Jacques Potus

Effect of Mixing Conditions on the Behavior of Lipoxygenase, Peroxidase, and Catalase in Wheat Flour Doughs

ABSTRACT The effect of mixing has been tested on the extractable activities of lipoxygenase, peroxidase, and catalase from dough after 2, 5, and 20 min of mixing, and 30 min of rest period after 20 min of mixing. Different mixing conditions have been studied including temperature, atmosphere, speed, amount of water added to the dough, buffer solutions between pH 3.6 and 7.5 added to the dough, and different additives (linoleic acid, guaiacol, hydrogen peroxide, ascorbic acid, cysteine, yeast, and sodium chloride). In all the mixing conditions tested, the dough peroxidase activity remains equivalent to the initial flour activity, whereas losses in lipoxygenase and catalase activities largely varied according to mixing conditions. The results show that a self-destruction mechanism as well as physicochemical denaturation are responsible for these losses. Lipoxygenase losses seem mainly associated with the former mechanism, whereas catalase losses are highly increased in acidic conditions (physicochemical den...

Effect of Exogenous Lipase on Dough Lipids During Mixing of Wheat Flours

ABSTRACT In control dough, endogenous wheat lipase was inactive, because the triacylglycerol (TAG), 1,2-diacylglycerol (DAG1,2), and 1,3-diacylglycerol (DAG1,3) fractions of nonpolar lipids were not affected by mixing. Conversely, the free fatty acid (FFA) and monoacylglycerol (MAG) fractions decreased, mainly due to the oxidation of polyunsaturated fatty acids (PUFA) catalyzed by wheat lipoxygenase. Addition of exogenous lipase to flour (15 lipase units [LU] per gram of dry matter) resulted in substantial modification of nonpolar lipids during dough mixing. Due to the 1,3 specificity of the lipase used in this experiment, the TAG and DAG1,3 fractions decreased, whereas the MAG and FFA fractions increased. The DAG1,2 fraction increased at the beginning of mixing and decreased after 40 min of mixing. Moreover, part of the PUFA released by lipase activity was oxidized by wheat lipoxygenase, resulting in major losses of PUFA. Conversely, the net content of the saturated and monounsaturated fatty acids (SMUFA...

Effect of Adding Exogenous Oxidative Enzymes on the Activity of Three Endogenous Oxidoreductases During Mixing of Wheat Flour Dough

ABSTRACT The behavior of different exogenous enzymes (soybean lipoxygenase [SLOX], horseradish peroxidase [HPOD], catalase from bovine liver [BCAT], and glucose oxidase [GOX] from Aspergillus niger) added to dough was studied during mixing. The effect of adding these exogenous oxidoreductases on the activity of three oxidative enzymes present in wheat flour (lipoxygenase [WLOX], peroxidase [WPOD], and catalase [WCAT]) was examined. Proper assay conditions were established to differentiate between added WLOX, WPOD, and WCAT and the corresponding activities present in wheat flour. For doughs with added SLOX, an immediate loss of extractable SLOX (≈40%) was observed which remained constant during further mixing. When compared with the control dough, addition of SLOX decreased the losses in WLOX and WCAT activities, whereas WPOD activity was unaffected. With doughs supplemented by HPOD, an immediate loss of 20% in the HPOD activity was observed which did not change after 20 min of mixing. Compared with contro...

Effect of Storage Temperature and Flour Water Content on Lipids, Lipoxygenase Activity, and Oxygen Uptake During Dough Mixing

ABSTRACT Flours differing in water content of 10% (F10), 12% (F12), and 14% (F14) were stored for 16 weeks at 22, 32, and 45°C. The major changes in lipids concerned the free fatty acids (increase) and the triglycerides (decrease). In all cases, the changes increased with increasing storage temperature and water content. After 16 weeks of storage, the losses in lipoxygenase (LOX) activity increased with increasing flour moisture and storage temperature from 10% for F10 at 22°C to 100% for F14 at 45°C. At the end of storage at 22 and 32°C, the bread volumes decreased by 10 and 25%, respectively, with no statistical differences (P < 0.05) between the samples. At 45°C, the volume losses were equal to 35, 46, and 61% for the F10, F12, and F14 samples, respectively. In the same time, the flour oxidative ability (oxygen uptake during dough mixing) increased for the F10 and F12 samples with increasing storage temperature, whereas it decreased for the F14 samples stored at 45°C. Therefore, provided the residual L...

Effects of Mixing Conditions and Wheat Flour Dough Composition on Lipid Hydrolysis and Oxidation Levels in the Presence of Exogenous Lipase

ABSTRACT The lipid profiles of wheat flour doughs containing exogenous lipase were studied under different mixing conditions using a microscale mixer. An experimental design comparing the effects of dough water content (52–68%), the speed of mixing (50–100 rpm), and the mixer temperature (18–32°C) showed that the hydrolysis levels were positively influenced by temperature and speed of mixing and negatively influenced by water content. The positive effect of temperature was enhanced both by highspeed mixing and low water content. The lipid oxidation levels were positively influenced by the speed of mixing and negatively influenced by the water content. The positive effect of temperature on the oxidation levels was less important. A series of experiments conducted with different types of industrial and semi-industrial mixers with equal concentrations of lipase added to the dough showed large differences among the rates of lipid hydrolysis and oxidation. However, the mixing conditions proposed by bakers to o...

Studies on the Glutathione-Dehydroascorbate Oxidoreductase (EC 1.8.5.1) from Wheat Flour

ABSTRACT Glutathione (GSH) dehydrogenase was partially purified from wheat flour after extraction, ammonium sulfate precipitation, and ionic-exchange chromatography on diethylaminoethyl (DEAE) Sepharose CL6B. Kinetic studies showed that the optimum pH was close to 7.5. The Km values varied between 0.15 and 0.28 mM for dehydroascorbic acid (DHA) and between 1.8 and 0.62 mM for GSH when pH was varied from 5.5 to 7.5. The kinetic pattern was consistent with a sequential mechanism for the binding of GSH and DHA. NaCl is a competitive inhibitor with respect to GSH and is uncompetitive with respect to DHA, which suggests that the enzyme combines with DHA before it does with GSH. IsoDHA can replace DHA as hydrogen acceptor but with a Km of 1.2 mM. γ-Glu-cys was enzymically oxidized but much less efficiently than GSH (Vm = 47 nkat/mL and Km = 5.5 mM compared to Vm = 362 nkat/mL and Km = 1.8 mM for GSH), whereas cysteine and cys-gly were not substrates. In the presence of DHA, addition of cysteine and cys-gly to s...

3D-front-face fluorescence spectroscopy and independent components analysis: a new way to monitor bread dough development

Following bread dough development can be a hard task as no reliable method exists to give the optimal mixing time. Dough development is linked to the evolution of gluten proteins, carbohydrates and lipids which can result in modifications in the spectral properties of the various fluorophores naturally present in the system. In this paper, we propose to use 3-D-front-face-fluorescence (3D-FFF) spectroscopy in the 250-550nm domain to follow the dough development as influenced by formulation (addition or not of glucose, glucose oxidase and ferulic acid in the dough recipe) and mixing time (2, 4, 6 and 8min). In all the 32 dough samples as well as in flour, three regions of maximum fluorescence intensities have been observed at 320nm after excitation at 295nm (Region 1), at 420nm after excitation at 360nm (Region 2) and 450nm after excitation at 390nm (Region 3). The principal components analysis (PCA) of the evolution of these maxima shows that the formulations with and without ferulic acid are clearly separated since the presence of ferulic acid induces a decrease of fluorescence in Region 1 and an increase in Regions 2 and 3. In addition, a kinetic effect of the mixing time can be observed (decrease of fluorescence in the Regions 1 and 2) mainly in the absence of ferulic acid. The analysis of variance (ANOVA) on these maximum values statistically confirms these observations. Independent components analysis (ICA) is also applied to the complete 3-D-FFF spectra in order to extract interpretable signals from spectral data which reflect the complex contribution of several fluorophores as influenced by their environment. In all cases, 3 signals can be clearly separated matching the 3 regions of maximal fluorescence. The signals corresponding to regions 1 and 2 can be ascribed to proteins and ferulic acid respectively, whereas the fluorophores associated with the 3rd signal (corresponding to region 3) remain unidentified. Good correlations are obtained between the IC score values of the 3 signals and the fluorescence intensities in Region 1, Region 2 and Region 3. Ferulic acid addition increases fluorescence in Region 2 and decreases fluorescence in Region 1, probably via a reabsorption of the protein fluorescence by ferulic acid. These phenomena are less pronounced when glucose oxidase is present. The enzymatic oxidation of ferulic acid by the glucose oxidase-peroxidase association could explain some of these effects.

Competition for Oxygen Among Oxidative Systems During Bread Dough Mixing: Consequences of Addition of Glucose Oxidase and Lipoxygenase on Yeasted Dough Rheology

ABSTRACT The effect on O2 uptake during the mixing of yeasted dough, either unsupplemented or supplemented with glucose oxidase (GOX), horsebean flour (HB), soybean flour (SB), or combinations thereof, was studied using an airtight mixer. Two wheat flours with a low (flour A) and a high (flour B) content of free polyunsaturated fatty acids were used. Addition of HB or SB provokes a similar increase of O2 uptake for both wheat flours, whereas addition of GOX causes a larger increase for flour A than for flour B. When the wheat flours were supplemented with HB or SB, addition of GOX caused a small but significant increase of O2 uptake for flour A. This increase was not observed for flour B. The mixing tolerance of dough A, determined with the Chopin Consistograph, is increased by GOX addition. However, this effect is less pronounced when flour A is supplemented with HB or SB. Similarly, the relaxation index of dough B is decreased by GOX addition, but the decrease is less distinct in the presence of HB or S...