Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 4.294
Filtrar
1.
J Pharm Biomed Anal ; 226: 115238, 2023 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-36645985

RESUMO

Ormosia hosiei Hemsl. et Wils. is an economical and medicinal plant, increasingly cultivated in China; however, its branches and leaves are often pruned as waste. This is the first study focused on the phytochemical profiles and antioxidant, anti-α-glucosidase, anti-tyrosinase, and anti-neuroinflammatory activities of the branches and leaves of O. hosiei. Herein, thirty-seven characteristic compounds were identified by UPLC-MS/MS and twelve were detected for the first time in O. hosiei. Twenty-seven phenolics were further quantified and significant differences in phenolic compositions between the branches and leaves of O. hosiei were observed. The ethanol extracts exhibited promising antioxidant, anti-α-glucosidase, anti-tyrosinase, and anti-neuroinflammatory effects, and the bioactivities significantly correlated with total phenolic content and twelve individual phenolics. Naringin, genistein, vitexin, vitexin-2-O-rhamnoside, syringaresinol and syringaresinol-4-O-ß-D-glucopyranoside can be considered potential quality markers of O. hosiei. Our results provided solid evidence that the branches and leaves of O. hosiei deserve more attention and exploitation, considering the potential to be developed as functional foods or herbal medicines.


Assuntos
Extratos Vegetais , Plantas Medicinais , Extratos Vegetais/química , Antioxidantes/química , Cromatografia Líquida , Espectrometria de Massas em Tandem , Compostos Fitoquímicos/análise , Fenóis/análise , Glucosidases , Folhas de Planta/química
2.
Sci Rep ; 12(1): 21707, 2022 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-36522378

RESUMO

Diabetic nephropathy is characterized by excessive accumulation of extracellular matrix (ECM) leading to renal fibrosis, progressive deterioration of renal function, and eventually to end stage renal disease. Matrix metalloproteinases (MMPs) are known to regulate synthesis and degradation of the ECM. Earlier, we demonstrated that imbalanced MMPs promote adverse ECM remodeling leading to renal fibrosis in type-1 diabetes. Moreover, elevated macrophage infiltration, pro-inflammatory cytokines and epithelial‒mesenchymal transition (EMT) are known to contribute to the renal fibrosis. Various bioactive compounds derived from the medicinal plant, Azadirachta indica (neem) are shown to regulate inflammation and ECM proteins in different diseases. Nimbidiol is a neem-derived diterpenoid that is considered as a potential anti-diabetic compound due to its glucosidase inhibitory properties. We investigated whether Nimbidiol mitigates adverse ECM accumulation and renal fibrosis to improve kidney function in type-1 diabetes and the underlying mechanism. Wild-type (C57BL/6J) and type-1 diabetic (C57BL/6-Ins2Akita/J) mice were treated either with saline or with Nimbidiol (0.40 mg kg-1 d-1) for eight weeks. Diabetic kidney showed increased accumulation of M1 macrophages, elevated pro-inflammatory cytokines and EMT. In addition, upregulated MMP-9 and MMP-13, excessive collagen deposition in the glomerular and tubulointerstitial regions, and degradation of vascular elastin resulted to renal fibrosis in the Akita mice. These pathological changes in the diabetic mice were associated with functional impairments that include elevated resistive index and reduced blood flow in the renal cortex, and decreased glomerular filtration rate. Furthermore, TGF-ß1, p-Smad2/3, p-P38, p-ERK1/2 and p-JNK were upregulated in diabetic kidney compared to WT mice. Treatment with Nimbidiol reversed the changes to alleviate inflammation, ECM accumulation and fibrosis and thus, improved renal function in Akita mice. Together, our results suggest that Nimbidiol attenuates inflammation and ECM accumulation and thereby, protects kidney from fibrosis and dysfunction possibly by inhibiting TGF-ß/Smad and MAPK signaling pathways in type-1 diabetes.


Assuntos
Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 1 , Nefropatias Diabéticas , Diterpenos , Camundongos , Animais , Nefropatias Diabéticas/patologia , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Camundongos Endogâmicos C57BL , Fibrose , Fator de Crescimento Transformador beta1/metabolismo , Rim/metabolismo , Diterpenos/metabolismo , Inflamação/patologia , Glucosidases
3.
Environ Monit Assess ; 194(12): 858, 2022 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-36208349

RESUMO

We examined 10 subsurface water, 5 benthic water and 19 sediment (02 cm) samples along a 518 km of the middle segment of the Ganga River to assess the possible improvements that resulted from the industrial shutdown during the COVID-19 pandemic. The sites included the main stem river, tributary confluences, and two point sources, one of which releases metal-rich effluents and the other flushes municipal sewage. We found significant declines in the carbon, nutrient and metal concentrations in both the water and sediment. Even the most polluted zones did not show hypoxia (dissolve oxygen; DO < 2.0 mg L-1) that had been observed in the previous year. Despite a significant decline in carbon and nitrogen as substrates, the activities of extracellular enzymes (EEs), such as ß-D-glucosidase, FDAase and protease in sediment (0-2 cm depth), increased significantly (p < 0.05) in response to the declining metal concentrations resulting from the industrial shutdown. We found strong negative correlations between EE activity and the concentrations of metal pollutants measured in 2019, but the correlations between these variables appeared poor in 2020 (lockdown period). Also, we found large variances (low stability coefficients) during the period of strong anthropogenic effects (2019). The study indicates that industrial sources are important contributors of metal pollution in the Ganga River and has relevance exploring river ecosystem recovery windows for management decisions.


Assuntos
COVID-19 , Metais Pesados , Poluentes Químicos da Água , Carbono , Controle de Doenças Transmissíveis , Ecossistema , Monitoramento Ambiental/métodos , Sedimentos Geológicos , Glucosidases , Humanos , Metais , Metais Pesados/análise , Nitrogênio , Oxigênio , Pandemias , Peptídeo Hidrolases , Rios , Esgotos , Água , Poluentes Químicos da Água/análise
4.
Arch Microbiol ; 204(11): 668, 2022 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-36220932

RESUMO

Glycogen is important for transmission of V. vulnificus undergoing disparate environments of nutrient-rich host and nutrient-limited marine environment. The malZ gene of V. vulnificus encoding a maltodextrin glucosidase was cloned and over-expressed in E. coli to investigate its roles in glycogen/maltodextrin metabolism in the pathogen. The malZ gene encoded a protein with a predicted molecular mass of 70 kDa. The optimal pH and temperature of MalZ was 7.0 and 37 °C, respectively. MalZ hydrolyzed maltodextrin to glucose and maltose most efficiently, while hydrolyzed other substrates such as starch, maltose, ß-cyclomaltodextrin, and glycogen less efficiently. The activity was enhanced greatly by Mn2+. It also exhibited transglycosylation activity toward excessive maltotriose. The malZ knock-out mutant accumulated 2.3-5.6-fold less glycogen than the wild type when excessive maltodextrin or glucose was added to LB medium, while it accumulated more glycogen than the wild type (3.5-fold) in the presence of excessive maltose. Growth and glycogen accumulation of the mutant were retarded most significantly in the M63 minimal medium supplemented with 0.5% maltodextrin. Side chain length distributions of glycogen molecules were varied by the malZ mutation and types of the excessive carbon source. Based on the results, MalZ of V. vulnificus was likely to be involved in maltose/maltodextrin metabolism, thereby balancing synthesis of glycogen and energy generation in the cell. The bacterium seemed to have multiple and unique pathways for glycogen metabolism according to carbon sources.


Assuntos
Proteínas de Escherichia coli , Vibrio vulnificus , Carbono/metabolismo , Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Glucose/metabolismo , Glucosidases/metabolismo , Glicogênio/metabolismo , Glicosídeo Hidrolases/genética , Maltose/metabolismo , Polissacarídeos , Amido/metabolismo
5.
Molecules ; 27(19)2022 Oct 04.
Artigo em Inglês | MEDLINE | ID: mdl-36235098

RESUMO

Amylase and glucosidase enzymes are the primary harmful source in the development of the chronic condition known as diabetes mellitus. The main function of these enzymes is to break the macromolecules into simple sugar units which are directly involved in the solubility of blood, hence increasing blood glucose levels. To overcome this effect, there is a need for a potent and effective inhibitor that inhibits the conversion of macromolecules of sugar into its smaller units. In this regard, we synthesized thiazolidinone-based indole derivatives (1-20). The synthesized derivatives were evaluated for α-amylase and α-glucosidase inhibitory activity. Different substituted derivatives were found with moderate to good potentials having IC50 values ranging, for α-amylase, from 1.50 ± 0.05 to 29.60 ± 0.40 µM and, for α-glucosidase, from IC50 = 2.40 ± 0.10 to 31.50 ± 0.50 µM. Among the varied substituted compounds, the most active analogs four (1.80 ± 0.70 and 2.70 ± 0.70), five (1.50 ± 0.05 and 2.40 ± 0.10, respectively) of the series showed few folds better inhibitory activity than standard drug acarbose (IC50 = 10.20 ± 0.10 and 11.70 ± 0.10 µM, respectively). Moreover, structure-activity relationship (SAR) was established and binding interactions were analyzed for ligands and proteins (α-amylase and α-glucosidase) through a molecular docking study.&nbsp.


Assuntos
Glucosidases , alfa-Glucosidases , Acarbose , Amilases/metabolismo , Glicemia , Glucosidases/metabolismo , Inibidores de Glicosídeo Hidrolases/química , Indóis/química , Indóis/farmacologia , Ligantes , Simulação de Acoplamento Molecular , Estrutura Molecular , Receptores de Droga , Relação Estrutura-Atividade , alfa-Amilases , alfa-Glucosidases/metabolismo
6.
Molecules ; 27(18)2022 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-36144535

RESUMO

In the present study, the extracts of Cytinus hypocistis (L.) L using both traditional solvents (hexane, ethyl acetate, dichloromethane, ethanol, ethanol/water, and water) and natural deep eutectic solvents (NADESs) were investigated in terms of their total polyphenolic contents and antioxidant and enzyme-inhibitive properties. The extracts were found to possess total phenolic and total flavonoid contents in the ranges of 26.47-186.13 mg GAE/g and 0.68-12.55 mg RE/g, respectively. Higher total phenolic contents were obtained for NADES extracts. Compositional differences were reported in relation to antioxidant potential studied by several assays (DPPH: 70.19-939.35 mg TE/g, ABTS: 172.56-4026.50 mg TE/g; CUPRAC: 97.41-1730.38 mg TE/g, FRAP: 84.11-1534.85 mg TE/g). Application of NADESs (choline chloride-urea 1:2, a so-called Reline) allowed one to obtain the highest number of extracts having antioxidant potential in the radical scavenging and reducing assays. NADES-B (protonated by HCl L-proline-xylitol 5:1) was the only extractant from the studied solvents that isolated a specific fraction without chelating activity. Reline extract exhibited the highest acetylcholinesterase inhibition compared to NADES-B and NADES-C (protonated by H2SO4 L-proline-xylitol 5:1) extracts, which showed no inhibition. The NADES extracts were observed to have higher tyrosinase inhibitory properties compared to extracts obtained by traditional organic solvents. Furthermore, the NADES extracts were relatively better inhibitors of the diabetic enzymes. These findings provided an interesting comparison in terms of total polyphenolic content yields, antioxidant and enzyme inhibitory properties (cholinesterase, amylase, glucosidase, and tyrosinase) between traditional solvent extracts and NADES extracts, used as an alternative. While the organic solvents showed better antioxidant activity, the NADES extracts were found to have some other improved properties, such as higher total phenolic content and enzyme-inhibiting properties, suggesting functional prospects for their use in phytonutrient extraction and fractionation. The obtained results could also be used to give a broad overview of the different biological potentials of C. hypocistis.


Assuntos
Antioxidantes , Solventes Eutéticos Profundos , Acetilcolinesterase , Amilases , Antioxidantes/farmacologia , Colina , Cromatografia Líquida , Etanol , Flavonoides/farmacologia , Glucosidases , Hexanos , Malvales , Cloreto de Metileno , Monofenol Mono-Oxigenase , Fenóis , Extratos Vegetais/farmacologia , Prolina , Solventes , Espectrometria de Massas em Tandem , Ureia , Água , Xilitol
7.
Chemosphere ; 308(Pt 2): 136385, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36096301

RESUMO

The dissimilatory nitrate (NO3-) reduction processes (DNRPs) play an important role in regulating the nitrogen (N) balance of aquatic ecosystem. Organic carbon (OC) and sulfur are important factors that influence the DNRPs. In this study, we investigated the effects of sulfur cycle and enzyme activity on DNRPs in the natural and human-modified heterotrophic sediments. Quarterly monitoring of anaerobic ammonium oxidation, denitrification (DNF), and dissimilatory NO3- reduction to ammonium (DNRA) in sediments was conducted using 15N isotope tracing method. qPCR and high-throughput sequencing were applied to characterize the DNF and DNRA microbial abundances and communities. Results showed that instead of the OC, the glucosidase activity (GLU) was the key driver of the DNRPs. Furthermore, instead of the ratio of OC to NO3-, the GLU and the ratio of OC to sulfide (C/S) correctly indicated the partitioning of DNRPs in this study. We deduced that the sulfur reduction processes competed with the DNRPs for the available OC. In addition, the inhibitory effect of sulfide (final product of the sulfur reduction processes) on the DNRPs bacterial community were observed, which suggested a general restrictive role of the sulfur cycle in the regulation and partitioning of the DNRPs in heterotrophic sediments.


Assuntos
Compostos de Amônio , Nitratos , Carbono , Desnitrificação , Ecossistema , Glucosidases , Humanos , Nitratos/análise , Nitrogênio/análise , Óxidos de Nitrogênio/análise , Oxirredução , Sulfetos , Enxofre
8.
Appl Microbiol Biotechnol ; 106(19-20): 6847-6859, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36121483

RESUMO

Surfactants are used to control microbial biofilms in industrial and medical settings. Their known toxicity on aquatic biota, and their longevity in the environment, has encouraged research on biodegradable alternatives such as rhamnolipids. While previous research has investigated the effects of biological surfactants on single species biofilms, there remains a lack of information regarding the effects of synthetic and biological surfactants in freshwater ecosystems. We conducted a mesocosm experiment to test how the surfactant sodium dodecyl sulfate (SDS) and the biological surfactant rhamnolipid altered community composition and metabolic activity of freshwater biofilms. Biofilms were cultured in the flumes using lake water from Lake Lunz in Austria, under high (300 ppm) and low (150 ppm) concentrations of either surfactant over a four-week period. Our results show that both surfactants significantly affected microbial diversity. Up to 36% of microbial operational taxonomic units were lost after surfactant exposure. Rhamnolipid exposure also increased the production of the extracellular enzymes, leucine aminopeptidase, and glucosidase, while SDS exposure reduced leucine aminopeptidase and glucosidase. This study demonstrates that exposure of freshwater biofilms to chemical and biological surfactants caused a reduction of microbial diversity and changes in biofilm metabolism, exemplified by shifts in extracellular enzyme activities. KEY POINTS: • Microbial biofilm diversity decreased significantly after surfactant exposure. • Exposure to either surfactant altered extracellular enzyme activity. • Overall metabolic activity was not altered, suggesting functional redundancy.


Assuntos
Leucil Aminopeptidase , Tensoativos , Biofilmes , Ecossistema , Água Doce/química , Glucosidases/farmacologia , Leucil Aminopeptidase/metabolismo , Leucil Aminopeptidase/farmacologia , Dodecilsulfato de Sódio , Tensoativos/farmacologia , Água/farmacologia
9.
Int J Mol Sci ; 23(18)2022 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-36142555

RESUMO

Salicylic acid (SA) is a stress hormone synthesized in phenylalanine ammonia-lyase (PAL) and the branching acid pathway. SA has two interconvertible forms in plants: SAG (SA O-ß-glucoside) and SA (free form). The molecular mechanism of conversion of SA to SAG had been reported previously. However, which genes regulate SAG to SA remained unknown. Here, we report a cytoplasmic ß-glucosidase (ß-Glu) which participates in the SA pathway and is involved in the brown hull pigmentation in rice grain. In the current study, an EMS-generated mutant brown hull 1 (bh1) displayed decreased contents of SA in hulls, a lower photosynthesis rate, and high-temperature sensitivity compared to the wild type (WT). A plaque-like phenotype (brown pigmentation) was present on the hulls of bh1, which causes a significant decrease in the seed setting rate. Genetic analysis revealed a mutation in LOC_Os01g67220, which encodes a cytoplasmic Os1ßGlu4. The knock-out lines displayed the phenotype of brown pigmentation on hulls and decreased seed setting rate comparable with bh1. Overexpression and complementation lines of Os1ßGlu4 restored the phenotype of hulls and normal seed setting rate comparable with WT. Subcellular localization revealed that the protein of Os1ßGlu4 was localized in the cytoplasm. In contrast to WT, bh1 could not hydrolyze SAG into SA in vivo. Together, our results revealed the novel role of Os1ßGlu4 in the accumulation of flavonoids in hulls by regulating the level of free SA in the cellular pool.


Assuntos
Celulases , Oryza , Celulases/metabolismo , Flavonoides , Regulação da Expressão Gênica de Plantas , Glucosidases/metabolismo , Glucosídeos , Hormônios , Oryza/genética , Oryza/metabolismo , Fenilalanina Amônia-Liase/metabolismo , Pigmentação/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Salicilatos , Ácido Salicílico/metabolismo
10.
Ying Yong Sheng Tai Xue Bao ; 33(7): 1791-1800, 2022 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-36052781

RESUMO

The formation and development of biological soil crusts (biocrusts) potentially affect the cycles and stoichiometric characteristics of soil carbon (C), nitrogen (N), and phosphorus (P). However, it is still unclear how soil microbes adapt to such changes. In this study, we examined the effects of moss-dominated biocrusts coverage (0, 1%-20%, 20%-40%, 40%-60%, 60%-80%, and 80%-100%) on soil physicochemical properties, soil microbial biomass, and ectoenzyme activities [ß-1, 4-glucosidase (BG), ß-1, 4-N-acetyl glucosidase (NAG), acid phosphatase (AP)] in two soil layers (0-5 and 5-10 cm) in the Three Gorges Reservoir area, as well as the covariations of soil-microbe-ectoenzyme C:N:P stoichiometry. The results showed that biocrust development significantly increased soil clay content, water stable aggregates, soil C, N, P contents, and significantly decreased soil bulk density and sand content. Microbial biomass C, N, P and ectoenzyme activities were significantly increased with increasing biocrust coverage. Soil depth did not affect soil physicochemical properties and C:N:P, but significantly affected microbial biomass, ectoenzyme activities, BG:AP and NAG:AP. Soil C, N and P contents were significantly positively correlated with microbial biomass and ectoenzyme activities, negatively correlated with BG:NAG, while positively correlated with NAG:AP, but had no significant correlation with microbial biomass C:N:P. There was no significant correlation between soil-microbe and microbial-ectoenzyme C:N:P. BG:NAG:AP decreased gradually with the increase of C:N:P stoichiometric imbalance between microbe and soil. This study indicated that the microbial metabolism was co-limited by N and P and with stronger P limitation. Microbes could maintain homeostasis by adjusting their own biomass and ectoenzyme C:N:P to adapt to changes in soil ecological stoichiometry driven by biocrust development.


Assuntos
Briófitas , Solo , Fosfatase Ácida , Carbono/química , China , Ecossistema , Glucosidases , Nitrogênio , Fósforo/química , Solo/química , Microbiologia do Solo
11.
Nat Commun ; 13(1): 5577, 2022 09 23.
Artigo em Inglês | MEDLINE | ID: mdl-36151080

RESUMO

In the barley ß-D-glucan glucohydrolase, a glycoside hydrolase family 3 (GH3) enzyme, the Trp286/Trp434 clamp ensures ß-D-glucosides binding, which is fundamental for substrate hydrolysis during plant growth and development. We employ mutagenesis, high-resolution X-ray crystallography, and multi-scale molecular modelling methods to examine the binding and conformational behaviour of isomeric ß-D-glucosides during substrate-product assisted processive catalysis that operates in GH3 hydrolases. Enzyme kinetics reveals that the W434H mutant retains broad specificity, while W434A behaves as a strict (1,3)-ß-D-glucosidase. Investigations of reactant movements on the nanoscale reveal that processivity is sensitive to mutation-specific alterations of the tryptophan clamp. While wild-type and W434H utilise a lateral cavity for glucose displacement and sliding of (1,3)-linked hydrolytic products through the catalytic site without dissociation, consistent with their high hydrolytic rates, W434A does not adopt processive catalysis. Phylogenomic analyses of GH3 hydrolases disclose the evolutionary advantage of the tryptophan clamp that confers broad specificity, high catalytic efficiency, and processivity.


Assuntos
Glicosídeo Hidrolases , Triptofano , Cristalografia por Raios X , Glucose , Glucosidases/química , Glucosídeos , Glicosídeo Hidrolases/metabolismo , Glicosídeos , Cinética , Plantas/metabolismo , Especificidade por Substrato
12.
BMC Cancer ; 22(1): 817, 2022 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-35879690

RESUMO

BACKGROUND: Urothelial carcinoma (UC) is among the most prevalent malignancies. The muscle-invasive bladder cancer (MIBC) shows an invasive feature and has poor prognosis, while the non-muscle invasive bladder cancer (NMIBC) shows a better prognosis as compared with the MIBC. However, a significant proportion (10%-30%) of NMIBC cases progress to MIBC. Identification of efficient biomarkers for the prediction of the course of UC remains challenging nowadays. Recently, there is an emerging study showed that post-translational modifications (PTMs) by glycosylation is an important process correlated with tumor angiogenesis, invasion and metastasis. Herein, we reported a data-driven discovery and experimental validation of GANAB, a key regulator of glycosylation, as a novel prognostic marker in UC. METHODS: In the present study, we conducted immunohistochemistry (IHC) assay to evaluate the correlation between the expression levels of GANAB protein and the prognosis of UC in our cohort of 107 samples using whole slide image (WSI) analysis. In vitro experiments using RNAi were also conducted to investigate the biological functions of GANAB in UC cell lines. RESULTS: We observed that positive GANAB protein expression was significantly correlated with poor prognosis of UC in our cohort, with p-value of 0.0017 in Log-rank test. Notably, tumor cells at the invasive front of the tumor margin showed stronger GANAB expression than the tumor cells inside the tumor body in UCs. We further validated that the elevated expression levels of GANAB were significantly correlated with high grade tumors (p-values of 1.72 × 10-10), advanced stages (6.47 × 10-6), and elevated in luminal molecular subtypes. Moreover, knocking-down GANAB using RNAi in UM-UC-3 and T24 cells inhibited cell proliferation and migration in vitro. Knockdown of GANAB resulted in cell cycle arrest at G1 phase. We demonstrated that GANAB mediated HIF1A and ATF6 transcriptional activation in the ER stress signaling, and regulated the gene expression of cell cycle-related transcriptional factors E2F7 and FOXM1. CONCLUSIONS: The elevated expression of GANAB is a novel indicator of poorer prognosis of UC. Our data suggests that GANAB is not only a new and promising prognostic biomarker for UC, but also may provide important cues for the development of PTM-based therapeutics for UC treatment.


Assuntos
Carcinoma de Células de Transição , Neoplasias da Bexiga Urinária , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/genética , Carcinoma de Células de Transição/patologia , Glucosidases , Glicoproteínas , Humanos , Invasividade Neoplásica , Prognóstico , Neoplasias da Bexiga Urinária/patologia , alfa-Glucosidases
13.
Food Res Int ; 156: 111344, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35651090

RESUMO

A new Aspergillus niger whole-cell catalyst was cultured for the cascade hydrolysis of hesperidin (HES) to produce high-value hesperetin-7-O-glucoside (HG) and hesperetin with high conversion (above 90%). Moreover, the inducers used were shown to be useful for cell growth and to induce cells to produce specific enzymes. Remarkably, the type of inducers determined whether the cells can hydrolyze HES. The product composition was also controllable by adjusting different inducers. Transcriptome analysis suggested that both naringin-vs-blank group and saccharose-vs-blank group had obviously difference in gene expression. The naringin-vs-blank group was mainly up-regulated differentially expressed genes (DEGs), while saccharose-vs-blank group was mainly down-regulated DEGs. The Gene Ontology (GO) analysis showed that whether naringin or saccharose was added as an inducer would greatly affect the catalytic activity of cells. Furthermore, 3 genes related to rhamnosidase, 14 genes related to glucosidase and 5 genes related to hydrolase activity were found. These genes were not only involved in rhamnosidase and glucosidase activities, but also spliceosome and the sucrose and starch metabolic pathways. The quantitative real-time polymerase chain reaction (qRT-PCR) analysis indicated that the results of transcriptome sequencing were reliable. This study gave a new approach to hydrolyze HES, and new perspectives to understand the mechanisms associated with the hydrolysis of whole-cell catalyst.


Assuntos
Citrus , Aspergillus , Aspergillus niger/genética , Flavonoides , Glucosidases , Hidrólise , Sacarose , Transcriptoma
14.
Nanoscale ; 14(12): 4495-4510, 2022 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-35254362

RESUMO

PRKCSH, also known as glucosidase II beta, functions as a contributor to lung tumorigenesis by regulating the cell cycle in a p53-dependent manner under severe environmental stress. However, the prognostic value and molecular mechanisms by which the level of PRKCSH is significantly increased in cancer cells are not clearly understood. Here, we first generated a biological profile of PRKCSH expression changes in cancers by analysing bioinformatic data from cancer databases. We found that higher PRKCSH expression was correlated with a poorer prognosis and greater infiltration of most immune cell types in patients with lung cancer. In particular, PRKCSH expression showed significant negative correlations with the level of STAT6 (r = -0.31, p < 0.001) in lung cancer tissues. We further found that PRKCSH deficiency promoted G2/M arrest in response to zinc oxide nanoparticle (Nano ZnO) treatment in A549 cells. With regard to the mechanism, PRKCSH deficiency may induce STAT6 translocation to the nucleus to activate p53 expression through binding to the p53 promoter region from -365 bp to +126 bp. Eventually, activated p53 contributed to Nano-ZnO-induced G2/M arrest in lung cancer cells. Taken together, our data provide new insights into immunotherapy target choices and the prognostic value of PRKCSH. Since the G2/M cell cycle checkpoint is crucial for lung cancer prognosis, targeting PRKCSH expression to suppress the activation of the STAT6/p53 pathway is a potential therapeutic strategy for managing lung cancer.


Assuntos
Neoplasias Pulmonares , Óxido de Zinco , Apoptose , Proteínas de Ligação ao Cálcio/uso terapêutico , Linhagem Celular Tumoral , Biologia Computacional , Pontos de Checagem da Fase G2 do Ciclo Celular , Glucosidases/metabolismo , Glucosidases/uso terapêutico , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo
15.
J Biosci Bioeng ; 133(6): 560-566, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35314117

RESUMO

Kitchen waste (KW) is a vast potential source of fermentable substrates. To bio-convert the KW into high-value chemicals, we used KW as substrate for the production of fengycin by an artificial consortium containing Bacillus amyloliquefaciens HM618 producing fengycin and the engineering Pichia pastoris producing amylase, glucosidase, or lipases. The maximal amylase activity of the constructed amylase-producing engineering strain (recombinant P. pastoris GS115-amy98) reached 385.4 U‧mL-1. The engineering strain GS115-α-glu53 producing glucosidase reached an enzyme activity titer of 247.3 U‧mL-1, while the lipase activities of the engineering strains GS115-lip2, GS115-α-lip2, and GS115-lip7 were around 90.0 U‧mL-1, with no significant differences among them. Liquid chromatography-mass spectrometry (LC-MS) analysis showed that the components of fengycin synthesized by B. amyloliquefaciens HM618 were complex, including C14-C18 fengycins A, C13-C14 fengycins B, C16-C18 fengycins B, C16 fengycin B2 and some fengycin homologues with unsaturated fatty acid chains. The levels of fengycin were 15.9 mg‧L-1 and 4.6 mg‧L-1 under the co-culture with strain HM618 and the recombinant strains producing amylase and lipase, respectively. The maximal titer of fengycin was 21.2 mg‧L-1 in the artificial consortia consisting of HM618 and the engineering strains producing glucosidase, amylase and lipase. Taken together, these results show that the co-culture of B. amyloliquefaciens HM618 and engineering strains producing amylase and lipase can promote the conversion of KW into fengycin. The work provides a new strategy for boosting the resource utilization of KW.


Assuntos
Bacillus amyloliquefaciens , Amilases , Bacillus amyloliquefaciens/genética , Técnicas de Cocultura , Glucosidases , Lipase/genética , Lipopeptídeos , Pichia/genética , Saccharomycetales
16.
Appl Biochem Biotechnol ; 194(10): 4424-4438, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35357664

RESUMO

The emergence of new technologies has led to the discovery of the biological properties of nanoparticles through green approach. In the present investigation, we report the potential antibacterial, antioxidant, and anti-diabetic properties of copper nanoparticle (CuNPs) synthesized by reducing 3 mM copper acetate solution with aqueous leaf extract of Cocculus hirsutus. A colour change from deep brown to dark greenish brown indicated the formation of copper nanoparticles. The so-formed CuNPs were characterized by employing UV spectroscopy, FTIR, SEM, and EDX analyses which described sheet-like structure morphology having typical size of 63.46 nm. Later, the synthesized CuNPs efficiency was evaluated against bacterial pathogens, and was found highly toxic to B. subtilis and S. aureus strains. The synthesized CuNPs were examined through H2O2 and PMA assays which demonstrated the highest free radical scavenging activity. Besides, the resulted CuNPs revealed the higher anti-diabetic efficacy in both the [Formula: see text]-amylase and [Formula: see text] -glucosidase inhibition assays (64.5% ± 0.11 and 68.5% ± 0.11, respectively). Finally, our findings report that C. hirsutus can be exploited as a source for green synthesis of CuNPs, having potent in vitro antioxidant, antibacterial, and anti-diabetic properties.


Assuntos
Cocculus , Menispermaceae , Nanopartículas Metálicas , Amilases , Antibacterianos/química , Antibacterianos/farmacologia , Antioxidantes/química , Antioxidantes/farmacologia , Cobre/química , Glucosidases , Peróxido de Hidrogênio , Nanopartículas Metálicas/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Staphylococcus aureus
17.
J Biol Chem ; 298(5): 101827, 2022 05.
Artigo em Inglês | MEDLINE | ID: mdl-35293315

RESUMO

Carbohydrate-active enzymes are involved in the degradation, biosynthesis, and modification of carbohydrates and vary with the diversity of carbohydrates. The glycoside hydrolase (GH) family 31 is one of the most diverse families of carbohydrate-active enzymes, containing various enzymes that act on α-glycosides. However, the function of some GH31 groups remains unknown, as their enzymatic activity is difficult to estimate due to the low amino acid sequence similarity between characterized and uncharacterized members. Here, we performed a phylogenetic analysis and discovered a protein cluster (GH31_u1) sharing low sequence similarity with the reported GH31 enzymes. Within this cluster, we showed that a GH31_u1 protein from Lactococcus lactis (LlGH31_u1) and its fungal homolog demonstrated hydrolytic activities against nigerose [α-D-Glcp-(1→3)-D-Glc]. The kcat/Km values of LlGH31_u1 against kojibiose and maltose were 13% and 2.1% of that against nigerose, indicating that LlGH31_u1 has a higher specificity to the α-1,3 linkage of nigerose than other characterized GH31 enzymes, including eukaryotic enzymes. Furthermore, the three-dimensional structures of LlGH31_u1 determined using X-ray crystallography and cryogenic electron microscopy revealed that LlGH31_u1 forms a hexamer and has a C-terminal domain comprising four α-helices, suggesting that it contributes to hexamerization. Finally, crystal structures in complex with nigerooligosaccharides and kojibiose along with mutational analysis revealed the active site residues involved in substrate recognition in this enzyme. This study reports the first structure of a bacterial GH31 α-1,3-glucosidase and provides new insight into the substrate specificity of GH31 enzymes and the physiological functions of bacterial and fungal GH31_u1 members.


Assuntos
Bactérias/enzimologia , Fungos/enzimologia , Glucosidases , Glicosídeo Hidrolases , Sequência de Aminoácidos , Bactérias/metabolismo , Microscopia Crioeletrônica , Cristalografia por Raios X , Fungos/metabolismo , Glucosidases/metabolismo , Glicosídeo Hidrolases/metabolismo , Lactococcus lactis/enzimologia , Lactococcus lactis/metabolismo , Modelos Moleculares , Oligossacarídeos/metabolismo , Filogenia , Especificidade por Substrato
18.
Mol Ecol ; 31(10): 2920-2934, 2022 05.
Artigo em Inglês | MEDLINE | ID: mdl-35344623

RESUMO

Soil microbiota increase their fitness to local habitats by adjusting their life history strategies. Yet, how such adjustments drive their ecological adaptations in xeric grasslands remains elusive. In this study, shifts in the traits that potentially represent microbial life history strategies were studied along two aridity gradients with different climates using metagenomic and trait-based approaches. The results indicated that resource acquisition (e.g., higher activities of ß-d-glucosidase and N-acetyl-ß-d-glucosidase, higher degradation rates of cellulose and chitin, as well as genes involved in cell motility, biodegradation, transportation and competition) and growth yield (e.g., higher biomass and respiration) strategies were depleted at higher aridity. However, maintenance of cellular and high growth potential (e.g., higher metabolic quotients and genes related to DNA replication, transcription, translation, central carbon metabolism and biosynthesis) and stress tolerance (e.g., genes involved in DNA damage repair, cation transportation, sporulation and osmolyte biosynthesis) strategies were enriched at higher aridity. This implied that microbiota have lower growth yields but are probably well primed for rapid responses to pulses of rainfall in more arid soils, whereas those in less arid soils may have stronger resource acquisition and growth yield abilities. By integrating a large amount of evidence from taxonomic, metagenomic, genomic and biochemical investigations, this study demonstrates that the ecological adaptations of soil microbiota to aridity made by adjusting and optimizing their life history strategies are universal in xeric grasslands and provides an underlying mechanistic understanding of soil microbial responses to climate changes.


Assuntos
Traços de História de Vida , Microbiota , Ecossistema , Glucosidases , Microbiota/genética , Solo , Microbiologia do Solo
19.
Food Chem ; 384: 132497, 2022 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-35219994

RESUMO

Current methods for vanilla bean curing are long and reduce the enzymatic activity necessary for flavor development. High hydrostatic pressure (HHP) at 50-600 MPa was used to improve phenolic compounds formation and ß-d-glucosidase activity in vanilla beans compared with scalded beans. Phenolics were analyzed by HPLC and ß-d-glucosidase activity by spectrophotometry. Vanillin was the main phenolic and it was formed by ß-d-glucovanillin hydrolysis and vanillyl alcohol oxidation. HHP improved vanillin content and influenced ß-d-glucosidase activity. At the beginning of the curing the highest increments of vanillin were produced at 400 MPa (up to 15%), while at the end, this was observed at 50 (138%) and 600 MPa (74%). Maximum increment of up to 400% in ß-d-glucosidase activity was observed from 100 to 300 MPa, which was attributed to tissue decompartmentalization, and conformational changes induced by pressure. HHP could be used during vanilla curing to improve vanillin content and ß-d-glucosidase activity.


Assuntos
Vanilla , Benzaldeídos/metabolismo , Cromatografia Líquida de Alta Pressão , Glucosidases/metabolismo , Pressão Hidrostática , Fenóis/metabolismo , Vanilla/metabolismo
20.
Food Funct ; 13(5): 2456-2464, 2022 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-35147627

RESUMO

Codonopsis pilosula (Franch.) Nannf. (CPN), mainly planted in the northwest region, is a traditional Chinese medicine/good health food for nourishing qi and promoting blood circulation. This study firstly evaluated the inhibitory effects of the CPN extraction (CPNE) on α-glucosidase in vitro and in vivo, and tentatively confirmed its chemical ingredients by employing UHPLC-Triple-TOF-MS/MS. The CPNE had strong inhibitory activities against mammalian α-glucosidase (sucrase and maltase) and yeast α-glycosidase with semi-inhibitory concentrations (IC50) of 0.241 mg mL-1, 0.326 mg mL-1 and 1.167 mg mL-1, respectively. In addition, the CPNE could significantly decrease the postprandial blood glucose (PBG) levels in the sucrose/maltose/starch tolerance assays of diabetic mice. Furthermore, a total of 29 compounds, including 3 alkaloids, 13 phenolic acids, 8 alcohol glycosides and 5 alkynosides, were assigned based on comparison with the standards and references, as well as the analysis of main fragments. These results demonstrated that CPN could be used as an adjuvant therapy or dietary supplements to effectively control the occurrence and development of diabetes.


Assuntos
Codonopsis , Diabetes Mellitus Experimental/prevenção & controle , Medicamentos de Ervas Chinesas/farmacologia , Hipoglicemiantes/farmacologia , Extratos Vegetais/farmacologia , Aloxano , Animais , Animais não Endogâmicos , Cromatografia Líquida de Alta Pressão , Medicamentos de Ervas Chinesas/química , Glucosidases/efeitos dos fármacos , Hipoglicemiantes/química , Concentração Inibidora 50 , Masculino , Camundongos , Fitoterapia , Extratos Vegetais/química , Raízes de Plantas
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...