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1.
Int J Mol Sci ; 22(18)2021 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-34575908

RESUMO

Ganoderma lucidum is a medicinal fungus abundant in triterpenoids, its primary bioactive components. Although numerous Ganoderma triterpenoids have already been identified, rare Ganoderma triterpenoid saponins were recently discovered. To create novel Ganoderma saponins, ganoderic acid G (GAG) was selected for biotransformation using four Bacillus glycosyltransferases (GTs) including BtGT_16345 from the Bacillus thuringiensis GA A07 strain and three GTs (BsGT110, BsUGT398, and BsUGT489) from the Bacillus subtilis ATCC 6633 strain. The results showed that BsUGT489 catalyzed the glycosylation of GAG to GAG-3-o-ß-glucoside, while BsGT110 catalyzed the glycosylation of GAG to GAG-26-o-ß-glucoside, which showed 54-fold and 97-fold greater aqueous solubility than that of GAG, respectively. To our knowledge, these two GAG saponins are new compounds. The glycosylation specificity of the four Bacillus GTs highlights the possibility of novel Ganoderma triterpenoid saponin production in the future.


Assuntos
Bacillus/metabolismo , Glicosiltransferases/metabolismo , Triterpenos/metabolismo , Proteínas de Bactérias , Biotransformação , Catálise , Cromatografia Líquida de Alta Pressão , Glicosilação , Estrutura Molecular , Solubilidade , Triterpenos/química
2.
Braz J Biol ; 83: e243874, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34378658

RESUMO

In recent days, cheapest alternative carbon source for fermentation purpose is desirable to minimize production cost. Xylanases have become attractive enzymes as their potential in bio-bleaching of pulp and paper industry. The objective of the present study was to identify the potential ability on the xylanase production by locally isolated Bacillus pumilus BS131 by using waste fiber sludge and wheat bran media under submerged fermentation. Culture growth conditions were optimized to obtain significant amount of xylanase. Maximum xylanase production was recorded after 72 hours of incubation at 30 °C and 7 pH with 4.0% substrate concentration. In the nutshell, the production of xylanase using inexpensive waste fiber sludge and wheat-bran as an alternative in place of expensive xylan substrate was more cost effective and environment friendly.


Assuntos
Bacillus pumilus , Bacillus , Bacillus/metabolismo , Bacillus pumilus/metabolismo , Fibras na Dieta , Endo-1,4-beta-Xilanases/metabolismo , Fermentação , Concentração de Íons de Hidrogênio , Esgotos , Temperatura
3.
Int J Food Microbiol ; 354: 109318, 2021 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-34246014

RESUMO

The presence of mesophilic and thermophilic spore-forming bacteria in UHT milk, as well as biofilm formation in dairy plants, are concerning. The current study explored the spore-forming bacilli diversity in 100 samples of UHT milk (skimmed and whole). Through this work, a total of 239 isolates from UHT milk samples were obtained. B. cereus s.s. was isolated from 7 samples, B. sporothermodurans from 19 and, G. stearothermophilus from 25 samples. Genes encoding hemolysin (HBL), and non-hemolytic (NHE) enterotoxins were detected in B. cereus s.s. isolates. All isolates of B. cereus s.s. (12) B. sporothermodurans (38), and G. stearothermophilus (47) were selected to verify the ability of biofilm formation in microtiter plates. The results showed all isolates could form biofilms. The OD595 values of biofilm formation varied between 0.14 and 1.04 for B. cereus, 0.20 to 1.87 for B. sporothermodurans, and 0.49 to 2.77 for G. stearothermophilus. The data highlights that the dairy industry needs to reinforce control in the initial quality of the raw material and in CIP cleaning procedures; avoiding biofilm formation and consequently a persistent microbiota in processing plants, which can shelter pathogenic species such as B. cereus s.s.


Assuntos
Bacillus cereus , Bacillus , Microbiologia de Alimentos , Geobacillus stearothermophilus , Temperatura Alta , Leite , Animais , Bacillus/genética , Bacillus/metabolismo , Bacillus cereus/genética , Bacillus cereus/metabolismo , Biofilmes , Geobacillus stearothermophilus/genética , Geobacillus stearothermophilus/metabolismo , Incidência , Leite/microbiologia
4.
Int J Mol Sci ; 22(13)2021 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-34208885

RESUMO

We previously demonstrated that flavonoid metabolites inhibit cancer cell proliferation through both CDK-dependent and -independent mechanisms. The existing evidence suggests that gut microbiota is capable of flavonoid biotransformation to generate bioactive metabolites including 2,4,6-trihydroxybenzoic acid (2,4,6-THBA), 3,4-dihydroxybenzoic acid (3,4-DHBA), 3,4,5-trihyroxybenzoic acid (3,4,5-THBA) and 3,4-dihydroxyphenylacetic acid (DOPAC). In this study, we screened 94 human gut bacterial species for their ability to biotransform flavonoid quercetin into different metabolites. We demonstrated that five of these species were able to degrade quercetin including Bacillus glycinifermentans, Flavonifractor plautii, Bacteroides eggerthii, Olsenella scatoligenes and Eubacterium eligens. Additional studies showed that B. glycinifermentans could generate 2,4,6-THBA and 3,4-DHBA from quercetin while F. plautii generates DOPAC. In addition to the differences in the metabolites produced, we also observed that the kinetics of quercetin degradation was different between B. glycinifermentans and F. plautii, suggesting that the pathways of degradation are likely different between these strains. Similar to the antiproliferative effects of 2,4,6-THBA and 3,4-DHBA demonstrated previously, DOPAC also inhibited colony formation ex vivo in the HCT-116 colon cancer cell line. Consistent with this, the bacterial culture supernatant of F. plautii also inhibited colony formation in this cell line. Thus, as F. plautii and B. glycinifermentans generate metabolites possessing antiproliferative activity, we suggest that these strains have the potential to be developed into probiotics to improve human gut health.


Assuntos
Ácido 3,4-Di-Hidroxifenilacético/farmacologia , Antineoplásicos/farmacologia , Bactérias/classificação , Bromobenzoatos/farmacologia , Ácido Gálico/farmacologia , Hidroxibenzoatos/farmacologia , Quercetina/química , Ácido 3,4-Di-Hidroxifenilacético/química , Actinobacteria/genética , Actinobacteria/isolamento & purificação , Actinobacteria/metabolismo , Antineoplásicos/química , Bacillus/genética , Bacillus/isolamento & purificação , Bacillus/metabolismo , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo , Proteínas de Bactérias , Bacteroides/genética , Bacteroides/isolamento & purificação , Bacteroides/metabolismo , Bromobenzoatos/química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Clostridiales/genética , Clostridiales/isolamento & purificação , Clostridiales/metabolismo , Eubacterium/genética , Eubacterium/isolamento & purificação , Eubacterium/metabolismo , Ácido Gálico/química , Microbioma Gastrointestinal , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Células HCT116 , Humanos , Hidroxibenzoatos/química , Filogenia , Análise de Sequência de RNA
5.
Biomed Res Int ; 2021: 6611657, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34195272

RESUMO

Two novel Algerian field-collected isolates were selected for their antifungal activity against Zymoseptoria tritici (teleomorph Mycosphaerella graminicola). The novel strains, termed Alg.24B1 and Alg.24B2, were identified as Bacillus subtilis and Bacillus simplex since their respective nucleotide sequences of the 16S rRNA gene were 100% and 99.93% identical to those of B. subtilis and B. simplex, respectively. The antifungal activities of Alg.24B1 and Alg.24B2 were evaluated by the well diffusion method and compared to those of other Bacillus species. The maximum activity was obtained after two days of confrontation of the bacterial strain supernatants with the fungus for Alg.24B1 and three days for Alg.24B2. Furthermore, the metabolites responsible for the antifungal activity of both strains were detected by the investigation of either gene presence (PCR) or molecule production (activity detection of lytic enzymes and HPLC detection of lipopeptides). Overall, this study showed that in addition to their ability to produce lytic enzymes (protease and ß-glucanase), both strains coproduce three types of lipopeptides viz. surfactin, iturin, and fengycin. Thus, the biofungicide activity of both strains may be a result of a combination of different mechanisms. Therefore, they had a great potential to be used as biocontrol agents to effectively manage septoria tritici blotch of wheat (STB).


Assuntos
Ascomicetos/metabolismo , Bacillus/metabolismo , Doenças das Plantas/prevenção & controle , Triticum/microbiologia , Antifúngicos , Ascomicetos/genética , Bacillus/genética , Bacillus subtilis/genética , Cromatografia Líquida de Alta Pressão , Técnicas In Vitro , Lipopeptídeos/química , Controle Biológico de Vetores , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S , Especificidade da Espécie , Triticum/genética
6.
PLoS One ; 16(7): e0254676, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270610

RESUMO

Microbially Induced Carbonate Precipitation (MICP) is currently viewed as one of the potential prominent processes for field applications towards the prevention of soil erosion, healing cracks in bricks, and groundwater contamination. Typically, the bacteria involved in MICP manipulate their environment leading to calcite precipitation with an enzyme such as urease, causing calcite crystals to form on the surface of grains forming cementation bonds between particles that help in reducing soil permeability and increase overall compressive strength. In this paper, the main focus is to study the MICP performance of three indigenous landfill bacteria against a well-known commercially bought MICP bacteria (Bacillus megaterium) using sand columns. In order to check the viability of the method for potential field conditions, the tests were carried out at slightly less favourable environmental conditions, i.e., at temperatures between 15-17°C and without the addition of urease enzymes. Furthermore, the sand was loose without any compaction to imitate real ground conditions. The results showed that the indigenous bacteria yielded similar permeability reduction (4.79 E-05 to 5.65 E-05) and calcium carbonate formation (14.4-14.7%) to the control bacteria (Bacillus megaterium), which had permeability reduction of 4.56 E-5 and CaCO3 of 13.6%. Also, reasonably good unconfined compressive strengths (160-258 kPa) were noted for the indigenous bacteria samples (160 kPa). SEM and XRD showed the variation of biocrystals formation mainly detected as Calcite and Vaterite. Overall, all of the indigenous bacteria performed slightly better than the control bacteria in strength, permeability, and CaCO3 precipitation. In retrospect, this study provides clear evidence that the indigenous bacteria in such environments can provide similar calcite precipitation potential as well-documented bacteria from cell culture banks. Hence, the idea of MICP field application through biostimulation of indigenous bacteria rather than bioaugmentation can become a reality in the near future.


Assuntos
Carbonato de Cálcio/química , Microbiota , Microbiologia do Solo , Bacillus/metabolismo , Carbonato de Cálcio/metabolismo , Precipitação Química , Conservação dos Recursos Naturais/métodos , Areia/química , Areia/microbiologia , Solo/química
7.
Sci Rep ; 11(1): 14316, 2021 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-34253740

RESUMO

Recently, the program INTMSAlign_HiSol for identifying aggregation hotspots in proteins only requiring secondary structure data was introduced. We explored the utility of this program further and applied it for engineering of the aldoxime dehydratase from Bacillus sp. OxB-1. Towards this end, the effect of inverting the hydropathy at selected positions of the amino acid sequence on the enzymatic activity was studied leading to 60% of our constructed variants, which showed improved activity. In part, this activity increase can be rationalised by an improved heme incorporation of the variants. For example, a single mutation gave a 1.8 fold increased enzymatic activity and 30% improved absolute heme incorporation.


Assuntos
Hidroliases/metabolismo , Engenharia de Proteínas/métodos , Bacillus/enzimologia , Bacillus/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biocatálise , Hidroliases/química , Alinhamento de Sequência , Especificidade por Substrato
8.
J Dairy Sci ; 104(9): 9465-9477, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34127264

RESUMO

A novel galactosidase gene (gal3149) was identified from Bacillus velezensis SW5 and heterologously expressed in Escherichia coli BL21 (DE3). The novel galactosidase, Gal3149, encoded by gal3149 in an open reading frame of 1,299 bp, was 433 amino acids in length. Protein sequence analysis showed that Gal3149 belonged to family 4 of glycoside hydrolases (GH4). Gal3149 displayed higher enzyme activity for the substrate 2-nitrophenyl-ß-d-galactopyranoside (oNPG) than for 4-nitrophenyl-α-d-galactopyranoside (pNPαG). This is the first time that an enzyme belonging to GH4 has been shown to exhibit ß-galactosidase activity. Gal3149 showed optimal activity at pH 8.0 and 50°C, and exhibited excellent thermal stability, with retention of 50% relative activity after incubation at a temperature range of 0 to 50°C for 48 h. Gal3149 activity was significantly improved by K+ and Na+, and was strongly or completely inhibited by Ag+, Zn2+, Tween-80, Cu2+, carboxymethyl cellulose, and oleic acid. The rate of hydrolyzed lactose in 1 mL of milk by 1 U of Gal3149 reached about 50% after incubation for 4 h. These properties lay a solid foundation for Gal3149 in application of the lactose-reduced dairy industry.


Assuntos
Bacillus , Galactosidases , Animais , Bacillus/genética , Bacillus/metabolismo , Clonagem Molecular , Concentração de Íons de Hidrogênio , Cinética , Lactose , Temperatura , beta-Galactosidase/genética , beta-Galactosidase/metabolismo
9.
J Mol Biol ; 433(15): 167100, 2021 07 23.
Artigo em Inglês | MEDLINE | ID: mdl-34119489

RESUMO

Bacterial NusG associates with RNA polymerase (RNAP) through its N-terminal domain, while the C-terminal domain (CTD) forms dynamic interactions with Rho, S10, NusB and NusA to affect transcription elongation. While virtually all bacteria encode for a core NusG, many also synthesize paralogs that transiently bind RNAP to alter expression of targeted genes. Yet, despite the importance of the genes they regulate, most of the subfamilies of NusG paralogs (e.g., UpxY, TaA, ActX and LoaP) have not been investigated in depth. Herein, we discover that LoaP requires a small RNA hairpin located within the 5' leader region of its targeted operons. LoaP binds the RNA element with nanomolar affinity and high specificity, in contrast to other NusG proteins, which have not been shown to exhibit RNA-binding activity. These data reveal a sequence feature that can be used to identify LoaP-regulated operons. This discovery also expands the repertoire of macromolecular interactions exhibited by the NusG CTD during transcription elongation to include an RNA ligand.


Assuntos
Bacillus/genética , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , RNA Bacteriano/metabolismo , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/metabolismo , Regiões 5' não Traduzidas , Bacillus/metabolismo , RNA Polimerases Dirigidas por DNA/metabolismo , Conformação Molecular , Óperon , Domínios Proteicos , RNA Bacteriano/química
10.
Arch Microbiol ; 203(7): 4337-4350, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34110482

RESUMO

This study targeted the production of exopolysaccharide from Bacillus sp. NRC5 grown in Egyptian seawater to use it as natural antitumor therapy. The biological activities of selected exopolysaccharide (BEPS) as an antioxidant, anti-inflammatory and anticancer have been studied. BEPS was evaluated as an anti-inflammatory in vitro against cyclooxygenase (COX-1 and COX-2) and evaluated as an anticancer on human breast and prostate cancer cell lines (MCF-7 and PC3). In addition, BEPS antitumor activity was tested against the Ehrlich Ascites Carcinoma (EAC) model. The BEPS presented potent antioxidant activities, consisted of glucose, mannose, and mannuronic acid in a molar ratio of 1.0:1.7:0.8 with a molecular weight of 3.59 × 105 g/mol. BEPS showed a promising COX-2 inhibitory effect in comparison with the reference drug celecoxib. BEPS appeared efficient anticancer property, where it killed 64.20 and 70.20% of MCF-7 and PC3 cells at 100 µg/ml, respectively (IC50, 76.70, and 70.40 µg/ml, respectively). BEPS exhibited antitumor ability as it prolonged the lifespan of mice to reach 75 days instead of 20 days in the tumor control, reduced viable cancer cells count, tumor volume and weight, modulated blood components, and white blood cells differentiation. BEPS produced from Bacillus sp. NRC5 showed its antioxidant and anti-inflammatory abilities and antitumor abilities, which may all be attributed to its unique composition containing sulfated moieties and uronic acids.


Assuntos
Bacillus , Polissacarídeos Bacterianos , Animais , Antineoplásicos/farmacologia , Antioxidantes/farmacologia , Bacillus/química , Bacillus/metabolismo , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Humanos , Longevidade/efeitos dos fármacos , Camundongos , Polissacarídeos Bacterianos/biossíntese , Polissacarídeos Bacterianos/química , Polissacarídeos Bacterianos/farmacologia
11.
J Agric Food Chem ; 69(26): 7409-7419, 2021 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-34180240

RESUMO

An increasing number of Bacillus strains have been identified, and the removal capacity of zearalenone (ZEN) was determined; however, they failed to reveal the detoxification mechanism and transformation product. Here, Bacillus subtilis Y816, which could transform 40 mg/L of ZEN within 7 h of fermentation, was identified and studied. First, the biotransformation products of ZEN and 17-ß-estradiol (E2) were identified as ZEN-14-phosphate and E2-3-phosphate by HPLC-TOF-MS and NMR, respectively. An intracellular zearalenone phosphotransferase (ZPH) was found through transcriptome sequencing analysis of B. subtilis Y816. The phosphorylated reaction conditions of ZEN by ZPH were further revealed in this work. Furthermore, the phosphorylated conjugates showed reduced estrogenic toxicity compared with their original substances (ZEN and α/ß-zearalenol) using an engineered yeast biosensor system. The first report on the phosphorylated conjugated mode of ZEN in B. subtilis Y816 will inspire new perspectives on the biotransformation of ZEN in Bacillus strains.


Assuntos
Bacillus , Zearalenona , Bacillus/metabolismo , Bacillus subtilis/genética , Bacillus subtilis/metabolismo , Biotransformação , Fermentação , Zearalenona/metabolismo
12.
Int J Mol Sci ; 22(9)2021 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-34066922

RESUMO

Human gut microbiota harbors numerous microbial species with molecular enzymatic potential that impact on the eubiosis/dysbiosis and health/disease balances. Microbiota species isolation and description of their specific molecular features remain largely unexplored. In the present study, we focused on the cultivation and selection of species able to tolerate or biodegrade the endocrine disruptor bisphenol A (BPA), a xenobiotic extensively found in food plastic containers. Chemical xenobiotic addition methods for the directed isolation, culturing, Whole Genome Sequencing (WGS), phylogenomic identification, and specific gene-encoding searches have been applied to isolate microorganisms, assess their BPA metabolization potential, and describe encoded catabolic pathways. BPA-tolerant strains were isolated from 30% of infant fecal microbial culture libraries analyzed. Most isolated strains were phylogenetically related to the operational taxonomic group Bacillus amyloliquefaciens spp. Importantly, WGS analysis of microbial representative strain, Bacillus sp. AM1 identified the four complete molecular pathways involved on BPA degradation indicating its versatility and high potential to degrade BPA. Pathways for Exopolysaccharide (EPS) and Polyhydroxyalkanates (PHA) biopolymer synthesis were also identified and phenotypically confirmed by transmission electronic microscopy (TEM). These microbial biopolymers could generally contribute to capture and/or deposit xenobiotics.


Assuntos
Bacillus/metabolismo , Compostos Benzidrílicos/metabolismo , Microbioma Gastrointestinal , Fenóis/metabolismo , Transdução de Sinais , Antibacterianos/farmacologia , Bacillus/citologia , Bacillus/genética , Bacillus/ultraestrutura , Compostos Benzidrílicos/química , Biodegradação Ambiental , Genoma Bacteriano , Humanos , Testes de Sensibilidade Microbiana , Fenóis/química , Filogenia , RNA Ribossômico 16S/genética
13.
Int J Mol Sci ; 22(9)2021 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-34068779

RESUMO

Bacillus volatiles to control plant nematodes is a topic of great interest among researchers due to its safe and environmentally friendly nature. Bacillus strain GBSC56 isolated from the Tibet region of China showed high nematicidal activity against M. incognita, with 90% mortality as compared with control in a partition plate experiment. Pure volatiles produced by GBSC56 were identified through gas chromatography and mass spectrometry (GC-MS). Among 10 volatile organic compounds (VOCs), 3 volatiles, i.e., dimethyl disulfide (DMDS), methyl isovalerate (MIV), and 2-undecanone (2-UD) showed strong nematicidal activity with a mortality rate of 87%, 83%, and 80%, respectively, against M. incognita. The VOCs induced severe oxidative stress in nematodes, which caused rapid death. Moreover, in the presence of volatiles, the activity of antioxidant enzymes, i.e., SOD, CAT, POD, and APX, was observed to be enhanced in M. incognita-infested roots, which might reduce the adverse effect of oxidative stress-induced after infection. Moreover, genes responsible for plant growth promotion SlCKX1, SlIAA1, and Exp18 showed an upsurge in expression, while AC01 was downregulated in infested plants. Furthermore, the defense-related genes (PR1, PR5, and SlLOX1) in infested tomato plants were upregulated after treatment with MIV and 2-UD. These findings suggest that GBSC56 possesses excellent biocontrol potential against M. incognita. Furthermore, the study provides new insight into the mechanism by which GBSC56 nematicidal volatiles regulate antioxidant enzymes, the key genes involved in plant growth promotion, and the defense mechanism M. incognita-infested tomato plants use to efficiently manage root-knot disease.


Assuntos
Bacillus/genética , Resistência à Doença/genética , Lycopersicon esculentum/genética , Tylenchoidea/patogenicidade , Animais , Antinematódeos/metabolismo , Bacillus/metabolismo , China , Cromatografia Gasosa-Espectrometria de Massas , Lycopersicon esculentum/microbiologia , Lycopersicon esculentum/parasitologia , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Tylenchoidea/genética , Compostos Orgânicos Voláteis/metabolismo
14.
World J Microbiol Biotechnol ; 37(7): 123, 2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34160683

RESUMO

Lipopeptides are important secondary metabolites produced by microbes. They find applications in environmental decontamination and in the chemical, pharmaceutical and food industries. However, their production is expensive. In the present work we propose three strategies to lower the production costs of surfactin. First, the coproduction of surfactin and arginase in a single growth. Second, extract the fraction of surfactin that adsorbs to the biomass and is removed from the growth medium through centrifugation. Third, use microbial biomass for the remediation of organic and inorganic contaminants. The coproduction of surfactin and arginase was evaluated by factorial design experiments using the LB medium supplemented with arginine. The best conditions for surfactin production were 22 h of growth at 37 °C using LB supplemented with arginine 7.3 g/L. Almost similar conditions were found to produce highest levels of arginase, 24 h and 6.45 g/L arginine. Decontamination of phenol and copper from artificial samples was attained by treatment with residues from lipopeptide production. Thus, cell suspensions and wash-waters used to extract surfactin from the biomass. Cell suspensions were used to successfully remove hydroquinone. Cell suspensions and wash-waters containing surfactin were successfully used to recover copper from solution. Specific monitoring methods were used for phenol and metal solutions, respectively a biosensor based on tyrosinase and either atomic absorption flame ionization spectrometry or absorbance coupled to the Arduino™ platform. Therefore, we report three alternative strategies to lower the production costs in lipopeptide production, which include the effective recovery of copper and phenol from contaminated waters using residues from surfactin production. Sustainable and profitable production of surfactin can be achieved by a coproduction strategy of lipopeptides and enzymes. Lipopeptides are collected in the supernatant and enzymes in the biomass. In addition, lipopeptides that coprecipitate with biomass can be recovered by washing. Lipopeptide wash-waters find applications in remediation and cells can also be used for environmental decontamination.


Assuntos
Arginase/biossíntese , Bacillus/enzimologia , Bacillus/crescimento & desenvolvimento , Bacillus/metabolismo , Lipopeptídeos/biossíntese , Peptídeos Cíclicos/biossíntese , Bacillus/genética , Proteínas de Bactérias/biossíntese , Biomassa , Reatores Biológicos , Cobre/metabolismo , Meios de Cultura , DNA Bacteriano , Microbiologia Ambiental , Recuperação e Remediação Ambiental , Hidroquinonas/metabolismo , Fenol/metabolismo
15.
J Basic Microbiol ; 61(8): 757-768, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34101885

RESUMO

The purpose of this study was to isolate lignin-degrading bacteria from buffalo rumen and to explore their interactions further. Using lignin as the carbon source, three bacteria, B-04 (Ochrobactrum pseudintermedium), B-11 (Klebsiella pneumoniae), and B-45 (Bacillus sonorensis), which have shown lignin degradation potential, were successfully isolated and identified from the rumen fluid of buffalo by colony morphology, 16S ribosomal RNA gene sequencing, and biochemical and physiological analyses. The degradation rates of lignin were determined, and the maximum values were 4.86%, 11.1%, and 7.68% for B-04, B-11, and B-45, respectively. The maximum laccase activities were 0.65, 0.93, and 1.15 U/ml, while the maximum lignin peroxidase activities were 5.72, 8.29, and 18.69 U/ml, respectively. Pairwise interaction studies showed inhibitory interaction between B-04 and B-45, inhibitory interaction between B-04 and B-11, and symbiotic interaction between B-11 and B-45. This is the first report on the lignin degradation ability of bacteria isolated from the buffalo's rumen, which provides a new understanding for revealing the mechanism of roughage tolerance of buffalo.


Assuntos
Bactérias/isolamento & purificação , Bactérias/metabolismo , Búfalos/microbiologia , Lignina/metabolismo , Rúmen/microbiologia , Animais , Bacillus/isolamento & purificação , Bacillus/metabolismo , Bactérias/classificação , Bactérias/genética , Klebsiella pneumoniae/isolamento & purificação , Klebsiella pneumoniae/metabolismo , Interações Microbianas , Ochrobactrum/isolamento & purificação , Ochrobactrum/metabolismo , Filogenia , RNA Ribossômico 16S/genética , Alinhamento de Sequência
16.
Sci Rep ; 11(1): 12243, 2021 06 10.
Artigo em Inglês | MEDLINE | ID: mdl-34112820

RESUMO

The bioremediation of beverage (treated and untreated) effluent was investigated in the current study by using the potential strains of Bacillus sp. (BK1) and Aspergillus sp. (BK2). Effluent was collected from the beverage industry (initial concentration of nitrogen were 3200 ± 0.5 mg/L and 4400 ± 0.6 mg/L whereas phosphorus were 4400 ± 2 mg/L and 2600 ± 1 mg/L in treated and untreated effluent correspondingly). Further, the BK1 and BK2 exhibited high removal competence after 1 week of incubation; BK1 removed phosphorus 99.95 ± 0.7% and BK2 95.69 ± 1% in treated effluent while nitrogen removed about 99.90 ± 0.4% by BK1 and 81.25 ± 0.8% by BK2 (initial concentration of phosphorus 4400 ± 2 mg/L and nitrogen 3200 ± 0.5 mg/L). Next, in the untreated effluent BK1 removed 99.81 ± 1% and BK2 99.85 ± 0.8% of phosphorus while removed nitrogen 99.93 ± 0.5% by BK1 and 99.95 ± 1.2% by BK2 correspondingly, (initial concentration of phosphorus 2600 ± 1 mg/L and nitrogen 4400 ± 0.6 mg/L). The physiochemical composition of sample such as pH, total carbohydrates, total proteins, total solids of treated and untreated effluent were also analysed before and after treatment of both the samples. BK1 and BK2 increased the pH by 8.94 ± 0.3 and 9.5 ± 0.4 correspondingly in treated effluent whereas 6.34 ± 0.5 and 7.5 ± 0.2 correspondingly in untreated effluent (initial pH of treated and untreated effluent 7.07 ± 0.8 and 4.85 ± 0.3 correspondingly). Total Carbohydrates removed about 17,440 ± 4.6 mg/L and 10,680 ± 3.2 mg/L by BK1 and BK2 correspondingly in treated effluent whereas 18,050 ± 3.5 mg/L and 18,340 ± 2.3 mg/L correspondingly in untreated effluent (initial concentration of treated and untreated effluent 25,780 ± 1.6 mg/L and 35,000 ± 1.5 mg/L correspondingly) while BK1 and BK2 removed total proteins by 30.336 ± 4.6 mg/L and 40.417 ± 2.3 mg/L correspondingly in treated effluent whereas 18.929 ± 1.2 mg/L and 17.526 ± 0.8 mg/L correspondingly in untreated effluent (initial concentration of treated and untreated effluent 49.225 ± 1.5 mg/L and 20.565 ± 1 mg/L correspondingly). Next, total solids removed by BK1 and BK2 2.5 ± 0.3 mg/L and 1.6 ± 0.6 mg/L correspondingly in treated effluent whereas 5.5 ± 0.8 mg/L and 4.6 ± 0.6 mg/L in untreated effluent (initial concentration of treated and untreated effluent 5.6 ± 1.5 mg/L and 9.48 ± 1.2 mg/L correspondingly). Both the strains BK1 and BK2 are highly efficient in the nitrogen and phosphorus removal therefore this strain may be applied for the potential remediation.


Assuntos
Aspergillus/metabolismo , Bacillus/metabolismo , Bebidas , Biodegradação Ambiental , Nitrogênio/metabolismo , Fósforo/metabolismo , Fenômenos Químicos , Indústria Alimentícia , Solo/química , Microbiologia do Solo
17.
Microb Cell Fact ; 20(1): 100, 2021 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-33992112

RESUMO

BACKGROUND: Owing to the excellent properties of photosensitization, cercosporin, one of naturally occurring perylenequinonoid pigments, has been widely used in photodynamic therapy, or as an antimicrobial agent and an organophotocatalyst. However, because of low efficiency of total chemical synthesis and low yield of current microbial fermentation, the limited production restricts its broad applications. Thus, the strategies to improve the production of cercosporin were highly desired. Besides traditional optimization methods, here we screened leaf-spot-disease-related endophytic bacteria to co-culture with our previous identified Cercospora sp. JNU001 to increase cercosporin production. RESULTS: Bacillus velezensis B04 and Lysinibacillus sp. B15 isolated from leaves with leaf spot diseases were found to facilitate cercosporin secretion into the broth and then enhance the production of cercosporin. After 4 days of co-culture, Bacillus velezensis B04 allowed to increase the production of cercosporin from 128.2 mg/L to 984.4 mg/L, which was 7.68-fold of the previously reported one. Lysinibacillus sp. B15 could also enhance the production of cercosporin with a yield of 626.3 mg/L, which was 4.89-fold higher than the starting condition. More importantly, we found that bacteria B04 and B15 employed two different mechanisms to improve the production of cercosporin, in which B04 facilitated cercosporin secretion into the broth by loosening and damaging the hyphae surface of Cercospora sp. JNU001 while B15 could adsorb cercosporin to improve its secretion. CONCLUSIONS: We here established a novel and effective co-culture method to improve the production of cercosporin by increasing its secretion ability from Cercospora sp. JNU001, allowing to develop more potential applications of cercosporin.


Assuntos
Cercospora/metabolismo , Endófitos/metabolismo , Interações Microbianas/fisiologia , Perileno/análogos & derivados , Doenças das Plantas/microbiologia , Bacillaceae/crescimento & desenvolvimento , Bacillaceae/metabolismo , Bacillus/crescimento & desenvolvimento , Bacillus/metabolismo , Cercospora/genética , Cercospora/crescimento & desenvolvimento , Endófitos/genética , Endófitos/crescimento & desenvolvimento , Regulação Fúngica da Expressão Gênica , Técnicas In Vitro , Perileno/análise , Perileno/metabolismo
18.
Food Funct ; 12(13): 5685-5702, 2021 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-34037049

RESUMO

Traditional fermented fish products are favored due to their unique flavors. The fermentation process of fish is accompanied by the formation of flavor substances through a complex metabolic reaction of microorganisms, especially lipolysis and lipid oxidation. However, it is difficult to precisely control the reaction of microorganisms during the fermentation process in modern industrial production, and fermented fish products have lost their traditional characteristic flavors. The purpose of this review is to summarize the different kinds of fermented fish, core microorganisms in it, and flavor formation mechanisms, providing guidance for industrial cultural starters. Future research on the flavor formation mechanism is necessary to confirm the relationship between flavor formation, lipid metabolism, and microorganisms to ensure stable flavor and safety, and to elucidate the mechanism directly toward industrial application.


Assuntos
Fermentação , Produtos Pesqueiros , Microbiologia de Alimentos/métodos , Metabolismo dos Lipídeos , Paladar , Animais , Bacillus/metabolismo , Reatores Biológicos , Peixes/metabolismo , Humanos , Lactobacillus/metabolismo , Lipólise , Micrococcus/metabolismo , Oxirredução , Leveduras/metabolismo
19.
PLoS One ; 16(5): e0251514, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33974665

RESUMO

Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have reached epidemic proportions globally. Therefore, there is an urgent need for a continuous supply of antibiotics to combat the problem. In this study, bacteria initially identified as species belonging to the Bacillus amyloliquefaciens operational group were re-identified based on the housekeeping gene, gyrB. Cell-free supernatants (CFS) from the strains were used for antimicrobial tests using the agar well diffusion assay against MRSA and various types of pathogenic bacteria. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and physicochemical characteristics of the CFS were determined. Based on gyrB sequence analysis, five strains (PD9, B7, PU1, BP1 and L9) were identified as Bacillus velezensis. The CFS of all B. velezensis strains showed broad inhibitory activities against Gram-negative and -positive as well as MRSA strains. Strain PD9 against MRSA ATCC 33742 was chosen for further analysis as it showed the biggest zone of inhibition (21.0 ± 0.4 mm). The MIC and MBC values obtained were 125 µl/ml. The crude antimicrobial extract showed bactericidal activity and was stable at various temperatures (40-80°C), pH (4-12), surfactants (Tween 20, Tween 80, SDS and Triton X-100) and metal ions (MgCI2, NaCI2, ZnNO3 and CuSO4) when tested. However, the crude extract was not stable when treated with proteinase K. All these properties resembled the characteristics of peptides. The antimicrobial compound from the selected strain was purified by using solvent extraction method and silica gel column chromatography. The purified compound was subjected to High Performance Liquid Chromatography which resulted in a single peak of the anti-MRSA compound being detected. The molecular weight of the anti-MRSA compound was determined by using SDS-PAGE and zymogram. The size of the purified antimicrobial peptide was approximately ~ 5 kDa. The antimicrobial peptide produced from B. velezensis strain PD9 is a promising alternative to combat the spread of MRSA infections in the future.


Assuntos
Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Bacillus/metabolismo , Abelhas/microbiologia , Staphylococcus aureus Resistente à Meticilina , 1-Butanol , Animais , Peptídeos Catiônicos Antimicrobianos/farmacologia , Bacillus/genética , Proteínas de Bactérias/genética , Fracionamento Celular , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Meios de Cultivo Condicionados/farmacologia , DNA Girase/genética , DNA Bacteriano/genética , Eletroforese em Gel de Poliacrilamida , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Negativas/crescimento & desenvolvimento , Bactérias Gram-Positivas/efeitos dos fármacos , Bactérias Gram-Positivas/crescimento & desenvolvimento , Concentração de Íons de Hidrogênio , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Filogenia , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico
20.
World J Microbiol Biotechnol ; 37(6): 97, 2021 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-33969441

RESUMO

Bacterial lipopeptides have become a research focus of many studies owing to their industrial and pharmaceutical importance. Although such studies focused on researching purification procedures and qualitative analysis, much remains to be explored and developed to improve the current methods. To enable thorough studies of lipopeptides, this paper describes a new method for purification and characterization of in-gel anionic lipopeptides. Specifically, lipopeptides attributed to the anti-staphylococcal activity of Bacillus mojavensis HF were separated using SDS-PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) and subsequently characterized using mass spectrometry. Lipopeptide band obtained by gel electrophoresis was first visualized using three different staining methods. Next, the lipopeptide isomers were efficiently recovered from the gel band and structural characterization of the extracted lipopeptides was carried out by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). MS analysis revealed that Bacillus mojavensis HF produced three types of lipopeptides including surfactin, fengycin, and kurstakin. 14 clusters of ion peaks were identified as fengycin A with fatty acid of C15-C17, fengycin B (C16, C17), surfactin (C13-C16), and kurstakin (C9-C12). Moreover, tandem mass spectrometric analysis (MS/MS) revealed the sequences of fengycin A and surfactin. In this study, we identified a high variety and number of surfactin and fengycin isomers, which previous reports lacked. To the best of our knowledge, we are the first to report the presence of kurstakin in Bacillus mojavensis species. Finally, we demonstrated that our gel-based study of lipopeptides allowed for a precise and reproducible investigation of these molecules.


Assuntos
Antibacterianos/análise , Bacillus/metabolismo , Lipopeptídeos/análise , Antibacterianos/química , Lipopeptídeos/química , Lipopeptídeos/farmacologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/crescimento & desenvolvimento , Espectrometria de Massas em Tandem
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