الفهرس 
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Abstract
Gluten free products such as gluten free Balady bread and gluten free pasta are an important stable foods consumed by celiac patients. Therefore, the objective of this study was to develop and evaluate such products. where the current study was to achieve this goal by using different ratios of gluten free materials such as rice flour, corn flour, potato starch, and soy protein isolate. Psyllium husk and carboxymethyl cellulose which used as binding agents due to the absence of gluten in these materials. Maltogenic α-amylase and xylanase were used to retard staling of gluten free Balady bread. Also glucose oxidase and transglutaminase were used as to improve the quality of gluten free Balady bread. Thus, other trial was conducted to degrade wheat gliadin by using fungal protease from A. niger at different conditions of incubation time, incubation temperature, pH and enzyme concentration.
The obtained results could be summarized as follows:
5.1. Physicochemical characteristics raw materials
5.1.1. Chemical composition of raw materials
• The obtained results refer to the percentage of protein, total lipid, ash, crude fiber and nitrogen free extract. It was 13.64, 1.59, 0.85, 1.48 and 82.44% respectively; for wheat flour 82% extraction, but it was 11.31, 0.95, 0.57, 0.66 and 86.51% respectively; for wheat flour 72% extraction.
• The obtained results also refer to the percentage of protein, total lipid, ash, crude fiber and nitrogen free extract. It was 9.76, 4.24, 1.27, 2.94 and 81.79% respectively for white corn flour, but it was 7.16, 1.50, 0.57, 1.21 and 89.56% respectively for rice flour.
5.1.2. Physical properties of raw materials
• Physical characteristics of wheat flour 82% extraction; falling number, liquefaction number and starch damage which was 287sec., 25.37% and 5.40% respectively; and it was 464sec., 14.50%, 3.60% respectively for white corn flour. While rice flour recorded 514sec., 12.93% and 8.17% respectively.
5.2. Physicochemical characteristics of Balady bread
5.2.1. Chemical composition of Balady bread
• The results revealed that wheat flour Balady bread contained 31.04% moisture, 16.24% protein, 1.74% lipids, 1.16% ash, 1.64% crude fiber and 78.85% NFE. But white corn flour Balady bread contained 32.06, 11.71, 4.46, 1.88, 3.10 and 78.85% respectively. While the chemical composition of gluten free Balady bread which prepared from different gluten free formula with 2.5% psyllium husk ranged from 33.16 to 35.97% moisture, 13.43 to 18.22% protein 75.58 to 82.78% NFE. Also gluten free Balady bread which prepared from different gluten free formula with 2.5% carboxymethyl cellulose were ranged from 32.11 to 34.92, 13.52 to 18.19% and 75.77 to 83.19% respectively.
5.2.2. Physical properties of Balady bread
• The physical properties of Balady bread were investigated and the results refer to wheat flour Balady bread recorded 37.55g, 265cm3 and 7.05cm3/g for weight, volume and specific volume respectively. Also, white corn flour Balady bread recorded 38.40g, 105cm3 and 2.75cm3/g respectively.
• Gluten free Balady bread which prepared from different gluten free formula with 2.5% psyllium husk ranged from 42.60 to 44.25g for weight, 295 to 325cm3 for volume and 6.70 to 7.65g/cm3 for specific volume. Also, the gluten free Balady bread which prepared from different gluten free formula with 2.5% carboxymethyl cellulose ranged from 39.55 to 4470g, 170 to 270cm3 and 3.80 to 6.78g/cm3 respectively.
5.2.3. Color measurement of Balady bread
• The color parameters represented in lightness, redness and yellowness (L*, a* and b* respectively) were investigated and the results refer to wheat flour Balady bread recorded 61.64, 11.39 and 25.65 respectively. Also, white corn flour Balady bread recorded 68.62, 8.26 and 28.70 respectively.
• Gluten free Balady bread which prepared from different gluten free formula with 2.5% psyllium husk ranged from 60.24 to 68.56 for L* parameter, 7.38 to 10.83 for a* parameter and 25.95 to 32.62 for b* parameter. But, the gluten free Balady bread which prepared from different gluten free formula with 2.5% carboxymethyl cellulose ranged from 52.52 to 68.18, 6.90 to 10.73 and 24.19 to 29.51 respectively.
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5.2.4. Alkaline water retention capacity of Balady bread
• The alkaline water retention capacity (during different storage periods) of Balady bread were estimated and the results indicated that wheat flour Balady bread recorded 268.46 after 72hr. and it was 197.73 for white corn flour Balady bread. While the gluten free Balady bread which prepared from different gluten free formula with 2.5% psyllium husk ranged from 217.36 to 223.63 of alkaline water retention capacity after 72hr. Also, the gluten free Balady bread which prepared from different gluten free formula with 2.5% carboxymethyl cellulose ranged from 200.66 to 217.96 after 72hr respectively.
5.2.5. Sensory evaluation of Balady bread
• Sensory evaluation of Balady bread was estimated and the results indicated that wheat flour Balady bread recorded the highest score of overall acceptability (8.80). While the gluten free Balady bread which prepared from different gluten free formula with 2.5% psyllium husk ranged from 8.0 to 8.70 for the same parameter, whereas, the gluten free Balady bread which prepared from different gluten free formula with 2.5% carboxymethyl cellulose ranged from 3.80 to 6.70 for the same parameter.
5.3. Development and evaluation of gluten free Balady bread by using maltogenic α-amylase and xylanase
5.3.1. Effect of maltogenic α-amylase and xylanase on falling number and liquefaction number of gluten free flour
• The effect of maltogenic α-amylase or xylanase at different concentrations (20, 40, 60 and 80ppm) on falling number and liquefaction number of gluten free flour at difference blends were studied, the results refer to the untreated gluten free flour recorded 383sec. and 18.02 for falling number and liquefaction number respectively. While the others which treated by maltogenic α-amylase ranged between 235sec. to 331sec. and 21.31 to 32.44 respectively; and it was 363sec. to 374sec. and 18.52 to 19.14 respectively in case of xylanase addition.
5.3.2. Effect of maltogenic α-amylase and xylanase on pasting properties of gluten free dough
• The results refer to the untreated gluten free flour recorded 4811, 1788 and 5191cP respectively. While the others which treated by different concentration maltogenic α-amylase ranged between 2065 to 3098cP for peak viscosity, 1002 to 1700cP for breakdown and 2018 to 3300cP for final viscosity. But it was 4337 to 4646cP, 1835 to 2113cP and 3943 to 4847cP, respectively in case of xylanase addition with different concentration.
5.3.3. Effect of matrixes of maltogenic α-amylase and xylanase enzymes on alkaline water retention capacity of gluten free Balady bread
• The effect of maltogenic α-amylase or xylanase on alkaline water retention capacity of gluten free Balady bread were determined. The results refer to the untreated gluten Balady bread recorded 212.66 of alkaline water retention capacity after 72hr of storage periods. While the others which treated by different concentration of maltogenic α-amylase ranged between 251.22 to 338.05 for the same parameter after 72hr. But it was 224.55 to 277.19 in case of xylanase addition with different concentration.
5.3.4. Effect of maltogenic α-amylase on alkaline water retention capacity of gluten free Balady bread (quadratic polynomial regression model)
• According to the quadratic polynomial regression model the maltogenic α-amylase enzyme can be used alone at a predicted 27.10ppm concentration according to the regression analysis equations where the predicted AWRC is 265.2 which is the closest value to the bread sample prepared from wheat flour, which had an actual value of 265 after 72hr.
5.3.5. Effect of maltogenic α-amylase and xylanase on water activity of gluten free Balady bread
• The untreated gluten Balady bread recorded 0.701 of water activity after 72hr. While the others which treated by different concentration of maltogenic α-amylase ranged between 0.838 to 0.914 for the same parameter after 72hr. But it was 0.810 to 0.857 in case of xylanase addition with different concentration.
5.3.6. Effect of maltogenic α-amylase on the quality attributes of gluten free Balady bread (quadratic polynomial regression model)
• According to the regression analysis equations the maltogenic α-amylase enzyme can be used at concentration of 20 ppm where the specific volume and volume at this concentration is similar to the control made from wheat flour as control
5.3.7. Effect of xylanase on alkaline water retention capacity of gluten free Balady bread (quadratic polynomial regression model)
• According to the quadratic polynomial regression model the xylanase enzyme can be used alone at a predicted concentration of 59.6 ppm according to the regression analysis equations where the AWRC of the predicted is 265.2. It is the closest value to the bread sample prepared from wheat flour, which had an actual value of 265 after 72 hr.
5.3.8. Three-dimension response surface of maltogenic α-amylase and xylanase combination on the alkaline water retention capacity of gluten free Balady bread
• The recommended dose according to the three-dimension regression plot are 10ppm of MA plus 45ppm of XY to obtain the loaf has AWRC (after 72hr) value closest to the control wheat Balady bread. Where, the GFBB sample which prepared by gluten free formula (under study) with 10ppm of MA plus 45ppm of XY it is expected to have a 266.90 of AWRC after 72hr compared to 265 which obtained by wheat flour Balady bread sample.
5.3.9. Effect of xylanase on the quality attributes of gluten free Balady bread (quadratic polynomial regression model)
• According to the regression analysis equations the xylanase enzyme can be used at concentration of 20 ppm where the specific volume and volume at this concentration is similar to the control made from wheat flour as control.
5.3.10. Effect of maltogenic α-amylase and xylanase on sensory evaluation of gluten free Balady bread
• The effect of maltogenic α-amylase or xylanase on sensory evaluation of gluten free Balady bread was investigated three times (fresh bread, after 24hr and after 48hr). As for the fresh bread, the results refer the untreated gluten Balady bread recorded 7.70 of overall acceptability. While after treatment by different concentration of maltogenic α-amylase, it ranged from 7.90 to 8.60 for the same parameter. But it ranged from 7.80 to 8.20 in case of xylanase addition with different concentration. While, after 24hr of storage the untreated gluten Balady bread recorded 2.60 of overall acceptability. But after treatment by different concentration of maltogenic α-amylase it ranged from 2.90 to 8.20 for the same parameter. But it ranged from 2.60 to 6.60 in case of xylanase addition with different concentration. While after 48hr of storage, the untreated gluten Balady bread recorded 1.40 of overall acceptability. While after treatment by different concentration of maltogenic α-amylase it ranged from 1.70 to 4.60 for the same parameter. But it ranged from 1.40 to 4.30 in case of xylanase addition with different concentration
5.4. Development and evaluation of gluten free Balady bread by using glucose oxidase and transglutaminase
5.4.1. Effect of glucose oxidase and transglutaminase enzymes on physical properties of gluten free Balady bread
• For the effect of glucose oxidase and transglutaminase at different concentration on weight, volume and specific volume of gluten free Balady bread, the results refer to the untreated gluten free flour recorded 43.62g, 285cm3 and 6.53g/cm3 respectively. While the others which treated by different concentration of glucose oxidase ranged between 43.16 to 44.03g, 287.50 to 320.00cm3 and 6.66 to 7.27g/cm3 for weight volume and specific volume respectively; and it was 41.51 to 43.72g, 295.00 to 327.50cm3 and 6.75 to 7.89g/cm3 respectively in case of addition of transglutaminase at different concentration.
5.4.2. Effect of glucose oxidase on weight, volume and specific volume of gluten free Balady bread (quadratic polynomial regression model)
• According to the regression analysis equations glucose oxidase can be used alone at a predicted 17.4ppm concentration according to the regression analysis equations in terms of expected quality attributes parameters (43.90g for weight, 315.30cm3 for volume and 7.16cm3/g for specific volume) similar to the control (prepared from 100% wheat flour) parameters.
5.4.3. Effect of transglutaminase on weight, volume and specific volume of gluten free Balady bread (quadratic polynomial regression model)
• According to the regression analysis equations transglutaminase enzyme can be used alone at a predicted concentration of 9.50ppm according to the regression analysis equations in terms of expected quality attributes parameters (43.10g for weight, 308.80cm3 for volume and 7.16cm3/g for specific volume).
5.4.4. Three-dimension response surface of glucose oxidase and transglutaminase combination on weight, volume and specific volume of gluten free Balady bread
• The recommended dose according to the three-dimension regression plot are 0ppm of GOX plus 25ppm of TG to obtain the loaf has quality attributes values closest to the control wheat Balady bread. Where, the GFBB sample which prepared by gluten free formula (under study) with 25ppm of transglutaminase it is expected to have a 41.7g of weight, 332.60cm3 of volume and 7.74cm3/g of specific volume.
5.4.5. Effect of glucose oxidase and transglutaminase on color parameters of gluten free Balady bread
• For the effect of glucose oxidase and transglutaminase at different concentration on color parameters of gluten free Balady breadwere studied, and the results refer to the untreated gluten free flour recorded 63.76, 9.51 and 32.24 for lightness, redness and yellowness respectively. While the others which treated by different concentration of glucose oxidase ranged between 64.63 to 73.57, 2.94 to 8.76 and 25.37 to 28.22 respectively for lightness, redness and yellowness respectively; and it was 62.57 to 62.66, 9.08 to 11.10 and 31.07 to 31.27 respectively in case of addition of transglutaminase at different concentration.
5.5. Amino acid content of gluten free Balady bread
• The results indicated that wheat flour Balady bread contained 4.33, 3.92, 6.09, 1.59, 3.06, 8.91 and 10.18 g/100g of valine, isoleucine, leucine, lysine, histidine, aspartic acid and proline respectively and it was 5.02, 4.87, 6.69, 5.85, 4.51, 11.25 and 3.57 g/100g respectively for gluten free Balady bread. On the other hand, wheat flour Balady bread recorded 35.26 g/100g and 52.55 g/100g of essential and non-essential amino acids, compared to 40.45 and 49.30 g/100g respectively which obtained by gluten free Balady bread.
5.6. Minerals content of gluten free Balady bread
• The obtained results indicated that wheat flour Balady bread contained 137, 141, 151, 26, 1.55 and 0.92mg/100g of sodium, 137.00mg/100g sodium, 137.00mg/100g sodium, 137.00mg/100g sodium, phosphorus, magnesium, Calcium, iron and zinc respectively. While the gluten free Balady bread sample recorded 202.70, 248.75, 62.40, 55.65, 2.81 and 1.62mg/100g respectively.
5.7. Physical properties of gluten free pasta
5.7.1. The hardness of uncooked pasta
• Uncooked pasta produced from wheat flour recorded the highest value of hardness (65.13N) compared to lower values which obtained by uncooked gluten free pasta, where the obtained hardness values of uncooked gluten free pasta with 2.5% psyllium husk were ranged from 44.97 to 15.16N. While, the uncooked gluten free pasta with 2.5% carboxymethyl cellulose ranged from 4.26 to 23.30N for the same parameter.
5.7.2. Optimum cooking time and cooking quality parameters of cooked gluten free pasta
• Optimum cooking time, cooking yield, swelling index, cooking loss and nitrogen loss were estimated for cooked pasta and the results refer to wheat flour pasta recorded 13.16min, 136.93%, 14283, 6.16% and 4% respectively. While the gluten free cooked pasta samples produced from different formulas with 2.5% psyllium husk were ranged from 10.16 to 12.83 min, 144.80 to 183.80%, 159.43 to 190.60, 6.83 to 9.80% and 6.50 to 10.20% respectively compared to 10 to 13 min, 120.80 to 166.40%, 150 to 186.66, 9.70 to 18.76% and 12.10 to 40.55% respectively for gluten free cooked pasta samples produced from different formulas with 2.5% carboxymethyl cellulose.
5.7.3. Texture profile analysis of cooked pasta
• Texture profile of cooked pasta was conducted on two cycle (first and second bit). During the first bit hardness cycle1, adhesiveness and resilience could be observed where the cooked pasta which produced from 100% wheat flour recorded 3.96 N, 0.1 mJ and 0.61 respectively for the same aforementioned parameters. While the gluten free pasta samples which produced from different formulas with 2.5% psyllium husk were ranged between 1.76 to 3.31N for hardness cycle1, 0.2 to 0.30mJ for adhesiveness and 0.32 to 0.57 for resilience compared to 0.69 to 1.73N, 0.3 to 0.7mJ and 0.1 to 0.55, respectively which obtained by gluten free pasta samples which produced from different formulas with 2.5% carboxymethyl cellulose.
• During the second bit hardness cycle2, cohesiveness, springiness, gumminess and chewiness could be observed where the cooked pasta which produced from 100% wheat flour recorded 3.74N, 0.9, 4.95mm, 3.56 and 17.64, respectively for the same aforementioned parameters. While the gluten free pasta samples which produced from different formulas with 2.5% psyllium husk were ranged between 0.84 to 3.16N for hardness cycle2, 0.38 to 0.75 for cohesiveness, 3.34 to 4.58mm for springiness, 0.67 to 2.48 for gumminess and 2.23 to 11.37mJ for chewiness. While the gluten free pasta samples which produced from different formulas with 2.5% carboxymethyl cellulose were ranged from 0.56 to 1.64N for hardness cycle2, 0.04 to 0.83 for cohesiveness, 1.80 to 4.30mm for springiness, 0.03 to 1.09 for gumminess and 0.05 to 4.69mJ for chewiness.
5.7.4. Sensory evaluation of gluten free cooked pasta
• Sensory evaluation of gluten free cooked pasta with 2.5% psyllium husk or 2.5% carboxymethyl cellulose compared to wheat flour pasta as control sample was investigated by hedonic scale.
• The results indicated that, the highest values of overall acceptability (5 and 4.8) were observed by the control sample and gluten free sample which consist from 37.5% rice flour + 37.5% corn flour + 15% potato starch + 10% soy protein isolate with 2.5% psyllium husk with no significant between them and with significant compared to the other gluten free samples. Generally, gluten free pasta containing psyllium husk showed higher quality in terms of all sensory parameters compared to the other gluten free pasta samples which containing carboxymethyl cellulose.
5.8. Degradation of wheat gliadin by fungal protease
• Fungal protease from Aspergillus niger was used to degrade wheat gliadin at different conditions (enzyme concentration, pH, incubation time, and incubation temperature) All protein fractions were analyzed by electrophoresis, where, the results indicated that, the best condition which completely degrade α-, β- and γ-gliadin and ω-gliadin are 0.3g/100g flour of enzyme concentration at pH 3, during 16hr, at 55°C.
Conclusion
Generally it could be conclude that, concerning the production of gluten free Balady bread psyllium husks can be used as a substitute for gluten in gluten-free products, as the study demonstrated a high ability of psyllium husks compared to carboxymethyl cellulose as a binding agent which one can obtain a separate layer and high quality attributes, whether in terms of the organoleptic or physical characteristics, the results also showed the possibility of obtaining gluten-free pasta with high quality by using psyllium husks compared to using carboxymethyl cellulose. Both maltogenic alpha-amylase and xylanase enzymes showed the ability to delay bread staling. Thus, transglutaminase showed a high ability to improve the quality of gluten-free Balady bread compared to the use of glucose oxidase. As for the nutritional value of gluten-free bread, it was characterized by its high protein and fiber content, as well as its high content of the amino acid lysine and some important mineral elements such as iron and zinc compared to the control sample which produced from wheat flour. On the other hand, the fungal protease enzyme can be used under appropriate conditions to degrade wheat gliadin to obtain safe wheat flour that can be used in gluten-free products for celiac patients.