Quantitative determination of amylase activity in germinating cereal grains using agar plates and ImageJ software

The research is devoted to the technological modes of germination of triticale grain of the Asiada variety, as well as factors affecting the germination of grain. The purpose of the study is the influence of technological modes of germination of triticale grain of the Asiada variety. In the article, studies have been carried out and regression equations have been obtained describing the dependences of the indicated quality indicators of the germination modes of triticale grain of the Asiada variety on the factors influencing them tв, τ, w and tз To optimize the technological modes of processing Asiada grain, the crude protein content was chosen as the target function. Optimization of the processing modes of triticale grain of the Asiada variety was carried out by the method of nonlinear programming. The following optimal technological modes of grain processing were obtained: water temperature tb = 15 °C; germination time τ = 72 h; grain moisture w = 13%; grain temperature tz = 10 °C. With these optimal grain processing modes, the target function was 19.11%. The practical significance of the technology for the production of compound feeds using sprouted triticale grain in order to improve quality and increase shelf life.

During storage, elevated temperature and humidity cause accelerated aging and deterioration of seeds of various plant species, including important cultivated cereals, such as wheat and triticale. Germination of old seeds can be facilitated by seed priming with physiologically active substances that correct the pro/antioxidant balance and reduce the development of oxidative stress. Gamma-aminobutyric acid (GABA) is a regulatory compound with direct and indirect antioxidant effects. However, its effect on the germination of grains of cereals with low germination has not been extensively studied. The objective of this study was to examine the impact of GABA priming on the germination of aged grains of winter bread wheat (Triticum aestivum ‘Scorpion’) and winter triticale (×Triticosecale ‘Raritet’). the extent of oxidative stress and the state of the antioxidant system in seedlings. The results indicated that a three-hour treatment of grains with GABA at the optimal concentration (1 mM) resulted in a significant (18–21%) increase in germination energy and seed germination rate, as well as an increase in the biomass of shoots and roots of seedlings of both species. Concomitantly, the influence of GABA resulted in a reduction in oxidative stress markers, including the generation of superoxide anion radicals, hydrogen peroxide content, and the product of lipid peroxidation malondialdehyde. In wheat seedlings, the total content of phenolic compounds increased, while in triticale seedlings, the content of anthocyanins increased almost twofold. In seedlings derived from GABA-primed grains, catalase activity was also significantly elevated in the absence of notable alterations in superoxide dismutase and peroxidase activity. It was postulated that GABA priming is a promising approach for enhancing the germination of cereal seeds with diminished sowing quality.

An example of effective crossbreeding of wheat and rye grain is the grain of classic triticale, which is currently gaining growing popularity among manufacturers of bakery products and cereals. As a result of further hybridization of the grain of classical triticale and common wheat, highly productive forms of four species triticale were obtained. Despite the similarities to wheat grains, four species triticale grains have differences in technological properties, and therefore require further study. One of the most problematic places is the identification of significant differences between the properties of the four species triticale and its parent forms. Therefore, it is important to conduct a comprehensive study and comparison of the technological properties of four species triticale, classic triticale and common wheat, which will justify its intended purpose. So, the object of research was selected four species triticale of Alkyd, Strateg, Tactic, line LP 195 varieties (Ukraine).In the course of the study, current methods for determining the technological properties of cereal grain were used, and the results were processed by analysis of variance. The grain of four species triticale has a large mass of 1000 grains compared to wheat and classical triticale, which is associated with its larger size. Four species triticale is characterized by less sphericity compared to wheat due to the longer length, affects the duty cycle of the grain mass and grain nature. The nature of the grain of four species triticale is significantly inferior (by 38-85 g/l) to the nature of wheat grain. Four species triticale grains are glassy (98-99%) and similar to traditional triticale grains according to this indicator. According to the gluten content (20-25%), the grain of four species triticale is significantly inferior to wheat, but its quality is satisfactory.The research results presented in the work demonstrate a high glassiness and a low amount of gluten. This makes it possible to efficiently use the grain of four species triticale for cereal production, or as part of composite mixtures in the production of bakery products.

Environments containing significant concentration of NaCl such as salt lakes harbor extremophiles microorganisms which have a great biotechnology interest. To explore the diversity of Bacteria in Chott Tinsilt (Algeria), an isolation program was performed. Water samples were collected from the saltern during the pre-salt harvesting phase. This Chott is high in salt (22.47% (w/v). Seven halophiles Bacteria were selected for further characterization. The isolated strains were able to grow optimally in media with 10–25% (w/v) total salts. Molecular identification of the isolates was performed by sequencing the 16S rRNA gene. It showed that these cultured isolates included members belonging to the Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus genera with less than 98% of similarity with their closest phylogenetic relative. The halophilic bacterial isolates were also characterized for the production of biosurfactant and industrially important enzymes. Most isolates produced hydrolases and biosurfactants at high salt concentration. In fact, this is the first report on bacterial strains (A4 and B4) which were a good biosurfactant and coagulase producer at 20% and 25% ((w/v)) NaCl. In addition, the biosurfactant produced by the strain B4 at high salinity (25%) was also stable at high temperature (30-100°C) and high alkalinity (pH 11).

Peroxidase and amylase activity in developing grains of triticale, wheat and rye

Seed dormancy is of particular importance in the cultivation of cereals, as it directly affects the quality of crop yield. If the dormancy period is too short, this may lead to pre-harvest sprouting, whereas a dormancy period that is too long may cause uneven germination; both of these scenarios are associated with economic losses. Most enzymes engaged in the metabolism of abscisic acid (ABA) have been identified, and significant progress has been made in understanding the role of this phytohormone in the induction and maintenance of dormancy, mainly as a result of research conducted in <em>Arabidopsis</em>. Much less is known about the metabolism and function of ABA in cereal grains, especially in relation to dormancy and germination. This review focuses on the regulation of ABA metabolism in dormant and non-dormant cereal grains, in both the dry state and upon imbibition. Moreover, this review describes the influence of factors such as after-ripening, light, temperature, nitric oxide, and reactive oxygen species (ROS) on the dormancy and germination of cereal grains. These factors, with the exception of ROS, appear to affect the level of dormancy and germination of grains through regulation of ABA metabolism.

Triticale grain crop is promising for cultivation in various regions of central Russia. Stable yield, high resistance to adverse conditions, allow us to consider this cereal plant as a reliable source of food production. The purpose of this study was to quantify the content of active α-amylase in triticale grain to provide a forecast of agrotechnical measures. The analysis of the quantitative content of α-amylase was carried out on a laboratory spectrofluorimeter using standard bioassays conducted on the basis of a set of reagents for determining the activity of α-amylase in blood serum and urine; the conversion factor for calculating enzyme activity was calculated using a multiparametric calibrator of biochemical analyzers. To improve the methodology for the determination of α-amylase in real samples of plant raw materials, flour of winter triticale varieties grown in the field was used. Based on the results of measuring the activity of α-amylase in triticale grain, a comparative assessment of the enzyme content in the flour of grain crops of 2020 and 2021 was carried out; the dependence of pre-harvest seed germination on the content of α-amylase in grain of various varieties, lines and hybrids was shown. Based on the data obtained on the content of α-amylase in triticale grain, a comparative assessment of the characteristics of pre-harvest seed germination was carried out. The indicator of the quantitative content of α-amylase can be recommended for use in the diagnosis of pre-harvest germination of grain, as well as in breeding work when creating new varieties of grain crops.

Bioethanol production from triticale by simultaneous saccharification and fermentation with magnesium or calcium ions addition

To our knowledge, there is no research on the molecular structure of triticale grain in comparison with other types of cereal grains and metabolic characteristics of the protein and energy in this grain and its coproducts, called dried distillers grains with solubles (DDGS), for dairy cattle. The objective of this study was to identify differences in molecular structures of proteins among grains and their DDGS using a molecular spectroscopy technique, namely, DRIFT, and to determine the nutrient profile and supply to dairy cattle. The protein molecular structure studies showed a difference (P < 0.01) in the amide I to amide II ratio and the α-helix to β-sheet ratio between grains and their DDGS. The energy content was similar for triticale grain and DDGS. There were differences in the protein and carbohydrate subfractions (P < 0.05) and the ruminal degradability of DM, CP, and NDF (P < 0.01) between triticale grain and DDGS. Triticale grain and DDGS had similar intestinal digestibility of rumen undegraded CP. However, triticale DDGS had higher (P < 0.01) predicted total metabolizable protein and degraded protein balance than triticale, indicating that triticale DDGS is a superior protein source for dairy cattle as compared with triticale grain. Bioethanol processing induced changes in the protein molecular structure.

Marijuana consumption is prevalent among young adults in Nigeria, and its effects on various physiological systems are of growing concern. This study investigates the impact of marijuana use on digestive enzymes, specifically serum amylase and lipase, to understand its potential implications on digestive health. This cross-sectional study was conducted in 120 participants comprising of 60 marijuana smokers and 60 non-smokers. Sociodemographic and lifestyle data were collected through structured questionnaires. Serum amylase and lipase activity levels were measured by spectrophotometric method and compared between the two groups. Statistical analysis was performed to assess differences and correlations. The study revealed that marijuana smokers had significantly higher serum amylase and lipase activities compared to non-smokers. Lipase activity correlated positively (r=0.425, p=0.019) with duration of marijuana use. Amylase activity was higher among males than females (r=-0.40, p=0.028). No significant correlation was found between the quantity of marijuana consumed and enzyme activities. Chronic marijuana consumption was associated with increased serum amylase and lipase activities, indicating potential alterations in pancreatic function. These findings suggest that marijuana use may have adverse effects on digestive health, warranting further investigation into the long-term implications. It is recommended that healthcare providers monitor digestive enzyme activities in marijuana users to detect early signs of pancreatic dysfunction.

The objective of this study was to evaluate the possibility of application of two triticale varieties, NS Paun and Odisej, as partial substitutes for barley malt in wort production, based on the triticale analyses, micromalting, and analyses of the produced triticale malts, with a highlight on the activity of amylases during the malting process. The analyses of two triticale varieties showed higher extract content and diastatic power in comparison to barley malt. In the varieties NS Paun and Odisej, the extract was by 11.4% and 6.3% higher than the standard values for barley malt, respectively. Diastatic power in varieties NS Paun and Odisej was by 100% and 111.9% higher than in barley malt, respectively. In both varieties, ?- and ?-amylases activities were also high. The ?-amylase activity increased during the micromalting 34 and 20 times, respectively, in comparison to native triticale grain. The ?-amylase activity also increased during micromalting, but to a lesser extent. The produced triticale malts had good technological parameters and could be used as a partial substitute for barley malt in wort production.

The underlying mechanisms of starch hydrolysis in cereal grains, as measured by the Falling Number procedure, were studied. Wheat starch, spiked with barley malt α-amylase, was used as a model experimental system. A non-invasive and real-time monitoring system was developed to simultaneously record the sample temperature and stirrer velocity profiles, which are critical parameters that affect Falling Number readings. Experimental results show that the repeatability of the Falling Number procedure can be improved by slowing the heating rate, which is accomplished by increasing the quantity of sample components (ie starch and water) while maintaining the same proportion as currently specified in standard methodology. Among the tested range of heating rates, the best repeatability of Falling Number readings was obtained with 8.4 g starch and 30.0 g water. The experimental findings of this study also serve as basic information for the development of mathematical models on the hydrolysis of starch. © 1999 Society of Chemical Industry

Gluten-free products from rice are gaining popularity because of its hypoallergenic characteristic. The absence of gluten results in inferior bread qualities such as hard texture, reduced volume, and shorter shelf-life. Hydrolytic enzymes are activated during germination to stimulate plant growth, and germinated brown rice (GBR) has been shown to improve gluten-free bread properties. However, the changes in hydrolytic enzyme activities under different germination conditions and their relationship with the properties of germinated rice flour and bread have not been reported. Therefore, the objectives of this work were to investigate the activities of amylases and protease in GBR under aerobic and anaerobic germination for 2 and 4 days and their impacts on starch hydrolysis, flour properties, and bread qualities. Greater enzyme activities were observed in GBR germinated under aerobic condition and a longer time, and correlated with increased sugar content and foaming capacity. Breads were prepared from GBR along with brown rice (control). GBR breads showed a greater specific volume (4% to 10%), a reduced hardness (34% to 90%), and a lower starch retrogradation (66% to 90%) compared with the control. Bread prepared from 4-day aerobic GBR had the largest reduction in starch molecular size and displayed the lowest hardness and starch retrogradation. After stored for 5 days, GBR breads exhibited no change in specific volume and less hardness and retrogradation than the control bread. In conclusion, greater activities of protease and amylases in GBR significantly increased foaming capacity and reduced starch molecule size, respectively, which were responsible for the improved GBR bread qualities. PRACTICAL APPLICATION: Rice flour is widely used as the main ingredient in gluten-free breads, which however tend to have poor texture and reduced shelf-life due to the absence of gluten. The qualities of gluten-free breads are usually improved by the addition of many ingredients such as tapioca and potato starches. Germination process naturally produces bioactive compounds and activates enzymes. Germination conditions that produce greater activities of amylases and protease can be used to produce gluten-free breads with better qualities and longer shelf-life without the addition of starch.

When fertilization of developing cereal grains occurs, the pericarp (ovary wall) already contains starch granules. Hence the starch content of developing grains of wheat, triticale, rye, oats and barley commences at 30 or 40% of the dry weight. It increases only to 60 or 70% at ripeness when the pericarp has degenerated to bran layers and the endosperm is filled with stored starch granules and protein. All these cereals showed similar patterns of starch increase, with an inflexion at 40 to 50% starch due to degeneration of pericarp starch before endosperm deposition.In wheats and triticale there was a second inflexion at 50 to 60% starch caused by endosperm cells proliferating faster than they filled with starch granules. The rise and fall of rate of starch deposition coincided with the rise and fall of percentage of metabolic (non‐storage) protein in the grain, calculated from total amino acid analyses. In the mature grain about 8% of the dry matter is not associated with starch and thus is non‐endosperm tissues. About 70% of the remaining dry matter (endosperm) is starch. We suggest that fluctuations of metabolites and enzymes result from development or breakdown of groups of cells constituting distinct tissues in the grain.

The aim of this study was to evaluate the effect of diets with different inclusion levels of high-moisture corn (HMC) and triticale grain (TG) on nutrient digestibility and utilization, the growth performance of lambs, and the fatty acid profile of the leg muscle. The experiment was performed on 24 young rams, divided into four groups based on grain type and inclusion rate (50% or 75% DM). The higher inclusion rate of cereal grain and a lower concentration of crude fiber (CF) in the feed ration decreased CF digestibility by 26% and 35% in diets containing HMC and TG, respectively. Diets containing HMC increased crude fat digestibility relative to animals receiving TG. Final body weight and average daily gain were highest in lambs fed a diet containing 50% HMC. Grain type had no effect on nitrogen (N) retention or the proportions of SFA and UFA in lamb meat. Diets containing HMC decreased total cholesterol levels in the leg muscle. HMC can be a valuable component of diets for growing lambs when included at up to 50% of the ration. An increase in the dietary inclusion rate of cereal grain to 75% can be recommended only in the case of TG.