Evaluation of functional properties of peptides obtained from hydrolysis of healthy wheat flour protein by sunn pest protease

Introduction: Wheat is the most important sustainable food product for more than a third of the world's population and has more calories and protein than other cereal products in the world diet.
The main property that distinguishes it from other products is the unique properties of the resulting dough, which allows it to appear in a variety of breads and other food products such as cakes, biscuits and pasta. These properties in these products depend on the structures and interactions of the grain storage proteins, which together constitute the gluten protein. Lutein is the predominant carotenoid in wheat. Wheat bran and sprouts contain a lot of carotenoids and antioxidant activity compared to endosperm. Lutein along with zeaxanthin is important for human skin and eye health. Heart disease protection may come from whole grains, antioxidants, vitamins, and fiber and minerals. Whole grains are also effective in preventing diabetes. Whole grains appear to protect against heart disease and cancer. Proteins in plant tissue are simple and consist of four main types, including: albumin (soluble in water and dilute buffer), globulins (soluble in salt water) and prolamine (soluble in 90-90% ethanol) and glutlin (soluble in Wheat gluten protein is classically composed of two parts: alcohol-soluble gliadin and alcohol-insoluble glutenin. Glutenins are known as the largest polymers in nature. Wheat proteins can be divided into structural proteins (gluten-free) and storage proteins or (gluten). Structural proteins include albumin, globulin, and amphiphilic. Non-membrane amphiphilic proteins have many effects on grain hardness and rheological properties of dough. Wheat storage proteins are known as prolamins due to their amino acids proline and glutamine. Aqueous salivary secretions act rapidly on the plant cell, first increasing cell respiration and then causing the protoplasm to flow, which is usually due to increased permeability of the cell membrane. They contain soluble amino acids that immediately after secretion on plant materials with the formation of a large number of hydrogen bonds as well as a number of disulfide bonds in the form of gels that gradually become solid. In this study, the effect of extracted protease from aged wheat, protease (serine) has been investigated. The aim of this study was to produce short chain peptides with beneficial functional and therapeutic properties such as antioxidant, antidiabetic, anti-hypertension by aged wheat protease.
Material and Methods: The tests include determination of enzymatic activity, degree of hydrolysi, determination of functional properties such as solubility, antioxidant, emulsification, and solvent retention capacity (SRC). Microstructure of the contaminated seed was also investigated by electron microscopy (SEM) ). Statistical analysis was performed using SPSS software at 95% confidence level. First, for extraction of aged wheat flour mix 2 gr of flour with 10 cc of 0.2 M acetate buffer with pH = 3.8 and centrifuge for 5 minutes. The supernatant was separated and filtered from a 0.45 microfilter and stored in the freezer. flour enzymatic activity was performed in the first, 5 cc of 0.75% casein solution in which 50 mM hydrodisodium phosphate buffer with pH= 7 is equilibrated for 10 minutes at 37 ° C by pH. Slow addition of 0.1 N hydrochloric acid is adjusted to this substrate. To this substrate a certain volume of enzyme is diluted with 1 cc of active buffer of 30 mM cysteine hydrochloride monohydrate in 6 mM EDTA and diluted in a bath for 10 minutes and water 37 °C is mixed gently. The reaction is then stopped by adding 5 cc of 30% v / v trichloroacetic acid. At room temperature, cool with Whatman 42 filter paper and measure the absorption at 280 nm visible-ultraviolet wavelengths . For hydrolyse degree, First, make an opa reagent that dissolves 3.81 g of disodium tetra borate + 0.1 g of SDS plus 75 cc of distilled water, then dissolve 80 mg of orthophthaldehyde in 2 cc of 96% ethanol and place it on the shaker with a magnet until to be solved. Add 250 microliters of mercaptoethanol to the main container and then increase the volume to 100 with distilled water. The next step is to add 0.5 g of flour in different proportions with the same enzymatic activity plus 10 cc of distilled water and in the control sample we used water instead of enzyme. Then we put the tubes on the shaker for 30 minutes and then centrifuge for 20 minutes at 10,000 rpm and 4 ° C and add 400 μl to 3 ml of opa and then zero with reagent and then absorb at 340 nm. we read.Also, the effect of this protease enzyme on functional properties and creation of free peptides was investigated.
Results and discussion: It was observed that the enzyme extracted from aged wheat performed better in the creation of free peptides and increased the functional properties such as antioxidant properties and solubility, but the emulsifying properties of healthy flour from flour with aged enzymatic extract increased due to lower hydrolysis of healthy flour than healthy flour with aged enzyme. Wheat SRC was carried out according to AACC no. 11-56 method with two solvents of water and lactic acid, the results of which indicated that by increasing hydrolysis, SRC decreased in lactic acid solvent and was ineffective in water solvent. In addition, electron microscopy (SEM) with a magnification of 100 and 50 μm was observed that in aged wheat, protein structure was degraded compared to healthy wheat and protein cells were partially destroyed. These peptides are very beneficial for human health and are used in food industry and various food products.
Conclusion: The results of this study showed that the use of protease enzyme of sunn pest flour wheat enzyme extract in the production of free peptides has worked well and also has been very effective in improving functional properties such as solubility, antioxidants. Continuous innovations in the food industry and their higher quality requirements force the food industry to produce flours with specific functional properties. It seems protease isolated from sunn pest wheat flour might be considered as a suitable source for creating peptides with several advantages. According to the results, it was observed that aged wheat protease is a suitable animal protease for peptides with beneficial health benefits. In this study, it was found that in general, the advantages of age protease are the cheapness and availability of aged wheat and its high hydrolysis.