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A New Approach of Extraction of ?-Amylase/trypsin Inhibitors from Wheat (Triticum aestivum L.), Based on Optimization Using Plackett-Burman and Box-Behnken Designs.

ABSTRACT: Wheat is one of the most consumed foods in the world and unfortunately causes allergic reactions which have important health effects. The ?-amylase/trypsin inhibitors (ATIs) have been identified as potentially allergen components of wheat. Due to a lack of data on optimization of ATI extraction, a new wheat ATIs extraction approach combining solvent extraction and selective precipitation is proposed in this work. Two types of wheat cultivars (Triticum aestivum L.), Julius and Ponticus were used and parameters such as solvent type, extraction time, temperature, stirring speed, salt type, salt concentration, buffer pH and centrifugation speed were analyzed using the Plackett-Burman design. Salt concentration, extraction time and pH appeared to have significant effects on the recovery of ATIs (p < 0.01). In both wheat cultivars, Julius and Ponticus, ammonium sulfate substantially reduced protein concentration and inhibition of amylase activity (IAA) compared to sodium chloride. The optimal conditions with desirability levels of 0.94 and 0.91 according to the Doehlert design were: salt concentrations of 1.67 and 1.22 M, extraction times of 53 and 118 min, and pHs of 7.1 and 7.9 for Julius and Ponticus, respectively. The corresponding responses were: protein concentrations of 0.31 and 0.35 mg and IAAs of 91.6 and 83.3%. Electrophoresis and MALDI-TOF/MS analysis showed that the extracted ATIs masses were between 10 and 20 kDa. Based on the initial LC-MS/MS analysis, up to 10 individual ATIs were identified in the extracted proteins under the optimal conditions. The positive implication of the present study lies in the quick assessment of their content in different varieties especially while considering their allergenic potential.

SUBMITTER: Tchewonpi Sagu S

PROVIDER: S-EPMC6803881 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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A New Approach of Extraction of α-Amylase/trypsin Inhibitors from Wheat (Triticum aestivum L.), Based on Optimization Using Plackett-Burman and Box-Behnken Designs.

Tchewonpi Sagu Sorel S Huschek Gerd G Bönick Josephine J Homann Thomas T Rawel Harshadrai M HM

Molecules (Basel, Switzerland) 20191005 19

Wheat is one of the most consumed foods in the world and unfortunately causes allergic reactions which have important health effects. The α-amylase/trypsin inhibitors (ATIs) have been identified as potentially allergen components of wheat. Due to a lack of data on optimization of ATI extraction, a new wheat ATIs extraction approach combining solvent extraction and selective precipitation is proposed in this work. Two types of wheat cultivars (Triticum aestivum L.), Julius and Po ...[more]

PMID: 31590396

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Project description:Juniper berry is an important medicinal plant used in pharmaceutical and petrochemical industries thanks to its strong antioxidant potential, which is attributed to the presence of phenolic compounds. In this study, four different solvents, namely, aqueous acetone, aqueous ethanol, aqueous NaOH, and water, were used in the extraction process with a view to optimize and determine the polyphenolic contents in the juniper berry using ultraviolet (UV) spectrophotometry. Many experiments were performed at different solvent concentrations, time, temperature, and liquid-solid ratio. The models to evaluate the effects and the optimum of these variables on the polyphenols extraction using the response surface methodology (RSM) were developed. The predicted values of the polyphenol content of juniper berry were thus highly correlated with costly measured values (SECV = 0.14 and R 2 = 0.97), and the optimal conditions of extraction were determined for the different solvents. Following the numerical optimization, the maximum predicted polyphenol contents obtained under the optimum extraction conditions are as follows: 17.57% for 58 °C extraction temperature, 78.5 min extraction time, 60% acetone concentration, and 29.8 liquid-solid ratio for the aqueous ethanol extraction; 20.68% for 71.46 °C extraction temperature, 79.2 min extraction time, 21.9% ethanol concentration, and 26.4:1 liquid-solid ratio for the aqueous acetone extraction; 34.51% for 96.4 °C extraction temperature, 37.7 min extraction time, 1.48% NaOH concentration, and 15.2:1 liquid-solid ratio for the aqueous NaOH extraction; and 9.8% was obtained under the optimum extraction conditions of 69 °C extraction temperature, 126 min extraction time, and 23:1 liquid-solid ratio for the water extraction. The GC-MS analysis of the chemical composition of juniper Berry revealed 60 identified components that represent 97.43% of the sample. The predominant fraction was monoterpene representing 80.87% especially for α-pinene (39.12%), β-pinene (12. 68%), and myrcene (12.92%). In the other fraction of sesquiterpene representing 16.54%, the predominant components were β-caryophyllene (4.41%) and germacrene D (4.23%).

Dataset Information

A New Approach of Extraction of ?-Amylase/trypsin Inhibitors from Wheat (Triticum aestivum L.), Based on Optimization Using Plackett-Burman and Box-Behnken Designs.

Publications

A New Approach of Extraction of α-Amylase/trypsin Inhibitors from Wheat (<i>Triticum aestivum</i> L.), Based on Optimization Using Plackett-Burman and Box-Behnken Designs.

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