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1.
Toxins (Basel) ; 15(1)2023 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-36668875

RESUMO

The beetle Anthonomus grandis Boheman, 1843, is the main cotton pest, causing enormous losses in cotton. The breeding of genetically modified plants with A. grandis resistance is seen as an important control strategy. However, the identification of molecules with high toxicity to this insect remains a challenge. The susceptibility of A. grandis larvae to proteins (Cry1Ba, Cry7Ab, and Mpp23Aa/Xpp37Aa) from Bacillus thuringiensis Berliner, 1915, with toxicity reported against Coleopteran, has been evaluated. The ingestion of different protein concentrations (which were incorporated into an artificial diet) by the larvae was tested in the laboratory, and mortality was evaluated after one week. All Cry proteins tested exhibited higher toxicity than that the untreated artificial diet. These Cry proteins showed similar results to the control Cry1Ac, with low toxicity to A. grandis, since it killed less than 50% of larvae, even at the highest concentration applied (100 µg·g-1). Mpp/Xpp proteins provided the highest toxicity with a 0.18 µg·g-1 value for the 50% lethal concentration. Importantly, this parameter is the lowest ever reported for this insect species tested with B. thuringiensis proteins. This result highlights the potential of Mpp23Aa/Xpp37Aa for the development of a biotechnological tool aiming at the field control of A. grandis.


Assuntos
Bacillaceae , Bacillales , Bacillus thuringiensis , Besouros , Inseticidas , Gorgulhos , Animais , Larva , Bacillus thuringiensis/genética , Bacillus thuringiensis/metabolismo , Inseticidas/toxicidade , Inseticidas/metabolismo , Melhoramento Vegetal , Gossypium
2.
Sci Rep ; 13(1): 1243, 2023 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-36690691

RESUMO

Aromatic compounds and metalloid oxyanions are abundant in the environment due to natural resources and industrial wastes. The high toxicity of phenol and tellurite poses a significant threat to all forms of life. A halotolerant bacterium was isolated and identified as Lysinibacillus sp. EBL303. The remediation analysis shows that 500 mg/L phenol and 0.5 mM tellurite can be remediated entirely in separate cultures within 74 and 56 h, respectively. In addition, co-remediation of pollutants resulted in the same phenol degradation and 27% less tellurite reduction within 98 h. Since phenol and tellurite exhibited inhibitory behavior, their removal kinetics fitted well with the first-order model. In the characterization of biosynthesized tellurium nanoparticles (TeNPs), transmission electron microscopy, dynamic light scattering, FE-SEM, and dispersive X-ray (EDX) showed that the separated intracellular TeNPs were spherical and consisted of only tellurium with 22-148 nm in size. Additionally, investigations using X-ray diffraction and Fourier-transform infrared spectroscopy revealed proteins and lipids covering the surface of these amorphous TeNPs. Remarkably, this study is the first report to demonstrate the simultaneous bioremediation of phenol and tellurite and the biosynthesis of TeNPs, indicating the potential of Lysinibacillus sp. EBL303 in this matter, which can be applied to environmental remediation and the nanotechnology industry.


Assuntos
Bacillaceae , Nanopartículas , Telúrio/química , Biodegradação Ambiental , Fenol , Nanopartículas/metabolismo , Bacillaceae/metabolismo , Fenóis
3.
Biomolecules ; 13(1)2023 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-36671516

RESUMO

Poly(3-hydroxybutyrate) (PHB) is a versatile thermoplastic with superior biodegradability and biocompatibility that is intracellularly accumulated by numerous bacterial and archaeal species. Priestia sp. strain JY310 that was able to efficiently biotransform reducing sugars in d-xylose-rich rice husk hydrolysate (reducing sugarRHH) to PHB was isolated from the soil of a rice paddy. Reducing sugarRHH including 12.5% d-glucose, 75.3% d-xylose, and 12.2% d-arabinose was simply prepared using thermochemical hydrolysis of 3% H2SO4-treated rice husk for 15 min at 121 °C. When cultured with 20 g/L reducing sugarRHH under optimized culture conditions in a batch bioreactor, Priestia sp. strain JY310 could produce PHB homopolymer up to 50.4% of cell dry weight (6.2 g/L). The melting temperature, heat of fusion, and thermal decomposition temperature of PHB were determined to be 167.9 °C, 92.1 J/g, and 268.1 °C, respectively. The number average and weight average molecular weights of PHB with a broad polydispersity index value (4.73) were estimated to be approximately 16.2 and 76.8 kg/mol, respectively. The findings of the present study suggest that Priestia sp. strain JY310 can be exploited as a good candidate for the low-cost production of low molecular weight PHB with improved biodegradability and reduced brittleness from inexpensive agricultural waste hydrolysates.


Assuntos
Bacillaceae , Oryza , Ácido 3-Hidroxibutírico , Xilose/metabolismo , Solo , Hidroxibutiratos/metabolismo , Oryza/metabolismo , Peso Molecular , Bacillaceae/metabolismo , Bactérias/metabolismo , Biotransformação
4.
Syst Appl Microbiol ; 46(1): 126389, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36577291

RESUMO

The strains designed PP-18T, JC-4 and JC-7 isolated from soils, were Gram-stain-positive rods, facultative anaerobe, endospore-forming bacteria. The strains produced l-lactic acid from glucose. They showed positive for catalase but negative for oxidase, nitrate reduction and arginine hydrolysis. Strains P-18T, JC-4 and JC-7 were closely related to Weizmannia coagulans LMG 6326T (97.27-97.64%) and W. acidiproducens KCTC 13078T (96.46-96.74%) based on 16S rRNA gene sequence similarity, respectively. They contained meso-diaminopimelic acid in cell wall peptidoglycan and had seven isoprene units (MK-7) as the predominant menaquinone. The major cellular fatty acids of strain PP-18T were iso-C15:0, anteiso-C17:0, iso-C16:0 and anteiso-C15:0. The ANIb and ANIm values among the genomes of strains PP-18T, JC-4 and JC-7 are above 99.4% while their ANIb and ANIm values among them and W. coagulans LMG 6326T and W. acidiproducens KCTC 13078T were ranged from 76.61 to 79.59%. These 3 strains showed the digital DNA-DNA hybridization (dDDH) values of 20.7-23.6% when compared with W. coagulans LMG 6326T and W. acidiproducens DSM 23148T. The DNA G + C contents of strains PP-18T, JC-4 and JC-7 were 45.82%, 45.86% and 45.86%, respectively. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphoglycolipids. The results of phenotypic and chemotaxonomic characteristics and whole-genome analysis indicated that the strains PP-18T, JC-4 and JC-7 should be represented as a novel species within the genus Weizmannia for which the name Weizmannia acidilactici sp. nov. is proposed. The type strain is PP-18T (=KCTC 33974T = NBRC 113028T = TISTR 2515T).


Assuntos
Bacillaceae , Fosfolipídeos , Fosfolipídeos/análise , Ácido Láctico , RNA Ribossômico 16S/genética , Solo , DNA Bacteriano/genética , Filogenia , Ácidos Graxos/análise , Hibridização de Ácido Nucleico , Análise de Sequência de DNA , Técnicas de Tipagem Bacteriana
5.
Artigo em Inglês | MEDLINE | ID: mdl-36554967

RESUMO

In recent years, PBAT (polybutylene adipate-co-terephthalate) mulch has become one of the most commonly used biodegradable mulching films. In this paper, five potential strains of PBAT film degrading bacteria were screened from the soil sample using PBAT film as the sole carbon source. A highly efficient PBAT degrading strain JZ1 was isolated by comparing the degradation performance of PBAT mulching film identified as Peribacillus frigoritolerans S2313 by 16S rDNA sequence analysis. The capacity of the strain to degrade PBAT film was optimized by adjusting the cultivation conditions such as nitrogen source, pH, and inoculum volume. After 8 weeks of cultivation, the actual degradation rate of the strain to PBAT mulch film reached 12.45%. SEM (scanning electron microscopy) coupled with EDX (energy dispersive spectroscopy) analysis showed that microbial degradation is an oxidation process and is mainly due to the amorphous regions of the PBAT film. The biodegradation of PBAT film by Peribacillus frigoritolerans may provide a promising method for regulating the degradation progress of PBAT film in the farmlands.


Assuntos
Bacillaceae , Poliésteres , Poliésteres/química , Microscopia Eletrônica de Varredura , Filmes Cinematográficos , Bactérias , Fazendas , Biodegradação Ambiental
6.
Ecotoxicol Environ Saf ; 248: 114294, 2022 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-36402075

RESUMO

Biochar provides a suitable microenvironment for the growth of microorganisms. It may directly or indirectly affect changes in the population of microorganisms, thus affecting heavy metal bioavailability. This study aims to explore the effects of microbiological inoculation with and without biochar on microorganisms and on the bioavailability of heavy metals during pig manure composting. Three composting experiments were conducted under various conditions including no treatment (CK), only microbiological inoculation (TA), and integration with biochar (TB). Compared with raw materials before compost, TA reduced the bioavailability of Cu by 25.1%, Zn by 25.64%, and both Pb and Cr by 1.75%. TB reduced the bioavailability of Cu by 35.38%, Zn by 19.34%, Pb by 0.81%, and Cr by 3.9%. Furthermore, correlation analysis demonstrated that Debaryomyces were the primary fungi, possibly controlling the passivation of Cr. Bacillus, Fusarium, Pseudogracilibacillus, Sinibacillus, and Botryotrichum were the primary bacteria and fungi potentially governing the passivation of Zn, Lastly, Debaryomyces and Penicillium were the primary bacteria and fungi potentially controlling the passivation of Pb and Cu, respectively. Overall, we demonstrated that pig manure added to the microbial inoculum and biochar effectively reduced the bioavailability of heavy metals, thereby offering an applicable technology for reducing heavy metal contamination during pig manure composting.


Assuntos
Bacillaceae , Compostagem , Metais Pesados , Suínos , Animais , Esterco , Disponibilidade Biológica , Chumbo
7.
Sci Rep ; 12(1): 18883, 2022 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-36344604

RESUMO

Pine wilt disease (PWD) is a destructive vector-borne forest disease caused by the nematode Bursaphelenchus xylophilus. To date, several options are available for the management of pine wilt disease; however constant development and search for natural products with potential nematicidal activity are imperative to diversify management options and to cope with the possible future emergence of resistance in parasitic nematodes. Here, a combined metabolomics and genomics approach was employed to investigate the chemical repertoire and biosynthetic potential of the bacterial endophyte Peribacillus frigoritolerans BE93, previously characterized to exhibit nematicidal activity against B. xylophilus. Feature-based molecular networking revealed the presence of diverse secondary metabolites. A cyclic imine heptapeptide, koranimine, was found to be among the most abundant secondary metabolites produced. Genome mining displayed the presence of several putative biosynthetic gene clusters (BGCs), including a dedicated non-ribosomal peptide synthase (NRPS) BGC for koranimine. Given the non-ribosomal peptide nature of koranimine, in silico molecular docking analysis was conducted to investigate its potential nematicidal activity against the target receptor ivermectin-sensitive invertebrate α glutamate-gated chloride channel (GluCl). Results revealed the binding of koranimine at the allosteric site of the channel-the ivermectin binding site. Moreover, the ligand-receptor interactions observed were mostly shared between koranimine and ivermectin when bound to the α GluCl receptor thus, suggesting a possibly shared mechanism of potential nematicidal activity. This study highlights the efficiency of combined metabolomics and genomics approach in the identification of candidate compounds.


Assuntos
Bacillaceae , Nematoides , Pinus , Animais , Ivermectina , Iminas , Simulação de Acoplamento Molecular , Antinematódeos/farmacologia , Antinematódeos/química , Nematoides/microbiologia , Pinus/parasitologia
8.
Int J Syst Evol Microbiol ; 72(11)2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36327328

RESUMO

A Gram-stain-positive, facultative anaerobic endospore-forming bacterium, which originated from roots/rhizosphere of maize (Zea mays), was investigated for its taxonomic position. On the basis of 16S rRNA gene sequence similarities, strain JJ-3T was grouped together with Neobacillus species showing the highest similarities to Neobacillus bataviensis (98.8 %) and the three species Neobacillus dendrensis, Neobacillus soli and Neobacillus cucumis (all 98.6 %). The 16S rRNA gene sequence similarities to the sequences of the type strains of other Neobacillus species were lower than 98.5 %. The average nucleotide identity, average amino acid identity and digital DNA-DNA hybridization values between the JJ-3T genome assembly and those of the other Neobacillus type strains were <83, <85 and <27 %, respectively. Chemotaxonomic features supported the grouping of the strain to the genus Neobacillus, e.g. the major fatty acids were C15 : 0 anteiso and C15 : 0 iso, the polar lipid profile contained the major components diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine, and the major quinone was menaquinone MK-7. Physiological and biochemical test results were slightly different from those of the most closely related species. For this reason, JJ-3T represents a novel species of the genus Neobacillus, for which we propose the name Neobacillus rhizosphaerae sp. nov., with JJ-3T (= CIP 111895T=LMG 32087T=DSM 111784T=CCM 9084T) as the type strain. We also propose to reclassify Bacillus dielmonensis as Neobacillus dielmonensis comb. nov. based mainly on the results of phylogenomic and conserved signature indel analyses.


Assuntos
Bacillaceae , Bacillus , Rizosfera , RNA Ribossômico 16S/genética , Filogenia , DNA Bacteriano/genética , Composição de Bases , Técnicas de Tipagem Bacteriana , Ácidos Graxos/química , Análise de Sequência de DNA , Fosfolipídeos/química , Zea mays/microbiologia
9.
Int J Syst Evol Microbiol ; 72(10)2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36223177

RESUMO

An aerobic, Gram-staining-positive, endospore-forming bacterium, isolated from the rhizosphere of roots of maize (Zea mays), was taxonomically studied. On the basis of 16S rRNA gene sequence similarity comparisons, strain JJ-125T clustered together with species of the genus Sutcliffiella and showed the highest similarities with Sutcliffiella zhanjiangensis (98.7 %). The 16S rRNA gene sequence similarities to the sequences of the type strains of other species of the genus Sutcliffiella were <98.4 %. The genome sequence of JJ-125T was 4 516 360 bp long and had a DNA G+C content of 37.3 %. A DNA-DNA hybridization with the type strain of S. zhanjiangensis DSM 23010T resulted in values of 42.3 and 43.9 % (reciprocal). The average nucleotide identity, average amino acid identity and digital DNA-DNA hybridization values between the JJ-125T genome assembly and those of the other type strains of species of the genus Sutcliffiella were <75%, <80 % and <21 %, respectively. Chemotaxonomic features supported the grouping of the strain with the genus Sutcliffiella, e.g. the major fatty acids included iso-C15 : 0, iso-C17 : 1 ω10c and iso-C17 : 0, the polar lipid profile contained the major components diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine, the only quinone was menaquinone MK-7 and the characteristic diamino acid was meso-diaminopimelic acid. Physiological and biochemical test results were also different from those of the most closely related species. As a consequence, JJ-125T represents a novel species of the genus Sutcliffiella, for which we propose the name Sutcliffiella rhizosphaerae sp. nov., with JJ-125T (= CIP 111883T = LMG 32156T = CCM 9046T) as the type strain.


Assuntos
Bacillaceae , Fosfatidiletanolaminas , Bacillaceae/genética , Técnicas de Tipagem Bacteriana , Composição de Bases , Cardiolipinas , DNA Bacteriano/genética , Ácido Diaminopimélico/química , Ácidos Graxos/química , Nucleotídeos , Peptidoglicano/química , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/química , Zea mays/microbiologia
10.
J Biosci ; 472022.
Artigo em Inglês | MEDLINE | ID: mdl-36222136

RESUMO

Capsicum chinense is the chilli species containing the highest amount of capsaicin, and is an important traditional spice crop of Northeast India. Capsaicinoids derived from C. chinense are used in anticancer and anti-obesity treatments, as temperature regulators, in pain therapy, and as antioxidants. The current production and yield are very low due to the lack of organized cultivation and scientific inputs, and various plant diseases. Synthetic pesticides are frequently applied to boost yields, which creates potential risks to the environment, crops, and humans. The use of plant growth-promoting rhizobacteria is an alternative strategy in crop disease management to reduce the dependency on agrochemicals, which have detrimental effects on the environment. Lysinibacillus xylanilyticus t26 isolated from the C. chinense rhizosphere has shown good prospects in plant growth promotion and biocontrol. It showed strong antagonistic activity against Pythium ultimum ITCC 1650, Rhizoctonia solani ITCC 6491, and Fusarium oxysporum ITCC 6246. The draft genome sequencing of L. xylanilyticus t26 yielded a total of 5.69 Mbp with a G+C content of 36.80%. Genome analysis revealed that L. xylanilyticus t26 is very similar to L. xylanilyticus MH683160.1, and is phylogenetically related to L. xylanilyticus IBBPo7. Bioinformatics analysis predicted that it harbored type III polyketides, non-ribosomal peptides, terpenes, and lantibiotics including cerecidin, bacteriocins, siderophores, and thiopeptides, which are important traits of rhizobacteria for the utilization of minerals and to compete with other microbes for food. The strain t26 is a potential biocontrol agent for soil-borne fungal diseases. In this study, we derived the possible siderophore production pathways through the analysis of L. xylanilyticus t26 draft genome and plant growth response bioassays. The availability of genome data provides information that this draft genome harbored a siderophore BGC, which is 33% similar to petrobactin.


Assuntos
Bacteriocinas , Capsicum , Praguicidas , Policetídeos , Agroquímicos/metabolismo , Bacillaceae , Bactérias/genética , Bacteriocinas/metabolismo , Capsaicina/metabolismo , Capsicum/metabolismo , Humanos , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Policetídeos/metabolismo , Rizosfera , Sideróforos , Solo , Microbiologia do Solo , Terpenos/metabolismo
11.
J Microbiol Methods ; 202: 106600, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36241006

RESUMO

Parageobacillus thermoglucosidasius is a promising chassis for producing chemicals and fuels. Here we designed, built and tested the pMTL60000 modular plasmids containing standardised Gram-positive and Gram-negative replicons, selectable markers and application-specific modules. The pMTL60000 modular plasmids were characterised with regard to transformation efficiency, segregational stability, copy number and compatibility.


Assuntos
Bacillaceae , Vetores Genéticos , Plasmídeos/genética , Replicon
12.
Artigo em Inglês | MEDLINE | ID: mdl-36107764

RESUMO

In the present study, the taxonomic position of Bacillus lacisalsi YSP-3T was evaluated using phylogenetic and genome-based comparison. B. lacisalsi YSP-3T showed the highest 16S rRNA gene sequence similarity to Alteribacter natronophilus M30T (98.4 %), followed by Alteribacter aurantiacus K1-5T (97.5 %) and Alteribacter populi FJAT-45347T (97.2 %). In phylogenetic (based on 16S rRNA gene sequences) and phylogenomic (based on 71 bacterial single-copy genes) trees, B. lacisalsi YSP-3T clustered with the members of the genus Alteribacter. The amino acid identity (AAI) values between B. lacisalsi YSP-3T and the members of the genus Alteribacter were >65 %, which is above the cut-off level (65-95 %) for genus delineation. The average nucleotide identity (ANI) values between B. lacisalsi YSP-3T and the members of the genus Alteribacter were <95 %, which is lower than the threshold value (95-96 %) for bacterial species delineation. The AAI value suggested that B. lacisalsi YSP-3T was a member of the genus Alteribacter while the ANIb value suggested it as a novel species of the genus Alteribacter. Based on the results, we propose to transfer Bacillus lacisalsi to the genus Alteribacter as Alteribacter lacisalsi comb. nov.


Assuntos
Bacillaceae , Bacillus , Aminoácidos , Bacillus/genética , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Nucleotídeos , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
13.
Microbiol Res ; 265: 127208, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36162147

RESUMO

Microbially induced carbonate precipitation (MICP) is a biomineralization process that has various applications in environmental pollution remediation and restoration of a range of building materials. In this study, a ureolytic bacterium, Lysinibacillus sp. GY3, isolated from an E-waste site, was found as a promising catalyst for remediation of heavy metals via the MICP process. This bacterial isolate produced significant amounts of urease and showed a great persistence in immobilization of potentially toxic elements. A reference ureolytic strain, Bacillus megaterium VS1, was selected in order to compare the efficiency of Lysinibacillus sp. GY3. Study on urease localization indicated 80 % more urease activity secreted extracellularly as for Lysinibacillus sp. GY3 compared to B. megaterium VS1. From the investigation on effects of metals on both intra- and extra-cellular urease, it was clear that Lysinibacillus sp. GY3 produced the most stable urease under conditions of metal pressure, especially retaining more than 70 % activity in the presence of 1 g/L Pb2+ and Zn2+. These results suggest that this isolated microorganism could be promisingly introduced in the MICP process to stabilize complex heavy metal pollutions, with reference to the regulating ability under harsh conditions to stabilize urease activity. This species is so important both for its biological features and environmental impacts. In addition, the present study will bring new insight in the field of metal remediation coupled with enzyme engineered biotechnology.


Assuntos
Bacillaceae , Bacillus megaterium , Metais Pesados , Bacillaceae/genética , Carbonato de Cálcio , Carbonatos , Chumbo , Urease
14.
Comput Intell Neurosci ; 2022: 4411876, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36093479

RESUMO

The focus of this research is to isolating and identifying bacteria that produce calcite precipitate, as well as determining whether or not these bacteria are suitable for incorporation into concrete in order to enhance the material's strength and make the environment protection better. In order to survive the high "potential of hydrogen" of concrete, microbes that are going to be added to concrete need to be able to withstand alkali, and they also need to be able to develop endospores so that they can survive the mechanical forces that are going to be put on the concrete while it is being mixed. In order to precipitate CaCO3 in the form of calcite, they need to have a strong urease activity. Both Bacillus sphaericus and the Streptococcus aureus bacterial strains were evaluated for their ability to precipitate calcium carbonate (CaCO3). These strains were obtained from the Department of Biotechnology at GLA University in Mathura. This research aims to solve the issue of augmenting the tension and compression strengths of concrete by investigating possible solutions for environmentally friendly concrete. The sterile cultures of the microorganisms were mixed with water, which was one of the components of the concrete mixture, along with the nutrients in the appropriate proportions. After that, the blocks were molded, and then pond-cured for 7, 28, 56, 90, 120, 180, 270, and 365 days, respectively, before being evaluated for compressibility and tensile strength. An investigation into the effect that bacteria have on compression strength was carried out, and the outcomes of the tests showed that bacterial concrete specimens exhibited an increase in mechanical strength. When compared to regular concrete, the results showed a maximum increase of 16 percent in compressive strength and a maximum increase of 12 percent in split tensile strength. This study also found that both bacterial concrete containing 106, 107, and 108 cfu/ml concentrations made from Bacillus sphaericus and Streptococcus aureus bacteria gave better results than normal concrete. Both cluster analysis (CA) and regression analysis (RA) were utilized in this research project in order to measure and analyze mechanical strength.


Assuntos
Bacillaceae , Materiais de Construção , Bacillaceae/metabolismo , Bactérias/metabolismo , Carbonato de Cálcio/metabolismo , Materiais de Construção/análise , Materiais de Construção/microbiologia , Humanos , Análise de Regressão
15.
World J Microbiol Biotechnol ; 38(12): 217, 2022 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-36070019

RESUMO

Cold-adapted esterases have potential industrial applications. To fulfil the global continuous demand for these enzymes, a cold-adapted esterase member of family VI from Lysinibacillus sp. YS11 was cloned on pET-28b (+) vector and expressed in E. coli BL21(DE3) Rosetta cells for the first time. The open reading frame (654 bp: GenBank MT120818.1) encodes a polypeptide (designated EstRag: 217 amino acid residues). EstRag amino acid sequence has conserved esterase signature motifs: pentapeptide (GFSQG) and catalytic triad Ser110-Asp163-His194. EstRag 3D predicted model, built with LOMETS3 program, showed closest structural similarity to PDB 1AUO_A (esterase: Pseudomonas fluorescens); TM-align score program inferences. Purified EstRag to 9.28-fold, using Ni2+affinity agarose matrix, showed a single protein band (25 kDa) on SDS-PAGE, Km (0.031 mM) and Kcat/Km (657.7 s-1 mM-1) on p-NP-C2. Temperature and pH optima of EstRag were 35 °C and 8.0, respectively. EstRag was fully stable at 5-30 °C for 120 min and at pH(s) 8.0-10.0 after 24 h. EstRag activity (391.46 ± 0.009%) was impressively enhanced after 30 min preincubation with 5 mM Cu2+. EstRag retained full stability after 30 min pre-incubation with 0.1%(v/v) SDS, Triton X-100, and Tween-80. EstRag promising characteristics motivate performing guided evolution and industrial applications prospective studies.


Assuntos
Bacillaceae , Esterases , Álcalis , Bacillaceae/genética , Bacillaceae/metabolismo , Temperatura Baixa , Detergentes/química , Detergentes/farmacologia , Estabilidade Enzimática , Escherichia coli/genética , Escherichia coli/metabolismo , Esterases/metabolismo , Estudos Prospectivos
16.
Arch Microbiol ; 204(10): 612, 2022 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-36087123

RESUMO

Using the culturomics approach, the previously unknown strain Marseille-P8953T, was isolated and classified within the Weizmannia genus. Strain Marseille-P8953T was isolated from the faeces of a healthy subject and consisted of Gram-stain positive, spore-forming, motile rod-shaped cells. A 99.2% similarity was observed between the 16S rRNA gene of strain Marseille-P8953T (accession number LR735539) and that of Weizmannia coagulans strain NBRC 12583T (accession number KX261624), its closest phylogenetic relative, while the genome of strain Marseille-P8953T (3.5 Mpb long, 46.5% GC content) shared the average nucleotide identity by Orthology and digital DNA-DNA Hybridisation values of 95 and 60.4%, respectively. Given the phylogenetic classification and phenotypic characteristics of strain Marseille-P8953T, we propose the creation of a new species within the Weizmannia genus named Weizmannia faecalis (= CSUR P8953T = CECT 9904 T).


Assuntos
Bacillaceae , Bacillaceae/genética , Composição de Bases , DNA Bacteriano/genética , Humanos , Filogenia , RNA Ribossômico 16S/genética
17.
Artigo em Inglês | MEDLINE | ID: mdl-35998028

RESUMO

In this study, we isolated a novel Gram-stain-positive, aerobic, motile, rod-shaped bacterium, named GX 13764T, from the rhizosphere soil of a decayed mangrove plant Kandelia candel collected from Beihai, Guangxi, PR China, and characterized using a polyphasic taxonomic approach. The strain exhibited yellow-orange, round, convex, shiny, smooth, opaque and 2-3 mm diameter colonies on marine agar 2216 media after 3 days of incubation at 30 °C and was capable of growth at 4-45 °C (optimum, 30 °C), pH 5.0-9.0 (optimum, pH 7.0) and 0-4 % NaCl (w/v; optimum, 2 %). The strain was positive for catalase and negative for the oxidase. The main cellular fatty acid was anteiso-C15:0, iso-C15:0, iso-C16:0 and iso-C14:0. The cell-wall peptidoglycan comprised meso-diaminopimelic acid and the main menaquinone was MK-7. The polar lipids included one diphosphatidylglycerol, one phosphatidylglycerol, two glycolipids, two unidentified phospholipids and three unidentified lipids. Based on 16S rRNA gene analysis, GX 13764T presented the highest sequence similarity to Metabacillus mangrovi KCTC 33872T (97.04 %). The DNA G+C content of the type strain was 44.2 mol%. The average nucleotide identity values between GX 13764T and M. mangrovi KCTC 33872T, Metabacillus idriensis DSM 19097T and Metabacillus indicus LMG 22858T were 69.39, 68.87 and 68.95 %, respectively, with digital DNA-DNA hybridization values of 19.9, 19.5 and 19.5 %, respectively. Based on the polyphasic data, strain GX 13764T should be nominated as a novel species of the genus Metabacillus, for which the name Metabacillus kandeliae sp. nov. is proposed. The type strain is GX 13764T (=MCCC 1K06654T=KCTC 43366T).


Assuntos
Bacillaceae , Rhizophoraceae , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , Rhizophoraceae/microbiologia , Rizosfera , Análise de Sequência de DNA , Solo , Microbiologia do Solo
19.
Arch Microbiol ; 204(9): 545, 2022 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-35939215

RESUMO

In the present study, the taxonomic position of Bacillus tepidiphilus was re-evaluated. Bacillus tepidiphilus (B. tepidiphilus) SYSU G01002T showed the highest 16S rRNA gene sequence similarity with the type strain Peribacillus alkalitolerans (P. alkalitolerans) (97.7%). In the phylogenetic (based on 16S rRNA sequence) and phylogenomic (based on 71 bacterial single-copy genes) trees, B. tepidiphilus SYSU G01002T clade with the members of the genus Peribacillus. The amino acid identity (AAI) value of B. tepidiphilus SYSU G01002T was highest with P. alkalitolerans KCTC 33631T (73.6%). The AAI value between B. tepidiphilus SYSU G01002T and P. alkalitolerans KCTC 33631T was above the cutoff level for genus delineation. The average nucleotide identity (ANI) between B. tepidiphilus and P. alkalitolerans KCTC 33631T was 74.1%, which was below the ANI value (95-96%) for species delineation. Based on the phylogenetic, phylogenomic, AAI, and ANI analysis, Bacillus tepidiphilus is proposed to transfer to the genus Peribacillus as Peribacillus tepidiphilus comb. nov. The type strain is SYSU G01002T (= KCTC 43131T = CGMCC 1.17491T).


Assuntos
Bacillaceae , Bacillus , Bacillaceae/genética , Bacillus/genética , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/análise , Hibridização de Ácido Nucleico , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
20.
Curr Microbiol ; 79(10): 301, 2022 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-36002665

RESUMO

A novel bacterium, designated strain APA_H-1(4)T, was isolated from the saline-alkaline soil, Zhaodong, Heilongjiang Province, China. Phenotypic and chemotaxonomic analyses, and whole-genome sequencing were used to determine the taxonomic position of the strain. Phylogenetic analysis indicated that the isolate belongs to the genus Oceanobacillus, and showed the highest sequence similarity to O. damuensis KCTC 33146T (98.35%, similarity) and 'O. massiliensis' DSM 24644 (98.32%). The average nucleotide identity values between strain APA_H-1(4)T and other members of the genus Oceanobacillus were lower than 82% recommended for distinguishing novel prokaryotic species. The digital DNA-DNA hybridization values of strain APA_H-1(4)T with O. damuensis KCTC 33146T and 'O. massiliensis' DSM 24644 were 13.60 and 17.60%, respectively. Cells of strain APA_H-1(4)T were Gram-staining positive, motile, aerobic, spore-forming rods (0.5-0.7 × 1.8-2.6 µm) with flagella. The growth was found to occur optimally at 37 °C. The whole-cell hydrolysate contained meso-diaminopimelic acid as the diagnostic cell wall diamino acid. The main detected polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, an unidentified phospholipid and an unidentified polar lipid. The predominant respiratory quinone was identified as menaquinone-7 (MK-7). The major cellular fatty acid (>10%) was anteiso-C15:0. The G + C content of the genomic DNA was determined to be 38.4% based on the draft genome sequence. Based on the comparative analysis of polyphasic taxonomic data, strain APA_H-1(4)T represents a novel species of the genus Oceanobacillus, for which the name Oceanobacillus saliphilus sp. nov. is proposed. The type strain is APA_H-1(4)T (=GDMCC 1.2239T = KCTC 43254T).


Assuntos
Bacillaceae , Solo , Álcalis , Técnicas de Tipagem Bacteriana , China , DNA Bacteriano/química , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
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