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1.
Brief Bioinform ; 24(2)2023 03 19.
Artigo em Inglês | MEDLINE | ID: mdl-36892153

RESUMO

Accurate and effective drug-target interaction (DTI) prediction can greatly shorten the drug development lifecycle and reduce the cost of drug development. In the deep-learning-based paradigm for predicting DTI, robust drug and protein feature representations and their interaction features play a key role in improving the accuracy of DTI prediction. Additionally, the class imbalance problem and the overfitting problem in the drug-target dataset can also affect the prediction accuracy, and reducing the consumption of computational resources and speeding up the training process are also critical considerations. In this paper, we propose shared-weight-based MultiheadCrossAttention, a precise and concise attention mechanism that can establish the association between target and drug, making our models more accurate and faster. Then, we use the cross-attention mechanism to construct two models: MCANet and MCANet-B. In MCANet, the cross-attention mechanism is used to extract the interaction features between drugs and proteins for improving the feature representation ability of drugs and proteins, and the PolyLoss loss function is applied to alleviate the overfitting problem and the class imbalance problem in the drug-target dataset. In MCANet-B, the robustness of the model is improved by combining multiple MCANet models and prediction accuracy further increases. We train and evaluate our proposed methods on six public drug-target datasets and achieve state-of-the-art results. In comparison with other baselines, MCANet saves considerable computational resources while maintaining accuracy in the leading position; however, MCANet-B greatly improves prediction accuracy by combining multiple models while maintaining a balance between computational resource consumption and prediction accuracy.


Assuntos
Desenvolvimento de Medicamentos , Descoberta de Drogas , Descoberta de Drogas/métodos , Proteínas/metabolismo , Sistemas de Liberação de Medicamentos , Domínios Proteicos
2.
J Pathol ; 262(4): 517-528, 2024 04.
Artigo em Inglês | MEDLINE | ID: mdl-38361487

RESUMO

This study aimed to provide more information for prognostic stratification for patients through an analysis of the T-cell spatial landscape. It involved analyzing stained tissue sections of 80 patients with stage III lung adenocarcinoma (LUAD) using multiplex immunofluorescence and exploring the spatial landscape of T cells and their relationship with prognosis in the center of the tumor (CT) and invasive margin (IM). In this study, multivariate regression suggested that the relative clustering of CT CD4+ conventional T cell (Tconv) to inducible Treg (iTreg), natural regulatory T cell (nTreg) to Tconv, terminal CD8+ T cell (tCD8) to helper T cell (Th), and IM Treg to tCD8 and the relative dispersion of CT nTreg to iTreg, IM nTreg to nTreg were independent risk factors for DFS. Finally, we constructed a spatial immunological score named the GT score, which had stronger prognostic correlation than IMMUNOSCORE® based on CD3/CD8 cell densities. The spatial layout of T cells in the tumor microenvironment and the proposed GT score can reflect the prognosis of patients with stage III LUAD more effectively than T-cell density. The exploration of the T-cell spatial landscape may suggest potential cell-cell interactions and therapeutic targets and better guide clinical decision-making. © 2024 The Pathological Society of Great Britain and Ireland.


Assuntos
Adenocarcinoma de Pulmão , Neoplasias Pulmonares , Humanos , Linfócitos T CD8-Positivos , Linfócitos T Reguladores , Prognóstico , Adenocarcinoma de Pulmão/patologia , Reino Unido , Microambiente Tumoral , Neoplasias Pulmonares/patologia
3.
Microb Pathog ; 192: 106709, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38810766

RESUMO

This study prepared a novel monoclonal antibody (MAb) against mink enteritis parvovirus (MEV) and identified its antigen epitope. The antibody subclass is identified as IgG1, the titers of the MAb is up to 1:1 × 106 and keeps stably after low-temperature storage for 9 months or 11 passages of the MAb cells. The MAb can specifically recognize MEV in the cells in IFA, but not Aleutian disease virus (ADV) or canine distemper virus (CDV). Its antigen epitope was identified as a polypeptide containing 5 key amino acids (378YAFGR382) and the homology in 20 MEV strains, 4 canine parvovirus strains, and 4 feline panleukopenia virus strains was 100%. This study supplies a biological material for developing new methods to detect MEV.


Assuntos
Anticorpos Monoclonais , Anticorpos Antivirais , Vírus da Cinomose Canina , Epitopos , Vírus da Enterite do Vison , Animais , Anticorpos Monoclonais/imunologia , Epitopos/imunologia , Vírus da Enterite do Vison/imunologia , Vírus da Cinomose Canina/imunologia , Anticorpos Antivirais/imunologia , Antígenos Virais/imunologia , Vison/imunologia , Imunoglobulina G/imunologia , Vírus da Doença Aleutiana do Vison/imunologia , Parvovirus Canino/imunologia , Vírus da Panleucopenia Felina/imunologia , Mapeamento de Epitopos , Camundongos , Camundongos Endogâmicos BALB C , Enterite Viral do Vison/imunologia
4.
Artigo em Inglês | MEDLINE | ID: mdl-38206131

RESUMO

A Gram-stain-negative, aerobic, rod-shaped, non-flagellated, non-gliding bacterial strain, designated MT50T, was isolated from a deep-sea sediment sample collected from the Mariana Trench. Optimal growth of strain MT50T was observed at 25 °C, pH 7.0-7.5 and in the presence of 3-5 % (w/v) NaCl. The strain was positive for oxidase and catalase. Phylogenetic analysis of 16S rRNA gene sequences revealed that strain MT50T is affiliated with the genus Mesonia, showing the highest sequence similarity (98.5 %) to the type strain of Mesonia ostreae. The digital DNA-DNA hybridization and average nucleotide identity values between strain MT50T and four closely related type strains of known Mesonia species (14.1-54.8 % and 72.7-86.8 %, respectively) were all below the threshold values to discriminate bacterial species, indicating that strain MT50T is affiliated with a novel species within the genus. The genomic G+C content deduced from the genome of strain MT50T was 36.2 mol%. The major fatty acids of strain MT50T were iso-C15 : 0, iso-C17 : 0 3-OH and anteiso-C15 : 0. The predominant respiratory quinone of the strain was MK-6. The polar lipids of strain MT50T included phosphatidylethanolamine and two unidentified lipids. Based on the polyphasic data presented in this study, strain MT50T represents a novel species of the genus Mesonia, for which the name Mesonia profundi sp. nov. is proposed. The type strain is MT50T (=MCCC 1K07833T=KCTC 92380T).


Assuntos
Ácidos Graxos , Filogenia , RNA Ribossômico 16S/genética , Composição de Bases , Ácidos Graxos/química , Análise de Sequência de DNA , DNA Bacteriano/genética , Técnicas de Tipagem Bacteriana
5.
Inorg Chem ; 63(27): 12658-12666, 2024 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-38916863

RESUMO

Phosphonate-based nerve agents, as a kind of deadly chemical warfare agent, are a persistent and evolving threat to humanity. Zirconium-based metal-organic frameworks (Zr-MOFs) are a kind of highly porous crystalline material that includes Zr-OH-Zr sites and imitates the active sites of the phosphotriesterase enzyme, representing significant potential for the adsorption and catalytic hydrolysis of phosphonate-based nerve agents. In this work, we present a new Zr-MOF, UiO-66-2I, which attaches two iodine atoms in the micropore of the MOF and exhibits excellent catalytic activity on the degradation of a nerve agent simulant, dimethyl 4-nitrophenyl phosphate (DMNP), as the result of the formation of halogen bonds between the phosphate ester bonds and iodine groups. Furthermore, various morphologies of UiO-66-2I, such as blocky-shaped nanoparticles (NPs), two-dimensional (2D) nanosheets, hexahedral NPs, stick-like NPs, colloidal microspheres, and colloidal NPs, have been obtained by adding acetic acid (AA), formic acid (FA), propionic acid (PA), valeric acid (VA), benzoic acid (BA), and trifluoroacetic acid (TFA) as modulators, respectively, and show different catalytic hydrolysis activities. Specifically, the catalytic activities follow the trend UiO-66-2I-FA (t1/2 = 1 min) > UiO-66-2I-AA-NP (t1/2 = 4 min) ≈ UiO-66-2I-VA (t1/2 = 4 min) > UiO-66-2I-BA (t1/2 = 5 min) > UiO-66-2I-PA (t1/2 = 15 min) > UiO-66-2I-TFA (t1/2 = 18 min). The experimental results show that the catalytic hydrolysis activity of Zr-MOF is regulated by the crystallinity, defect quantity, morphologies, and hydrophilicity of these samples, which synergistically affect the accessibility of catalytic sites and the diffusion of phosphate in the pores of Zr-MOFs.

6.
Environ Res ; 251(Pt 1): 118626, 2024 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-38467358

RESUMO

The research was carried out to examine the pollution characteristics, ecological risk, and origins of seven heavy metals (Hg, As, Pb, Cu, Cd, Zn, and Ni) in 51 sediment samples gathered from 8 rivers located on the Qinghai-Tibet Plateau (QTP) in China. The contents of Hg and Cd were 5.0 and 1.1 times higher than their background values, respectively. The mean levels of other measured heavy metals were below those found naturally in the local soil. The enrichment factor showed that the study area exhibited significantly enriched Hg with 70.6% sampling sites. The Cd contents at 19.6% of sampling sites were moderately enriched. The other sampling sites were at a less enriched level. The sediments of all the rivers had a medium level of potential ecological risk. Hg was the major ecological risk factor in all sampling sites, followed by Cd. The findings from the positive matrix factorization (PMF) analysis shown agricultural activities, industrial activities, traffic emissions, and parent material were the major sources. The upper, middle, and low reaches of the Quanji river had different Hg isotope compositions, while sediments near the middle reaches were similar to the δ202Hg of the industrial source. At the upstream sampling sites, the Hg isotope content was very close to the background level. The results of this research can establish a strong scientific sound to improve the safety of the natural circumstances of rivers on the QTP.


Assuntos
Monitoramento Ambiental , Sedimentos Geológicos , Metais Pesados , Rios , Poluentes Químicos da Água , Metais Pesados/análise , Sedimentos Geológicos/química , Sedimentos Geológicos/análise , Rios/química , Poluentes Químicos da Água/análise , Tibet , Medição de Risco , China , Isótopos/análise
7.
Environ Res ; 252(Pt 1): 118753, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38527718

RESUMO

Most lakes in the world are permanently or seasonally covered with ice. However, little is known about the distribution of microbes and their influencing factors in ice-covered lakes worldwide. Here we analyzed the microbial community composition in the waters of 14 ice-covered lakes in the Hoh Xil region of northern Qing-Tibetan Plateau (QTP), and conducted a meta-analysis by integrating published microbial community data of ice-covered lakes in the tripolar regions (the Arctic, Antarctica and QTP). The results showed that there were significant differences in microbial diversity, community composition and distribution patterns in the ice-covered tripolar lakes. Microbial diversity and richness were lower in the ice-covered QTP lakes (including the studied lakes in the Hoh Xil region) than those in the Arctic and Antarctica. In the ice-covered lakes of Hoh Xil, prokaryotes are mainly involved in S-metabolic processes, making them more adaptable to extreme environmental conditions. In contrast, prokaryotes in the ice-covered lakes of the Arctic and Antarctica were predominantly involved in carbon/nitrogen metabolic processes. Deterministic (salinity and nutrients) and stochastic processes (dispersal limitation, homogenizing dispersal and drift) jointly determine the geographical distribution patterns of microorganisms in ice-covered lakes, with stochastic processes dominating. These results expand the understanding of microbial diversity, distribution patterns, and metabolic processes in polar ice-covered lakes.


Assuntos
Camada de Gelo , Lagos , Lagos/microbiologia , Camada de Gelo/microbiologia , Regiões Antárticas , Regiões Árticas , Microbiota , Bactérias/classificação , Bactérias/isolamento & purificação , Bactérias/genética , Biodiversidade , China
8.
Biochem Biophys Res Commun ; 661: 10-20, 2023 06 18.
Artigo em Inglês | MEDLINE | ID: mdl-37084488

RESUMO

There is an increasing interest in combining immune checkpoint inhibitors (ICIs) with anti-angiogenic drugs to enhance their anti-tumor effects. In this study, three anti-angiogenic agents, DC101 (acting on VEGFR2), SAR131675 (acting on VEGFR3), and fruquintinib (a small-molecule inhibitor acting on multiple targets) were administered to B16F1-OVA-loaded C57BL/6 mice. Immune cells infiltration in the tumor tissues, vascular normalization, and high-endothelial venule (HEV) formation were assessed to provide evidence for drug combination. Both DC101 and fruquintinib significantly slowed the melanoma growth and increased the proportion of CD3+ and CD8+ T cells infiltration compared with SAR131675, of note, the effect of DC101 was more pronounced. Moreover, DC101 and fruquintinib increased the interferon-γ and perforin levels, meanwhile, DC101 increased the granzyme B levels, whereas fruquintinib and SAR131675 did not. Only the fruquintinib-treated group showed decreased regulatory T cells infiltration. We found upregulation of PD-L1 expression in tumor cells and CD45+ immune cells in DC101-treated group as well as upregulation of PD-1 expression on CD3+ T cells. However, fruquintinib only increased PD-L1 expression in tumors. Both DC101 and fruquintinib reduced the proportion of CD31+ vessels, while DC101 increased the ratio of α-SMA +/CD31+ cells and reduced the expression of HIF-1α more than fruquintinib. Moreover, DC101 enhanced the infiltration of dendritic cells and B cells, and local HEV formation. In conclusion, our data indicate that DC101 may be a better choice for the combined clinical application of ICIs and anti-angiogenic agents.


Assuntos
Antígeno B7-H1 , Melanoma , Camundongos , Animais , Vênulas , Linfócitos T CD8-Positivos , Camundongos Endogâmicos C57BL , Inibidores da Angiogênese/farmacologia , Melanoma/tratamento farmacológico
9.
Artigo em Inglês | MEDLINE | ID: mdl-37350580

RESUMO

A Gram-stain-negative, aerobic, flagellated, and long rod-shaped bacterium, designated strain SM1973T, was isolated from an intertidal sediment sample collected from the coast of Qingdao, PR China. Strain SM1973T grew at 15-37 °C and with 0-5.5 % NaCl. It reduced nitrate to nitrite and hydrolysed aesculin but did not hydrolyse casein and gelatin. The strain showed the highest 16S rRNA gene sequence similarity (98.2 %) to the type strain of Spartinivicinus ruber. The phylogenetic trees based on the 16S rRNA genes and single-copy orthologous clusters showed that strain SM1973T clustered with S. ruber, forming a separate lineage within the family Zooshikellaceae. The major cellular fatty acids were summed feature 3 (C16 : 1 ω7с and/or C16 : 1 ω6с) and C16 : 0. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The main respiratory quinone was ubiquinone-9. The genomic DNA G+C content of strain SM1973T was 40.4 mol%. Based on the polyphasic evidence presented in this paper, strain SM1973T is considered to represent a novel species within the genus Spartinivicinus, for which the name Spartinivicinus marinus sp. nov. is proposed. The type strain is SM1973T (=MCCC 1K04833T=KCTC 72846T).


Assuntos
Ácidos Graxos , Gammaproteobacteria , Ácidos Graxos/química , Fosfolipídeos , Filogenia , RNA Ribossômico 16S/genética , DNA Bacteriano/genética , Composição de Bases , Análise de Sequência de DNA , Técnicas de Tipagem Bacteriana , Gammaproteobacteria/genética
10.
Artigo em Inglês | MEDLINE | ID: mdl-37103434

RESUMO

A Gram-stain-negative, aerobic, rod-shaped, non-gliding bacterial strain, designated as MT39T, was isolated from a deep-sea sediment sample collected from the Mariana Trench. Strain MT39T grew optimally at 35°C and pH 7.0, and could tolerate up to 10% (w/v) NaCl. The strain was positive for catalase and negative for oxidase. The genome of strain MT39T was 4 033 307 bp, with a 41.1 mol % genomic G+C content and 3514 coding sequences. Phylogenetic analysis based on 16S rRNA gene sequences placed strain MT39T within the genus Salinimicrobium, showing the highest 16S rRNA gene sequence similarity to Salinimicrobium terrea CGMCC 1.6308T (98.1%). The average nucleotide identity and in silico DNA-DNA hybridization values between strain MT39T and the type strains of seven Salinimicrobium species were all less than the threshold values to discriminate bacterial species, indicating that strain MT39T is affiliated with a novel species within the genus. The major cellular fatty acids of strain MT39T were iso-C15 : 0, anteiso-C15 : 0 and iso-C17 : 0 3-OH. Polar lipids of strain MT39T included phosphatidylethanolamine, one unidentified aminolipid and four unidentified lipids. Menaquinone-6 was the only respiratory quinone in strain MT39T. On the basis of the polyphasic data present in this study, strain MT39T represents a novel species of the genus Salinimicrobium, for which the name Salinimicrobium profundisediminis sp. nov. is proposed, with type strain being MT39T (=MCCC 1K07832T=KCTC 92381T).


Assuntos
Ácidos Graxos , Flavobacteriaceae , Ácidos Graxos/química , Sedimentos Geológicos/microbiologia , Água do Mar/microbiologia , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Composição de Bases , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Vitamina K 2/química
11.
Microb Cell Fact ; 22(1): 179, 2023 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-37689719

RESUMO

BACKGROUND: Alginate oligosaccharides (AOs) are the degradation products of alginate, a natural polysaccharide abundant in brown algae. AOs generated by enzymatic hydrolysis have diverse bioactivities and show broad application potentials. AOs production via enzymolysis is now generally with sodium alginate as the raw material, which is chemically extracted from brown algae. In contrast, AOs production by direct degradation of brown algae is more advantageous on account of its cost reduction and is more eco-friendly. However, there have been only a few attempts reported in AOs production from direct degradation of brown algae. RESULTS: In this study, an efficient Laminaria japonica-decomposing strain Pseudoalteromonas agarivorans A3 was screened. Based on the secretome and mass spectrum analyses, strain A3 showed the potential as a cell factory for AOs production by secreting alginate lyases to directly degrade L. japonica. By using the L. japonica roots, which are normally discarded in the food industry, as the raw material for both fermentation and enzymatic hydrolysis, AOs were produced by the fermentation broth supernatant of strain A3 after optimization of the alginate lyase production and hydrolysis parameters. The generated AOs mainly ranged from dimers to tetramers, among which trimers and tetramers were predominant. The degradation efficiency of the roots reached 54.58%, the AOs production was 33.11%, and the AOs purity was 85.03%. CONCLUSION: An efficient, cost-effective and green process for AOs production directly from the underutilized L. japonica roots by using strain A3 was set up, which differed from the reported processes in terms of the substrate and strain used for fermentation and the AOs composition. This study provides a promising platform for scalable production of AOs, which may have application potentials in industry and agriculture.


Assuntos
Alginatos , Laminaria , Análise Custo-Benefício , Oligossacarídeos
12.
Sensors (Basel) ; 23(4)2023 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-36850507

RESUMO

Agricultural droughts cause a great reduction in winter wheat productivity; therefore, timely and precise irrigation recommendations are needed to alleviate the impact. This study aims to assess drought stress in winter wheat with the use of an unmanned aerial system (UAS) with multispectral and thermal sensors. High-resolution Water Deficit Index (WDI) maps were derived to assess crop drought stress and evaluate winter wheat actual evapotranspiration rate (ETa). However, the estimation of WDI needs to be improved by using more appropriate vegetation indices as a proximate of the fraction of vegetation cover. The experiments involved six irrigation levels of winter wheat in the harvest years 2019 and 2020 at Luancheng, North China Plain on seasonal and diurnal timescales. Additionally, WDI derived from several vegetation indices (VIs) were compared: near-infrared-, red edge-, and RGB-based. The WDIs derived from different VIs were highly correlated with each other and had similar performances. The WDI had a consistently high correlation to stomatal conductance during the whole season (R2 between 0.63-0.99) and the correlation was the highest in the middle of the growing season. On the contrary, the correlation between WDI and leaf water potential increased as the season progressed with R2 up to 0.99. Additionally, WDI and ETa had a strong connection to soil water status with R2 up to 0.93 to the fraction of transpirable soil water and 0.94 to the soil water change at 2 m depth at the hourly rate. The results indicated that WDI derived from multispectral and thermal sensors was a reliable factor in assessing the water status of the crop for irrigation scheduling.


Assuntos
Solo , Triticum , Estações do Ano , China , Água
13.
J Environ Manage ; 339: 117933, 2023 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-37080099

RESUMO

Sewage sludge (SS) has been extensively used as an alternative fertilizer in forest plantations, which are beneficial in supplying timbers and mitigating climate change. However, whether the extra nitrogen (N) applied by SS would enhance the soil nitrous oxide (N2O) emission, an important greenhouse gas, in forest plantations have not been well understood. The objective of this study is to evaluate the ecological effects of SS application on soils, by investigating the soil N2O emission and the toxicity of the potentially toxic elements (PTEs) in soil. A field fertilization experiment was conducted in Eucalyptus plantations with four fertilization rates (0 kg m-2, 1.5 kg m-2, 3.0 kg m-2, and 4.5 kg m-2). The soil N2O emissions were monitored at a soil depth of 0-10 cm using static chamber method, soil chemical properties, and PTEs were determined at soil depths of 0-10 cm, 10-20 cm, and 20-40 cm. The average soil N2O emission rate was 8.1 µg N2O-N h-1 m-2 in plots without SS application (control). The application of SS significantly increased the soil N2O emissions by 7-10 times as to control. The increased N2O emissions were positively related to the soil total phosphorus and nitrogen and negatively correlated with copper and zinc, which increased with the SS application. However, the potential ecological risk index (Ei) and the comprehensive potential ecological risk index (RI) of PTEs were lower than 40 and 150 respectively, which indicating a low toxicity of PTEs to soil health. After seven months of SS application, the priming effects of SS on soil N2O emissions gradually diminished. These findings suggest that the application of SS may increase N2O emissions at the initial stages of application (<7 months) and may have a low PTEs pollution risk, even at a high SS addition rate (4.5 kg m-2).


Assuntos
Eucalyptus , Metais Pesados , Solo/química , Esgotos , Nitrogênio/análise , Fertilizantes/análise , Óxido Nitroso/análise , Agricultura , China
14.
J Biol Chem ; 297(1): 100841, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34058201

RESUMO

SGNH-type acetyl xylan esterases (AcXEs) play important roles in marine and terrestrial xylan degradation, which are necessary for removing acetyl side groups from xylan. However, only a few cold-adapted AcXEs have been reported, and the underlying mechanisms for their cold adaptation are still unknown because of the lack of structural information. Here, a cold-adapted AcXE, AlAXEase, from the Arctic marine bacterium Arcticibacterium luteifluviistationis SM1504T was characterized. AlAXEase could deacetylate xylooligosaccharides and xylan, which, together with its homologs, indicates a novel SGNH-type carbohydrate esterase family. AlAXEase showed the highest activity at 30 °C and retained over 70% activity at 0 °C but had unusual thermostability with a Tm value of 56 °C. To explain the cold adaption mechanism of AlAXEase, we next solved its crystal structure. AlAXEase has similar noncovalent stabilizing interactions to its mesophilic counterpart at the monomer level and forms stable tetramers in solutions, which may explain its high thermostability. However, a long loop containing the catalytic residues Asp200 and His203 in AlAXEase was found to be flexible because of the reduced stabilizing hydrophobic interactions and increased destabilizing asparagine and lysine residues, leading to a highly flexible active site. Structural and enzyme kinetic analyses combined with molecular dynamics simulations at different temperatures revealed that the flexible catalytic loop contributes to the cold adaptation of AlAXEase by modulating the distance between the catalytic His203 in this loop and the nucleophilic Ser32. This study reveals a new cold adaption strategy adopted by the thermostable AlAXEase, shedding light on the cold adaption mechanisms of AcXEs.


Assuntos
Acetilesterase/química , Acetilesterase/metabolismo , Adaptação Fisiológica , Temperatura Baixa , Acetilesterase/antagonistas & inibidores , Acetilesterase/genética , Sequência de Aminoácidos , Bactérias/enzimologia , Domínio Catalítico , Inibidores Enzimáticos/farmacologia , Estabilidade Enzimática/efeitos dos fármacos , Cinética , Metais/farmacologia , Modelos Moleculares , Simulação de Dinâmica Molecular , Mutação/genética , Filogenia , Multimerização Proteica , Especificidade por Substrato/efeitos dos fármacos , Temperatura
15.
Appl Environ Microbiol ; 88(7): e0167721, 2022 04 12.
Artigo em Inglês | MEDLINE | ID: mdl-35285716

RESUMO

Vibrio collagenases of the M9A subfamily are closely related to Vibrio pathogenesis for their role in collagen degradation during host invasion. Although some Vibrio collagenases have been characterized, the collagen degradation mechanism of Vibrio collagenase is still largely unknown. Here, an M9A collagenase, VP397, from marine Vibrio pomeroyi strain 12613 was characterized, and its fragmentation pattern on insoluble type I collagen fibers was studied. VP397 is a typical Vibrio collagenase composed of a catalytic module featuring a peptidase M9N domain and a peptidase M9 domain and two accessory bacterial prepeptidase C-terminal domains (PPC domains). It can hydrolyze various collagenous substrates, including fish collagen, mammalian collagens of types I to V, triple-helical peptide [(POG)10]3, gelatin, and 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-o-Arg (Pz-peptide). Atomic force microscopy (AFM) observation and biochemical analyses revealed that VP397 first assaults the C-telopeptide region to dismantle the compact structure of collagen and dissociate tropocollagen fragments, which are further digested into peptides and amino acids by VP397 mainly at the Y-Gly bonds in the repeating Gly-X-Y triplets. In addition, domain deletion mutagenesis showed that the catalytic module of VP397 alone is capable of hydrolyzing type I collagen fibers and that its C-terminal PPC2 domain functions as a collagen-binding domain during collagenolysis. Based on our results, a model for the collagenolytic mechanism of VP397 is proposed. This study sheds light on the mechanism of collagen degradation by Vibrio collagenase, offering a better understanding of the pathogenesis of Vibrio and helping in developing the potential applications of Vibrio collagenase in industrial and medical areas. IMPORTANCE Many Vibrio species are pathogens and cause serious diseases in humans and aquatic animals. The collagenases produced by pathogenic Vibrio species have been regarded as important virulence factors, which occasionally exhibit direct pathogenicity to the infected host or facilitate other toxins' diffusion through the digestion of host collagen. However, our knowledge concerning the collagen degradation mechanism of Vibrio collagenase is still limited. This study reveals the degradation strategy of Vibrio collagenase VP397 on type I collagen. VP397 binds on collagen fibrils via its C-terminal PPC2 domain, and its catalytic module first assaults the C-telopeptide region and then attacks the Y-Gly bonds in the dissociated tropocollagen fragments to release peptides and amino acids. This study offers new knowledge regarding the collagenolytic mechanism of Vibrio collagenase, which is helpful for better understanding the role of collagenase in Vibrio pathogenesis and for developing its industrial and medical applications.


Assuntos
Colágeno Tipo I , Vibrio , Sequência de Aminoácidos , Aminoácidos , Animais , Colágeno/metabolismo , Colágeno Tipo I/genética , Colagenases/genética , Colagenases/metabolismo , Mamíferos , Peptídeos/metabolismo , Tropocolágeno , Vibrio/metabolismo
16.
New Phytol ; 236(4): 1267-1280, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-35945699

RESUMO

Determining whether isotope fractionation occurs during root water uptake is a prerequisite for using stem or xylem water isotopes to trace water sources. However, it is unclear whether isotope fractionation occurs during root water uptake in gramineous crops. We conducted prevalidation experiments to estimate the isotope measurement bias associated with cryogenic vacuum distillation (CVD). Next, we assessed isotope fractionation during root water uptake in two common agronomic crops, wheat (Triticum aestivum L.) and maize (Zea mays L.), under flooding after postdrought stress conditions. Cryogenic vacuum distillation caused significant depletion of 2 H but negligible effects on 18 O for both soil and stem water. Surprisingly CVD caused depletion of 2 H and enrichment of 18 O in root water. Stem and root water δ18 O were more than soil water δ18 O, even considering the uncertainty of CVD. Soil water 18 O was depleted compared with irrigation water 18 O in the pots with plants but enriched relative to irrigation water 18 O in the pots without plants. These results indicate that isotope fractionation occurred during wheat and maize root water uptake after full irrigation and led to a heavy isotope enrichment in stem water. Therefore, the xylem/stem water isotope approach widely used to trace water sources should be carefully evaluated.


Assuntos
Doenças Cardiovasculares , Água , Isótopos de Oxigênio/análise , Deutério/análise , Poaceae , Solo , Triticum , Produtos Agrícolas , Zea mays , Hidratação
17.
Arch Microbiol ; 204(6): 325, 2022 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-35575833

RESUMO

A Gram-stain-negative, aerobic and rod-shaped bacterium, designated strain SM 2104T, was isolated from a deep-sea sediment sample collected from the Southwest Indian Ocean. Strain SM 2104T grew at 10-37 °C (optimum at 25 °C), and with 1.0-9.0% (w/v, optimum with 2-4%) NaCl. It hydrolyzed starch, tween 80 and gelatin but did not reduced nitrate to nitrite. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain SM 2104T was affiliated with the genus Alteromonas, sharing the highest 16S rRNA gene sequence similarities with type strains of Alteromonas flava (97.5%) and Alteromonas facilis (97.4%) and forming a distinct clade together with the two Alteromonas species. The digital DNA-DNA hybridization and average nucleotide identity values between strain SM 2104 T and type strains of Alteromonas flava and Alteromonas facilis were below 14.5%, and 71.0%, respectively. The major fatty acids of strain SM 2104T were summed feature 3 (C16:1ω6c/C16:1ω7c), C16:0 and summed feature 8 (C18:1ω7c/C18:1ω6c). The major polar lipids of strain SM 2104T were phosphatidylethanolamine and phosphatidylglycerol and the only respiratory quinone of strain SM 2104T was ubiquinone-8. The genomic DNA G + C content of strain SM 2104T was 48.0%. On the basis of the phylogenetic, phenotypic, chemotaxonomic and genomic analyses presented in this study, strain SM 2104T is considered to represent a novel species within the genus Alteromonas, for which the name Alteromonas oceansediminis sp. nov. is proposed. The type strain is SM 2104T (= CCTCC AB 2021121T = KCTC 82867T).


Assuntos
Alteromonas , Alteromonas/genética , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Ácidos Graxos , Fosfolipídeos , Filogenia , RNA Ribossômico 16S/genética , Água do Mar/microbiologia , Análise de Sequência de DNA , Ubiquinona
18.
Artigo em Inglês | MEDLINE | ID: mdl-35852492

RESUMO

A Gram-stain-negative, aerobic, flagellated and rod-shaped bacterium, designated strain SM2107T, was isolated from a deep-sea sediment sample collected from the Southwest Indian Ocean. Strain SM2107T grew at 4-40 °C and with 0-10.0 % (w/v) NaCl. It reduced nitrate to nitrite and hydrolysed casein, gelatin, chitin and DNA. The phylogenetic trees based on the 16S rRNA genes and single-copy orthologous clusters showed that strain SM2107T, together with Rheinheimera tuosuensis, Rheinheimera perlucida and Arsukibacterium ikkense, formed a separate clade, having the highest similarity to the type strain of Rheinheimera tuosuensis (98.3%). The major polar lipids were phosphatidylethanolamine and phosphatidylglycerol and the major cellular fatty acids were summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c), C16 : 0, C17 : 1 ω8с and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c). The only respiratory quinone was Q-8. The genomic DNA G+C content of strain SM2107T was 48.8 %. The digital DNA-DNA hybridization values between strain SM2107T and type strains of Rheinheimera tuosuensis, Rheinheimera perlucida and Arsukibacterium ikkense were 41.16, 37.70 and 31.80 %, while the average amino acid identity values between them were 87.59, 86.76 and 83.64 %, respectively. Based on the polyphasic evidence presented in this study, strain SM2107T was considered to represent a novel species within the genus Arsukibacterium, for which the name Arsukibacterium indicum was proposed. The type strain is SM2107T (=MCCC M24986T=KCTC 82921T). Moreover, the transfer of Rheinheimera tuosuensis and Rheinheimera perlucida to the genus Arsukibacterium as Arsukibacterium tuosuense comb. nov. (type strain TS-T4T=CGMCC 1.12461T=JCM 19264T) and Arsukibacterium perlucidum comb. nov. (type strain BA131T=LMG 23581T=CIP 109200T) is also proposed.


Assuntos
Ácidos Graxos , Fosfolipídeos , Técnicas de Tipagem Bacteriana , Composição de Bases , Chromatiaceae , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ubiquinona/química
19.
Artigo em Inglês | MEDLINE | ID: mdl-35100102

RESUMO

Two novel Gram-stain-negative, facultative anaerobic, non-flagellated, rod-shaped bacterial strains, designated MT13T and MT32, were isolated from sediment samples collected from the Mariana Trench at a depth of 8300 m. The two strains grew at -2-30 °C (optimum, 25 °C), at pH 5.5-10.0 (optimum, pH 7.5-8.0) and with 0-15 % (w/v) NaCl (optimum, 3-6 %). They did not reduce nitrate to nitrite nor hydrolyse Tweens 40 and 80, aesculin, casein, starch and DNA. The genomic G+C contents of draft genomes of strain MT13T and MT32 were 52.2 and 54.1 m ol%, respectively. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strains MT13T and MT32 were affiliated with the genus Halomonas, with the highest similarity to the type strain of Halomonas olivaria. The values of average nucleotide identity and in silico DNA-DNA hybridization between strain MT13T and MT32, and between strain MT13T and five closely related type strains of Halomonas species indicated that strains MT13T and MT32 belonged to the same species, but represented a novel species in the genus of Halomonas. The major cellular fatty acids of strains MT13T and MT32 were C16 : 0, summed feature 3(C16 : 1 ω7c/ω6c) and summed feature 8 (C18 : 1 ω7c/ω6c). Major polar lipids of strains MT13T and MT32 included phosphatidylglycerol, phosphatidylethanolamine and diphosphatidylglycerol. Ubiquinone-9 was the predominant respiratory quinone. Based on data from the present polyphasic study, strains MT13T and MT32 represent a novel species of the genus Halomonas, for which the name Halomonas profundi sp. nov. is proposed. The type strain is MT13T (=MCCC 1K06389T=KCTC 82923T).


Assuntos
Sedimentos Geológicos/microbiologia , Halomonas , Filogenia , Água do Mar/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Halomonas/classificação , Halomonas/isolamento & purificação , Hibridização de Ácido Nucleico , Oceano Pacífico , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ubiquinona/química
20.
Artigo em Inglês | MEDLINE | ID: mdl-35156916

RESUMO

A Gram-negative, aerobic, non-flagellated and rod-shaped bacterium, strain ASW11-22T, was isolated from an intertidal sediment collected from a coastal area of Qingdao, PR China. The strain grew at 15-40 °C (optimum, 37 °C), at pH 6.0-9.0 (optimum, pH 7.0) and with 0.5-10 % (w/v) NaCl (optimum, 1.0 %). It hydrolysed gelatin and aesculin but did not reduce nitrate to nitrite. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain ASW11-22T belonged to the genus Celeribacter, showing the highest sequence similarity to the type strains of Celeribacter halophilus MCCC 1A06432T (98.20 %) and Celeribacter ethanolicus NH195T (97.84 %). The genomic DNA G+C content was 59.1 mol%. The major cellular fatty acid (>10 %) of the strain was summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and its main polar lipids were phosphatidylglycerol and one unidentified aminolipid. The sole respiratory quinone of strain ASW11-22T was ubiquinone-10. On the basis of the polyphasic evidence presented in this paper, strain ASW11-22T represents a novel Celeribacter species, for which the name Celeribacter litoreus sp. nov. is proposed. The type strain is ASW11-22T (=KCTC 82495T=MCCC 1K05584T).


Assuntos
Alphaproteobacteria/classificação , Sedimentos Geológicos , Filogenia , Água do Mar , Alphaproteobacteria/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Sedimentos Geológicos/microbiologia , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Água do Mar/microbiologia , Análise de Sequência de DNA , Ubiquinona/análogos & derivados , Ubiquinona/química
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