Tyrosine Supplementation Research Articles

Characterization and Enhanced Production of Fungal Melanin from Aspergillus Species Isolated from Soil

This study aimed to isolate and identify fungi from soil samples, focusing on their ability to produce melanin. Eleven different genera of fungi were isolated and identified using lactophenol cotton blue staining and microscopic examination, referencing H.L. Barnett and Barry B. Hunter. Among these, only Aspergillus was found to produce melanin. Optimal melanin production conditions were determined to be a 1.5% tyrosine supplement in the growth medium, under shaking conditions (120 rpm) and dark incubation for three weeks, resulting in a yield of 21.08 mg/100 ml. Physiochemical characterization revealed that the extracted melanin was insoluble in organic solvents but soluble in alkaline solutions (NaOH, KOH), and partially soluble in DMSO. Spectrometric analysis using UV-Visible spectroscopy showed characteristic absorption peaks. FTIR indicated functional groups and Scanning Electron Microscopy (SEM) images displayed a granular and heterogeneous surface topology. The study also assessed the effects of different carbon and nitrogen sources, and trace elements on melanin production. Maltose and sucrose were the most effective carbon sources, while peptone was the most effective nitrogen source. Among trace elements, calcium significantly enhanced melanin yield, while copper and zinc had moderate effects. These findings provide valuable insights into optimizing fungal melanin production and its potential industrial applications. The study highlights the sustainable and scalable production of melanin from Aspergillus, contributing to the broader understanding of fungal metabolites and their commercial exploitation. Future research should focus on genetic and metabolic pathways to further enhance melanin biosynthesis and explore its diverse applications.

Evolutionary history of tyrosine-supplementing endosymbionts in pollen-feeding beetles.

Many insects feeding on nutritionally challenging diets like plant sap, leaves, or wood engage in ancient associations with bacterial symbionts that supplement limiting nutrients or produce digestive or detoxifying enzymes. However, the distribution, function, and evolutionary dynamics of microbial symbionts in insects exploiting other plant tissues or relying on a predacious diet remain poorly understood. Here, we investigated the evolutionary history and function of the intracellular gamma-proteobacterial symbiont "Candidatus Dasytiphilus stammeri" in soft-winged flower beetles (Coleoptera, Melyridae, Dasytinae) that transition from saprophagy or carnivory to palynivory (pollen-feeding) between larval and adult stage. Reconstructing the distribution of the symbiont within the Dasytinae phylogeny unraveled not only a long-term coevolution, originating from a single acquisition event with subsequent host-symbiont codiversification, but also several independent symbiont losses. The analysis of 20 different symbiont genomes revealed that their genomes are severely eroded. However, the universally retained shikimate pathway indicates that the core metabolic contribution to their hosts is the provisioning of tyrosine for cuticle sclerotization and melanization. Despite the high degree of similarity in gene content and order across symbiont strains, the capacity to synthesize additional essential amino acids and vitamins and to recycle urea is retained in some but not all symbionts, suggesting ecological differences among host lineages. This report of tyrosine-provisioning symbionts in insects with saprophagous or carnivorous larvae and pollen-feeding adults expands our understanding of tyrosine supplementation as an important symbiont-provided benefit across a broad range of insects with diverse feeding ecologies.

Tyrosine catabolism enhances genotoxic chemotherapy by suppressing translesion DNA synthesis in epithelial ovarian cancer

Amino acid metabolism has been actively investigated as a potential target for antitumor therapy, but how it may alter the response to genotoxic chemotherapy remains largely unknown. Here, we report that the depletion of fumarylacetoacetate hydrolase (FAH), an enzyme that catalyzes the final step of tyrosine catabolism, reduced chemosensitivity in epithelial ovarian cancer (EOC). The expression level of FAH correlated significantly with chemotherapy efficacy in patients with EOC. Mechanistically, under genotoxic chemotherapy, FAH is oxidized at Met308 and translocates to the nucleus, where FAH-mediated tyrosine catabolism predominantly supplies fumarate. FAH-produced fumarate binds directly to REV1, resulting in the suppression of translesion DNA synthesis (TLS) and improved chemosensitivity. Furthermore, invivo tyrosine supplementation improves sensitivity to genotoxic chemotherapeutics and reduces the occurrence of therapy resistance. Our findings reveal a unique role for tyrosine-derived fumarate in the regulation of TLS and may be exploited to improve genotoxic chemotherapy through dietary tyrosine supplementation.

Effects of Dietary Cystine and Tyrosine Supplementation on Melanin Synthesis in the Pacific Oyster (Crassostrea gigas).

Melanogenesis is a multistep process to produce melanin for dark pigmentation in skin coloration. Previous studies in vertebrates demonstrated that cystine and tyrosine amino acids are involved in the melanin synthesis. However, very little is known about the melanogenesis in bivalve. In this study, cystine supplementation for 30days significantly upregulated the expression of CgB-aat1, CgCbs and CgTyr and pheomelanin content in the Pacific oyster Crassostrea gigas. Transmission electron microscope (TEM) results revealed more melanosomes in the connective tissue and melanin granules were secreted in epithelium of mantle. In contrast, tyrosine supplementation had no clear effect on melanogenesis except the gene expression changes of CgB-aat1 and CgCbs. In addition, prolonged supplementation of cystine or tyrosine for 60days had a negative impact on melanogenesis. Indeed, after 60days, expression of most of the melanin synthesis-related genes under study was decreased, and melanin content was significantly reduced, indicating that cystine and tyrosine might inhibit production of eumelanin and pheomelanin, respectively. In addition, in vitro analysis using primary cell culture from mantle tissue indicated that incubation with cystine, tyrosine, or B-AAT1 polypeptide, CBS/TYR recombinant proteins induced the increase of CgB-aat1 and CgCbs expression in a dose-dependent manner, suggesting the presence of a regulatory network in response to cystine and tyrosine amino acids intakes in pheomelanin synthesis-related gene expression. Taken together, these data indicate that cystine-CgB-aat1-CgCbs-CgTyr axis is a potential regulator of the pheomelanin biosynthesis pathway, and thus plays an important role in the mantle pigmentation in C. gigas. This work provides a new clue for selective cultivation of oyster strains with specific shell colors in bivalve breeding.

Influence Of Tyrosine On Body Temperature And Perceptual Responses To Cold Water Immersion In Military Cold-weather Medicine Trainees

Cold stress often results in physical and cognitive performance decrements. Tyrosine supplementation may attenuate deficits due to its ability to synthesize and maintain appropriate levels of brain catecholamines. However, the influence of tyrosine on physiological and perceptual responses to cold water immersion has not been reported. PURPOSE: Determine the influence of tyrosine supplementation on physiological and perceptual responses to cold water immersion during a military field training exercise. METHODS: Military personnel (n = 14; age: 26 ± 7 y; body fat: 20.1 ± 8.5%) completed a field training exercise that included a 10-min, head-out, cold water (1.5 °C) immersion in cold air (4 °C). In a randomized, double-blind design, prior to immersion (range 30-120 min), participants ingested either a food bar with 150 mg/kg of body weight of supplemental tyrosine (TYR; n = 7) or placebo (PLA; n = 7). TYR and PLA food bars were matched for taste, texture, and color. Prior to, during, and after immersion, participants’ heart rate (HR), hand temperature (Thand), skin temperature (Tsk), and core temperature (Tc) were measured while ratings of thermal sensation (TS) and shivering sensation (SS) were obtained throughout the exercise. Comparisons between TYR and PLA were made using an analysis of variance with significance set at p < 0.05. RESULTS: Oral tyrosine supplementation improved Tc maintenance, with the greatest difference between TYR and PLA evidenced at 10 min after completion of immersion (TYR: 36.7 ± 0.5, PLA 35.7 ± 0.8 °C, p < 0.05). No differences were observed between TYR and PLA for HR (p = 0.284), Thand (p = 0.693), Tsk (p = 0.947), TS (p = 0.262), or SS (p = 0.068). CONCLUSION: Findings suggest that tyrosine supplementation improved Tc maintenance in response to cold water immersion. However, tyrosine supplementation had minimal influence on thermal perceptions and did not modify HR or Tsk prior to, during, or after immersion. The conservation of core temperature associated with tyrosine supplementation is consistent with other reports, and its mechanisms of action warrant further investigation. This information is critical for deeper understanding of the complex physiological responses to environmental extremes in order to optimize strategies to sustain or enhance human performance in cold environments.

What are the effects of tyrosine supplementation for people with phenylketonuria? A Cochrane Review summary with commentary.

What are the effects of tyrosine supplementation for people with phenylketonuria? A Cochrane Review summary with commentary.

Effects of Dietary Phenylalanine and Tyrosine Supplements on the Chronic Stress Response in the Seabream (Sparus aurata).

The increase of aquaculture production is associated with a growing interest in improving physiological status and welfare in fish. For this reason, the search for strategies for mitigating stress has been intensified, with one of these strategies being food supplementation with different amino acids (AA). The objective of this study was to evaluate the effects of dietary phenylalanine (Phe) and tyrosine (Tyr) supplements on the endocrine and physiological state of seabreams (Sparus aurata) subjected to chronic stress. The fish were stocked at 30 fish/tank in a recirculation aquatic system, fed one control diet and two diets supplemented with 5% Phe or Tyr for 90 days. Blood was drawn from 10 fish per tank every 30 days, and the weight and length were measured every 15 days. At the end of the experiment, length/weight of the fish were measured, and they were sacrificed for the extraction of blood, head kidney, liver, and brain. Classic plasma stress markers (glucose, lactate, proteins, and cortisol), as well as hormones derived from Phe and Tyr (adrenaline, norepinephrine, and dopamine) and the accumulation of AA were analyzed. Fish fed with diets supplemented with Phe or Tyr showed a reduction in various stress markers and physiological parameters. In addition, the stress condition favored a mobilization of AA toward the tissues, especially in supplemented diets, so this excess of AA could be used as an energy substrate to cope with stress.

Nutritional symbionts enhance structural defence against predation and fungal infection in a grain pest beetle.

ABSTRACTMany insects benefit from bacterial symbionts that provide essential nutrients and thereby extend the hosts’ adaptive potential and their ability to cope with challenging environments. However, the implications of nutritional symbioses for the hosts’ defence against natural enemies remain largely unstudied. Here, we investigated whether the cuticle-enhancing nutritional symbiosis of the saw-toothed grain beetle Oryzaephilus surinamensis confers protection against predation and fungal infection. We exposed age-defined symbiotic and symbiont-depleted (aposymbiotic) beetles to two antagonists that must actively penetrate the cuticle for a successful attack: wolf spiders (Lycosidae) and the fungal entomopathogen Beauveria bassiana. While young beetles suffered from high predation and fungal infection rates regardless of symbiont presence, symbiotic beetles were able to escape this period of vulnerability and reach high survival probabilities significantly faster than aposymbiotic beetles. To understand the mechanistic basis of these differences, we conducted a time-series analysis of cuticle development in symbiotic and aposymbiotic beetles by measuring cuticular melanisation and thickness. The results reveal that the symbionts accelerate their host's cuticle formation and thereby enable it to quickly reach a cuticle quality threshold that confers structural protection against predation and fungal infection. Considering the widespread occurrence of cuticle enhancement via symbiont-mediated tyrosine supplementation in beetles and other insects, our findings demonstrate how nutritional symbioses can have important ecological implications reaching beyond the immediate nutrient-provisioning benefits.

Metabolic engineering of Saccharomyces cerevisiae for hydroxytyrosol overproduction directly from glucose.

SummaryHydroxytyrosol (HT) is one of the most powerful dietary antioxidants with numerous applications in different areas, including cosmetics, nutraceuticals and food. In the present work, heterologous hydroxylase complex HpaBC from Escherichia coli was integrated into the Saccharomyces cerevisiae genome in multiple copies. HT productivity was increased by redirecting the metabolic flux towards tyrosol synthesis to avoid exogenous tyrosol or tyrosine supplementation. After evaluating the potential of our selected strain as an HT producer from glucose, we adjusted the medium composition for HT production. The combination of the selected modifications in our engineered strain, combined with culture conditions optimization, resulted in a titre of approximately 375 mg l−1 of HT obtained from shake‐flask fermentation using a minimal synthetic‐defined medium with 160 g l−1 glucose as the sole carbon source. To the best of our knowledge, this is the highest HT concentration produced by an engineered S. cerevisiae strain.

Natural history of children and adults with phenylketonuria in the NBS-PKU Connect registry

PurposePhenylalanine hydroxylase deficiency, or phenylketonuria (PKU), is a rare autosomal recessive metabolic disorder. Early diagnosis via newborn screening (NBS) and initiation of treatment prevent the development of cognitive impairment and other co-morbidities. The purpose of this study is to describe the natural history of PKU in the United States, including prevalence of co-morbidities and predictors of outcomes. MethodsWe analyzed data from a self-report survey in the NBS-PKU Connect online registry. We describe the participants' nutrition management strategies, barriers to management, outcomes of bone disorders, skin, and psychological co-morbidities, and the use of special education or other special services. Predictors of outcomes were identified and assessed, including the impact of sex, age, age at diagnosis, blood phenylalanine concentration, use of sapropterin, use of medical food, adherence to prescribed diet, use of low protein modified foods, whether they had ever been off-diet, and use of tyrosine supplementation. ResultsThe 219 respondents included individuals with PKU or hyperphenylalanemia (n = 78), or their caregivers (n = 141). Most (84.3%) started treatment before the age of two weeks. About one-third indicated that they had been off-diet at some point in their lives, and 81.4% reported that they currently adhered to their prescribed diet, with adherence to prescribed diet decreasing with age. Blood phenylalanine concentration was under the recommended threshold of 360 μmol/L for 68.5% of participants. One-quarter of respondents reported psychological co-morbidities, with anxiety and ADD/ADHD being the most common. The incidence of psychological co-morbidities increased with age and with ever having been off diet. Special education or other special services were more likely to be reported by individuals who were diagnosed after one week of age. Skin disorders such as acne and eczema were more common in females than males, and a minority of participants reported bone disorders. ConclusionsDespite recommendations to maintain blood phenylalanine concentrations in the therapeutic range throughout life, it is not uncommon for adults with PKU to discontinue dietary management of their disorder. Early diagnosis was associated with reduced need for special education or other special services, and continuous treatment was associated with decreased psychological co-morbidities.

Tyrosine Supplementation A Nutraceutical Approach to Counter Heat Stress Induced Cognitive Decline

Supplementation of tyrosine, non-essential amino acid, and precursor of catecholamine was found to ameliorate the heat-induced alterations in latencies of event-related potential P300 and contingent negative variation. Here we present the effect of tyrosine supplementation on heat stress (exposure to ambient temperature 45 oC and relative humidity 30 %) induced alterations in behavior (attention, mood) and levels of plasma monoamines. Ten healthy male participants received a placebo food bar or tyrosine-containing food bar (6.5 g in 50 g) 90 min before heat stress exposure of 90 min. Plasma and urine samples were assayed for catecholamine levels, their precursor, and metabolites using high-performance liquid chromatography. A computer-based automated test battery was used to assess attention and mood by profile of mood states questionnaire. A significantly higher plasma tyrosine (p&lt;0.001) leading to an increased norepinephrine (p&lt;0.05) levels in the tyrosine supplemented group was observed. Selective (p&lt;0.001) and sustained attention (p&lt;0.02) in the tyrosine group were significantly better compared to the placebo group. Reaction time and anger scores decreased (p&lt;0.001) with tyrosine supplementation. It may be concluded that tyrosine supplementation improves heat stress-induced decrement in attention by maintaining the synthesis and turnover of norepinephrine.

Inhibition of a nutritional endosymbiont by glyphosate abolishes mutualistic benefit on cuticle synthesis in Oryzaephilus surinamensis

Glyphosate is widely used as a herbicide, but recent studies begin to reveal its detrimental side effects on animals by targeting the shikimate pathway of associated gut microorganisms. However, its impact on nutritional endosymbionts in insects remains poorly understood. Here, we sequenced the tiny, shikimate pathway encoding symbiont genome of the sawtoothed grain beetle Oryzaephilus surinamensis. Decreased titers of the aromatic amino acid tyrosine in symbiont-depleted beetles underscore the symbionts’ ability to synthesize prephenate as the precursor for host tyrosine synthesis and its importance for cuticle sclerotization and melanization. Glyphosate exposure inhibited symbiont establishment during host development and abolished the mutualistic benefit on cuticle synthesis in adults, which could be partially rescued by dietary tyrosine supplementation. Furthermore, phylogenetic analyses indicate that the shikimate pathways of many nutritional endosymbionts likewise contain a glyphosate sensitive 5-enolpyruvylshikimate-3-phosphate synthase. These findings highlight the importance of symbiont-mediated tyrosine supplementation for cuticle biosynthesis in insects, but also paint an alarming scenario regarding the use of glyphosate in light of recent declines in insect populations.

Tyrosine supplementation for phenylketonuria.

Phenylketonuria is an inherited disease for which the main treatment is the dietary restriction of the amino acid phenylalanine. The diet has to be initiated in the neonatal period to prevent or reduce mental handicap. However, the diet is very restrictive and unpalatable and can be difficult to follow. A deficiency of the amino acid tyrosine has been suggested as a cause of some of the neuropsychological problems exhibited in phenylketonuria. Therefore, this review aims to assess the efficacy of tyrosine supplementation for phenylketonuria. This is an update of previously published versions of this review. To assess the effects of tyrosine supplementation alongside or instead of a phenylalanine-restricted diet for people with phenylketonuria, who commenced on diet at diagnosis and either continued on the diet or relaxed the diet later in life. To assess the evidence that tyrosine supplementation alongside, or instead of a phenylalanine-restricted diet improves intelligence, neuropsychological performance, growth and nutritional status, mortality rate and quality of life. We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group's Trials Register which is comprised of references identified from comprehensive electronic database searches, handsearches of relevant journals and abstract books of conference proceedings. Additional studies were identified from handsearches of the Journal of Inherited Metabolic Disease (from inception in 1978 to 1998). The manufacturers of prescribable dietary products used in the treatment of phenylketonuria were also contacted for further references. Date of the most recent search of the Group's Inborn Errors of Metabolism Trials Register: 07 December 2020. All randomised or quasi-randomised trials investigating the use of tyrosine supplementation versus placebo in people with phenylketonuria in addition to, or instead of, a phenylalanine-restricted diet. People treated for maternal phenylketonuria were excluded. Two authors independently assessed the trial eligibility, methodological quality and extracted the data. Six trials were found, of which three trials reporting the results of a total of 56 participants, were suitable for inclusion in the review. The blood tyrosine concentrations were significantly higher in the participants receiving tyrosine supplements than those in the placebo group, mean difference 23.46 (95% confidence interval 12.87 to 34.05). No significant differences were found between any of the other outcomes measured. The trials were assessed as having a low to moderate risk of bias across several domains. From the available evidence no recommendations can be made about whether tyrosine supplementation should be introduced into routine clinical practice. Further randomised controlled studies are required to provide more evidence. However, given this is not an active area of research, we have no plans to update this review in the future.

Downregulation of the tyrosine degradation pathway extends Drosophila lifespan.

Aging is characterized by extensive metabolic reprogramming. To identify metabolic pathways associated with aging, we analyzed age-dependent changes in the metabolomes of long-lived Drosophila melanogaster. Among the metabolites that changed, levels of tyrosine were increased with age in long-lived flies. We demonstrate that the levels of enzymes in the tyrosine degradation pathway increase with age in wild-type flies. Whole-body and neuronal-specific downregulation of enzymes in the tyrosine degradation pathway significantly extends Drosophila lifespan, causes alterations of metabolites associated with increased lifespan, and upregulates the levels of tyrosine-derived neuromediators. Moreover, feeding wild-type flies with tyrosine increased their lifespan. Mechanistically, we show that suppression of ETC complex I drives the upregulation of enzymes in the tyrosine degradation pathway, an effect that can be rescued by tigecycline, an FDA-approved drug that specifically suppresses mitochondrial translation. In addition, tyrosine supplementation partially rescued lifespan of flies with ETC complex I suppression. Altogether, our study highlights the tyrosine degradation pathway as a regulator of longevity.

Tyrosine supplement ameliorates murine aGVHD by modulation of gut microbiome and metabolome.

BackgroundMicrobial communities and their metabolic components in the gut are of vital importance for immune homeostasis and have an influence on the susceptibility of the host to a number of immune-mediated diseases like acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, little is known about the functional connections between microbiome and metabolome in aGVHD due to the complexity of the gastrointestinal environment.MethodInitially, gut microbiota and fecal metabolic phenotype in aGVHD murine models were unleashed by performing 16S ribosomal DNA gene sequencing and ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS)-based metabolomics.FindingsThe group with aGVHD experienced a significant drop in Lachnospiraceae_unclassified but an increase in the relative abundance of Clostridium XI, Clostridium XIVa and Enterococcus. Meanwhile, a lower content of tyrosine was observed in the gut of aGVHD mice. The correlation analysis revealed that tyrosine-related metabolites were inversely correlated with Clostridium XIVa, besides, Blautia and Enterococcus also displayed the negative tendency in aGVHD condition. Apart from exploring the importance and function of tyrosine, different tyrosine diets were offered to mice during transplantation. Additional tyrosine supplements can improve overall survival, ameliorate symptoms at the early stage of aGVHD and change the structure and composition of gut microbiota and fecal metabolic phenotype. In addition, aGVHD mice deprived from tyrosine displayed worse manifestations than the vehicle diet group.InterpretationThe results demonstrated the roles and mechanisms of gut microbiota, indispensable metabolites and tyrosine in the progression of aGVHD, which can be an underlying biomarker for aGVHD diagnosis and treatment.FundingThis research was funded by the International Cooperation and Exchange Program (81520108002), the National Key R&D Program of China, Stem Cell and Translation Research (2018YFA0109300), National Natural Science Foundation of China (81670169, 81670148, 81870080 and 91949115) and Natural Science Foundation of Zhejiang Province (LQ19H080006).

Catecholaminergic modulation of the cost of cognitive control in healthy older adults.

Catecholamines have long been associated with cognitive control and value-based decision-making. More recently, we have shown that catecholamines also modulate value-based decision-making about whether or not to engage in cognitive control. Yet it is unclear whether catecholamines influence these decisions by altering the subjective value of control. Thus, we tested whether tyrosine, a catecholamine precursor altered the subjective value of performing a demanding working memory task among healthy older adults (60-75 years). Contrary to our prediction, tyrosine administration did not significantly increase the subjective value of conducting an N-back task for reward, as a main effect. Instead, in line with our previous study, exploratory analyses indicated that drug effects varied as a function of participants' trait impulsivity scores. Specifically, tyrosine increased the subjective value of conducting an N-back task in low impulsive participants, while reducing its value in more impulsive participants. One implication of these findings is that the over-the-counter tyrosine supplements may be accompanied by an undermining effect on the motivation to perform demanding cognitive tasks, at least in certain older adults. Taken together, these findings indicate that catecholamines can alter cognitive control by modulating motivation (rather than just the ability) to exert cognitive control.

Transcriptome Analysis of the Breast Muscle of Xichuan Black-Bone Chickens Under Tyrosine Supplementation Revealed the Mechanism of Tyrosine-Induced Melanin Deposition

The Xichuan black-bone chicken, which is a rare local chicken species in China, is an important genetic resource of black-bone chickens. Tyrosine can affect melanin production, but the molecular mechanism underlying tyrosine-induced melanin deposition in Xichuan black-bone chickens is poorly understood. Here, the blackness degree and melanin content of the breast muscle of Xichuan black-bone chickens fed a basic diet with five levels of added tyrosine (i.e., 0.2, 0.4, 0.6, 0.8, and 1.0%; these groups were denoted test groups I-V, respectively) were assessed, and the results showed that 0.8% tyrosine was the optimal level of added tyrosine. Moreover, the effects of tyrosine supplementation on the proliferation and tyrosinase content of melanocytes in Xichuan black-bone chickens were evaluated. The results revealed a dose-dependent relationship between tyrosine supplementation and melanocyte proliferation. In addition, 417 differentially expressed genes (DEGs), including 160 upregulated genes and 257 downregulated genes, were identified in a comparative analysis of the transcriptome profiles constructed using the pooled total RNA from breast muscle tissues of the control group and test group IV, respectively (fold change ≥2.0, P < 0.05). These DEGs were mainly involved in melanogenesis, the calcium signaling pathway, the Wnt signaling pathway, the mTOR signaling pathway, and vascular smooth muscle contraction. The pathway analysis of the DEGs identified some key genes associated with pigmentation, such as DCT and EDNRB2. In summary, the melanin content of breast muscle could be markedly enhanced by adding an appropriate amount of tyrosine to the diet of Xichuan black-bone chickens, and the EDNRB2-mediated molecular regulatory network could play a key role in the biological process of tyrosine-induced melanin deposition. These results have deepened the understanding of the molecular regulatory mechanism of melanin deposition in black-bone chickens and provide a basis for the regulation of nutrition and genetic breeding associated with melanin deposition in Xichuan black-bone chickens.

Effects of acute caffeine, theanine and tyrosine supplementation on mental and physical performance in athletes

BackgroundA limited amount of research has demonstrated beneficial effects of caffeine and theanine supplementation for enhancement of mental performance. The purpose of this investigation was to determine whether the acute ingestion of a supplement containing caffeine, theanine and tyrosine improves mental and physical performance in athletes.MethodsTwenty current or former male collegiate athletes (age: 20.5 ± 1.4 y; height: 1.82 ± 0.08 m; weight: 83.9 ± 12.6 kg; body fat: 13.8 ± 5.6%) completed this randomized, double-blind, placebo-controlled crossover trial. After familiarization, each participant completed two identical testing sessions with provision of a proprietary dietary supplement (SUP) containing caffeine theanine and tyrosine or a placebo (PL). Within each testing session, participants completed assessments of mental and physical performance before and after provision of SUP or PL, as well as after two rounds of exercise. Assessments were performed using a performance testing device (Makoto Arena) that evaluated multiple aspects of mental and physical performance in response to auditory and visual stimuli. Testing was performed both with the body in a static position and during dynamic movement. General linear models were used to evaluate the effects of SUP and PL on performance.ResultsChanges in movement accuracy during performance assessment were greater following SUP ingestion as compared to PL for both static and dynamic testing (SUP: + 0.4 to 7.5%; PL: − 1.4 to 1.4% on average; p < 0.05). For dynamic testing, the change in number of targets hit was higher and the change in average hit time was lower with SUP as compared to PL (p < 0.05). However, there were no differences between conditions for the changes in number of targets hit or average hit time during static testing. There were no differences in changes of subjective variables during either condition, and performance measures during the two rounds of exercise did not differ between conditions (p > 0.05).DiscussionThe present results indicate that a combination of a low-dose of caffeine with theanine and tyrosine may improve athletes’ movement accuracy surrounding bouts of exhaustive exercise without altering subjective variables. Based on this finding, supplementation with caffeine, theanine and tyrosine could potentially hold ergogenic value for athletes in sports requiring rapid and accurate movements.Trial registrationNCT03019523. Registered 24 January 2017.

Determination of free tyrosine in equestrian supplements by LC–MS/MS and comparison of its quantity with total free amino acids content in view of doping control

The reports on the probable beneficial impact of tyrosine (TYR) supplementation on performance enhancement have contributed to the growth of interest in TYR in equestrian sports field and related industries, such as the manufacture of dietary and nutritional supplements. In this study, the first attempt to the assessment of horses exposure to TYR during nutritional supplementation was demonstrated by quantification of unbound TYR and a total free amino acids (AAs), and subsequent determination of the ratio of TYR to a total free AAs content in equestrian supplement samples. Within the scope of this study, a rapid and sensitive LC–MS/MS-based method for the determination of unbound TYR and a simple spectrophotometric protocol for the quantification of a total free AAs have been developed and validated according to the international guidelines for bioanalytical methods. For sample preparation, an ultrasound-assisted solid-liquid extraction (USLE; for solid samples) and ultrasound-assisted liquid-liquid extraction (ULLE; for liquid samples) have been optimized. The comprehensive approach on the simultaneous assessment of matrix effect, recovery, process efficiency, accuracy, precision, curve factor and internal standard association demonstrated satisfactory validation parameters for LC–MS/MS-based procedure, such as global matrix effect in the range of 89.9–91.9%, good linearity (R2 > 0.9929), limit of detection (0.6 ng/mL) and recovery within 89.2–108.0%. For spectrophotometric method, limit of detection was 1100 ng/mL, recovery ranged from 103.2 to 108.8%, both, intra- and inter-day accuracy was within 89.5–108.7%, and both, intra- and inter-assay precision was below 8.5%. In view of the satisfactory validation parameters obtained for both methods, the procedures can be utilized for routine analysis of dietary and nutritional supplements.

Process development for the continuous production of heterologous proteins by the industrial yeast, Komagataella phaffii

The current trend in industrial biotechnology is to move from batch or fed‐batch fermentations to continuous operations. The success of this transition will require the development of genetically stable production strains, the use of strong constitutive promoters, and the development of new medium formulations that allow an appropriate balance between cell growth and product formation. We identified genes that showed high expression in Komagataella phaffii during different steady‐state conditions and explored the utility of promoters of these genes (Chr1–4_0586 and FragB_0052) in optimizing the expression of two different r‐proteins, human lysozyme (HuLy), and the anti‐idiotypic antibody fragment, Fab‐3H6, in comparison with the widely used glyceraldehyde‐3‐phosphate dehydrogenase promoter. Our results showed that the promoter strength was highly dependent on the cultivation conditions and thus constructs should be tested under a range of conditions to determine both the best performing clone and the ideal promoter for the expression of the protein of interest. An important benefit of continuous production is that it facilitates the use of the genome‐scale metabolic models in the design of strains and cultivation media. In silico flux distributions showed that production of either protein increased the flux through aromatic amino acid biosynthesis. Tyrosine supplementation increased the productivity for both proteins, whereas tryptophan addition did not cause any significant change and, phenylalanine addition increased the expression of HuLy but decreased that of Fab‐3H6. These results showed that a genome‐scale metabolic model can be used to assess the metabolic burden imposed by the synthesis of a specific r‐protein and then this information can be used to tailor a cultivation medium to increase production.