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1.
Braz. j. biol ; 84: e250936, 2024. graf
Article in English | MEDLINE, LILACS, VETINDEX | ID: biblio-1345557

ABSTRACT

Abstract This study was carried out to evaluate the effect of Glutamine, as a dipeptide or a free amino acid form, on the progression of burn injuries in rats. Thirty male Wistar rats were burned with a comb metal plate heated in boiling water (98 °C) for three minutes, creating four rectangular full-thickness burn areas separated by three unburned interspaces (zone of stasis) in both dorsum sides. The animals were randomized into three groups (n=10): saline solution (G1-Control) and treated groups that orally received Glutamine as dipeptide (G2-Dip) or free amino acid (G3-FreeAA). Two and seven days after burn injury, lesions were photographed for unburned interspaces necrosis evolution assessment. Seven days after injury, glutathione seric was measured and histopathological analysis was performed. By photographs, there was a significant reduction in necrosis progression in G3-Free-AA between days two and seven. Histopathological analysis at day 7 showed a significantly higher stasis zone without necrosis and a higher number of fibroblasts in G2-Dip and G3-FreeAA compared with G1-Control. Also, glutathione serum dosage was higher in G2-Dip. The plasmatic glutathione levels were higher in the G2-Dip than the G1-Control, and there was a trend to higher levels in G3-FreeAA. The reduction in histological lesions, greater production of fibroblasts, and greater amounts of glutathione may have benefited the evolution of burn necrosis, which showed greater preservation of interspaces.


Resumo Este estudo foi realizado para avaliar o efeito da Glutamina, como um dipeptídeo ou forma de aminoácido livre, na progressão de queimaduras em ratos. Trinta ratos Wistar machos foram queimados com um pente de metal aquecido em água fervente (98 °C) por três minutos, criando quatro áreas retangulares queimadas separadas por três interesespaços não queimados (zona de estase) em ambos os lados do dorso. Os animais foram randomizados em três grupos (n = 10): solução salina (G1-Controle) e grupos tratados que receberam glutamina via oral como dipeptídeo (G2-Dip) ou aminoácido livre (G3-FreeAA). Dois e sete dias após a queimadura, as lesões foram fotografadas para avaliação da evolução da necrose entre os espaços não queimados. Sete dias após a lesão, foi dosada a glutationa sérica e realizada análise histopatológica. Pelas fotografias, houve uma redução significativa na progressão da necrose no G3-Free-AA entre os dias dois e sete. A análise histopatológica no dia 7 mostrou uma zona de estase significativamente maior sem necrose e número mais elevado de fibroblastos em G2-Dip e G3-FreeAA em comparação com G1-Controle. Os níveis plasmáticos de glutationa foram maiores no G2-Dip em relação ao G1-Controle, e houve tendência a níveis mais elevados no G3-FreeAA. A redução das lesões histológicas, maior produção de fibroblastos, maior quantidade de glutationa podem ter beneficiado a evolução da necrose da queimadura, que mostrou maior preservação dos interespaços.


Subject(s)
Animals , Male , Rats , Burns/drug therapy , Glutamine , Rats, Wistar , Dipeptides , Disease Models, Animal , Amino Acids
2.
Braz. j. biol ; 83: e249422, 2023. tab
Article in English | MEDLINE, LILACS, VETINDEX | ID: biblio-1339393

ABSTRACT

Abstract Ninety days study was conducted in hapas installed in earthen ponds. Fish of an average initial weight (220g) were evenly distributed in triplicate groups within fifteen hapas. Five experimental diets labeled as T1 (25% CP and NRC recommended amino acid level) as control diet, T2 (with 2% low protein and 5% amino acid supplementation), T3 (with 2% low protein and 10% amino acid supplementation), T4 (with 4% low protein and 10% amino acid supplementation) and T5 (with 4% low protein and 20% amino acid supplementation) were prepared. Fish were fed with @3% of their body weight twice a day at 10.00 & 16:00 hour. Significantly higher percent weight gain (420.18 ± 66.84a) and specific growth rate (13499.33±1273.54a) along with improved feed conversion ratio (1.29 ± 0.09b) and hundred percent survivals were recorded during the trial. Furthermore proximate analysis of meat showed significant improvement in the crude protein level (81.77 ± 0.19a) served with diet containing 20% limiting amino acids mixture. Therefore, limiting amino acids can be a source of cost effective feed and use safely in L. rohita diet.


Resumo O estudo de 90 dias foi realizado em hapas instalados em tanques de terra. Peixes com peso inicial médio (220 g) foram distribuídos uniformemente em grupos triplicados em 15 hapas. Cinco dietas experimentais rotuladas como T1 (25% de CP e NRC recomendado nível de aminoácidos) como dieta controle, T2 (com 2% de proteína baixa e 5% de suplementação de aminoácidos), T3 (com 2% de proteína baixa e 10% de suplementação de aminoácidos), T4 (com 4% de baixa proteína e 10% de suplementação de aminoácidos) e T5 (com 4% de baixa proteína e 20% de suplementação de aminoácidos) foram preparadas. Os peixes foram alimentados com 3% do seu peso corporal duas vezes por dia às 10h00 e 16h00. Ganho de peso significativamente maior (420,18 ± 66,84a) e taxa de crescimento específico (13499,33 ± 1273,54a) juntamente com taxa de conversão alimentar melhorada (1, 29 ± 0,09b) e sobrevivência de cem por cento foram registrados durante o ensaio. Além disso, a análise aproximada da carne mostrou melhora significativa no nível de proteína bruta (81,77 ± 0,19a) servida com dieta contendo 20% de mistura de aminoácidos limitantes. Portanto, a limitação de aminoácidos pode ser uma fonte de alimentação econômica e usada com segurança na dieta de L. rohita.


Subject(s)
Animals , Cyprinidae , Animal Feed/analysis , Dietary Supplements , Diet/veterinary , Amino Acids
3.
Braz. j. biol ; 82: e240133, 2022. tab
Article in English | MEDLINE, LILACS, VETINDEX | ID: biblio-1278516

ABSTRACT

Abstract A field study was carried out to determine the influence of foliage applied plant growth promoter and retardant in improving soil applied sulphur fertilizer use efficiency in cotton during two consecutive summers 2014 and 2015. Experimental trial comprised of three different sources of sulphur (ammonium sulphate, potassium sulphate and elemental sulphur) and foliar spray of plant growth promoter and growth retardant including tap water was taken as control. Among treatments soil applied ammonium sulphate with foliage applied amino acid produced maximum plant height, sympodial branches, pods per plant, seed cotton yield, fiber yield, biological yield, protein contents, oil contents and leaf nitrogen uptake as compared to the other treatments. Whereas, soil applied potassium sulphate with foliar spray of mepiquat chloride on cotton significantly improved the boll weight and leaf potassium uptake. We conclude that soil applied ammonium sulphate and foliage spray of amino acid was more effective in improving the productivity and quality attributes of cotton.


Resumo Foi realizado um estudo de campo para determinar a influência do promotor de crescimento das plantas e retardador da folhagem em algodão, para melhora da eficiência do uso de fertilizantes à base de enxofre aplicados no solo durante dois verões consecutivos (2014 e 2015). O ensaio experimental foi composto de três fontes diferentes de enxofre (sulfato de amônio, sulfato de potássio e enxofre elementar) e pulverização foliar do promotor de crescimento de plantas e retardador de crescimento, incluindo água da torneira que foi tomada como controle. Entre os tratamentos, o sulfato de amônio aplicado no solo com aminoácido aplicado na folhagem produziu o máximo na altura da planta, ramos simodiais, capulhos por planta, rendimento de algodão em caroço, rendimento de fibra, rendimento biológico, conteúdo de proteínas, conteúdo de óleo e absorção de nitrogênio nas folhas quando comparado a outros tratamentos. Enquanto o solo fertilizado com sulfato de potássio e aplicação foliar de cloreto de mepiquat no algodão melhorou, significativamente, o peso do capulho e a absorção de potássio nas folhas. Sulfato de amônio aplicado no solo e a aplicação foliar de aminoácidos foram mais eficazes na melhora dos atributos de produtividade e qualidade do algodão.


Subject(s)
Amino Acids , Nitrogen , Soil , Fertilizers , Ammonium Sulfate
4.
Arq. bras. med. vet. zootec. (Online) ; 73(4): 929-937, Jul.-Aug. 2021. tab, graf, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1285261

ABSTRACT

The objective of the present study was to investigate the different plasma metabolites between anestrus and estrus postpartum dairy cows and to provide a theoretical basis for prevention of anestrus in dairy farm cows. In the experiment, one hundred and sixty-seven Holstein dairy cows were selected with similar age and parity. According to the concentration of ß-hydroxybutyric acid, non-esterified fatty acids and glucose in plasma during 14 to 21 days in milk, all dairy cows were determined as having a status of energy balance. According to the results of clinical symptom, rectal and B ultrasound examination at 60 to 90 days postpartum, these cows were divided into twenty estrus and twenty-four anestrus group, other dairy cows were removed. 1H nuclear magnetic resonance technology was utilized to detect the plasma metabolites changes and screen different plasma metabolites between anestrus and estrus cows. Ten different metabolites including alanine, glutamic acid, asparagine, creatine, choline, phosphocholine, glycerophosphocholine, low-density lipoprotein, and very-low-density lipoprotein were significantly decreased in anestrous cows compared with estrous cows. Metabolic pathway analyses indicated that differential metabolites were primarily involved in amino acid and glycerophospholipid metabolism. These metabolites and their enrichment pathways indicate that reduced steroid hormone synthesis precursors result in lower levels of estradiol and progesterone and cause anestrus in negative energy balance. These data provide a better understanding of the changes that may affect estrus of postpartum dairy cows at NEB status and lay the ground for further research.(AU)


O objetivo do presente estudo foi investigar os diferentes metabolitos do plasma entre o cio e o cio pós-parto de vacas leiteiras e fornecer uma base teórica para a prevenção do cio de vacas em fazendas de leite. No experimento, foram selecionadas 127 vacas leiteiras Holstein com idade e paridade similares. De acordo com a concentração de ß- ácido hidroxibutírico, ácidos graxos não esterificados e glicose no plasma entre 14 e 21 dias no leite, todas as vacas leiteiras foram determinadas em estado de equilíbrio energético. De acordo com os resultados dos sintomas clínicos, do exame de ultra-som retal e B aos 60 a 90 dias pós-parto, estas vacas foram divididas em vinte cios e vinte e quatro grupos de cio, outras vacas leiteiras foram removidas. A tecnologia de ressonância magnética nuclear 1H foi utilizada para detectar as alterações dos metabólitos plasmáticos e para triar diferentes metabólitos plasmáticos entre as vacas do cio e do cio. Dez diferentes metabólitos incluindo alanina, ácido glutâmico, asparagina, creatina, colina, fosfocholina, glicerofosfocolina, lipoproteína de baixa densidade e lipoproteína de muito baixa densidade foram significativamente diminuídos nas vacas antróficas em comparação com as vacas estro. As análises da via metabólica indicaram que os metabólitos diferenciais estavam principalmente envolvidos no metabolismo de aminoácidos e glicerofosfolipídios. Estes metabólitos e suas vias de enriquecimento indicam que a redução dos precursores da síntese de hormônios esteróides resulta em níveis mais baixos de estradiol e progesterona e causa anestros no balanço energético negativo. Estes dados fornecem uma melhor compreensão das mudanças que podem afetar o cio das vacas leiteiras pós-parto no estado de NEB e preparam o terreno para mais pesquisas.(AU)


Subject(s)
Animals , Female , Cattle , Progesterone/analysis , Anestrus/blood , Estrus/blood , Postpartum Period/blood , Estradiol/analysis , Glycerophospholipids , Fatty Acids, Nonesterified , Amino Acids , Glucose , Hematologic Tests/veterinary
5.
Braz. j. biol ; 81(1): 44-52, Feb. 2021. tab
Article in English | LILACS | ID: biblio-1153322

ABSTRACT

Abstract Three trials were executed to examine the nutritive profile, metabolisable energy and digestible amino acid (AA) contents of four indigenous feed ingredients including wheat (W), wheat middling (WM), canola meal (CM) and rapeseed meal (RSM) in Nile tilapia. Three samples of each test ingredient were collected from three different locations of Multan (MUL) and Sukkar (SKR), of Pakistan. The collected three samples were pooled thereafter to make a homogenous/ representative sample of each test ingredient from a particular study site. Nutrients composition, AA and energy digestibility of these indigenous ingredients were evaluated by using laboratory analyses and fish studies. Proximate analysis indicated variations in some of the nutrients due to location (p < 0.05). Differences were also observed in some AA including arginine, lysine, serine, cysteine, glutamic and aspartic acids, histidine, valine and glycine contents of these ingredients (p < 0.05). Digestibility of leucine, glycine and glutamic acid was higher (p < 0.05) in RSM from MUL. Among W samples from MUL, AA digestibility for lysine, threonine, and aspartic acid was higher (p < 0.05). Crude protein, arginine, alanine, serine, and aspartic acid had higher digestibility (p < 0.05), whereas digestibility was lower (p < 0.05) for threonine, valine and tyrosine in RSM from MUL. Metaboliseable energy contents did not differ among W, WM, CM and RSM regarding their origin (p > 0.05). The results indicated that nutritional profiles and their digestibility indices vary with the location for Nile tilapia.


Resumo Três experimentos foram executados para examinar o perfil nutritivo, a energia metabolizável e o conteúdo de aminoácidos digestíveis (AA) de quatro ingredientes alimentícios, incluindo trigo (W), farelo de trigo (WM), farelo de canola (CM) e farelo de colza (RSM) em tilápia do Nilo. Três amostras de cada ingrediente do teste foram coletadas de dois locais diferentes (Multan (MUL) e Sukkar (SKR), do Paquistão) e assim agrupadas. A composição nutricional, AA e digestibilidade energética desses ingredientes indígenas foram avaliadas por meio de análises laboratoriais e estudos de peixes. A análise imediata indicou variações (p <0,05) em alguns dos nutrientes devido à localização. Variações (p <0,05) também foram observadas em alguns teores de AA desses ingredientes. A digestibilidade da leucina, glicina e ácido glutâmico foi maior (p <0,05) em RSM de MUL. Entre as amostras de W da MUL, a digestibilidade de AA para Lys, Thr e Asp foi maior (p <0,05). Proteína Crud, arginina, alanina, serina e ácido aspártico apresentaram maior digestibilidade (p <0,05), e menor (p <0,05) para treonina, valina e tirosina em MRS. Nenhuma diferença (p> 0,05) relacionada a energia metabolizável foi observada entre esses ingredientes em relação à sua origem. Os resultados indicaram que os perfis nutricionais e sua digestibilidade variam com a localização.


Subject(s)
Animals , Cichlids , Amino Acids , Pakistan , Diet , Digestion , Ileum , Animal Feed/analysis , Animal Nutritional Physiological Phenomena
6.
Arch. argent. pediatr ; 119(1): e49-e53, feb. 2021. ilus
Article in Spanish | LILACS, BINACIS | ID: biblio-1147262

ABSTRACT

El raquitismo afecta la diferenciación y mineralización del cartílago de crecimiento como consecuencia, en última instancia, de una alteración en los niveles de fósforo y/o calcio. El secundario a la deficiencia de vitamina D es la forma más frecuente (raquitismo carencial). Las manifestaciones clínicas durante los primeros años de vida suelen comprometer en forma más marcada las epífisis de los huesos.Se describe el caso de un lactante de 8 meses con diagnóstico de alergia a la proteína de la leche de vaca que presentó múltiples fracturas patológicas mientras se encontraba bajo tratamiento con fórmulas lácteas a base de aminoácidos. Se efectuó el diagnóstico de raquitismo hipofosfatémico por deficiencia de fósforo y, tras 3 meses de tratamiento con sales de fosfato, calcio, calcitriol, el abandono paulatino de la leche elemental y el descenso gradual de la medicación antiácida, el paciente evolucionó con curación clínico-radiológica del cuadro


The rickets is a disease that affects the differentiation and mineralization of the growth cartilage, as an ultimate consequence of a balance loss in calcium and phosphate levels. Vitamin D deficiency is the most common cause of the rickets (nutritional rickets). Its clinical manifestation during the first years of life involves long bones epiphysis in a more severe way.We report an 8-month-old infant who was diagnosed with cow ́s milk protein allergy and suffered from multiple fractures while receiving elemental formula as part of his treatment. The final etiology was hypophosphatemic rickets secondary to phosphate deficiency, and after 3 months of phosphate, calcium and calcitriol supplementation, in addition to the gradually reduction of the proportion of elemental formula intake and the decline of the antacid doses, clinical and radiological heal was achieved.


Subject(s)
Humans , Male , Infant , Rickets, Hypophosphatemic/diagnostic imaging , Vitamin D Deficiency , Milk Hypersensitivity , Infant Formula , Rickets, Hypophosphatemic/therapy , Amino Acids
7.
Article in Chinese | WPRIM | ID: wpr-922057

ABSTRACT

OBJECTIVE@#To establish an amino acid assay for the determination of β-lactoglobulin in Anti-HPV biological protein dressing.@*METHODS@#Under acidic conditions, β-lactoglobulin is hydrolyzed into free amino acids, separated by cation exchange chromatography, and derivatived after ninhydrin column. The chromatogram at 570 nm is collected. The content of β-lactoglobulin in the sample is indirectly determined by measuring the lysine content obtained by hydrolysis.@*RESULTS@#β-lactoglobulin has a good linear relationship in the concentration range of 77.28~309.12 μg/mL (@*CONCLUSIONS@#The method is simple, specific, accurate and reproducible, which is suitable for the quantitative analysis of β-lactoglobulin in anti-HPV biological protein dressing.


Subject(s)
Amino Acids , Bandages , Lactoglobulins
8.
Chinese Journal of Biotechnology ; (12): 4215-4230, 2021.
Article in Chinese | WPRIM | ID: wpr-921500

ABSTRACT

Threonine aldolases catalyze the aldol condensation of aldehydes with glycine to furnish β-hydroxy-α-amino acid with two stereogenic centers in a single reaction. This is one of the most promising green methods for the synthesis of optically pure β-hydroxy-α-amino acid with high atomic economy and less negative environmental impact. Several threonine aldolases from different origins have been identified and characterized. The insufficient -carbon stereoselectivity and the challenges of balancing kinetic versus thermodynamic control to achieve the optimal optical purity and yield hampered the application of threonine aldolases. This review summarizes the recent advances in discovery, catalytic mechanism, high-throughput screening, molecular engineering and applications of threonine aldolases, with the aim to provide some insights for further research in this field.


Subject(s)
Amino Acids , Catalysis , Glycine , Glycine Hydroxymethyltransferase/metabolism , Kinetics , Substrate Specificity , Threonine
9.
Chinese Journal of Biotechnology ; (12): 3231-3241, 2021.
Article in Chinese | WPRIM | ID: wpr-921420

ABSTRACT

The source of recombinant collagen is clean, and it has the advantages of flexible sequence design, high yield and high purity, so it has a wide application prospect as biomaterials in tissue engineering and other fields. However, how to promote the cross-linking of recombinant collagen molecules and make them form a more stable spatial structure is the difficulty to be overcome in the design of recombinant collagen nanomaterials. Unnatural amino acid O-(2-bromoethyl)-tyrosine was incorporated into collagen by two-plasmid expression system. The results showed that high-purity collagen incorporated with unnatural amino acid could be obtained by induction with final concentration of 0.5 mmol/L IPTG and 0.06% arabinose at 25 °C for 24 hours. The intermolecular cross-linking through thioether bond was formed between collagen molecule incorporated with unnatural amino acid and collagen molecule with cysteine mutation in pH 9.0 NH4HCO3 buffer, which formed aggregates with the largest molecular size up to 1 micrometre. The results pave the way for the design of recombinant collagen biomaterials.


Subject(s)
Amino Acids , Biocompatible Materials , Collagen/genetics , Sulfides
10.
Acta Physiologica Sinica ; (6): 723-733, 2021.
Article in Chinese | WPRIM | ID: wpr-921275

ABSTRACT

Amino acids are essential nutrients for humans and have a wide range of biological functions. They are the constituent units of protein and energy metabolites. In addition, they are also widely involved in the maintenance and regulation of various physiological functions, and play a role in transcription, translation, post-translational modification and other levels. The liver is a key metabolic organ, and it acts as a hub that connects the metabolism of various tissues. Amino acid sensing plays a very important role in the regulation of hepatic glucose and lipid metabolism. Therefore, accurately sensing the levels of intracellular and extracellular amino acids is the key to maintaining cell homeostasis. There are several well-known amino acid sensors in eukaryotic cells, such as general control non-derepressible-2 (GCN2), mammalian target of rapamycin (mTOR) and taste receptors, which play an important role in maintaining metabolic homeostasis. This article gives a detailed introduction to the role and mechanism of amino acids in regulating hepatic glucose and lipid metabolism, laying a foundation for further exploration of amino acid sensing mechanism and treatment of hepatic glucose and lipid metabolism disorders.


Subject(s)
Amino Acids , Glucose/metabolism , Homeostasis , Humans , Lipid Metabolism , Liver
11.
Chinese Acupuncture & Moxibustion ; (12): 1095-1102, 2021.
Article in Chinese | WPRIM | ID: wpr-921016

ABSTRACT

OBJECTIVE@#To observe the effect of acupuncture combined with rehabilitation training on cognitive function and amino acid metabolism in children with cerebral palsy.@*METHODS@#Twenty children with cerebral palsy (cerebral palsy group) were randomly divided into an acupuncture and rehabilitation group (11 cases) and a rehabilitation group (9 cases), and 10 healthy children were included as the normal group. The rehabilitation group was treated with rehabilitation training, 30 min each time; on the basis of rehabilitation training, the acupuncture and rehabilitation group was treated with acupuncture at Sishenzhen, Zhisanzhen, Naosanzhen, Niesanzhen, Dazhui (GV 14), Shenzhu (GV 12), Mingmen (GV 4), etc. The Sishenzhen (left and right points) and the ipsilateral Niesanzhen were respectively connected with a group of electrodes, intermittent wave, frequency of 2 Hz for 30 min. Both groups were treated once every other day, three times a week, totaling for 3 months. The Gesell developmental diagnostic scale (GESELL) was used to evaluate the developmental quotient (DQ) scores before and after treatment, and the blood samples of children with cerebral palsy before and after treatment and normal children were collected for amino acid metabolomics.@*RESULTS@#Compared before treatment, the DQ scores of adaptation, fine motor and social in the acupuncture and rehabilitation group was increased after treatment (@*CONCLUSION@#Acupuncture combined with rehabilitation training could improve cognitive function in children with cerebral palsy, and the effect may be related to the down-regulation of L-histidine and L-citrulline metabolism.


Subject(s)
Acupuncture Points , Acupuncture Therapy , Amino Acids , Cerebral Palsy/therapy , Child , Cognition , Humans
12.
Article in Chinese | WPRIM | ID: wpr-879157

ABSTRACT

A comprehensive analytical method based on ultra-fast liquid chromatography coupled with triple quadrupole/linear ion trap tandem mass spectrometry(UFLC-QTRAP-MS/MS) was established for simultaneous determination of the content of 38 active components in Abelmoschi Corolla, including flavonoids, organic acids, nucleosides and amino acids, so as to investigate the effects of different harvesting and processing methods on multi-active components in Abelmoschi Corolla. The chromatographic separation was performed on a XBridg®C_(18) column(4.6 mm×100 mm, 3.5 μm) with(0.1% formic acid water) methanol-acetonitrile(1∶1) as the mobile phase for gradient elution at 30 ℃. The flow rate was 0.5 mL·min~(-1). The components were detected in a multiple-reaction monitoring(MRM) mode. The gray relational analysis(GRA) was used to comprehensively evaluate the multiple active components of Abelmoschi Corolla at different harvesting times and drying temperatures. The results showed that 38 components had a good linearity with correlation coefficients all above 0.999 0. The method featured a good precision, repeatability and stability with the relative stan-dard deviations(RSDs) of less than 5.0%. Recoveries ranged from 98.06% to 104.4% with RSD between 0.22% and 4.9%. The results of GRA indicated that a better quality in the samples collected on September 9 th. Samples dried at 90 ℃ had a better quality. The established method is accurate and reliable, and can be used to assess the internal quality of Abelmoschi Corolla. This study can provide basic materials for determining appropriate harvesting time and processing method of Abelmoschi Corolla.


Subject(s)
Amino Acids , Chromatography, High Pressure Liquid , Chromatography, Liquid , Nucleosides , Tandem Mass Spectrometry
13.
Chinese Journal of Biotechnology ; (12): 1677-1696, 2021.
Article in Chinese | WPRIM | ID: wpr-878661

ABSTRACT

Fermentative production of amino acids is one of the pillars of the fermentation industry in China. Recently, with the fast development of metabolic engineering and synthetic biology technologies, the metabolic engineering for production of amino acids has been flourishing. Conventional forward metabolic engineering, reversed metabolic engineering based on omics data and in silico simulation, and evolutionary metabolic engineering mimicking the natural evolution, have shown increasingly promising applications. A series of highly efficient and robust amino acids-producing strains have been developed and applied in the industrial production of amino acids. The increasingly fierce market competition has put forward new requirements for strain breeding and selection, such as developing high value-added amino acids, dynamic regulation of cellular metabolism, and adapting to the requirements of new process. This review summarizes the advances and prospects in metabolic engineering for the production of amino acids.


Subject(s)
Amino Acids , China , Corynebacterium glutamicum/genetics , Metabolic Engineering , Synthetic Biology
14.
Chinese Journal of Biotechnology ; (12): 1603-1618, 2021.
Article in Chinese | WPRIM | ID: wpr-878657

ABSTRACT

Corynebacterium glutamicum is an important workhorse of industrial biotechnology, especially for amino acid bioindustry. This bacterium is being used to produce various amino acids at a level of over 6 million tons per year. In recent years, enabling technologies for C. glutamicum metabolic engineering have been developed and improved, which accelerated construction and optimization of microbial cell factoriers, expanding spectra of substrates and products, and facilitated basic researches on C. glutamicum. With these technologies, C. glutamicum has become one of the ideal microbial chasses. This review summarizes recent key technological developments of enabling technologies for C. glutamicum metabolic engineering and focuses on establishment and applications of CRISPR-based genome editing, gene expression regulation, adaptive laboratory evolution, and biosensor technologies.


Subject(s)
Amino Acids , Biotechnology , Corynebacterium glutamicum/genetics , Gene Editing , Metabolic Engineering
15.
Chinese Journal of Biotechnology ; (12): 1510-1525, 2021.
Article in Chinese | WPRIM | ID: wpr-878652

ABSTRACT

¹³C metabolic flux analysis (¹³C-MFA) enables the precise quantification of intracellular metabolic reaction rates by analyzing the distribution of mass isotopomers of proteinogenic amino acids or intracellular metabolites through ¹³C labeling experiments. ¹³C-MFA has received much attention as it can help systematically understand cellular metabolic characteristics, guide metabolic engineering design and gain mechanistic insights into pathophysiology. This article reviews the advances of ¹³C-MFA in the past 30 years and discusses its potential and future perspective, with a focus on its application in industrial biotechnology and biomedicine.


Subject(s)
Amino Acids , Carbon Isotopes , Isotope Labeling , Metabolic Engineering , Metabolic Flux Analysis , Models, Biological
16.
Chinese Journal of Biotechnology ; (12): 831-845, 2021.
Article in Chinese | WPRIM | ID: wpr-878599

ABSTRACT

As a model industrial host and microorganism with the generally regarded as safe (GRAS) status, Corynebacterium glutamicum not only produces amino acids on a large scale in the fermentation industry, but also has the potential to produce various new products. C. glutamicum usually encounters various stresses in the process of producing compounds, which severely affect cell viability and production performance. The development of synthetic biology provides new technical means for improving the robustness of C. glutamicum. In this review, we discuss the tolerance mechanisms of C. glutamicum to various stresses in the fermentation process. At the same time, we highlight new synthetic biology strategies for boosting C. glutamicum robustness, including discovering new stress-resistant elements, modifying transcription factors, and using adaptive evolution strategies to mine stress-resistant functional modules. Finally, prospects of improving the robustness of engineered C. glutamicum strains ware provided, with an emphasis on biosensor, screening and design of transcription factors, and utilizing the multiple regulatory elements.


Subject(s)
Amino Acids/metabolism , Corynebacterium glutamicum/metabolism , Fermentation , Metabolic Engineering , Synthetic Biology
17.
Chinese Journal of Biotechnology ; (12): 2803-2812, 2021.
Article in Chinese | WPRIM | ID: wpr-887843

ABSTRACT

Bacitracin is a broad-spectrum antibiotics mainly produced by Bacillus, and is used as veterinary medicine in the fields of livestock and poultry breeding. Insufficient supply of precursor amino acids might be an important factor that hinders high-level microbial production of bacitracin. We investigated the effect of strengthening L-cysteine supply on bacitracin production by an industrial bacitracin producer, Bacillus licheniformis DW2. Overexpression of cysK encoding L-cysteine synthase led to a 9.17% increase of the bacitracin titer. Moreover, overexpression of cysE encoding L-serine acetyltransferase and cysP encoding thiosulfate/sulfate intracellular transporter increased the bacitracin titers by 7.23% and 8.52%, respectively. Moreover, overexpression of a putative cystine importer TcyP led to a 29.19% increase of intracellular L-cysteine, and bacitracin titer was increased by 7.79%. Subsequently, the strong promoter PbacA was used to replace the promoters of genes cysP, cysE and tcyP in strain DW2::ysK, respectively. The resulted strain CYS4 (DW2::cysK-PbacA-(cysP)-PbacA(cysE)- PbacA(tcyP) produced 910.02 U/mL bacitracin, which was 21.10% higher than that of the original strain DW2 (747.71 U/mL). Together with the experiments in 3 L fermenters, this research demonstrated that enhancing intracellular L-cysteine supply is an effective strategy to increase bacitracin production of B. licheniformis.


Subject(s)
Amino Acids , Bacillus licheniformis/genetics , Bacitracin , Cysteine , Metabolic Engineering
18.
Chinese Journal of Biotechnology ; (12): 2435-2442, 2021.
Article in Chinese | WPRIM | ID: wpr-887809

ABSTRACT

The stability of virus-like particles (VLPs) is currently the main factor affecting the quality of foot-and-mouth disease VLPs vaccines. In order to further improve the quality of the VLPs vaccine of foot-and-mouth disease (FMD), three amino acid modification sites were designed and screened through kinetic analysis software, based on the three-dimensional structure of FMDV. The three mutant recombinant plasmids were successfully prepared by the point mutation kit, transformed into Escherichia coli strain BL21 and expressed in vitro. After purification by Ni ion chromatography column, SDS-PAGE proved that the three amino acid mutations did not affect the expression of the target protein. The results of the stability study of three FMD mutant VLPs obtained by in vitro assembly show that the introduction of internal hydrophobic side chain amino acids made the morphology of VLPs more uniform (N4017W), and their stability was significantly improved compared to the other two VLPs. The internal hydrophobic force of the capsid contributes to the formation of VLPs and helps to maintain the stability of the capsid, providing new experimental ideas for improving the quality of VLPs vaccines, and helping to promote the development of VLPs vaccines.


Subject(s)
Amino Acids , Animals , Capsid Proteins/genetics , Foot-and-Mouth Disease/prevention & control , Foot-and-Mouth Disease Virus/genetics , Kinetics , Vaccines, Virus-Like Particle/genetics , Viral Vaccines/genetics
19.
Actual. osteol ; 17(1): 8-17, 2021. graf, tab
Article in English | LILACS, BINACIS, UNISALUD | ID: biblio-1291888

ABSTRACT

Objective: The main purpose of this study was to evaluate serum 25-hydroxyvitamin D (25OHD) levels and its association with in"ammatory markers in patients with rheumatologic diseases (RD). Methods: A cross-sectional study in 154 women with RD (rheumatoid arthritis, spondyloarthritis and other connective tissue diseases) and 112 healthy individuals as a control group (CG) was carried out. Results: No differences in serum and urine calcium, serum phosphate, and urinary deoxypyridinoline were found. RD group had lower 25OHD and higher PTH compared to CG. RD group had higher C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) compared to CG. The overall mean level of 25OHD (ng/ml) was 26.3±12.0 in the CG and 19.4±6.8 in the RD group (p<0.0001). Moreover, CG had lower percentage of individuals with 25OHD de!ciency compared to RD (29.9% vs 53.2%). The femoral neck BMD was signi!cantly lower in postmenopausal RD women compared to CG. 25OHD levels signi!cantly correlated with ESR and CRP as in"ammatory markers. Age, BMI, presence of RD, and CRP were signi!cantly and negatively associated with 25OHD levels through linear regression analysis. According to univariate logistic regression analysis for 25OHD deficiency (<20 ng/ml), a significant and negative association with BMI, presence of RD, ESR and CRP were found. Conclusion: Patients with RD had lower 25OHD levels than controls and the presence of a RD increases by 2.66 the risk of vitamin D de!ciency. In addition, 25OHD has a negative correlation with ESR and CRP as in"ammatory markers. (AU)


Objetivo El objetivo principal de este estudio fue evaluar los niveles séricos de 25-hidroxivitamina D (25OHD) y su asociación con marcadores inflamatorios en enfermedades reumatológicas. Materiales y métodos: Se realizó un estudio transversal en 154 mujeres con enfermedades reumatológicas (artritis reumatoide, espondiloartritis y otras enfermedades del tejido conectivo) y 112 individuos sanos como grupo control (GC). Resultados: No se encontraron diferencias en el calcio sérico y urinario, el fosfato sérico y la desoxipiridinolina urinaria entre el GC y los sujetos con enfermedades reumatológicas. El grupo de pacientes con enfermedades reumatológicas tenía 25OHD más bajo y PTH más alto en comparación con el GC. Asimismo, el grupo de individuos con enfermedades reumatológicas tenía proteína C reactiva (PCR) y velocidad de eritrosedimentación (VES) más altas en comparación con el GC. El nivel de 25OHD (ng/ml) fue 26,3±12,0 en el GC y 19,4±6,8 en el grupo con enfermedades reumatológicas (p<0,0001). Además, el GC presentó un porcentaje menor de deficiencia de 25OHD en comparación con el grupo con enfermedades reumatológicas (29,9% vs 53,2%). La DMO del cuello femoral fue significativamente menor en las mujeres posmenopáusicas con enfermedades reumatológicas en comparación con el GC. La 25OHD correlacionó significativamente con la VES y la PCR como marcadores inflamatorios. El análisis de regresión lineal mostró que la edad, el IMC, la presencia de una enfermedad reumatológica y la PCR se asociaron significativa y negativamente con los niveles de 25OHD. Mientras que el análisis de regresión logística univariada mostró que la deficiencia de 25OHD (<20 ng/ml), se asoció significativa y negativamente con el IMC, la presencia de una enfermedad reumatológica, la VES y los niveles de PCR. Conclusiones: Los pacientes con enfermedades reumatológicas tenían niveles de 25OHD más bajos que los controles y la presencia de una enfermedad reumatológica aumenta en 2.66 el riesgo de deficiencia de vitamina D. Además, la 25OHD mostró correlación negativa con la VES y la PCR como marcadores inflamatorios. (AU)


Subject(s)
Humans , Female , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Young Adult , Vitamin D/analogs & derivatives , Vitamin D/blood , Vitamin D Deficiency/etiology , Biomarkers , Rheumatic Diseases/complications , Inflammation/blood , Phosphates/blood , Blood Sedimentation , C-Reactive Protein , Body Mass Index , Bone Density , Logistic Models , Calcium/urine , Calcium/blood , Rheumatic Diseases/blood , Risk , Cross-Sectional Studies , Postmenopause , Amino Acids/urine
20.
Article in English | WPRIM | ID: wpr-888495

ABSTRACT

To investigate the intestinal amino acids pathway in depression-like offspring rats induced by maternal separation. Sprague-Dawley (SD) female rats were randomly divided into a control group (=8) and a maternal separation group (=8). After normal delivery, the maternal rats were separated from offsprings for 14 consecutive days and 3 h per day in maternal separation group; while rats in the control group was received no interventions in postpartum. Depression-like behaviors of offspring rats were evaluated using the sucrose preference test, novelty suppressed feeding test, and forced swimming test. Amino acid analyzer was used to detect the changes of amino acid contents in the small intestine, and the expressions of alanine-serine-cysteine transporter 2 (ASCT2), solute carrier superfamily 6 member 19 (BAT1) and L-type amino acid transporter 1(LAT1) were detected by Western blot. The weight of the offspring rats in the maternal separation group was significantly lower than that of the control group at 21 and 28 d (=4.925 and 5.766, all <0.01). Compared with the control group, the percentage of sucrose preference of the offspring rats in the maternal separation group was significantly reduced (=2.709, <0.05), and the feeding latency was significantly prolonged (=-13.431, <0.01). The immobility time in FST of maternal separation group was significantly longer (=-3.616, <0.01).Increased concentration of aspartic acid (=-6.672, <0.01) and down-regulation of glutamine (=3.107, <0.01) and glycine (=9.781, <0.01) were observed in maternal separation group. Western blot analysis revealed that the protein expressions of ASCT2 (=6.734, <0.01) and BAT1 (=9.015, <0.01) in maternal separation group were reduced, while the expression of LAT1 was increased (=-8.942, <0.01). Maternal separation can induce the depression-like behavior in offspring rats; the amino acid contents and the amino acid transporter expression in the small intestine are reduced, which may be related to depression-like behavior induced by maternal separation.


Subject(s)
Amino Acids , Animals , Depression/etiology , Female , Hippocampus , Maternal Deprivation , Rats , Rats, Sprague-Dawley
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