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1.
J Sci Food Agric ; 102(2): 557-566, 2022 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-34145902

RESUMO

BACKGROUND: Fermentation efficiency of thermophiles of Bacillus licheniformis YYC4 and Geobacillus stearothermophilus A75, and mesophilic Bacillus subtilis 10 160 on soybean meal (SBM), was evaluated by examining the nutritional and protein structural changes. RESULTS: SBM fermentation by B. licheniformis YYC4, B. subtilis 10 160 and G. stearothemophilus A75 increased significantly the crude and soluble protein from 442.4 to 524.8, 516.1 and 499.9 g kg-1 , and from 53.9 to 203.3, 291.3 and 74.6 g kg-1 , and decreased trypsin inhibitor from 8.19 to 3.19, 2.14 and 5.10 mg g-1 , respectively. Bacillus licheniformis YYC4 and B. subtilis 10 160 significantly increased phenol and pyrazine content. Furthermore, B. licheniformis YYC4 fermentation could produce abundant alcohols, ketones, esters and acids. Surface hydrophobicity, sulfhydryl groups and disulfide bond contents of SBM protein were increased significantly from 98.27 to 166.13, 173.27 and 150.71, from 3.26 to 4.88, 5.03 and 4.21 µmol g-1 , and from 20.77 to 27.95, 29.53 and 25.5 µmol g-1 after their fermentation. Fermentation induced red shifts of the maximum absorption wavelength (λmax ) of fluorescence spectra from 353 to 362, 376 and 361 nm, while significantly reducing the fluorescence intensity of protein, especially when B. subtilis 10 160 was used. Moreover, fermentation markedly changed the secondary structure composition of SBM protein. Analyses by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and atomic force microscopy showed that macromolecule protein was degraded into small-sized protein or peptide during fermentation of SBM. CONCLUSION: Bacillus licheniformis YYC4 fermentation (without sterilization) improved nutrition and protein structure of SBM as B. subtilis 10 160, suggesting its potential application in the SBM fermentation industry. © 2021 Society of Chemical Industry.


Assuntos
Bacillus licheniformis/metabolismo , Bacillus subtilis/metabolismo , Geobacillus stearothermophilus/metabolismo , Proteínas de Soja/química , Soja/microbiologia , Fermentação , Conformação Proteica , Proteínas de Soja/metabolismo , Soja/química , Soja/metabolismo
2.
Food Chem ; 368: 130868, 2022 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-34438173

RESUMO

A novel lipase from Bacillus licheniformis NCU CS-5 was expressed in different Escherichia coli cells. The recombinant enzyme achieved a high activity (161.74 U/mL) with protein concentration of 0.27 mg/mL under optimal conditions at the large-scale expression of 12 h. The recombinant lipase showed optimal activity at 40 ℃ and pH 10.0, and maintained more than 80% relative activity after 96 h of incubation at pH 9.0-10.0. This typical alkaline lipase was activated under medium temperature conditions (30 and 45 ℃ for 96 h). The lipase exhibited a degree of adaptability in various organic solvents and metal ions, and showed high specificity towards triglycerides with short and medium chain fatty acids. Among different substrates, the lipase showed the strongest binding affinity towards pNPP (Km = 0.674 mM, Vmax = 950.196 µM/min). In the experiments of its application in enhancing fatty acids flavor release for low-fat cheeses, the lipase was found to hydrolyze cheeses and mainly increase the contents of butyric acid, hexanoic acid, caprylic acid and decanoic acid. The results from NMR and GC provided the possibility of enhancing fatty acids flavor released from low-fat cheeses by the lipolysis method.


Assuntos
Bacillus licheniformis , Queijo , Bacillus licheniformis/genética , Ácidos Graxos , Concentração de Íons de Hidrogênio , Lipase/genética , Paladar
3.
Sheng Wu Gong Cheng Xue Bao ; 37(8): 2803-2812, 2021 Aug 25.
Artigo em Chinês | MEDLINE | ID: mdl-34472298

RESUMO

Bacitracin is a broad-spectrum antibiotics mainly produced by Bacillus, and is used as veterinary medicine in the fields of livestock and poultry breeding. Insufficient supply of precursor amino acids might be an important factor that hinders high-level microbial production of bacitracin. We investigated the effect of strengthening L-cysteine supply on bacitracin production by an industrial bacitracin producer, Bacillus licheniformis DW2. Overexpression of cysK encoding L-cysteine synthase led to a 9.17% increase of the bacitracin titer. Moreover, overexpression of cysE encoding L-serine acetyltransferase and cysP encoding thiosulfate/sulfate intracellular transporter increased the bacitracin titers by 7.23% and 8.52%, respectively. Moreover, overexpression of a putative cystine importer TcyP led to a 29.19% increase of intracellular L-cysteine, and bacitracin titer was increased by 7.79%. Subsequently, the strong promoter PbacA was used to replace the promoters of genes cysP, cysE and tcyP in strain DW2::ysK, respectively. The resulted strain CYS4 (DW2::cysK-PbacA-(cysP)-PbacA(cysE)- PbacA(tcyP) produced 910.02 U/mL bacitracin, which was 21.10% higher than that of the original strain DW2 (747.71 U/mL). Together with the experiments in 3 L fermenters, this research demonstrated that enhancing intracellular L-cysteine supply is an effective strategy to increase bacitracin production of B. licheniformis.


Assuntos
Bacillus licheniformis , Aminoácidos , Bacillus licheniformis/genética , Bacitracina , Cisteína , Engenharia Metabólica
4.
Molecules ; 26(18)2021 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-34577096

RESUMO

To adapt to various ecological niches, the members of genus Bacillus display a wide spectrum of glycoside hydrolases (GH) responsible for the hydrolysis of cellulose and lignocellulose. Being abundant and renewable, cellulose-containing plant biomass may be applied as a substrate in second-generation biotechnologies for the production of platform chemicals. The present study aims to enhance the natural cellulase activity of two promising 2,3-butanediol (2,3-BD) producers, Bacillus licheniformis 24 and B. velezensis 5RB, by cloning and heterologous expression of cel8A and cel48S genes of Acetivibrio thermocellus. In B. licheniformis, the endocellulase Cel8A (GH8) was cloned to supplement the action of CelA (GH9), while in B. velezensis, the cellobiohydrolase Cel48S (GH48) successfully complemented the activity of endo-cellulase EglS (GH5). The expression of the natural and heterologous cellulase genes in both hosts was demonstrated by reverse-transcription PCR. The secretion of clostridial cellulases was additionally enhanced by enzyme fusion to the subtilisin-like signal peptide, reaching a significant increase in the cellulase activity of the cell-free supernatants. The results presented are the first to reveal the possibility of genetic complementation for enhancement of cellulase activity in bacilli, thus opening the prospect for genetic improvement of strains with an important biotechnological application.


Assuntos
Bacillus licheniformis/enzimologia , Bacillus licheniformis/genética , Bacillus/enzimologia , Bacillus/genética , Celulases/genética , Celulases/metabolismo , Clostridium/genética , Proteínas de Bactérias/biossíntese , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Celulose/metabolismo , Clonagem Molecular , Hidrólise , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
5.
J Chem Inf Model ; 61(9): 4554-4570, 2021 09 27.
Artigo em Inglês | MEDLINE | ID: mdl-34423980

RESUMO

Bacterial glycoside hydrolase 1 (GH1) enzymes with 6-phospho-ß-galactosidase and 6-phospho-ß-glucosidase activities have the important task of releasing phosphorylated and nonphosphorylated monosaccharides into the cytoplasm. Curiously, dual 6-phospho-ß-galactosidase/6-phospho-ß-glucosidase (dual-phospho) enzymes have broad specificity and are able to hydrolyze galacto- and gluco-derived substrates. This study investigates the structure and substrate specificity of a GH family 1 enzyme from Bacillus licheniformis, hereafter known as BlBglC. The enzyme structure has been solved, and sequence analysis, molecular dynamics simulations, and binding free energy calculations offered evidence of dual-phospho activity. Both test ligands p-nitrophenyl-ß-d-galactoside-6-phosphate (PNP6Pgal) and p-nitrophenyl-ß-d-glucoside-6-phosphate (PNP6Pglc) demonstrated strong binding to BlBglC although the pose and interactions of the PNP6Pglc triplicates were slightly more consistent. Interestingly, known specificity-inducing residues, Gln23 and Trp433, bind strongly to the ligand O3 hydroxyl group in the PNP6Pgal-BlBglC complex and to the ligand O4 hydroxyl group in the PNP6Pglc-BlBglC complex. Additionally, the BlBglC-His124 residue is a major contributor of hydrogen bonds to the PNP6Pgal O3 hydroxyl group but does not form any hydrogen bonds with PNP6Pglc. On the other hand, BlBglC residues Tyr173, Tyr301, Gln302, and Thr321 form hydrogen bonds with PNP6Pglc but not PNP6Pgal. These findings provide important details of the broad specificity of dual-phospho activity GH1 enzymes.


Assuntos
Bacillus licheniformis , Glucosidases , Bacillus licheniformis/metabolismo , Galactosidases , Glucosidases/metabolismo , Glicosídeo Hidrolases/metabolismo , Especificidade por Substrato
6.
Poult Sci ; 100(9): 101358, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34358955

RESUMO

This study investigated the effects of dietary supplementation with Bacillus subtilis (B. subtilis) or Bacillus licheniformis (B. licheniformis) on growth performance, immunity, antioxidant capacity, short chain fatty acid (SCFA) production, and the cecal microflora in broiler chickens. In total, 360 male, 1-day-old Cobb 500 birds were randomly divided into 3 groups: the control group was fed a basal diet; the B. subtilis group was fed a basal diet supplemented with 1.5 × 109 CFU/kg B. subtilis; the B. licheniformis group was fed a basal diet supplemented with 1.5 × 109 CFU/kg B. licheniformis. Results showed that chickens supplemented with either B. subtilis or B. licheniformis had comparatively higher (P < 0.05) body weight and average daily gain, whereas no difference (P > 0.05) was observed in feed efficiency. Concentrations of serum IgA, IgY, and IgM, as well as anti-inflammatory IL-10 were significantly increased (P < 0.05), and proinflammatory IL-1ß and IL-6 were significantly decreased (P < 0.05) by B. subtilis or B. licheniformis supplementation. Moreover, chickens fed with diets supplemented by either B. subtilis or B. licheniformis had greater antioxidant capacity, indicated by the notable increases (P < 0.05) in glutathione peroxidase, superoxide dismutase, and catalase, along with decrease (P < 0.05) in malondialdehyde. Compared to the control group, levels of SCFA, excluding acetic and propionic acid, in cecal content had improved (P < 0.05) by adding B. licheniformis, and significant increase (P < 0.05) in acetic and butyric acid was observed with B. subtilis supplementation. Microbial analysis showed that both B. subtilis or B. licheniformis supplementation could increase butyrate-producing bacteria such as Alistipes and Butyricicoccus, and decrease pathogenic bacteria such as the Synergistetes and Gammaproteobacteria. In summary, dietary supplemented with B. subtilis or B. licheniformis improved growth performance, immune status, and antioxidant capacity, increased SCFA production, and modulated cecal microbiota in chickens. Moreover, B. licheniformis was more effective than B. subtilis with the same supplemental amount.


Assuntos
Bacillus licheniformis , Microbioma Gastrointestinal , Probióticos , Ração Animal/análise , Animais , Antioxidantes , Bacillus subtilis , Galinhas , Dieta/veterinária , Suplementos Nutricionais , Ácidos Graxos Voláteis , Masculino
7.
Ecotoxicol Environ Saf ; 222: 112505, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34273849

RESUMO

Recently, slurry phase bioremediation as a simple and economical method is shown to be a successful technique for remediation of clayey soils. Besides, the use of microbial cell immobilization as a promising technique has drawn the attention of some researchers. The primary objective of this survey is to examine the synergistic adsorption and biodegradation performance of heavy crude oil by an isolated Bacillus licheniformis immobilized in a novel hybrid matrix (PUF/alginate/microbial cell) in aqueous phase. Isotherm studies and adsorption kinetics of crude oil on PUF matrix were carried out and their results revealed a good correlation between experimental data and Langmuir's isotherm and maximum monolayer coverage was found out to be 1.25 g/g PUF. The other objective of this research is examination of hybrid matrix in slurry phase bioremediation of heavy crude oil polluted clayey soil as a reluctant model soil. In order to model, optimize, and investigate the factors affecting the total organic carbon (TOC) reduction, response surface methodology (RSM) was applied. For this purpose, the effect of three variables including crude oil concentration (5000-25,000 mg/kg dry soil), soil salinity (0-10%), and water to soil ratio (WSR: 2-10) have been studied. In this study, TOC reduction was achieved in ranging from 39% to 80% in crude oil polluted soil after 21 days. Additionally, experiments by polyurethane foam (PUF)-immobilized cell, alginate-immobilized cell, and freely cell suspended systems were conducted to compare the performance of hybrid-immobilized cell with other systems. Our results showed the superiority of immobilized cells in hybrid matrix of PUF/alginate compared to other immobilized cell (IC) and free cell (FC) systems. Overall, the results indicated that the hybrid matrix with simultaneous adsorption-biodegradation capacity is potentially suitable for further development for oil spill treatment and it can be used as an efficient cleaning method in TOC removal from actual polluted soils.


Assuntos
Bacillus licheniformis , Petróleo , Poluentes do Solo , Adsorção , Biodegradação Ambiental , Solo , Poluentes do Solo/análise , Água
8.
Sheng Wu Gong Cheng Xue Bao ; 37(7): 2453-2462, 2021 Jul 25.
Artigo em Chinês | MEDLINE | ID: mdl-34327910

RESUMO

The ban on addition of antibiotics in animal feed in China has made the search for new antibiotics substitutes, e.g. bacteriocin, a hot topic in research. The present study successfully isolated an antibacterial substance producing strain of Bacillus sp. from alpaca feces by agar diffusion method, using Escherichia coli, Salmonella enterica, Staphylococcus aureus, Staphylococcus epidermidis, Micrococcus luteus and Listeria monocytogenes as indicator bacteria. The isolated strain was named as B. licheniformis SXAU06 based on colony morphology, Gram staining and 16S rRNA gene sequence. The antibacterial substance was isolated and purified through a series of procedures including (NH4)2SO4 precipitation, chloroform extraction, molecular interception and SDS-PAGE analysis. Bioinformatics analysis of the LC-MS/MS data indicated that the antibacterial substance was a bacteriocin-like substance (BLIS) with an approximate molecular weight of 14 kDa, and it was designated as BLIS_SXAU06. BLIS_SXAU06 exhibited high resistance to treatment of proteinase K, high temperature, high acidity and alkalinity. BLIS_SXAU06 was heterologously expressed in E. coli and the recombinant BLIS_SXAU06 exhibited effective antibacterial activity against S. aureus, S. epidermidis, M. luteus, and L. monocytogenes, showing potential to be investigated further.


Assuntos
Bacillus licheniformis , Bacteriocinas , Listeria monocytogenes , Animais , Antibacterianos/farmacologia , Bacteriocinas/genética , Bacteriocinas/farmacologia , China , Cromatografia Líquida , Escherichia coli/genética , RNA Ribossômico 16S , Staphylococcus aureus , Espectrometria de Massas em Tandem
9.
Bioresour Technol ; 337: 125467, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34320747

RESUMO

Bacillus licheniformis α-amylase is a thermostable enzyme used in industrial starch hydrolysis. However, difficulties in the genetic manipulation of B. licheniformis hamper further enhancement of α-amylase production. In this regard, adaptive evolution is a useful strategy for promoting the productivity of microbial hosts, although the success of this approach requires the application of suitable evolutionary stress. In this study, we designed a growth-coupled adaptive evolution model to enrich B. licheniformis strains with enhanced amylase productivity and utilization capacity of starch substrates. Single cells of high α-amylase-producing B. licheniformis were isolated using a droplet-based microfluidic platform. Clones with 67% higher α-amylase yield were obtained and analyzed by genome resequencing. Our findings confirmed that growth-coupled evolution combined with high-throughput screening is an efficient strategy for enhanced α-amylase production. In addition, we identified several potential target genes to guide further modification of the B. licheniformis host for efficient protein expression.


Assuntos
Bacillus licheniformis , Bacillus , Amilases , Bacillus/genética , Bacillus licheniformis/genética , Ensaios de Triagem em Larga Escala , Amido , alfa-Amilases
10.
J Environ Manage ; 295: 112969, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34146779

RESUMO

In this study, the influence of a static magnetic field (SMF) on a Chlorella vulgaris-Bacillus licheniformis consortium and the subsequent effect of this algal-bacterial consortium on sewage treatment were explored. Accordingly, the algal density, Fv/Fm, algal aggregation percentage, extracellular polymeric substances (EPS) content, dissolved organic matter distribution, enzymatic activity, metabolites, microbial community diversity and nutrient removal were investigated. For the treatment group exposed to an SMF of 150 mT, the total phosphorus removal rate reached 82.21%, which was 19.10% higher than the control group. On the last day, the algal density of the 150 mT group was the highest, being 56.01% greater than the control group. The high intensity SMF promoted the anti-oxidative stress response in C. vulgaris. It also affected EPS secretion, subsequently influencing the algal aggregation percentage and bacterial growth. Bacillus accounted for the largest proportion of the overall microbial community in the 150 mT group, which was conducive to rapid formation of the C. vulgaris-B. licheniformis consortium. In short, the SMF was conducive to the rapid formation of a C. vulgaris-B. licheniformis consortium. The use of an SMF can promote the efficiency of the algal-bacterial consortium, thereby shortening the processing time.


Assuntos
Bacillus licheniformis , Chlorella vulgaris , Campos Magnéticos , Fósforo , Esgotos
11.
Int J Biol Macromol ; 185: 277-286, 2021 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-34147526

RESUMO

Members of group Bacillus are most widely occurring microbes in agricultural soil and they affect crop health in various ways. They directly stimulate plant growth either by augmenting nutrients availability, invigorating plants' defence mechanisms; repressing soil-borne phytopathogens or by producing growth-regulating hormones like auxins and cytokinins. It is a well known fact that indole-3- acetic acid (a type of auxin) is a vital biologically active phytohormone excreted by certain Bacillus species, but its molecular mechanism has not yet been described. In this study, the auxin efflux carrier gene is isolated from the metagenome of the Tapta Kund hot spring, Uttrakhand, India. In addition, auxin efflux carrier (AEC) transporter protein of Bacillus licheniformis is modeled and the 318 amino acid residues long protein was found homologous to the apical sodium-dependent bile acid transporter (ASBT) of Yersinia frederiksnii, with 10 transmembrane segments (TM1-10) split into different domains: a panel domain defined by TM1, 2, 6 and 7; and a core domain defined by TM3-5 and 8-10. Finally, the predicted Bacillus licheniformis AEC protein has also been phylogenetically evaluated and its detailed molecular transport mechanism was worked out using molecular dynamics simulation analysis. Conclusively, this study demonstrates the efflux mechanism of the substrate, Indole 3- acetic acid by AEC transporter protein.


Assuntos
Bacillus licheniformis/isolamento & purificação , Fontes Termais/microbiologia , Ácidos Indolacéticos/metabolismo , Proteínas de Membrana Transportadoras/química , Proteínas de Membrana Transportadoras/genética , Sequência de Aminoácidos , Bacillus licheniformis/genética , Bacillus licheniformis/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Transporte Biológico , Índia , Proteínas de Membrana Transportadoras/metabolismo , Metagenômica , Modelos Moleculares , Simulação de Dinâmica Molecular , Domínios Proteicos , Estrutura Terciária de Proteína
12.
Int Microbiol ; 24(3): 455-470, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34100180

RESUMO

The reemergence of infectious diseases and resistant pathogens represents a serious problem for human life. Hence, the screening for new or alternative antimicrobial compounds is still urgent. Unusual ecosystems such as saline habitats are considered promising environments for the purposes of isolating bacterial strains able to produce potent natural products. The aim of this study is the identification of bioactive compounds biosynthesized by three halotolerant strains isolated from the Sebkha of Oran (Algeria) using gas chromatography coupled to mass spectrometry. Primary screening investigation of antimicrobial activities were performed against reference bacterial and fungal strains and revealed a broad-spectrum activity. Secondary metabolite extraction was carried out using ethyl acetate and chloroform. Crude extracts were tested for bioactivity using the disc diffusion method and subjected to GC-MS analysis. The extracts showed an important inhibitory effect against all tested strains. Fifty-six compounds were identified; they include tert-butyl phenol compounds, fatty acid methyl esters due to the methylation procedure, hydrocarbons, aldehydes, benzoquinones, pyrrols, and terpenes. Literature reports such compounds to have wide biological and pharmaceutical applications. The molecular identification of the three isolates was achieved using the 16S-23S rRNA gene intergenic spacer region (ITS) and 16S rRNA sequencing. Partial 16S rRNA gene sequencing showed very high similarity with many species of Bacillus. This study provided insights on the potential of halotolerant Bacillus as drug research target for bioactive metabolites. The findings suggest that the Great Sebkha of Oran is a valuable source of strains exhibiting variety of beneficial attributes that can be utilized in the development of biological antibiotics.


Assuntos
Anti-Infecciosos/metabolismo , Anti-Infecciosos/farmacologia , Bacillus licheniformis/metabolismo , Bactérias/efeitos dos fármacos , Fungos/efeitos dos fármacos , Argélia , Bacillus licheniformis/classificação , Bacillus licheniformis/genética , Bacillus licheniformis/isolamento & purificação , DNA Bacteriano , Ecossistema , Cromatografia Gasosa-Espectrometria de Massas/métodos , Lagos/microbiologia , Testes de Sensibilidade Microbiana , RNA Ribossômico 16S , Tolerância ao Sal , Metabolismo Secundário , Microbiologia do Solo
13.
J Ind Microbiol Biotechnol ; 48(5-6)2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-34124759

RESUMO

Ammonium hydroxide is conventionally used as an alkaline reagent and cost-effective nitrogen source in enzyme manufacturing processes. However, few ammonia-inducible enzyme expression systems have been described thus far. In this study, genomic-wide transcriptional changes in Bacillus licheniformis CBBD302 cultivated in media supplemented with ammonia were analyzed, resulting in identification of 1443 differently expressed genes, of which 859 genes were upregulated and 584 downregulated. Subsequently, the nucleotide sequences of ammonia-inducible promoters were analyzed and their functionally-mediated expression of amyL, encoding an α-amylase, was shown. TRNA_RS39005 (copA), TRNA_RS41250 (sacA), TRNA_RS23130 (pdpX), TRNA_RS42535 (ald), TRNA_RS31535 (plp), and TRNA_RS23240 (dfp) were selected out of the 859 upregulated genes and each showed higher transcription levels (FPKM values) in the presence of ammonia and glucose than that of the control. The promoters, PcopA from copA, PsacA from sacA, PpdpX from pdpX, Pald from ald, and Pplp from plp, except Pdfp from dfp, were able to mediate amyL expression and were significantly induced by ammonia. The highest enzyme expression level was mediated by Pplp and represented 23% more α-amylase activity after induction by ammonia in a 5-L fermenter. In conclusion, B. licheniformis possesses glucose-independent ammonia-inducible promoters, which can be used to mediate enzyme expression and therefore enhance the enzyme yield in fermentations conventionally fed with ammonia for pH adjustment and nitrogen supply.


Assuntos
Amônia/metabolismo , Bacillus licheniformis/metabolismo , Regiões Promotoras Genéticas , alfa-Amilases/metabolismo , Bacillus licheniformis/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sequência de Bases , DNA Bacteriano , Fermentação , Perfilação da Expressão Gênica , Regulação Bacteriana da Expressão Gênica , Concentração de Íons de Hidrogênio , Microbiologia Industrial , Nitrogênio/metabolismo , Estresse Fisiológico , alfa-Amilases/genética
14.
J Dairy Sci ; 104(9): 9627-9644, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34127263

RESUMO

Heat treatment is one of the most widely used processing technologies in the dairy industry. Its primary purpose is to destroy microorganisms, both pathogenic and spoilage, to ensure the product is safe and has a reasonable shelf life. In this study microwave volumetric heating (MVH) was compared with a conventional tubular heat exchanger (THE), in terms of the effects of each at a range of temperatures (75°C, 85°C, 95°C, 105°C, 115°C, and 125°C) on indigenous microflora viability and the germination of inoculated Bacillus licheniformis endospores in reconstituted skim milk. To assess the heat treatment-related effects on microbial viability, classical agar-based tests were applied to obtain the counts of 4 various microbiological groups including total bacterial, thermophilic bacterial, mesophilic aerobic bacterial endospore, and thermophilic aerobic bacterial endospore counts, and additional novel insights into cell permeability and spore germination profiles post-heat treatment were obtained using real-time flow cytometry (FC) methods. No significant differences in the plate counts of the indigenous microorganisms tested, the plate counts of the inoculated B. licheniformis, or the relative percentage of germinating endospores were observed between MVH- and THE-treated samples, at equal temperatures in the range specified above, indicating that both methods inactivated inoculated endospores to a similar degree (up to 70% as measured by FC and 5 log reduction as measured by plate counting for some treatments of inoculated endospores). Furthermore, increased cell permeability of indigenous microflora was observed by FC after MVH compared with THE treatment of uninoculated skim milk, which was reflected in lower total bacterial count at a treatment temperature of 105°C. This work demonstrates the utility of FC as a rapid method for assessing cell viability and spore inactivation for postthermal processing in dairy products and overall provides evidence that MVH is at least as effective at eliminating native microflora and inoculated B. licheniformis endospores as THE.


Assuntos
Bacillus licheniformis , Leite , Animais , Citometria de Fluxo/veterinária , Calefação , Temperatura Alta , Micro-Ondas , Esporos Bacterianos
15.
Folia Microbiol (Praha) ; 66(5): 787-795, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34128186

RESUMO

Bacillus licheniformis HJ4 showing strong fibrinolytic activity was isolated from Hwangseokae jeotgal. aprEHJ4, a major fibrinolytic gene, was cloned by PCR, and an ORF consisting of 379 amino acids was located. The mature enzyme was expected to be 27 kDa in size after processing, but a 24-kDa protein was observed by SDS-PAGE and fibrin zymography, indicating additional processing. RT-qPCR showed that expression level of aprEHJ4 in culture with 0% salt (control) was the highest followed by culture with 8% salt (89.7% of control) and 5% salt (74.2%) at 84 h. The expression level in culture with 15% salt was 46.9%. The results matched with the fibrinolytic activity measurements of cultures and indicated that AprEHJ4 maintained significant activity in the presence of salt up to 15% (w/v). AprEHJ4 was overproduced in Escherichia coli, and mature 27 kDa protein was purified after in vitro renaturation. The optimum pH and temperature of AprEHJ4 were pH 8 and 40 ℃, respectively.


Assuntos
Bacillus licheniformis , Alimentos Marinhos , Bacillus licheniformis/enzimologia , Ativação Enzimática/efeitos dos fármacos , Concentração de Íons de Hidrogênio , Peptídeo Hidrolases/metabolismo , República da Coreia , Alimentos Marinhos/microbiologia , Cloreto de Sódio/farmacologia
16.
Toxins (Basel) ; 13(6)2021 05 21.
Artigo em Inglês | MEDLINE | ID: mdl-34064255

RESUMO

The present study was designed to determine the efficacy of a novel multicomponent mycotoxin detoxifying agent (MMDA) containing modified zeolite (Clinoptilolite), Bacillus subtilis, B. licheniformis, Saccharomyces cerevisiae cell walls and silymarin against the deleterious effects of Aflatoxin B1 (AFB1) and Ochratoxin A (OTA) in broiler chicks. A total of 160 one-day-old Ross 308® broiler chicks were randomly allocated in four treatment groups, with four replicates, according to the following experimental design for 42 days. Group A received a basal diet; Group B received a basal diet contaminated with AFB1 and OTA at 0.1 mg/kg and 1 mg/kg, respectively; Group C received a basal diet contaminated with AFB1 and OTA and MMDA at 1 g/kg feed, and Group D received a basal diet contaminated with AFB1 and OTA and MMDA at 3 g/kg feed. Results showed that ingested mycotoxins led to significant (p ≤ 0.05) reduction in body weight and feed conversion from 25 days of age, induced histopathological changes, increased the pH of the intestinal content, and altered the biochemical profile of birds with significantly (p ≤ 0.05) increased aspartate aminotransferase (AST) values (p ≤ 0.05). On the other hand, the supplementation of MMDA significantly (p ≤ 0.05) improved the feed conversion ratio (FCR) during the second part of the study, diminished biochemical alterations, reduced pH in jejunal and ileal content, and E. coli counts in the caeca of birds (p ≤ 0.05). It may be concluded that the dietary supplementation of the MMDA partially ameliorated the adverse effects of AFB1 and OTA in broilers and could be an efficient tool in a mycotoxin control program.


Assuntos
Aflatoxina B1/envenenamento , Micotoxicose/tratamento farmacológico , Ocratoxinas/envenenamento , Silimarina/administração & dosagem , Zeolitas/administração & dosagem , Ração Animal , Animais , Bacillus licheniformis , Bacillus subtilis , Galinhas , Micotoxicose/metabolismo , Micotoxicose/patologia , Distribuição Aleatória , Saccharomyces cerevisiae
17.
J Plant Physiol ; 262: 153437, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34034041

RESUMO

Detrimental effects caused by the overuse of synthetic agrochemicals have led to the development of natural biostimulants such as seaweed extracts and plant growth-promoting rhizobacteria (PGPR) being used as an alternative, environmentally-friendly technology to improve crop growth and increase agricultural yields. The present study aimed to investigate the interactions between PGPR and a commercial seaweed extract on the growth and biochemical composition of onion (Allium cepa). A pot trial was conducted under greenhouse conditions where onion plants were treated individually with the two PGPR, namely Bacillus licheniformis (BL) and Pseudomonas fluorescens (PF) and a seaweed extract Kelpak® (KEL) and combinations of KEL + BL and KEL + PF. Growth and yield parameters were measured after 12 weeks. KEL-treated plants showed the best growth response and overcame the inhibitory effects of BL treatment. KEL-treated plants also had the highest chlorophyll content. PGPR application improved the mineral nutrition of onion with these plants having the highest mineral content in the leaves and bulb. All biostimulant treatments increased the endogenous cytokinin and auxin content with the highest concentrations generally detected in the PF-treated plants. These results suggest that co-application of different biostimulant classes with different modes of action could further increase crop productivity with an improvement in both growth and nutrition content being achieved in onion with the co-application of a seaweed extract and PGPR.


Assuntos
Bacillus licheniformis , Cebolas/crescimento & desenvolvimento , Extratos Vegetais/farmacologia , Pseudomonas fluorescens , Alga Marinha/química , Bacillus licheniformis/metabolismo , Carotenoides/metabolismo , Clorofila/metabolismo , Produção Agrícola/métodos , Cebolas/efeitos dos fármacos , Cebolas/microbiologia , Cebolas/fisiologia , Reguladores de Crescimento de Plantas/metabolismo , Pseudomonas fluorescens/metabolismo
18.
J Phys Chem B ; 125(22): 5766-5774, 2021 06 10.
Artigo em Inglês | MEDLINE | ID: mdl-34047564

RESUMO

Levan-type fructooligosaccharides (LFOs) and levan can potentially be used as ingredients in prebiotics, skincare products, and antitumor agents. The Y246S mutant of Bacillus licheniformis RN-01 levansucrase (oligosaccharide-producing levansucrase, OPL) was reported to productively synthesize LFOs; however, OPL's thermostability is low at high temperatures. To enhance OPL structural stability, this study employed molecular dynamics (AMBER) to identify a highly flexible region, as measured by its average root-mean-square fluctuation (RMSF) value, on the OPL surface and computational protein design (Rosetta) to rigidify and increase favorable interactions to increase its structural stability. AMBER identified region nine (residues 277-317) as a highly flexible region that was selected for design because it has the highest number of residues and the second-highest average RMSF, and it is farthest from the active site. Rosetta designed 14 mutants with the best ΔΔG value in each position, where three mutants have better ΔG than OPL. To determine whether their flexibilities and stabilities are lower than those of OPL, all 14 designed mutants were simulated at high temperature (500 K), and we found that K296E, G309S, and A310W mutants were predicted to be more stable and could retain their native structures better than OPL. Our results suggest that enhanced structural stabilities of these mutants are caused by increased hydrogen bond strengths of the designed residues and their neighboring residues. This study designed K296E, G309S, and A310W mutants of OPL with high potential for stability improvement, and they could potentially be used for the effective production of LFOs.


Assuntos
Bacillus licheniformis , Hexosiltransferases , Bacillus licheniformis/genética , Bacillus licheniformis/metabolismo , Estabilidade Enzimática , Hexosiltransferases/genética , Hexosiltransferases/metabolismo , Oligossacarídeos , Temperatura
19.
Arch Microbiol ; 203(7): 4081-4089, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34052893

RESUMO

Bacillus licheniformis DAS-1 and DAS-2 were found as potent tool for removal/uptake of arsenic [As(V) and As(III)] and reduction [(As(V) to As(III)] of arsenic from the liquid growth medium in our previous studies. Present work gives light on modulation of arsenic remediation (in terms of uptake and reduction) by two selected essential soil nutrients, phosphate (PO4)3- and nitrate (NO3)-. PO43- has structural analogy with arsenate [AsO43-/As(V)] that compete with cell uptake of As(V). It was found that enrichment of 0.75 mM of PO43- had significantly moderated the As(V) toxicity in liquid broth culture by retarding As(V) uptake. Lowering level of PO43- can lead to increase in As(V) removal from medium and vice versa. NO3- has strong oxidant properties which controls As(III) oxidation into As(V) in medium that resulted less toxicity favouring growth of bacteria and also more uptake via phosphate transporters. Hence, accelerated As(III) uptake has shown on enrichment of 0.5 mM of NO3- in medium. All the results of work give evidence that appropriate enrichment of PO43- and NO3- into liquid growth medium, can significantly contribute in alteration of efficiency for arsenic removal/uptake and reduction by bacteria from the medium.


Assuntos
Arsênio , Bacillus licheniformis , Arsênio/metabolismo , Bacillus licheniformis/metabolismo , Biodegradação Ambiental , Nitratos/química , Nitratos/metabolismo , Fosfatos/metabolismo , Solo
20.
Appl Microbiol Biotechnol ; 105(9): 3601-3610, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33937931

RESUMO

The food industry has developed a wide range of products with reduced lactose to allow people with intolerance to consume dairy products. Although ß-galactosidase has extensive applications in the food, pharma, and biotechnology industries, the enzymes are high-cost catalysts, and their use makes the process costly. Immobilization is a viable strategy for enzyme retention inside a reactor, allowing its reuse and application in continuous processes. Here, we studied the immobilization of ß-galactosidase from Bacillus licheniformis in ion exchange resin. A central composite rotational design (CCRD) was proposed to evaluate the immobilization process in relation to three immobilization solution variables: offered enzyme activity, ionic strength, and pH. The conditions that maximized the response were offered enzyme activity of 953 U, 40 mM ionic strength, and pH 4.0. Subsequently, experiments were performed to provide additional stabilization for biocatalyst, using a buffer solution pH 9.0 at 25 °C for 24 h, and crosslinking with different concentrations of glutaraldehyde. The stabilization step drastically impacted the activity of the immobilized enzyme, and the reticulation with different concentrations of glutaraldehyde showed significant influence on the activity of the immobilized enzyme. In spite of substantially affecting the initial activity of the immobilized enzyme, higher reagent concentrations (3.5 g L-1) were effective for maintaining stability related to the number of cycles of the enzyme immobilized. The ß-galactosidase from Bacillus licheniformis immobilized in Duolite A568 is a promising technique to produce reduced or lactose-free dairy products, as it allows reuse of the biocatalyst, decreasing operational costs.Key Points• Immobilization of ß-galactosidase from Bacillus licheniformis in batch reactor• Influence of buffer pH and ionic concentration and offered enzyme activity on immobilization• Influence of glutaraldehyde on operational stability.


Assuntos
Bacillus licheniformis , Bacillus licheniformis/metabolismo , Indústria de Laticínios , Estabilidade Enzimática , Enzimas Imobilizadas/metabolismo , Humanos , Concentração de Íons de Hidrogênio , Lactose , Temperatura , beta-Galactosidase/metabolismo
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