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1.
Int J Syst Evol Microbiol ; 71(10)2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34617881

RESUMO

A Gram-stain-positive, strictly aerobic, motile, endospore-forming, milk-white, indole acetic acid-producing, rod-shaped bacterial strain, designated as HU2P27T, was isolated from a shoot of Kalidium cuspidatum collected in Tumd Right Banner, Inner Mongolia, PR China. Strain grew at 10-40 °C (optimum, 30 °C), at pH 6.0-9.0 (optimum, pH 7.0) and with 0-14.0 % NaCl (optimum, 5.0-8.0 %). The strain tested positive for oxidase, catalase and nitrate reductase. The phylogenetic trees based on the 16S rRNA gene sequence and the core genome both showed that strain HU2P27T clustered with Aquibacillus koreensis BH30097T, sharing 97.7 % and <97.0 % of 16S rRNA gene similarity with A. koreensis BH30097T and any other type strain. Strain HU2P27T contained MK-7 as the major respiratory quinone. Its major fatty acids were anteiso-C15 : 0 and iso-C15 : 0, and the major polar lipids were phosphatidylglycerol, diphosphatidylglycerol and four unidentified phospholipids. The genomic DNA G+C content was 36.0 mol%. The average nucleotide identity, amino acid identity and digital DNA-DNA hybridization values of strain HU2P27T with A. koreensis BH30097T were 71.7, 69.2 and 19.4%, respectively. The phylogenetic, physiological and phenotypic results allowed the discrimination of strain HU2P27T from its phylogenetic relatives. The name Aquibacillus kalidii sp. nov. is therefore proposed. The type strain is strain HU2P27T (=CGMCC 1.18646T=KCTC 43248T). Based on the results of 16S rRNA gene and genome analyses, we propose the reclassification of Virgibacillus campisalis Lee et al. 2012 as a later heterotypic synonym of Virgibacillus alimentarius Kim et al. 2011.


Assuntos
Endófitos , Ácidos Graxos , Bacillaceae , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Endófitos/genética , Ácidos Graxos/química , Ácidos Indolacéticos , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Virgibacillus
2.
World J Microbiol Biotechnol ; 37(7): 121, 2021 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-34143291

RESUMO

We performed several experiments using three strains of Virgibacillus salexigens, namely, P2, NT N53, and C-20MoT (DSM 11483T), which were isolated from completely different sources, in relation to bacteriocin production ability. Results of whole-genome sequencing analysis revealed that all strains have very similar sequences encoding class IId bacteriocin. Although a partial amino acid sequence of the purified bacteriocin produced by strain P2 isolated from fermented food was previously reported, whole-genome sequencing and the N-terminal sequencing results in this study showed that its complete amino acid sequence consisted of 48 residues, which corresponded to that of the hypothetical bacteriocin encoded by the gene in Virgibacillus massiliensis strain Vm-5T (DSM 28587T) isolated from the human gut. From the results of 16S rRNA gene sequencing and whole-genome sequencing analyses, we taxonomically confirmed Vm-5T to be a strain of V. salexigens, and its broth culture showed antibacterial activity. Strain NT N53 isolated from the deep-sea floor produced two bacteriocins, namely, NTN-A and NTN-B. The results of N-terminal sequencing, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and whole-genome sequencing analyses showed that their amino acid sequences differed in only one residue, and NTN-A showed the same sequence as the bacteriocin produced by strain P2. Although strain C-20MoT isolated from a solar saltern had the coding sequence very similar to that of NTN-A, its broth culture showed no antibacterial activity. This finding suggests that class IId bacteriocin-producing or bacteriocin-gene-encoding V. salexigens strains are widely distributed in distinct environment sources with different geographical and material properties.


Assuntos
Bacteriocinas/genética , Virgibacillus/classificação , Virgibacillus/genética , Sequência de Aminoácidos , Antibacterianos/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Bacteriocinas/metabolismo , Microbiologia Ambiental , Humanos , RNA Ribossômico 16S , Água do Mar/microbiologia , Análise de Sequência de DNA , Virgibacillus/metabolismo , Sequenciamento Completo do Genoma
3.
Molecules ; 25(3)2020 02 09.
Artigo em Inglês | MEDLINE | ID: mdl-32050419

RESUMO

Root-knot nematode diseases cause severe yield and economic losses each year in global agricultural production. Virgibacillus dokdonensis MCCC 1A00493, a deep-sea bacterium, shows a significant nematicidal activity against Meloidogyne incognita in vitro. However, information about the active substances of V. dokdonensis MCCC 1A00493 is limited. In this study, volatile organic compounds (VOCs) from V. dokdonensis MCCC 1A00493 were isolated and analyzed through solid-phase microextraction and gas chromatography-mass spectrometry. Four VOCs, namely, acetaldehyde, dimethyl disulfide, ethylbenzene, and 2-butanone, were identified, and their nematicidal activities were evaluated. The four VOCs had a variety of active modes on M. incognita juveniles. Acetaldehyde had direct contact killing, fumigation, and attraction activities; dimethyl disulfide had direct contact killing and attraction activities; ethylbenzene had an attraction activity; and 2-butanone had a repellent activity. Only acetaldehyde had a fumigant activity to inhibit egg hatching. Combining this fumigant activity against eggs and juveniles could be an effective strategy to control the different developmental stages of M. incognita. The combination of direct contact and attraction activities could also establish trapping and killing strategies against root-knot nematodes. Considering all nematicidal modes or strategies, we could use V. dokdonensis MCCC 1A00493 to set up an integrated strategy to control root-knot nematodes.


Assuntos
Antinematódeos/isolamento & purificação , Doenças das Plantas/prevenção & controle , Tylenchoidea/efeitos dos fármacos , Virgibacillus/química , Compostos Orgânicos Voláteis/isolamento & purificação , Acetaldeído/isolamento & purificação , Acetaldeído/farmacologia , Animais , Antinematódeos/farmacologia , Organismos Aquáticos , Derivados de Benzeno/isolamento & purificação , Derivados de Benzeno/farmacologia , Butanonas/isolamento & purificação , Butanonas/farmacologia , Quimiotaxia/efeitos dos fármacos , Dissulfetos/isolamento & purificação , Dissulfetos/farmacologia , Cromatografia Gasosa-Espectrometria de Massas , Lycopersicon esculentum/efeitos dos fármacos , Lycopersicon esculentum/parasitologia , Contagem de Ovos de Parasitas , Doenças das Plantas/parasitologia , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/parasitologia , Microextração em Fase Sólida , Tylenchoidea/crescimento & desenvolvimento , Compostos Orgânicos Voláteis/farmacologia
4.
Int J Biol Macromol ; 149: 552-561, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32006575

RESUMO

Virgibacillus salarius BM02 was identified as a highly exopolysaccharide (EPS) producing bacterium. The EPS production and its physico-chemical properties (intrinsic viscosity and total sugars/protein (TS/P) ratio) were optimized using Box-Behnken experimental design. Maximum EPS production of 5.87 g L-1 with TS/P ratio of 12.56 and intrinsic viscosity of 0.13 dL g-1 was obtained at optimal conditions of sucrose (4.0% w/v), peptone (0.75% w/v) and incubation period of 4.69 day. The monosaccharide composition of EPS was mannose, arabinose and glucose at a molar ratio of 1.0:0.26:0.08. The EPS showed high water solubility (38.5%), water holding capacity (514.46%) and foaming capacity (55.55%). The EPS showed moderate antioxidant activity in vitro and good emulsion stabilizing properties against several hydrophobic compounds. The emulsifying activity was stable at different temperatures, pH and ionic strength. Additionally, the acid hydrolysate of the EPS was evaluated as a carbon source for the mixotrophic cultivation of industrially important Spirulina platensis. It induced an enhancement of not only biomass production of S. platensis, but also cellular contents (pigments, proteins and lipids) leading to higher nutritional value.


Assuntos
Antioxidantes/farmacologia , Polissacarídeos/química , Spirulina/efeitos dos fármacos , Virgibacillus/química , Antioxidantes/química , Arabinose/química , Emulsificantes/química , Emulsificantes/farmacologia , Glucose/química , Interações Hidrofóbicas e Hidrofílicas/efeitos dos fármacos , Manose/química , Proteínas/química , Spirulina/crescimento & desenvolvimento , Spirulina/metabolismo , Açúcares/química , Viscosidade/efeitos dos fármacos , Água/química
5.
Arch Virol ; 165(3): 737-741, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31875246

RESUMO

The novel bacterial virus Mimir87, infecting the salt-tolerant bacterium Virgibacillus halotolerans, was isolated from worker honey bees. Mimir87 has an elongated head and a long non-contractile tail consistent with members of the Siphoviridae phage family. The phage genome comprises 48,016 base pairs and encodes 68 predicted proteins, to 34 of which a function could be assigned from homology analysis. The phage encodes two metabolism-related transporter proteins previously not observed in bacteriophage genomes. Mimir87 displays some relatedness to several Bacillus and Paenibacillus viruses; however, the overall sequence dissimilarity suggests Mimir87 to be a representative of a new phage genus.


Assuntos
Siphoviridae/classificação , Siphoviridae/genética , Proteínas Virais/genética , Virgibacillus/virologia , Sequência de Bases , DNA Viral/genética , Genoma Viral/genética , Análise de Sequência de DNA , Siphoviridae/isolamento & purificação
6.
Sci Rep ; 9(1): 19633, 2019 12 23.
Artigo em Inglês | MEDLINE | ID: mdl-31873136

RESUMO

Studies have demonstrated that microbes facilitate the incorporation of Mg2+ into carbonate minerals, leading to the formation of potential dolomite precursors. Most microbes that are capable of mediating Mg-rich carbonates have been isolated from evaporitic environments in which temperature and salinity are higher than those of average marine environments. However, how such physicochemical factors affect and concur with microbial activity influencing mineral precipitation remains poorly constrained. Here, we report the results of laboratory precipitation experiments using two mineral-forming Virgibacillus strains and one non-mineral-forming strain of Bacillus licheniformis, all isolated from the Dohat Faishakh sabkha in Qatar. They were grown under different combinations of temperature (20°, 30°, 40 °C), salinity (3.5, 7.5, 10 NaCl %w/v), and Mg2+:Ca2+ ratios (1:1, 6:1 and 12:1). Our results show that the incorporation of Mg2+ into the carbonate minerals is significantly affected by all of the three tested factors. With a Mg2+:Ca2+ ratio of 1, no Mg-rich carbonates formed during the experiments. With a Mg2+:Ca2+ ratios of 6 and 12, multivariate analysis indicates that temperature has the highest impact followed by salinity and Mg2+:Ca2+ ratio. The outcome of this study suggests that warm and saline environments are particularly favourable for microbially mediated formation of Mg-rich carbonates and provides new insight for interpreting ancient dolomite formations.


Assuntos
Carbonato de Cálcio/metabolismo , Temperatura Alta , Magnésio/metabolismo , Salinidade , Virgibacillus , Virgibacillus/crescimento & desenvolvimento , Virgibacillus/isolamento & purificação
7.
Biomed Res Int ; 2019: 6470897, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31886235

RESUMO

A new peptidase designated as SAPV produced from a moderately halophilic Virgibacillus natechei sp. nov., strain FarDT was investigated by purification to homogeneity followed by biochemical and molecular characterization purposes. Through optimization, it was determined that the optimum peptidase activity was 16,000 U/mL. It was achieved after 36 h incubation at 35°C in the optimized enzyme liquid medium (ELM) at pH 7.4 that contains only white shrimp shell by-product (60 g/L) as sole energy and carbon sources. The SAPV enzyme is a monomer protein with a molecular mass of 31 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and high-performance liquid chromatography (HPLC) gel filtration chromatography. The sequence of its NH2-terminal amino-acid residues showed homology with those of Bacillus peptidases S8/S53 superfamily. The SAPV showed optimal activity at pH 9 and 60°C. Irreversible inhibition of enzyme activity by diiodopropyl fluorophosphates (DFP) and phenylmethanesulfonyl fluoride (PMSF) confirmed its belonging to the serine peptidases. Considering its interesting biochemical characterization, the sapV gene was cloned, sequenced, and heterologously overexpressed in the extracellular fraction of E. coli BL21(DE3)pLysS. The biochemical properties of the recombinant peptidase (rSAPV) were similar to those of the native one. The highest sequence identity value (97.66%) of SAPV was obtained with peptidase S8 from Virgibacillus massiliensis DSM 28587, with 9 amino-acid residues of difference. Interestingly, rSAPV showed an outstanding and high resistance to several organic solvents than SPVP from Aeribacillus pallidus VP3 and Thermolysin type X. Furthermore, rSAPV exhibited an excellent detergent stability and compatibility than Alcalase 2.4 L FG and Bioprotease N100L. Considering all these remarkable properties, rSAPV has attracted the interest of industrialists.


Assuntos
Proteínas de Bactérias , Serina Proteases , Virgibacillus , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Detergentes , Escherichia coli/genética , Concentração de Íons de Hidrogênio , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Análise de Sequência de Proteína , Serina Proteases/química , Serina Proteases/genética , Serina Proteases/metabolismo , Virgibacillus/enzimologia , Virgibacillus/genética
8.
BMC Genomics ; 20(1): 696, 2019 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-31481022

RESUMO

BACKGROUND: Biosynthetic gene clusters produce a wide range of metabolites with activities that are of interest to the pharmaceutical industry. Specific interest is shown towards those metabolites that exhibit antimicrobial activities against multidrug-resistant bacteria that have become a global health threat. Genera of the phylum Firmicutes are frequently identified as sources of such metabolites, but the biosynthetic potential of its Virgibacillus genus is not known. Here, we used comparative genomic analysis to determine whether Virgibacillus strains isolated from the Red Sea mangrove mud in Rabigh Harbor Lagoon, Saudi Arabia, may be an attractive source of such novel antimicrobial agents. RESULTS: A comparative genomics analysis based on Virgibacillus dokdonensis Bac330, Virgibacillus sp. Bac332 and Virgibacillus halodenitrificans Bac324 (isolated from the Red Sea) and six other previously reported Virgibacillus strains was performed. Orthology analysis was used to determine the core genomes as well as the accessory genome of the nine Virgibacillus strains. The analysis shows that the Red Sea strain Virgibacillus sp. Bac332 has the highest number of unique genes and genomic islands compared to other genomes included in this study. Focusing on biosynthetic gene clusters, we show how marine isolates, including those from the Red Sea, are more enriched with nonribosomal peptides compared to the other Virgibacillus species. We also found that most nonribosomal peptide synthases identified in the Virgibacillus strains are part of genomic regions that are potentially horizontally transferred. CONCLUSIONS: The Red Sea Virgibacillus strains have a large number of biosynthetic genes in clusters that are not assigned to known products, indicating significant potential for the discovery of novel bioactive compounds. Also, having more modular synthetase units suggests that these strains are good candidates for experimental characterization of previously identified bioactive compounds as well. Future efforts will be directed towards establishing the properties of the potentially novel compounds encoded by the Red Sea specific trans-AT PKS/NRPS cluster and the type III PKS/NRPS cluster.


Assuntos
Mineração de Dados , Genômica , Família Multigênica/genética , Virgibacillus/genética , Virgibacillus/metabolismo , Genoma Bacteriano/genética , Ilhas Genômicas/genética , Ribossomos/metabolismo
9.
World J Microbiol Biotechnol ; 35(9): 133, 2019 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-31432254

RESUMO

There is a significant increase in the discovery of new antimicrobial compounds in recent past to combat drug resistant pathogens. Members of the genus Bacillus and related genera have been screened extensively due to their ability to produce wide range of antimicrobial compounds. In this study, we have isolated and characterized a new antimicrobial peptide from a marine bacterium identified as Virgibacillus species. The low molecular mass and stability of the antimicrobial substance pointed towards the bacteriocinogenic nature of the compound. The RAST analysis of genome sequence showed presence of a putative bacteriocin biosynthetic cluster containing genes necessary for synthesis of a lanthipeptide. Translated amino acid sequence of mature C-terminal propeptide showed identity with salivaricin A (52.2%) and lacticin A (33.3%). Accordingly, the mass (2417 Da) obtained by MALDI analysis was in agreement with posttranslational modifications of the leader peptide to yield three methyl lanthionine rings and a disulfide bond between two free cysteine residues. The lanthipeptide was named as virgicin, which selectively inhibited the growth of Gram-positive bacteria and biofilm formation by Enterococcus faecalis. Inhibition of biofilm formation by E. faecalis was also observed in in vitro model experiments using hydroxyapatite discs. Thus, virgicin appears to be a promising new bacteriocin to control oral biofilm formation by selective pathogens.


Assuntos
Bacteriocinas/isolamento & purificação , Bacteriocinas/farmacologia , Enterococcus faecalis/efeitos dos fármacos , Enterococcus faecalis/crescimento & desenvolvimento , Peptídeos/isolamento & purificação , Peptídeos/farmacologia , Virgibacillus/metabolismo , Bacteriocinas/química , Bacteriocinas/genética , Biofilmes/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Vias Biossintéticas/genética , Genoma Bacteriano , Peso Molecular , Família Multigênica , Peptídeos/química , Peptídeos/genética , Água do Mar/microbiologia , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Virgibacillus/classificação , Virgibacillus/isolamento & purificação
10.
Sci Rep ; 8(1): 16441, 2018 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-30401905

RESUMO

A new transaminase (VbTA) was identified from the genome of the halotolerant marine bacterium Virgibacillus 21D. Following heterologous expression in Escherichia coli, it was located entirely in the insoluble fraction. After a single mutation, identified via sequence homology analyses, the VbTA T16F mutant was successfully expressed in soluble form and characterised. VbTA T16F showed high stability towards polar organic solvents and salt exposure, accepting mainly hydrophobic aromatic amine and carbonyl substrates. The 2.0 Å resolution crystal structure of VbTA T16F is here reported, and together with computational calculations, revealed that this mutation is crucial for correct dimerisation and thus correct folding, leading to soluble protein expression.


Assuntos
Proteínas de Bactérias/química , Mutação Puntual , Cloreto de Sódio/química , Solventes/química , Transaminases/química , Virgibacillus/enzimologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Cristalografia por Raios X , Conformação Proteica , Solubilidade , Transaminases/genética , Transaminases/metabolismo , Virgibacillus/classificação
11.
FEMS Microbiol Lett ; 365(19)2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30124875

RESUMO

This study identified marine microorganisms from Mexican coasts that had antimicrobial activity against Staphylococcus aureus and Vibrio parahaemolyticus, which are known worldwide to be food-poisoning agents. Representative specimens of algae, saline sediment, crustaceans and mollusks were collected. Of the 42 tested strains, 15 inhibited these pathogens. Bacillus and Virgibacillus strains were identified by 16S rRNA gene sequencing. The strains with the highest inhibitory activity against S. aureus were PCRS1-07 (B. aerius), BLCG-05 and GUO-01 (B. pumilus). The strains GUHC-04, BLCG-05, GUHC-03 (B. altitudinis) and BLBSe-05 (B. oryzicola) showed higher antimicrobial activity against V. parahaemolyticus. Biofilm production by all strains was moderate, but B. altitudinis produced a stronger biofilm. This is the first study to isolate B. aerius, B. oryzicola, B. safensis, B. boroniphilus, B. altitudinis and V. senegalensis from marine ecosystems in Mexico as well as the first study to report their inhibitory effects against both S. aureus and V. parahaemolyticus. Bioactivity of spent media from the antagonistic strains cultured as biofilm also demonstrated high antimicrobial activity. The active compounds of the antagonists are currently being studied and tested. Marine ecosystems have the highest bacterial diversity associated with invertebrates and seaweed; however, this bacterial diversity has not been well-studied on Mexican coasts.


Assuntos
Bacillus/isolamento & purificação , Staphylococcus aureus , Vibrio parahaemolyticus , Virgibacillus/isolamento & purificação , Anti-Infecciosos , Bacillus/fisiologia , Biofilmes , México , Água do Mar , Alga Marinha/microbiologia , Virgibacillus/fisiologia , Microbiologia da Água
12.
PLoS One ; 13(8): e0201346, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30091990

RESUMO

We previously used whole-genome sequencing and Tn5 transposon mutagenesis to identify 16 critical genes involved in the halotolerance of Halomonas beimenensis, a species in the phylum Proteobacteria. In this present study, we sought to determine if orthologous genes in another phylum are also critical for halotolerance. Virgibacillus spp. are halotolerant species that can survive in high-saline environments. Some Virgibacillus species are used in different aspects of food processing, compatible solute synthesis, proteinase production, and wastewater treatment. However, genomic information on Virgibacillus chiguensis is incomplete. We assembled a draft V. chiguensis strain NTU-102 genome based on high-throughput next-generation sequencing (NGS) and used transcriptomic profiling to examine the high-saline response in V. chiguensis. The V. chiguensis draft genome is approximately 4.09 Mbp long and contains 4,166 genes. The expression profiles of bacteria grown in 5% and 20% NaCl conditions and the corresponding Gene Ontology (GO) and clusters of orthologous groups (COG) categories were also analyzed in this study. We compared the expression levels of these 16 orthologs of halotolerance-related genes in V. chiguensis and H. beimenensis. Interestingly, the expression of 7 of the 16 genes, including trkA2, smpB, nadA, mtnN2, rfbP, lon, and atpC, was consistent with that in H. beimenensis, suggesting that these genes have conserved functions in different phyla. The omics data were helpful in exploring the mechanism of saline adaptation in V. chiguensis, and our results indicate that these 7 orthologs may serve as biomarkers for future screening of halotolerant species in the future.


Assuntos
DNA Bacteriano/genética , Genes Bacterianos/fisiologia , Halomonas/genética , Tolerância ao Sal/fisiologia , Virgibacillus/genética , Técnicas de Tipagem Bacteriana/métodos , Perfilação da Expressão Gênica/métodos , Halomonas/fisiologia , Filogenia , Análise de Sequência de DNA/métodos , Virgibacillus/fisiologia
13.
Int J Syst Evol Microbiol ; 68(6): 2015-2023, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29688169

RESUMO

Three Gram-variable, moderately halophilic, motile, endospore-forming rods, designated P2-C2T, P3-H5T and P3-B8, were isolated from marine sediment of the Southwest Indian Ocean by using the microfluidic streak plate method. Phylogeny based on 16S rRNA gene sequences showed that strains P2-C2T and P3-H5T formed a robust cluster within the genus Virgibacillus and exhibited 16S rRNA gene similarity levels of 95.3-96.8 and 94.9-96.3 % to the type strains of Virgibacillus species, respectively. The 16S rRNA gene similarity between P2-C2T and P3-H5T was 97.6 %. Strain P3-B8 has an identical 16S rRNA gene sequence to strain P3-H5T. For all the novel strains, the predominant cellular fatty acids were anteiso-C15 : 0 and anteiso-C17 : 0, the main menaquinone was MK-7, and the polar lipid profiles contained diphosphatidylglycerol and phosphatidylglycerol. The genomic DNA G+C contents of strains P2-C2T, P3-H5T and P3-B8 were 38.3, 37.3 and 37.5 mol%, respectively. Combined data from phenotypic and genotypic studies demonstrated that strains P2-C2T and P3-H5T/P3-B8 are representatives of two different novel species of the genus Virgibacillus, for which the name Virgibacillus indicus sp. nov. and Virgibacillusprofundi are proposed. The type strains are P2-C2T (=CGMCC 1.16138T=NBRC 113014T) and P3-H5T (=CGMCC 1.16139T=NBRC 113015T).


Assuntos
Sedimentos Geológicos/microbiologia , Filogenia , Água do Mar/microbiologia , Virgibacillus/classificação , Archaea/genética , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Oceano Índico , Microfluídica , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Virgibacillus/genética , Virgibacillus/isolamento & purificação , Vitamina K 2/análogos & derivados , Vitamina K 2/química
14.
Anal Biochem ; 548: 109-114, 2018 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-29499175

RESUMO

The Fe(II)/2-oxoglutarate-dependent dioxygenases (2-OGDs) catalyze the oxidation of substrates ranging from small molecules to large biomolecules with concomitant oxidation of co-substrate (2-oxoglutarate) into succinate. In the present study, we reported a coupled colorimetric assay that can be generally applied to measure the activities of all members of 2-OGDs family. Succinyl-CoA synthetase is employed as the coupling enzyme to transform succinate produced from 2-OGDs catalysis to form succinyl-CoA with concomitant hydrolysis of ATP to form ADP and orthophosphate. Orthophosphate can be quantitated by reacting it with molybdic acid forming a blue pigment. As a proof of concept, kinetic parameters of ectoine hydroxylase obtained using this method are compared to a traditional time- and labor-consuming HPLC based method. As 2-OGDs family enzymes are important drug targets due to their impressive versatility in catalyzing numerous oxidative reactions that are still very challenging using synthetic chemistry, colorimetric method detailed in the manuscript has the potential to enable the practice of high throughput drug screening for 2-OGDs.


Assuntos
Proteínas de Bactérias/análise , Dioxigenases/análise , Virgibacillus/enzimologia , Colorimetria/métodos , Dioxigenases/metabolismo , Ácidos Cetoglutáricos/metabolismo , Succinato-CoA Ligases/análise , Succinato-CoA Ligases/metabolismo , Ácido Succínico/metabolismo
15.
Int J Syst Evol Microbiol ; 68(4): 1190-1196, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29465338

RESUMO

A rod-shaped, Gram-stain-positive, motile and aerobic bacterium, designated LM2416T, was isolated from faeces of Lophuras winhoii living in Seoul Grand Park, Gyeonggi-do, Republic of Korea. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain LM2416T belonged to the genus Virgibacillus, sharing high 16S rRNA gene sequence similarities to Virgibacillus necropolis LMG 19488T (99.0 %), Virgibacillus carmonensis LMG 20964T (98.4 %), Virgibacillus arcticus Hal 1T (98.3 %) and Virgibacillus flavescens S1-20T (97.9 %). The isolate grew at 10-30 °C, pH 6-7 and 0-20 % (w/v) NaCl. Optimal growth was observed at 30 °C, pH 6-7 and 10 % (w/v) NaCl. The major fatty acid was anteiso-C15 : 0. Polar lipids were composed of phosphatidylglycerol, diphosphatidylglycerol, three unknown phospholipids and two unknown aminophospholipids. The main menaquinone was MK-7. Strain LM2416T had alanine, lysine, glutamic acid, glycine and aspartic acid as cell-wall amino acids and ribose as a cell-wall sugar. The whole genome sequences of strain LM2416T and V. necropolis KCTC 3820T were sequenced by PacBio RS II sequencing. The genome sequence-based G+C content of strain LM2416T was 39.5 mol%. The orthologous average nucleotide identity value, showing genetic relatedness between strain LM2416T and V. necropolis KCTC 3820T, was 78.3 %. Based on the phylogenetic, biochemical, chemotaxonomic and genotypic data presented in this study, strain LM2416T is considered to represent a novel species of the genus Virgibacillus, for which the name Virgibacillus phasianinus is proposed. The type strain is LM2416T (=KCTC 33927T=JCM 32144T).


Assuntos
Fezes/microbiologia , Galliformes/microbiologia , Filogenia , Virgibacillus/classificação , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , DNA Ribossômico/genética , Ácidos Graxos/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Virgibacillus/genética , Virgibacillus/isolamento & purificação , Vitamina K 2/análogos & derivados , Vitamina K 2/química
16.
Mar Drugs ; 16(2)2018 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-29414856

RESUMO

Xanthomonas oryzae pv. oryzae, which causes rice bacterial blight, is one of the most destructive pathogenic bacteria. Biological control against plant pathogens has recently received increasing interest. 1-Deoxy-N-acetylglucosamine (1-DGlcNAc) was extracted from the supernatant of Virgibacillus dokdonensis MCCC 1A00493 fermentation through antibacterial bioassay-guided isolation. Its structure was elucidated by LC/MS, NMR, chemical synthesis and time-dependent density functional theory (TD-DFT) calculations. 1-DGlcNAc specifically suppressed X. oryzae pv. oryzae PXO99A (MIC was 23.90 µg/mL), but not other common pathogens including Xanthomonas campestris pv. campestris str.8004 and Xanthomonas oryzae pv. oryzicola RS105. However, its diastereomer (2-acetamido-1,5-anhydro-2-deoxy-d-mannitol) also has no activity to X. oryzae pv. oryzae. This result suggested that activity of 1-DGlcNAc was related to the difference in the spatial conformation of the 2-acetamido moiety, which might be attributed to their different interactions with a receptor. Eighty-four unique proteins were found in X. oryzae pv. oryzae PXO99A compared with the genome of strains8004 and RS105 by blastp. There may be unique interactions between 1-DGlcNAc and one or more of these unique proteins in X. oryzae pv. oryzae. Quantitative real-time PCR and the pharmMapper server indicated that proteins involved in cell division could be the targets in PXO99A. This research suggested that specificity of active substance was based on the active group and spatial conformation selection, and these unique proteins could help to reveal the specific mechanism of action of 1-DGlcNAc against PXO99A.


Assuntos
Acetilglucosamina/análogos & derivados , Antibacterianos/química , Antibacterianos/farmacologia , Virgibacillus/química , Acetilglucosamina/química , Acetilglucosamina/farmacologia , Fermentação , Regulação Bacteriana da Expressão Gênica , Testes de Sensibilidade Microbiana , Oryza/microbiologia , Doenças das Plantas/microbiologia , Água do Mar/microbiologia , Estereoisomerismo , Relação Estrutura-Atividade , Virgibacillus/genética , Xanthomonas/efeitos dos fármacos , Xanthomonas/genética
17.
J Microbiol ; 55(12): 933-938, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29214493

RESUMO

A Gram-stain-positive, halophilic, rod-shaped, non-motile, spore forming bacterium, strain NKC1-2T, was isolated from kimchi, a Korean fermented food. Comparative analysis based on 16S rRNA gene sequence demonstrated that the isolated strain was a species of the genus Virgibacillus. Strain NKC1-2T exhibited high level of 16S rRNA gene sequence similarity with the type strains of Virgibacillus xinjiangensis SL6-1T (96.9%), V. sediminis YIM kkny3T (96.8%), and V. salarius SA-Vb1T (96.7%). The isolate grew at pH 6.5-10.0 (optimum, pH 8.5-9.0), 0.0-25.0% (w/v) NaCl (optimum, 10-15% NaCl), and 15-50°C (optimum, 37°C). The major menaquinone in the strain was menaquinone-7, and the main peptidoglycan of the strain was meso-diaminopimelic acid. The predominant fatty acids of the strain were iso-C14:0, anteisio-C15:0, iso- C15:0, and iso-C16:0 (other components were < 10.0%). The polar lipids consisted of diphosphatidylglycerol and phosphatidylglycerol. The genomic DNA G + C content of NKC1-2T was 42.5 mol%. On the basis of these findings, strain NKC1-2T is proposed as a novel species in the genus Virgibacillus, for which the name Virgibacillus kimchii sp. nov. is proposed (=KACC 19404T =JCM 32284T). The type strain of Virgibacillus kimchii is NKC1-2T.


Assuntos
Brassica/microbiologia , Cloreto de Sódio/metabolismo , Virgibacillus/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Ácidos Graxos/metabolismo , Filogenia , RNA Ribossômico 16S/genética , Cloreto de Sódio/análise , Virgibacillus/classificação , Virgibacillus/genética , Virgibacillus/metabolismo
18.
Food Chem ; 224: 320-328, 2017 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-28159274

RESUMO

Modulation of inflammation-related immune response on THP-1 macrophages of protein hydrolysates derived from tilapia mince, casein and pea protein, were investigated. The protein substrates were hydrolyzed by Virgibacillus halodenitrificans SK1-3-7 proteinase. The degree of hydrolysis (DH) of casein was observed to be the highest throughout the course of hydrolysis. When challenging THP-1 macrophages, tilapia mince hydrolysate (TMH) enhanced innate immunity through induction of IL-1ß and COX-2 expression. Anti-inflammatory activity was observed in casein hydrolysate (CH) and pea protein hydrolysate (PPH) by attenuating lipopolysaccharide- (LPS) induced pro-inflammatory gene expression in THP-1 macrophages. CH suppressed IL-1ß, IL-6, IL-8, TNF-α and COX-2, while PPH reduced LPS-induced IL-6 and TNF-α responses. In addition, CH and PPH showed stronger suppression of LPS-induced pro-inflammatory gene expression compared with non-hydrolyzed casein and pea protein. These results suggest that TMH, CH and PPH prepared from V. halodenitrificans SK1-3-7 proteinase are potential functional food ingredients with immunomodulatory activity.


Assuntos
Caseínas/farmacologia , Proteínas de Peixes/farmacologia , Peptídeo Hidrolases/metabolismo , Hidrolisados de Proteína/farmacologia , Virgibacillus/enzimologia , Animais , Anti-Inflamatórios/farmacologia , Células Cultivadas , Ciclídeos , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Regulação da Expressão Gênica , Humanos , Hidrólise , Imunidade Inata , Imunomodulação , Inflamação , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Interleucina-8/genética , Interleucina-8/metabolismo , Lipopolissacarídeos/toxicidade , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Ervilhas/química , Proteínas de Plantas/farmacologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo
19.
Curr Microbiol ; 74(2): 219-223, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27957631

RESUMO

A Gram-positive, moderately halophilic, endospore-forming bacterium, designated MerVT, was isolated from a sediment sample of a saline lake located in Ain Salah, south of Algeria. The cells were rod shaped and motile. Isolate MerVT grew at salinity interval of 0.5-25% NaCl (optimum, 5-10%), pH 6.0-12.0 (optimum, 8.0), and temperature between 10 and 40 °C (optimum, 30 °C).The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, a glycolipid, a phospholipid, and two lipids, and MK-7 is the predominant menaquinone. The predominant cellular fatty acids were anteiso C15:0 and anteiso C17:0. The DNA G+C content was 45.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain MerVT was most closely related to Virgibacillus halodenitrificans (gene sequence similarity of 97.0%). On the basis of phenotypic, chemotaxonomic properties, and phylogenetic analyses, strain MerVT (=DSM = 28944T) should be placed in the genus Virgibacillus as a novel species, for which the name Virgibacillus ainsalahensis is proposed.


Assuntos
Sedimentos Geológicos/microbiologia , Virgibacillus/classificação , Virgibacillus/isolamento & purificação , Argélia , Composição de Bases , Análise por Conglomerados , Citosol/química , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Ácidos Graxos/análise , Glicolipídeos/análise , Lagos , Locomoção , Fosfolipídeos/análise , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Cloreto de Sódio/metabolismo , Temperatura , Virgibacillus/genética , Virgibacillus/fisiologia , Vitamina K 2/análise
20.
Food Chem ; 220: 190-197, 2017 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-27855889

RESUMO

The angiotensin I-converting enzyme (ACE) inhibitory activity of protein hydrolysates from tilapia muscle fractions, namely mince (M), washed mince (WM), and sarcoplasmic protein (SP), were investigated. Each fraction was hydrolyzed by Virgibacillus halodenitrificans SK1-3-7 proteinases for up to 24h. After 8h of hydrolysis, the M hydrolysate (48% degree of hydrolysis (DH)) showed the highest ACE inhibitory activity, with an IC50 value of 0.54mg/ml, while the SP hydrolysate exhibited the lowest DH and ACE inhibition. In vitro gastrointestinal digestion reduced the ACE inhibitory activity of the M hydrolysate but enhanced its transport across Caco-2 cell monolayers. The transported peptides were found to contain 3-4 amino acid residues showing strong ACE inhibition. The novel ACE inhibitory peptide with the highest inhibition was found to be MCS, with an IC50 value of 0.29µM. Therefore, tilapia mince hydrolyzed by V. halodenitrificans proteinases contained ACE inhibitory peptides that are potentially bioavailable.


Assuntos
Inibidores da Enzima Conversora de Angiotensina/farmacologia , Proteínas Musculares/farmacologia , Peptídeo Hidrolases/metabolismo , Peptídeos/farmacologia , Hidrolisados de Proteína/farmacologia , Virgibacillus/enzimologia , Animais , Disponibilidade Biológica , Células CACO-2 , Humanos , Tilápia
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