RESUMO
Renal ischemia-reperfusion injury (IRI) is an integral process in renal transplantation, which results in compromised graft survival. Macrophages play an important role in both the early inflammatory period and late fibrotic period in response to IRI. In this study, we investigated whether scutellarin (SCU) could protect against renal IRI by regulating macrophage polarization. Mice were given SCU (5-50 mg/kg) by gavage 1 h earlier, followed by a unilateral renal IRI. Renal function and pathological injury were assessed 24 h after reperfusion. The results showed that administration of 50 mg/kg SCU significantly improved renal function and renal pathology in IRI mice. In addition, SCU alleviated IRI-induced apoptosis. Meanwhile, it reduced macrophage infiltration and inhibited pro-inflammatory macrophage polarization. Moreover, in RAW 264.7 cells and primary bone marrow-derived macrophages (BMDMs) exposed to SCU, we found that 150 µM SCU inhibited these cells to polarize to an inflammatory phenotype induced by lipopolysaccharide (LPS) and interferon-γ (IFN-γ). However, SCU has no influence on anti-inflammatory macrophage polarization in vivo and in vitro induced by in interleukin-4 (IL-4). Finally, we explored the effect of SCU on the activation of the mitogen-activated protein kinase (MAPK) pathway both in vivo and in vitro. We found that SCU suppressed the activation of the MAPK pathway, including the extracellular signal-regulated kinase (ERK), Jun N-terminal kinase (JNK), and p38. Our results demonstrated that SCU protects the kidney against IRI by inhibiting macrophage infiltration and polarization toward pro-inflammatory phenotype via the MAPK pathway, suggesting that SCU may be therapeutically important in treatment of IRI.
Assuntos
Apigenina , Glucuronatos , Sistema de Sinalização das MAP Quinases , Macrófagos , Traumatismo por Reperfusão , Animais , Masculino , Camundongos , Apigenina/farmacologia , Apoptose/efeitos dos fármacos , Glucuronatos/farmacologia , Glucuronatos/uso terapêutico , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Inflamação/prevenção & controle , Inflamação/patologia , Rim/metabolismo , Rim/efeitos dos fármacos , Rim/patologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Células RAW 264.7 , Traumatismo por Reperfusão/tratamento farmacológico , Traumatismo por Reperfusão/metabolismoRESUMO
An uncharacterized gene encoding a glycoside hydrolase family 43-like enzyme from Clostridium boliviensis strain E-1 was identified from genomic sequence data, and the encoded enzyme, CbE1Xyn43-l, was produced in Escherichia coli. CbE1Xyn43-l (52.9 kDa) is a two-domain endo-ß-xylanase consisting of a C-terminal CBM6 and a GH43-like catalytic domain. The positions of the catalytic dyad conserved in GH43, the catalytic base (Asp74), and proton donor (Glu240) were identified in alignments including GH43-enzymes of known 3D-structure from different subfamilies. CbE1Xyn43-l is active at pH 7.0-9.0, with optimum temperature at 65°C, and a more than 7 days' half-life in irreversible deactivation studies at this temperature. The enzyme hydrolyzed birchwood xylan, quinoa stalks glucuronoarabinoxylan, and wheat arabinoxylan with xylotriose and xylotetraose as major hydrolysis products. CbE1Xyn43-l also released xylobiose from pNPX2 with low turnover (kcat of 0.044 s-1) but was inactive on pNPX, showing that a degree of polymerization of three (DP3) was the smallest hydrolyzable substrate. Divalent ions affected the specific activity on xylan substrates, which dependent on the ion could be increased or decreased. In conclusion, CbE1Xyn43-l from C. boliviensis strain E-1 is the first characterized member of a large group of homologous hypothetical proteins annotated as GH43-like and is a thermostable endo-xylanase, producing xylooligosaccharides of high DP (xylotriose and xylotetraose) producer. IMPORTANCE: The genome of Clostridium boliviensis strain E-1 encodes a number of hypothetical enzymes, annotated as glycoside hydrolase-like but not classified in the Carbohydrate Active Enzyme Database (CAZy). A novel thermostable GH43-like enzyme is here characterized as an endo-ß-xylanase of interest in the production of prebiotic xylooligosaccharides (XOs) from different xylan sources. CbE1Xyn43-l is a two-domain enzyme composed of a catalytic GH43-l domain and a CBM6 domain, producing xylotriose as main XO product. The enzyme has homologs in many related Clostridium strains which may indicate a similar function and be a previously unknown type of endo-xylanase in this evolutionary lineage of microorganisms.
Assuntos
Glucuronatos , Glicosídeo Hidrolases , Oligossacarídeos , Xilanos , Xilanos/metabolismo , Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/metabolismo , Especificidade por Substrato , Clostridium/genética , Clostridium/metabolismo , Endo-1,4-beta-Xilanases/metabolismo , Hidrólise , Estabilidade Enzimática , Concentração de Íons de HidrogênioRESUMO
The search for affordable enzymes with exceptional characteristics is fundamental to overcoming industrial and environmental constraints. In this study, a recombinant GH10 xylanase (Xyn10-HB) from the extremely alkaliphilic bacterium Halalkalibacterium halodurans C-125 cultivated at pH 10 was cloned and expressed in E. coli BL21(DE3). Removal of the signal peptide improved the expression, and an overall activity of 8 U/mL was obtained in the cell-free supernatant. The molecular weight of purified Xyn10-HB was estimated to be 42.6 kDa by SDS-PAGE. The enzyme was active across a wide pH range (5-10) with optimal activity recorded at pH 8.5 and 60 °C. It also presented good stability with a half-life of 3 h under these conditions. Substrate specificity studies showed that Xyn10-HB is a cellulase-free enzyme that conventionally hydrolyse birchwood and oat spelts xylans (Apparent Km of 0.46 mg/mL and 0.54 mg/mL, respectively). HPLC analysis showed that both xylans hydrolysis produced xylooligosaccharides (XOS) with a degree of polymerization (DP) ranging from 2 to 9. The conversion yield was 77% after 24 h with xylobiose and xylotriose as the main end-reaction products. When assayed on alkali-extracted wheat straw heteroxylan, the Xyn10-HB produced active XOS with antioxidant activity determined by the DPPH radical scavenging method (IC50 of 0.54 mg/mL after 4 h). Owing to its various characteristics, Xyn10-HB xylanase is a promising candidate for multiple biotechnological applications.
Assuntos
Endo-1,4-beta-Xilanases , Proteínas Recombinantes , Xilanos , Especificidade por Substrato , Hidrólise , Xilanos/metabolismo , Endo-1,4-beta-Xilanases/metabolismo , Endo-1,4-beta-Xilanases/genética , Endo-1,4-beta-Xilanases/química , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Escherichia coli/genética , Escherichia coli/metabolismo , Concentração de Íons de Hidrogênio , Clonagem Molecular , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/química , Glucuronatos/metabolismo , Estabilidade Enzimática , Cinética , Peso Molecular , Oligossacarídeos/metabolismo , DissacarídeosRESUMO
Herein we utilise automated glycan assembly to complete solid-phase synthesis of defined heparan sulfate oligosaccharides, employing challenging D-glucuronate disaccharide donors. Using an orthogonally protected D-GlcN-α-D-GlcA donor, milligram-scale synthesis of a heparan sulfate tetrasaccharide is completed in 18% yield over five steps. Furthermore, orthogonal protecting groups enabled regiospecific on-resin 6-O-sulfation. This methodology provides an important benchmark for the rapid assembly of biologically relevant heparan sulfate sequences.
Assuntos
Heparitina Sulfato , Oligossacarídeos , Dissacarídeos , Glucuronatos , Ácido GlucurônicoRESUMO
The xylanolytic enzymes Clocl_1795 and Clocl_2746 from glycoside hydrolase (GH) family 30 are highly abundant in the hemicellulolytic system of Acetivibrio clariflavus (Hungateiclostridium, Clostridium clariflavum). Clocl_1795 has been shown to be a xylobiohydrolase AcXbh30A releasing xylobiose from the non-reducing end of xylan and xylooligosaccharides. In this work, biochemical characterization of Clocl_2746 is presented. The protein, designated AcXyn30B, shows low sequence similarity to other GH30 members and phylogenetic analysis revealed that AcXyn30B and related proteins form a separate clade that is proposed to be a new subfamily GH30_12. AcXyn30B exhibits similar specific activity on glucuronoxylan, arabinoxylan, and aryl glycosides of linear xylooligosaccharides suggesting that it is a non-specific xylanase. From polymeric substrates, it releases the fragments of degrees of polymerization (DP) 2-6. Hydrolysis of different xylooligosaccharides indicates that AcXyn30B requires at least four occupied catalytic subsites for effective cleavage. The ability of the enzyme to hydrolyze a wide range of substrates is interesting for biotechnological applications. In addition to subfamilies GH30_7, GH30_8, and GH30_10, the newly proposed subfamily GH30_12 further widens the spectrum of GH30 subfamilies containing xylanolytic enzymes. KEY POINTS: Bacterial GH30 endoxylanase from A. clariflavus (AcXyn30B) has been characterized AcXyn30B is non-specific xylanase hydrolyzing various xylans and xylooligosaccharides Phylogenetic analysis placed AcXyn30B in a new GH30_12 subfamily.
Assuntos
Clostridiales , Endo-1,4-beta-Xilanases , Xilanos , Dissacarídeos/metabolismo , Endo-1,4-beta-Xilanases/metabolismo , Endo-1,4-beta-Xilanases/genética , Endo-1,4-beta-Xilanases/química , Glucuronatos/metabolismo , Hidrólise , Oligossacarídeos/metabolismo , Filogenia , Especificidade por Substrato , Xilanos/metabolismo , Clostridiales/enzimologia , Clostridiales/genéticaRESUMO
Abnormalities in vascular smooth muscle cells (VSMCs) are pivotal in the pathogenesis of cardiovascular pathologies such as atherosclerosis and hypertension. Scutellarin (Scu), a flavonoid derived from marigold flowers, exhibits a spectrum of biological activities including anti-inflammatory, antioxidant, antitumor, immunomodulatory and antimicrobial effects. Notably, Scu has demonstrated the capacity to mitigate vascular endothelial damage and prevent atherosclerosis via its antioxidative properties. Nevertheless, the influence of Scu on the formation of VSMC-derived foam cells remains underexplored. In this study, Scu was evidenced to efficaciously attenuate oleic acid (OA)-induced lipid accumulation and the upregulation of adipose differentiation-associated protein Plin2 in a dose- and time-responsive manner. We elucidated that Scu effectively diminishes OA-provoked VSMC foam cell formation. Further, it was established that Scu pretreatment augments the protein expression of LC3B-II and the mRNA levels of Map1lc3b and Becn1, concurrently diminishing the protein levels of the NLRP3 inflammasome compared to the OA group. Activation of autophagy through rapamycin attenuated NLRP3 inflammasome protein expression, intracellular lipid droplet content and Plin2 mRNA levels. Scu also counteracted the OA-induced decrement of LC3B-II levels in the presence of bafilomycin-a1, facilitating the genesis of autophagosomes and autolysosomes. Complementarily, in vivo experiments revealed that Scu administration substantially reduced arterial wall thickness, vessel wall cross-sectional area, wall-to-lumen ratio and serum total cholesterol levels in comparison to the high-fat diet model group. Collectively, our findings suggest that Scu attenuates OA-induced VSMC foam cell formation through the induction of autophagy and the suppression of NLRP3 inflammasome activation.
Assuntos
Apigenina , Aterosclerose , Glucuronatos , Inflamassomos , Humanos , Inflamassomos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Células Espumosas/metabolismo , Células Espumosas/patologia , Músculo Liso Vascular/metabolismo , Ácido Oleico/farmacologia , Ácido Oleico/metabolismo , Aterosclerose/metabolismo , Autofagia , RNA Mensageiro/metabolismo , Miócitos de Músculo Liso/metabolismoRESUMO
PURPOSE: Glucuronoyl esterases (GE, family CE15) catalyse the cleavage of ester linkages in lignin-carbohydrate complexes (LCCs), and this study demonstrate how transesterification reactions with a fungal GE from Cerrena unicolor (CuGE) can reveal the enzyme's preference for the alcohol-part of the ester-bond. METHODS: This alcohol-preference relates to where the ester-LCCs are located on the lignin molecule, and has consequences for how the enzymes potentially interact with lignin. It is unknown exactly what the enzymes prefer; either the α-benzyl or the γ-benzyl position. By providing the enzyme with a donor substrate (the methyl ester of either glucuronate or 4-O-methyl-glucuronate) and either one of two acceptor molecules (benzyl alcohol or 3-phenyl-1-propanol) we demonstrate that the enzyme can perform transesterification and it serves as a method for assessing the enzyme's alcohol preferences. CONCLUSION: CuGE preferentially forms the γ-ester from the methyl ester of 4-O-methyl-glucuronate and 3-phenyl-1-propanol and the enzyme's substrate preferences are primarily dictated by the presence of the 4-O-methylation on the glucuronoyl donor, and secondly on the type of alcohol.
Assuntos
Esterases , Lignina , Polyporales , Propanóis , Esterases/química , Carboidratos , Ésteres , Glucuronatos , Especificidade por SubstratoRESUMO
Parkinson's disease (PD) is a prevalent neurodegenerative disorder, and accumulating evidence suggests a link between dysbiosis of the gut microbiota and the onset and progression of PD. In our previous investigations, we discovered that intraperitoneal administration of glucuronomannan oligosaccharides (GMn) derived from Saccharina japonica exhibited neuroprotective effects in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mouse model. However, the complicated preparation process, difficulties in isolation, and remarkably low yield have constrained further exploration of GMn. In this study, we optimized the degradation conditions in the preparation process of GMn through orthogonal experiments. Subsequently, an MPTP-induced PD model was established, followed by oral administration of GMn. Through a stepwise optimization, we successfully increased the yield of GMn, separated from crude fucoidan, from 1~2/10,000 to 4~8/1000 and indicated the effects on the amelioration of MPTP-induced motor deficits, preservation of dopamine neurons, and elevation in striatal neurotransmitter levels. Importantly, GMn mitigated gut microbiota dysbiosis induced by MPTP in mice. In particular, GM2 significantly reduced the levels of Akkermansia, Verrucomicrobiota, and Lactobacillus, while promoting the abundance of Roseburia and Prevotella compared to the model group. These findings suggest that GM2 can potentially suppress PD by modulating the gut microbiota, providing a foundation for the development of a novel and effective anti-PD marine drug.
Assuntos
Modelos Animais de Doenças , Microbioma Gastrointestinal , Camundongos Endogâmicos C57BL , Oligossacarídeos , Animais , Microbioma Gastrointestinal/efeitos dos fármacos , Camundongos , Oligossacarídeos/farmacologia , Masculino , Fármacos Neuroprotetores/farmacologia , Disbiose/tratamento farmacológico , 1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/farmacologia , Neurônios Dopaminérgicos/efeitos dos fármacos , Doença de Parkinson/tratamento farmacológico , Manose/farmacologia , Manose/química , Manose/análogos & derivados , Glucuronatos/farmacologiaRESUMO
We presented a strategy utilizing 2D NMR-based metabolomic analysis of crude extracts, categorized by different pharmacological activities, to rapidly identify the primary bioactive components of TCM. It was applied to identify the potential bioactive components from Scutellaria crude extracts that exhibit anti-non-small cell lung cancer (anti-NSCLC) activity. Four Scutellaria species were chosen as the study subjects because of their close phylogenetic relationship, but their crude extracts exhibit significantly different anti-NSCLC activity. Cell proliferation assay was used to assess the anti-NSCLC activity of four species of Scutellaria. 1H-13C HSQC spectra were acquired for the chemical profiling of these crude extracts. Based on the pharmacological classification (PCA, OPLS-DA and univariate hypothesis test) were performed to identify the bioactive constituents in Scutellaria associated with the anti-NSCLC activity. As a result, three compounds, baicalein, wogonin and scutellarin were identified as bioactive compounds. The anti-NSCLC activity of the three potential active compounds were further confirmed via cell proliferation assay. The mechanism of the anti-NSCLC activity by these active constituents was further explored via flow cytometry and western blot analyses. This study demonstrated 2D NMR-based metabolomic analysis of pharmacologically classified crude extracts to be an efficient approach to the identification of active components of herbal medicine.
Assuntos
Carcinoma Pulmonar de Células não Pequenas , Proliferação de Células , Espectroscopia de Ressonância Magnética , Metabolômica , Extratos Vegetais , Scutellaria , Scutellaria/química , Humanos , Proliferação de Células/efeitos dos fármacos , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Apigenina/farmacologia , Apigenina/química , Apigenina/isolamento & purificação , Apigenina/análise , Flavanonas/farmacologia , Flavanonas/química , Flavanonas/isolamento & purificação , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Glucuronatos/farmacologia , Glucuronatos/isolamento & purificação , Glucuronatos/química , Antineoplásicos Fitogênicos/farmacologia , Antineoplásicos Fitogênicos/química , Antineoplásicos Fitogênicos/isolamento & purificação , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos AntitumoraisRESUMO
Phenolics produced during xylooligosaccharide production might inhibit xylanases and enhance the antioxidant and antimicrobial activities of XOS. The effects of phenolic compounds on xylanases may depend on the type and concentration of the compound, the plant biomass used, and the enzyme used. Understanding the effects of phenolic compounds on xylanases and their impact on XOS is critical for developing viable bioconversion of lignocellulosic biomass to XOS. Understanding the complex relationship between phenolic compounds and xylanases can lead to the development of strategies that improve the efficiency and cost-effectiveness of XOS manufacturing processes and optimise enzyme performance.
Assuntos
Glucuronatos , Oligossacarídeos , Fenóis , Prebióticos , Oligossacarídeos/química , Oligossacarídeos/farmacologia , Glucuronatos/farmacologia , Glucuronatos/química , Fenóis/química , Fenóis/farmacologia , Estrutura Molecular , Antioxidantes/farmacologia , Antioxidantes/química , Endo-1,4-beta-Xilanases/metabolismoRESUMO
OBJECTIVE: The aim of this study was to investigate the effects of scutellarin on the activation of astrocytes into the A1 type following cerebral ischemia and to explore the underlying mechanism. METHODS: In vivo, a mouse middle cerebral artery wire embolism model was established to observe the regulation of astrocyte activation to A1 type by scutellarin, and the effects on neurological function and brain infarct volume. In vitro, primary astrocytes were cultured to establish an oxygen-glucose deprivation model, and the mRNA and protein expression of C3, a specific marker of A1-type astrocytes pretreated with scutellarin, were examined. The neurons were cultured in vitro to detect the toxic effects of ischemia-hypoxia-activated A1 astrocyte secretion products on neurons, and to observe whether scutellarin could reduce the neurotoxicity of A1 astrocytes. To validate the signaling pathway-related proteins regulated by scutellarin on C3 expression in astrocytes. RESULTS: The results showed that scutellarin treatment reduced the volume of cerebral infarcts and attenuated neurological deficits in mice caused by middle cerebral artery embolism. Immunofluorescence and Western blot showed that treatment with scutellarin down-regulated middle cerebral artery embolism and OGD/R up-regulated A1-type astrocyte marker C3. The secretory products of ischemia-hypoxia-activated A1-type astrocytes were toxic to neurons and induced an increase in neuronal apoptosis, and astrocytes treated with scutellarin reduced the toxic effects on neurons. Further study revealed that scutellarin inhibited the activation of NF-κB signaling pathway and thus inhibited the activation of astrocytes to A1 type.
Assuntos
Apigenina , Isquemia Encefálica , Embolia , Glucuronatos , AVC Isquêmico , Acidente Vascular Cerebral , Ratos , Camundongos , Animais , Astrócitos/metabolismo , AVC Isquêmico/metabolismo , Ratos Sprague-Dawley , Isquemia/metabolismo , Hipóxia , Infarto da Artéria Cerebral Média/tratamento farmacológico , Infarto da Artéria Cerebral Média/metabolismo , Acidente Vascular Cerebral/tratamento farmacológico , Acidente Vascular Cerebral/metabolismoRESUMO
In this study, the α-glucosidase (maltase-glucoamylase: MGAM) and α-amylase inhibitory properties elicited by xylooligosaccharides (XOSs) prepared from dulse xylan were analysed as a potential mechanism to control postprandial hyperglycaemia for type-2 diabetes prevention and treatment. Xylan was purified from red alga dulse powder and used for enzymatic hydrolysis using Sucrase X to produce XOSs. Fractionation of XOSs produced xylobiose (X2), ß-(1â3)-xylosyl xylobiose (DX3), xylotriose (X3), ß-(1â3)-xylosyl-xylotriose (DX4), and a dulse XOS mixture with n ≥ 4 xylose units (DXM). The different fractions exhibited moderate MGAM (IC50 = 11.41-23.44 mg/mL) and α-amylase (IC50 = 18.07-53.04 mg/mL) inhibitory activity, which was lower than that of acarbose. Kinetics studies revealed that XOSs bound to the active site of carbohydrate digestive enzymes, limiting access to the substrate by competitive inhibition. A molecular docking analysis of XOSs with MGAM and α-amylase clearly showed moderate strength of interactions, both hydrogen bonds and non-bonded contacts, at the active site of the enzymes. Overall, XOSs from dulse could prevent postprandial hyperglycaemia as functional food by a usual and continuous consumption.
Assuntos
Algas Comestíveis , Glucuronatos , Hiperglicemia , Rodófitas , alfa-Amilases , Humanos , alfa-Glucosidases , Hipoglicemiantes/farmacologia , Xilanos/farmacologia , Simulação de Acoplamento Molecular , Oligossacarídeos/farmacologiaRESUMO
The primary aim of this study was to examine the impact of xylooligosaccharide (XOS) in rice protein concentrate (RPC) based diets on the growth performance, body composition, digestive enzymes, intestinal morphology and blood biochemistry of Labeo rohita fingerlings. Four different XOS levels (0%, 0.5%, 1% and 2%) were used at each RPC (75% and 100%) level. Twenty-five fish per tank with an average initial weight of 25 ± 0.05 g were randomly assigned (Randomised complete block design) to each of the 8 groups in triplicate aquaria (36 × 16 × 12â³) and then fed with respective diets @ 3% body weight for 90 days. The results showed significant improvements in growth performance, such as increased weight gain %, specific growth rate, and protein efficiency ratio and improved feed conversion ratio in 1% XOS supplemented diet at 75% RPC. A significant decrease in serum alkaline phosphatase activity (ALP) and plasma melanodialdehyde (MDA) were observed at 1% XOS level in 75% RPC based diets, respectively. Meanwhile, the lowest total cholesterol and highest lysozyme activity were observed in 1% XOS supplemented diet at 75% RPC levels. Moreover, the serum (alanine aminotransferase and aspartate transaminase) and plasma (superoxide dismutase, triglyceride, high density and low density lipoprotein) activities showed nonsignificant effects among the treatments. Furthermore, the digestive enzymes (protease & lipase) and intestinal morphology were significantly influenced at 1% XOS in the 75% RPC-based diet. Polynomial regression analysis showed that 1.25% XOS is the optimum requirement for the growth of rohu fingerlings when fed at 75% RPC based diets. Overall, it was concluded that the 75% RPC diet was efficiently replaced by fishmeal along with 1% XOS addition in L. rohita fingerlings without any negative effect on growth performance and intestinal health.
Assuntos
Ração Animal , Fenômenos Fisiológicos da Nutrição Animal , Cyprinidae , Dieta , Suplementos Nutricionais , Glucuronatos , Oligossacarídeos , Oryza , Animais , Oligossacarídeos/farmacologia , Oligossacarídeos/administração & dosagem , Ração Animal/análise , Dieta/veterinária , Glucuronatos/administração & dosagem , Glucuronatos/farmacologia , Cyprinidae/crescimento & desenvolvimentoRESUMO
Quantitative determination of ethyl glucuronide (EtG) in different biological objects in recent years has been positioned as one of the most reliable biomarkers of unconditional alcohol consumption. The aim of the study is to summarize the analytical methods of alcohol consumption testing with the use of EtG currently available in domestic and foreign literature and to present a schematic overview of possible errors in reproducibility and interpretation of research on EtG results, which may limit their use in forensic medical practice. The main objective is to increase the reliability and validity of EtG as a marker of ethanol consumption.
Assuntos
Consumo de Bebidas Alcoólicas , Detecção do Abuso de Substâncias , Reprodutibilidade dos Testes , Detecção do Abuso de Substâncias/métodos , Glucuronatos , Biomarcadores , EtanolRESUMO
Globally, the demands for sustainably sourced functional foods like prebiotic oligosaccharides have been constantly increasing. This study assessed the potential of pineapple leaves (PL) as lignocellulosic feedstock for sustainable production of cellulose and hemicellulose-derived oligosaccharides through its hydrothermal pretreatment (HT) followed by controlled enzymatic hydrolysis. PL was subjected to HT at 160, 175, and 190 °C for 20, 30, 60, and 90 min without any catalyst for xylooligosaccharide (XOS) production, whereas, the resulting solid content after HT was subjected to controlled enzymatic hydrolysis by commercial cellulase using conduritol B epoxide (0.5-5 mM) for glucooligosaccharides (GOS) production. HT at 160 °C for 60 min resulted in maximum yield of XOS and GOS at 23.7 and 18.3 %, respectively, in the liquid phase. Controlled enzymatic hydrolysis of HT treated (160 °C) PL solids for 20 and 30 min yielded â¼ 174 mg cellobiose/g dry biomass within 24 h, indicating overall high oligosaccharide production.
Assuntos
Ananas , Celulose , Polissacarídeos , Hidrólise , Oligossacarídeos , GlucuronatosRESUMO
The construction of high-performance microbial cell factories (MCFs) is the centerpiece of biomanufacturing. However, the complex metabolic regulatory network of microorganisms poses great challenges for the efficient design and construction of MCFs. The genome-scale metabolic network models (GSMs) can systematically simulate the metabolic regulation process of microorganisms in silico, providing effective guidance for the rapid design and construction of MCFs. In this review, we summarized the development status of 16 important industrial microbial GSMs, and further outline the technologies or methods that continuously promote high-quality GSMs construction from five aspects: I) Databases and modeling tools facilitate GSMs reconstruction; II) evolving gap-filling technologies; III) constraint-based model reconstruction; IV) advances in algorithms; and V) developed visualization tools. In addition, we also summarized the applications of GSMs in guiding metabolic engineering from four aspects: I) exploring and explaining metabolic features; II) predicting the effects of genetic perturbations on metabolism; III) predicting the optimal phenotype; IV) guiding cell factories construction in practical experiment. Finally, we discussed the development of GSMs, aiming to provide a reference for efficiently reconstructing GSMs and guiding metabolic engineering.
Assuntos
Dissacarídeos , Glucuronatos , Engenharia Metabólica , Redes e Vias Metabólicas , Engenharia Metabólica/métodos , Redes e Vias Metabólicas/genética , FenótipoRESUMO
The efficient utilization of biomass resources has gained widespread attention in current research. This study focused on the conversion of hemicellulose into xylo-oligosaccharides and furfural, as well as enhanced cellulose saccharification and lignin removal from residual biomass. The solid acid catalyst AT-Sn-MMT was prepared by sulfonation and tin ion loading of montmorillonite K-10. In a mixture of deep eutectic solvent and γ-valerolactone (3:7, v/v), AT-Sn-MMT was used to catalyze Phyllostachys edulis (PE) at 160 °C for 20 min, obtaining a furfural yield of 85.7 % and 1.5 g/L xylo-oligosaccharides. The delignification of pretreated PE was 59.5 %, reaching an accessibility of 221.3 g dye/g material. While the enzymatic saccharification efficiency was increased to 73.1 %. This work drew on the merits of solid acid catalysts and mixed solvent systems, and this constructed pretreatment method could be efficiently applied for co-production of reducing sugars, xylooligosaccharide and furfural, realizing the efficient valorization of PE.
Assuntos
Furaldeído , Glucuronatos , Açúcares , Solventes , Oligossacarídeos , Lignina , Poaceae , Biomassa , HidróliseRESUMO
We propose a closed-loop pretreatment process, wherein volatiles produced during steam explosion pretreatment were recovered and reintroduced as acid catalysts into the pretreatment system. The volatiles were separated through a drastic decompression process followed by a steam explosion process and recovered as a liquified catalyst (LFC) through a heat exchanger. The LFC effectively served as an acid catalyst for hemicellulose hydrolysis, significantly decreasing residence time from 90 min to 30 min to achieve 80 % conversion yield at 170 °C. Hydrolysates with high content of lower molecular weight oligomeric sugars were obtained using LFC, and were considered advantageous for application as prebiotics. These results are attributed to the complementary features of acetic acid and furfural contained within the LFC. Computational simulation using Aspen Plus was used to investigate the effects of recycling on LFC, and it demonstrated the feasibility of the catalyst-recirculating system. A validation study was conducted based on simulation results to predict the actual performance of the proposed pretreatment system. Based on these results, the recirculating system was predicted to improve the conversion yield and low-molecular weight oligomers yield by 1.5-fold and 1.6-fold, respectively.
Assuntos
Avena , Glucuronatos , Oligossacarídeos , Vapor , Catálise , Hidrólise , Oligossacarídeos/química , Avena/química , Glucuronatos/química , Polissacarídeos/químicaRESUMO
Rice husks are rich in xylan, which can be hydrolyzed by xylanase to form xylooligosaccharides (XOS). XOS are a functional oligosaccharide such as improving gut microbiota and antioxidant properties. In this study, the structure and functional characteristics of XOS were studied. The optimal xylanase hydrolysis conditions through response surface methodology (RSM) were: xylanase dosage of 3000â¯U/g, hydrolysis time of 3â¯h, hydrolysis temperature of 50⯰C. Under this condition, the yield of XOS was 150.9â¯mg/g. The TG-DTG curve showed that XOS began to decompose at around 200⯰C. When the concentration of XOS reached 1.0â¯g/L, the clearance rate of DPPH reached 65.76â¯%, and the scavenging rate of OH reached 62.10â¯%, while the clearance rate of ABTS free radicals reached 97.70â¯%, which was equivalent to the clearance rate of VC. XOS had a proliferative effect on four probiotics: Lactobacillus plantarum, Lactobacillus brucelli, Lactobacillus acidophilus, and Lactobacillus rhamnosus. However, the further experiments are needed to explore the improvement effect of XOS on human gut microbiota, laying a foundation for the effective utilization of XOS. XOS have a wide range of sources, low price, and broad development prospects. The reasonable utilization of XOS can bring greater economic benefits.
Assuntos
Antioxidantes , Glucuronatos , Oligossacarídeos , Oryza , Probióticos , Oligossacarídeos/farmacologia , Oligossacarídeos/química , Oryza/química , Glucuronatos/farmacologia , Glucuronatos/química , Antioxidantes/farmacologia , Antioxidantes/química , Hidrólise , Endo-1,4-beta-Xilanases/metabolismo , LactobacillusRESUMO
XOS production from lignocellulose using organic carboxylic acids and alkyd acids has been widely reported. However, it still faces harsh challenges such as high energy consumption, high cost, and low purity. Pyruvic acid (PYA), a carbonyl acid with carbonyl and carboxyl groups, was used to produce XOS due to its stronger catalytic activity. In this work, XOS was efficiently prepared from COS in an autoclave under the condition of 0.21 M PYA-121 °C-35 min. The total yield of XOS reached 68.72 % without producing any toxic by-products, including furfural (FF) and 5-hydroxymethylfurfural (5-HMF). The yield of xylobiose (X2), xylotriose (X3), xylotetraose (X4), and xylopentaose (X5) were 20.58 %, 12.47 %, 15.74 %, and 10.05 %, respectively. Meanwhile, 89.05 % of lignin was retained in the solid residue, which provides a crucial functional group for synthesizing layered carbon materials (SRG-a). It achieves excellent electromagnetic shielding (EMS) performance through graphitization, reaching -30 dB at a thickness of 2.0 mm. The use of a PYA catalyst in the production of XOS has proven to be an efficient method due to lower temperature, lower acid consumption, and straightforward operation.