. Changes in the PA content could be explained by the action of phospholipase D during the processing step and aqueous extraction. Total phospholipids in the oil recovered from the creams varied from 0.09 to 0.75%, and free oil yield, which is an indicator of cream stability, varied from 6 to 78%. Total phospholipid did not correlate with emulsion stability when it was lower than 0.20%. Inactivation of phospholipase D prior to aqueous extraction of flour resulted in a cream emulsion less stable toward enzymatic demulsification and containing less PA and total phospholipids than untreated flour. The phospholipid distributions in the cream, skim, and insolubles obtained from AEP flour were 7, 51, and 42%, respectively.

Textural changes in bovine meat treated with high pressure

Abstract The effect of high pressure (HP) treatment on mechanical resistance of post-rigor bovine meat (muscles Biceps femoris and Longissimus dor si) pressurised at low temperature (10°C) was studied. Two levels of pressure were used, 130 and 520 MPa for 260 s. The results show that HP treatment affects significantly (p < 0.05) the mechanical resistance of both raw and cooked meat (1 h at 65°C). Mechanical resistance of pressurised samples was higher than that of untreated samples. Among treated samples, those samples pressurised at 520 MPa showed higher toughness than those pressurised at 130 MPa. Increase of mechanical resistance is due to changes that occurs on the myofibrillar fraction rather than on the connective component. Based on these results, these treatment conditions seem to be ineffective to improved beef tenderness.

Comparison and optimization of enzymatic saccharification of soybean fibers recovered from aqueous extractions

Soybean insoluble fractions recovered from aqueous extraction processing (AEP) and enzyme-assisted AEP (EAEP) of full-fat soybean flakes (FFSF) and extruded FFSF were evaluated as a feedstock for the production of fermentable sugars using enzymes. Among the four insoluble fractions (AEP FFSF, EAEP FFSF, AEP extruded FFSF and EAEP extruded FFSF), the composition analysis revealed that the one recovered from EAEP of extruded FFSF had the highest glucan content, 16% [dry basis (db)], as compared to about 10% (db) for the other fractions. Thirty-three percent of the initial glucan of the insoluble recovered from AEP and EAEP of FFSF were converted into glucose using 33 FPU of Accellerase 1000/g-glucan. This saccharification yield was increased to 44% with extruded fibers. The higher saccharification yield of 49% was obtained at 45 °C, 1% glucan loading, and 101 FPU/g-glucan enzymes loading after 27 h of hydrolysis.

Effect of high pressure treatment on ovotransferrin.

High pressure processing of ovotransferrin was carried out to study the structural and physiochemical changes of ovotransferrin under various pressure levels. At pH 8 and pressures higher than 200 MPa, a decrease in total sulfhydryl groups and an increase in surface hydrophobicity were observed along with a partial aggregation. A gradual shift of denaturation peak towards higher temperature was noticed up to 500 MPa, leading to a total loss of the enthalpy of denaturation at pressures of 600 and 700 MPa, where a significant decrease in intrinsic fluorescence was also observed. At pH 3, the ovotransferrin adopted a molten globule state, associated with a significant increase in surface hydrophobicity and reactive sulfhydryl content; structurally, no clear denaturation peaks in differential scanning calorimetry (DSC) were detected at any level of pressure treatment whereas a noticeable decrease in intrinsic fluorescence was evidenced up to 600 MPa and then increased at 700 MPa pressure treatment. Fourier transform infrared spectroscopy (FT-IR) revealed that the conformational structure were changed from helices, sheets, turns, and aggregated strand to mostly intermolecular β-sheets or aggregated strands at pH 8 at 200 MPa but switched back to original structure at higher pressures.

Impact of high-pressure processing on microbial shelf-life and protein stability of refrigerated soymilk.

The effects of pressure (400, 500 and 600 MPa), dwell time (1 and 5 min) and temperature (25 and 75 degrees C) on microbial quality and protein stability of soymilk during 28 days of storage (4 degrees C) were evaluated under aerobic and anaerobic conditions. After processing and during storage, there were significant differences in total bacterial count (TBC), numbers of psychrotrophs (PSY) and Enterobacteriaceae (ENT), and protein stability between untreated (control) and pressurized samples (P < 0.05). Pressure applied at an initial temperature of 75 degrees C resulted in a greater suppression in growth of PSY compared to TBC. No ENT was detected in pressurized samples throughout the storage period tested. Dwell time had no significant effect on log reduction of TBC at 25 or 75 degrees C (P > 0.05). Pressure at 400 MPa (5 min), 500 and 600 MPa (1 and 5 min) produced 100% sub-lethal injury in surviving bacterial populations irrespective of temperature. After 28 days of refrigerated storage, both aerobic and anaerobic pressurized samples had better or similar stability as the control on day one of storage. Soymilk control samples were spoiled after 7 days whereas pressurization increased soymilk shelf-life by at least 2 weeks. Pressure (600 MPa) at 75 degrees C for 1 min not only significantly reduced initial microbial populations and increased the microbial shelf-life but also extended the protein stability of soymilk (P < 0.05).

Effects of calcium and pressure treatment on thermal gelation of soybean protein.

The effect of calcium and high-pressure (HP) treatment on the heat gelation of soybean proteins was investigated. In the presence of calcium (2 to 25 mM), the gelation of dispersions of soybean protein isolate (SPI), a beta-conglycinin-enriched fraction (7SEF), and a glycinin-enriched fraction (11SEF) started with protein having a lower degree of denaturation. The gels from these dispersions had greater stiffness than the samples without added calcium. HP treatment had different effects on heat-induced gelation depending on the presence of calcium and on the nature of the proteins. In the absence of calcium, gels with low stiffness were formed after HP treatment, compared with untreated samples, and regardless of the sample type (SPI, 7SEF, 11SEF). In the presence of calcium, gel stiffness was increased after HP treatment of dispersions containing beta-conglycinin (SPI and 7SEF), while the opposite effect was observed for 11SEF. In the presence of calcium, HP treatment promoted a greater contribution of hydrophobic interactions in SPI and 7SEF. In the dispersions containing beta-conglycinin, these conditions also promoted the appearance of a heterogeneous distribution of molecular sizes, from enormous aggregates to dissociated species. Our results suggest that, in the presence of calcium, HP treatment has an opposite effect on the ability of glycinin and beta-conglycinin to participate in the formation of a 3-dimensional network upon heating.