Differential scanning calorimetry and circular dichroism spectrometry of walleye pollack myosin and light meromyosin

The thermodynamic properties of myosin and its C-terminal fragment, light meromyosin (LMM), from walleye pollack, a typical cold-water fish efficiently utilized on an industrial scale, were analyzed by using differential scanning calorimetry (DSC) and circular dichroism (CD) spectrometry. Recombinant walleye pollack LMM expressed in Escherichia coli was also subjected to DSC and CD measurements for reference. The two proteins prepared from frozen surimi showed three endothermic peaks, the transition temperatures (T(m)) of which were quite similar, although overall DSC patterns differed considerably from one another. Their alpha-helical contents determined by CD were low compared to values reported before for other species. On the other hand, recombinant LMM gave four endothermic peaks at 27.4, 30.8, 36.5, and 43.4 degrees C in DSC and showed an alpha-helical content of approximately 80%. The peak at 27.4 degrees C could not be observed in walleye pollack LMM prepared from frozen surimi and thus was possibly attributed to its C terminus, because this extreme C-terminal region is supposedly truncated during preparation of LMM by tryptic digestion.     

Differences in polymer formation through disulfide bonding of recombinant light meromyosin between white croaker and walleye pollack and their possible relation to species specific differences in thermal unfolding

Fast skeletal light meromyosins (LMMs) of white croaker and walleye pollack were prepared in our expression system using Escherichia coli and determined for their polymer-forming ability and thermodynamic properties by using sodium dodecyl sulfate polyacrylamide gel electrophoresis and differential scanning calorimetry (DSC), respectively. White croaker LMM formed dimer by heating at 80 degrees C and showed only a single peak at 32.1 degrees C of temperature transition in DSC. On the other hand, walleye pollack LMM hardly formed polymer and showed four peaks at 27.7, 30.5, 35.8, and 43.9 degrees C. When Cys525 of white croaker LMM was replaced by alanine, this point-mutated LMM showed no change in its DSC profile but formed no dimer upon heating, suggesting a possible role of Cys525 in dimer formation. On the other hand, walleye pollack LMM where Cys491 was substituted by alanine changed its DSC profile, showing four peaks at 27.9, 29.1, 38.4, and 43.9 degrees C. However, this point-mutated LMM formed no dimer upon heating as in the case of native LMM. These results suggest that cysteine residue(s) participates in thermal gel formation of LMM when it locates in a suitable position of the sequence.     

Thermal denaturation and aggregation properties of Atlantic salmon myofibrils and myosin from white and red muscles

Thermal denaturation and aggregation abilities of salmon myofibrils and myosin were studied measuring turbidity, intrinsic fluorescence, 8-anilino-1-naphthalene sulfonic acid binding, and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide cross-linking. The thermal behaviors of protein preparation from white and red muscles were compared, and the relationship with thermal gelation properties is discussed. The low gelation ability of salmon muscle proteins was related to a limited extent of protein denaturation and aggregation upon heating. These properties seemed to be carried by myosin molecules as a similar behavior was observed for both myofibrils and myosin preparations. The higher thermal stability observed for red muscle proteins with higher transition temperatures in rheological profiles was related to a shift to higher temperature in denaturation and aggregation processes. The extent of denaturation and aggregation was very similar for both muscle types as was the final rigidity of the gels formed.     

Heat-induced gels of egg white/ovalbumins from five avian species: thermal aggregation, molecular forces involved, and rheological properties

Product processing (heating, pH change, etc.) usually alters protein structure, improves rheological properties, and gives a unique texture to foods. The thermal aggregation and structural properties of ovalbumins from five avian species were studied at different pH values by polyacrylamide gel electrophoresis (PAGE) and determinations of sulfhydryl group content and surface hydrophobicity. The results showed that sulfhydryl group content changed insignificantly in heat-denatured ovalbumins other than hen ovalbumin (pH-independent), and surface hydrophobicity markedly increased (pH-dependent) after heating, with a significant difference among species. Furthermore, it was demonstrated that the hydrophobic interaction and sulfhydryl-disulfide interchange reaction were necessary in the aggregation and cross-linking of gel networks. Creep tests were also used to characterize the gel network structures of various egg white/ovalbumins upon heating. The viscoelastic behavior of the ovalbumins of all species was dependent on pH values, and changed significantly with the phylogeny of these species. With increases in pH value (7.0-8.5), the heat-induced gels of ovalbumins gradually changed from turbid to translucent, the instantaneous modulus (E(0)) increased slightly and reached a nearly constant value, and the Newtonian modulus (etaN) increased significantly in each sample. The heated egg white from these five avian species also formed highly viscoelastic gels, with a good correlation of viscoelastic behavior between ovalbumin and egg white in corresponding species.     

Determination of free and bound phenolic compounds in buckwheat spaghetti by RP-HPLC-ESI-TOF-MS: effect of thermal processing from farm to fork

Nowadays there is considerable interest in the consumption of alternative crops as potential recipes for gluten-free products production. Therefore, the use of buckwheat for the production of gluten-free pasta has been investigated in the present study. RP-HPLC-ESI-TOF-MS has been applied for the separation and characterization of free and bound phenolic compounds in buckwheat flour and buckwheat spaghetti. Thus, 32 free and 24 bound phenolic compounds in buckwheat flour and spaghetti have been characterized and quantified. To the authors' knowledge, protochatechuic-4-O-glucoside acid and procyanidin A have been detected in buckwheat for the first time. The results have demonstrated a decrease of total free phenolic compounds from farm to fork (from flour to cooked spaghetti) of about 74.5%, with a range between 55.3 and 100%, for individual compounds. The decrease in bound phenols was 80.9%, with a range between 46.2 and 100%. The spaghetti-making process and the cooking caused losses of 46.1 and 49.4% of total phenolic compounds, respectively. Of the total phenolic compounds present in dried spaghetti, 11.6% were dissolved in water after cooking.     

Effect of green revolution technology from 1970 to 2003 on sawah soil properties in Java,Indonesia: I. Carbon and nitrogen distribution under different land management and soil types

Abstract

Many countries have reported that green revolution (GR) technology caused some adverse effects on agricultural lands, but there is no research on the effects of GR in Indonesia. To evaluate the effect of GR technology on sawah soil in Indonesia, a comparative study between seedfarms, where GR technology has been continuously applied, and non-seedfarms was conducted in Java as a pioneer place of GR technology in Indonesia and, in particular, on the carbon and nitrogen status in sawah soils. The term sawah refers to a leveled and bounded rice field with an inlet and outlet for irrigation and drainage. Soil samples collected by Kawaguchi and Kyuma in 1970 and new samples taken in 2003 from the same sites or sites close to the 1970 sampling sites were analyzed and compared. During the period 1970–2003 the land-use pattern of sawah in seedfarms and non-seedfarms did not change, but cultivation intensity increased. The results showed that total carbon (TC) and total nitrogen (TN) contents significantly increased from 31.90 to 40.42 Mg ha^?1 and from 3.04 to 3.97 Mg ha^?1, respectively, and TC and TN were mostly found accumulated in the surface soil layer. The difference in land management practices between seedfarms and non-seedfarms affected the change in TC and TN content in the 0–20 cm soil layer from 1970 to 2003. In seedfarms, where rice had been planted in a monoculture system, TC and TN contents in the 0–20 cm soil layer increased from 34.50 to 39.24 Mg ha^?1 and from 3.16 to 3.95 Mg ha^?1, respectively. In non-seedfarms, TC and TN increased more than in seedfarms from 29.77 to 41.37 Mg ha^?1 and from 2.94 to 3.98 Mg ha^?1, respectively. Within the 0–100 cm soil layer, TC and TN increased from 92.68 to 112.83 Mg ha^?1 and from 9.34 to 12.03 Mg ha^?1 and from 79.60 to 114.86 Mg ha^?1 and from 8.93 to 11.44 Mg ha^?1 for seedfarms and non-seedfarms, respectively. No significant difference was observed between the two main soil types, Inceptisols and Vertisols, in Java. Intensive use of sawah over a long time may eliminate the original difference in the properties of these two soil types.