Increase In Methionine Content Research Articles

Evaluation of forage quality in various soybean varieties and high-yield cultivation techniques

ContextThe rising demand for animal protein has intensified forage shortages and restricted pasture availability, underscoring the critical need for effective dual-purpose animal feeds. ObjectiveThis study aims to evaluate the potential of vegetative soybeans as animal feed by assessing their nutritional value and productivity. Specifically, it explores the feasibility of using late-maturing soybean varieties, adapted to low latitudes, as forage in high-latitude regions and examines the impact of different planting dates on their growth and effectiveness during limited growing seasons. MethodsTo achieve these objectives, a series of field and controlled experiments were conducted. The first experiment assessed 418 mid-maturing soybean varieties from the Huang-Huai-Hai Basin in China for their nutritional value as forage during 2020 and 2021. The second experiment tested late-maturing and photoperiod-sensitive soybean varieties, including "ZGDD" and the methionine-enhanced material "Ox-CGS," under tropical conditions with extended daylight. The third experiment evaluated forage soybeans sown on June 19th, July 14th, and August 8th, 2022, to determine their yield and protein content. ResultsResults showed that the first experiment yielded an average protein content of 24.7 %, ether extract content of 1.9 %, neutral detergent fiber content of 41.9 %, and acid detergent fiber content of 27.3 %, with a relative feeding value of 153, surpassing local standards for high-quality leguminous forage. The second experiment demonstrated that extended daylight increased crude protein yield by 3.7 times compared to Chinese soybean seed yields, and "Ox-CGS" showed a 24.2 % increase in methionine content. The third experiment revealed that forage soybeans achieved a crude protein yield of 0.95 t/ha within 60 days, exceeding the 2022 average of 0.68 t/ha for seed soybeans in China. ConclusionsThe findings indicate that photoperiod-sensitive soybeans can be effectively used as forage over extended periods, utilizing artificial lighting or cultivation in higher latitudes, and that short growth cycles enhance crude protein accumulation. Implications or significanceThis research highlights the potential of soybeans to alleviate forage shortages, improve land use efficiency, and contribute to food security and agricultural development, especially on marginal lands and in regions with limited growing seasons.

Changes in the amino acid pool of cerebral hemispheres in rats with total cerebral ischemia

Objective: assessment of changes in the pool of amino acids (AA) in rats with total cerebral ischemia (TCI). Material and Methods. Experiments were performed on 16 male outbred white rats weighing 260±20 g. TCI was modeled by decapitation of animals. Brain tissue was sampled 1 hour after decapitation. Results. In the parietal lobe (PL) and hippocampus (HC) of TCI group animals 1 hour after their decapitation, we detected an increase in the content of tyrosine (by 43%, p=0.044, and 40%, p=0.044, respectively) and tryptophan (by 24%, p=0.036, and 23%, p=0.046, respectively). Similar trend was observed for methionine that increased by 32% in PL(p=0.046) and by 27% in HC (p=0.046). Analogous increase in the content of L-arginine was noted in PL and HC (by 20%, p=0.037, and 33%, p=0.037, correspondingly). Isoleucine content increased by 12% in PL (p=0.054), while valine content decreased by 15% in HC (p=0.053). The ratio of the combined total content of branched-chain amino acids (BCAA)to the combined content of aromatic AA in TCI significantly declined from 1.4 to 1.0 in PL (p=0.053) and from 1.6 to 1.0 in GC (p=0.053). We observed an increase in methionine content by 33% (p=0.046) in PL and an increase in tryptophan content by 24% (p=0.046) in HC. Conclusion. One-hour TCI caused the following changes in the AA pool: an increase in the content of aromatic AA (tyrosine and tryptophan) and methionine; an increase in the content of L-arginine; and also, an increase in the concentration of the inhibitory neurotransmitter glycine.

Ріст телят - молочників за додаткового введення метіоніну і лізину у замінник незбираного молока

The relevance of the study is due to the need to optimize the content of lysine and methionine in milk replacer (MR) for calves. The aim of the work is to study the effect of additives of synthetic L-lysine and DL-methionine on the growth of calf calves. The objectives of the study were to study the growth rates of calves by live weight, average daily and relative gain. The study was conducted on six groups of calves (20 heads each) according to the scheme: Group I – control, which received a MR with a content of 1.95 % lysine and 0.49 % methionine; In groups II and III the lysine content was 2.24 and 2.44 %, respectively, the methionine content in these groups did not change and was 0.49 %; In MR for calves of group IV-V, 0.56 and 0.61 % of methionine were contained, respectively, and the lysine content did not change from the actual (1.95 %); The VI group received lysine and methionine supplements, the content of these amino acids in the MR was 2.34 and 0.59 %, respectively. The calves' age is 22-62 days in the experiment. The experimental calves were kept in individual cages, the microclimate parameters of the premises where the animals were kept corresponded to the accepted zoohygienic and veterinary-sanitary standards. Calves were fed three times a day using a MR, starting granulated feed and hay. At the beginning of the experiment, the difference in the BW between the groups did not exceed 5 % (39.4-41.2 kg). On the 10th day of the experiment, in all groups except II, results were obtained above the control group. The calves of group II had a BW of 2.11 kg less than the control group calves, while the animals of group VI, on the contrary, exceeded the control group by 1.75 kg (P> 0.1). The tendency to increase BW of calves, relative to control, remained for the calves of the VI group and for 42 days. They had an BW greater than the control animals, an average of 2.89 kg. In all other groups, this indicator was less by 2.8-9.4 %. On days 52 and 62, in all the test groups of calves, in contrast to the control, a decrease in the BW was observed. At the end of the experiment, the animals of the V and VI groups had close indicators to the control group (67.85, 69.76 and 70.38 kg, respectively). Analysis of the results of the study showed that the additional introduction of synthetic amino acids (lysine and methionine) into the composition of the MR for calves aged 22-62 days from 1.95 and 0.49 % to levels of 2.24- 2.44 and 0.56-0.61 % respectively, noticeably affects the growth rates. According to the BW, ADG, RG of calves enrichment of the MR with synthetic lysine and methionine does not give positive changes in the growth of calves during the dairy period under study. Changes in the growth of calves for 22-42 days indicate an increased need for lysine and methionine (content in the MR, respectively, 2.34 and 0.59 %), the provision of which contributes to the growth of BW by 5.3 % (P < 0.01). There was a slight increase in the calves' need for methionine during the growing period of 42-62 days. The increase in methionine content during this period from 0.49 to 0.61 % in the MR contributed to an increase in the relative growth of body weight by 1.4 %. Prospects for further research are to study the changes in the rates of linear growth in calves, the effectiveness of nutrient use in the diet, and the study of the impact on the milk productivity of first-calf cows. Keywords: calves, milk period, lysine, methionine, milk replacer, growth, body weight, gains

S–carboxyethylcysteine (a constituent of Acacia seed) negatively affects casein protein utilization by rats

ObjectiveTwo rat bioassay experiments are reported. The first investigated the first limiting amino acid in Acacia colei and the second experiment investigated the effect of S-carboxyethylcysteine (CEC; a compound present in acacia seed) on protein use. MethodsIn the first experiment, Wistar rats were fed A. colei seed supplemented with three levels of methionine, cysteine, and tryptophan (0.1%, 0.2%, and 0.4%). In the second experiment, the Wistar rats were fed CEC-incorporated casein diets. ResultsSupplementation of A. colei with tryptophan had no significant effect on the protein efficiency ratio, cysteine showed the highest protein efficiency ratio value at the 0.4% level, and the protein efficiency ratio increased significantly with the increase in methionine content, making methionine the first limiting amino acid. The methionine-induced growth rate was suppressed by the incorporation of CEC, which also had a negative effect on the plasma amino acid levels. ConclusionThe results indicated that methionine is the first limiting amino acid in A. colei and that CEC could affect the seed’s protein use. Acacia colei seed can be used effectively as famine food only if it is complemented with other cereals known to be rich in sulfur amino acids.

Bioengineering approaches to improve the nutritional values of seeds by increasing their methionine content

Methionine is a nutritionally essential, sulfur-containing amino acid found at low levels in plants and in their seeds. Methionine levels often limit the plant’s value as a source of dietary protein for humans and animals. Despite recent accumulated knowledge of methionine metabolism in vegetative tissues, there is still little knowledge of methionine metabolism in seeds. In this review, we summarize the efforts made to increase the levels of methionine in seeds using genetic engineering methods. Two main approaches were tested: the first was the expression of methionine-rich storage proteins in a seed-specific manner, with the goal of trapping the soluble methionine into protein form and competing with the catabolism of methionine to its essential metabolites. However, in many cases this approach does not lead to a significant increase in total methionine content. The second approach aimed to increase the soluble content of methionine in seeds. Despite the nutritional significance of methionine, the factors regulating soluble methionine content in seeds are not fully known. Evidence shows that two biosynthetic pathways, the aspartate family pathway and the S-methylmethionine pathway, contribute to soluble methionine content in seeds. However, their roles in soluble methionine synthesis and accumulation are not fully understood. In recent years, combinations of these two approaches have been tested; however, they have not yet succeeded in elevating total methionine content in seeds. More emphasis should be applied to gaining knowledge of the biosynthesis pathways that could contribute to an increase in methionine content in seeds.

Expression of an 11 kDa methionine‐rich delta‐zein in transgenic soybean results in the formation of two types of novel protein bodies in transitional cells situated between the vascular tissue and storage parenchyma cells

Soybean (Glycine max (L.) Merr.) is an important protein source in human diets and animal feeds. The sulphur content of soybean seed proteins, however, is not optimal for ration formulations. Thus, increasing the methionine and cysteine content of soybean seed proteins would enhance the nutritional quality of this widely utilized legume. We have earlier reported the isolation of an 11 kDa delta-zein protein rich in methionine from the endosperm of the maize (Zea mays L.) inbred line W23a1 [Kim, W.-S. and Krishnan, H.B. (2003) Allelic variation and differential expression of methionine-rich-delta-zeins in maize inbred lines B73 and W23a1. Planta, 217, 66-74]. Using Agrobacterium-mediated transformation, a construct consisting of the coding region of the cloned delta-zein gene under regulation of the beta-conglycinin alpha'-promoter was introduced into the soybean genome. The 11 kDa delta-zein gene was expressed in the seeds of transgenic soybeans, although low-level expression was also detected in the leaves. In situ hybridization indicated that the 11 kDa delta-zein mRNA was expressed predominantly in transitional cells located between the vascular tissue and storage parenchyma cells. Immunohistochemistry of developing transgenic soybeans revealed that the accumulation of the 11 kDa delta-zein occurred primarily in these transitional cells. Expression of the 11 kDa delta-zein gene in transgenic soybean resulted in the formation of two endoplasmic reticulum-derived protein bodies that were designated as either spherical or complex. Immunocytochemical localization demonstrated that both the spherical and complex protein bodies accumulated the 11 kDa delta-zein. Although expression of the 11 kDa delta-zein gene elevated the methionine content of the alcohol-soluble protein fraction 1.5-1.7-fold above that of the non-transgenic line, the overall methionine content of seed flour was not increased. Our results suggest that the confined expression of the 11 kDa delta-zein gene in transitional cells could be limiting the increase in methionine content in transgenic soybean seeds.

Increased sulfur amino acids in soybean plants overexpressing the maize 15 kDa zein protein

Four transgenic soybean [Glycine max (L.) Merrill] lines were generated containing the maize 15 kDa zein protein gene using somatic embryogenic protocols. The zein gene was inserted behind the β-phaseolin promoter for seed-specific expression. All four lines represent separate transformation events as they were generated in different experiments at different locations. Two of the transformation events produced multiple plants, and these produced identical Southern hybridization patterns (UKY/Z1, UKY/Z2 and UKY/Z3 from the first; and OSU/Z4, OSU/Z8 and OSU/Z10 from the second). Molecular characterization revealed that multiple copies of the zein gene were present in all of the transgenic lines. Immunoblot analysis confirmed the accumulation of the zein protein product in the seeds of the UKY/Z1, UKY/Z2, UKY/Z3, OSU/Z4, OSU/Z8 and OSU/Z10 transgenic lines. However, there was no accumulation of zein protein in the UGA/Z1 line and Northern analysis confirmed that the zein transgene was silenced in this line. It was not possible to analyze the zein expression in the seeds of the UKY/Z4 line, as it was sterile. Amino acid analysis of the UKY and OSU lines confirmed that there was a 12–20% increase in methionine, and 15–35% increase in cysteine content in these lines compared to the control. There were no consistent changes in the content of the other amino acids in the transgenic lines. These results suggest that while the increase in methionine content in these lines is modest, it is possible to increase the methionine content without adversely affecting the protein composition of soybean.

Enhanced methionine levels and increased nutritive value of seeds of transgenic lupins (Lupinus angustifolius L.) expressing a sunflower seed albumin gene.

With the aim of improving the nutritive value of an important grain legume crop, a chimeric gene specifying seed-specific expression of a sulfur-rich, sunflower seed albumin was stably transformed into narrow-leafed lupin (Lupinus angustifolius L.). Sunflower seed albumin accounted for 5% of extractable seed protein in a line containing a single tandem insertion of the transferred DNA. The transgenic seeds contained less sulfate and more total amino acid sulfur than the nontransgenic parent line. This was associated with a 94% increase in methionine content and a 12% reduction in cysteine content. There was no statistically significant change in other amino acids or in total nitrogen or total sulfur contents of the seeds. In feeding trials with rats, the transgenic seeds gave statistically significant increases in live weight gain, true protein digestibility, biological value, and net protein utilization, compared with wild-type seeds. These findings demonstrate the feasibility of using genetic engineering to improve the nutritive value of grain crops.

BIOCHEMICAL STUDIES ON BLACK GRAM (PHASEOLUS MUNGO L.) 111. FERMENTATION OF THE BLACK GRAM AND RICE BLEND AND ITS INFLUENCE ON THE IN VITRO DIGESTIBILITY OF THE PROTEINS

Fermentation of black gram and rice blend was investigated for the microbiological, physicochemical, and biochemical changes. Total soluble solids, soluble nitrogen, and soluble acids increased during fermentation whereas soluble sugars were decreased. Trypsin inhibiting activity was unaffected although chymotrypsin inhibiting activity was progressively reduced during fermentation process. Amino acid contents of the batter were increased with the increase in fermentation time with the exceptions of isoleucine and cysteine. Increase in methionine content was 60 and 420%, respectively at 20 and 45 h fermentation. Enzymic digestion with pepsin and pancreatin indicated improvement in the availability of essential amino acids during the fementation. Fermentation, thus, appeared to improve the nutritional quality of the proteins in the black gram and rice blend.

Sulfite: Preferential sulfur source and modifier of CO2 fixation in Chlorella vulgaris

Sulfite was added at the time of inoculation to a standard and to a sulfate deficient medium of Chlorella vulgaris. It was not only used as a sulfur source, but besides this, at concentrations <1.0 mmol l(-1), the growth yield was enhanced up to 30% compared to sulfate saturated conditions. Higher sulfite concentrations increasingly inhibited cell growth. Growth rate determinations indicated that the enhancement, and the inhibition respectively, were confined to the very beginning of culture growth; the time period during which the sulfite was not yet oxidized (5-10 h). In contrast, an increased CO2 fixation rate/unit of protein, occurring up to 5.0 mmol l(-1) sulfite and a shift towards the β-carboxylation pathway, are persisting at least during the growth period of 4 days. The preferential uptake of sulfite, also indicated by a marked increase in methionine content of algal protein, presumably causes an increase in thylakoidal sulfolipids, and is such modifying the CO2 fixation.

Formation of Methionine from α-Amino-n-Butyric Acid and 5′-Methylthioadenosine in the Rat

Adult rats may utilize two metabolites of methionine for the biosynthesis of this essential amino acid. In separate experiments methionine, labeled with 14C or with 35S was observed in plasma and urine following the administration of [2-14C]-alpha-amino-n-butyric acid or [35S]-5'-methylthioadenosine by stomach tube. Although alpha-amino-n-butyric acid (ABA) or homoserine, alone or with dietary sodium sulfate, choline, and/or S-methylcysteine, was not utilized for growth, weight loss in weanling rats was decreased by dietary cysteine when fed as an additive to a basal methionine-free, cysteine-free, labile methyl-free, sulfur-free diet. Following the addition of 10 mg ABA and 28 mg 5'-methylthioadenosine/day to the basal diet, growth response was equivalent to that occurring in rats receiving 27 mg of methionine/day with the basal diet. The implications of these findings for adaptation to protein restriction and a discussion of equilibrium and steady state conditions related to the increase in methionine content in the blood are presented.

Hereditary Trichodysplasia: Marie Unna'S Hypotrichosis

This is a study of eight members of one family with Marie Unna hypotrichosis occurring in five generations. All affected individuals were born with widespread facial milia, sparse scalp hair and decreased body hair. Laboratory studies including urinary amino acids and plasma testosterone were normal. On histologic examination the hair follicles of the scalp showed proliferation of the internal root sheath and horn pearl formation in the lower third of the follicle. Abnormal hairs were flat and ribbon-like and twisted at irregular intervals. Extensive peeling of the cuticle was demonstrated by scanning electron microscopy. Electron microscopy of the hair shafts revealed intracellular fractures of the cuticular cells, increased interfibrillar matrix and fractures of the cortical cell fibrils and fractures of the medullary cells. X-ray diffraction studies were normal. On amino acid analysis of affected hairs a small decrease in cysteine-cystine and an increase in methionine content was noted. Since the hair shafts are clearly abnormal in this disorder, the name hereditary trichodysplasia is suggested. The condition is inherited as an autosomal dominant trait.