Project description:In this study, the upstream and downstream production processes of curdlan from Priestia megaterium were optimized to enhance its yield. Additionally, a novel extraction method was developed for curdlan recovery. Optimization studies were conducted using Central composite design (CCD). Curdlan yield improved from 0.15 g/L (unoptimized) to 0.46 g/L (3-fold increase) when fermentation was carried out in CCD-optimized media of (w/v) sucrose 20%, urea 0.1%, KH2PO4 0.02%, agitation speed 250 rpm. To further enhance curdlan yield during extraction, ultrasonication was incorporated as a novel step into the conventional method of acid/alkali-assisted curdlan recovery. A two-step optimization was chosen for extraction, namely, one-factor-at-a-time (OFAT) and CCD, wherein the optimized extraction parameters were determined to be 25 s sonication, 1 N NaOH, and 2 h solubilization time. The curdlan yield improved by 1.5-fold (0.70 g/L) post optimization, in comparison with unoptimized conventional extraction step. Finally, the biopolymer was validated through characterization by nuclear magnetic resonance (NMR) which showed characteristic curdlan spectra in the13C and1H NMR studies. To the best of our knowledge, this study represents the first documented report on curdlan extraction using this novel method of ultrasonication.
Project description:Comparison of chitosan-treated B. cereus ATCC 14579 cells with non-treated B. cereus ATCC 14579 cells. 2 chitosans with similar molecular weight (Mw) but different degrees of acetylation (Fa) were used: chitosan B (Mw: 28.4 kDa, Fa: 0.16) (samples 1-3) and chitosan A (Mw: 36.0 kDa, Fa: 0.01) (samples 4-6).
Project description:An enzymic assay for individual isomers (meso-, LL- and DD-) of 2,6-diaminopimelate was developed. The enzyme 2,6-diaminopimelate decarboxylase specifically attacked meso-diaminopimelate and was used to measure this isomer manometrically. The meso- and LL-isomers were measured together manometrically in a coupled assay with diaminopimelate decarboxylase and diaminopimelate epimerase (which converts LL-diaminopimelate into meso-diaminopimelate). The DD-isomer was not attacked by either enzyme and was measured, as residual diaminopimelate after the coupled assay, by a colorimetric method, which was also used to measure total diaminopimelate before enzymic treatments. The coupled enzymes were also used to prepare pure DD-isomer from chemically synthesized diaminopimelate. A mixture of diaminopimelate isomers was present in walls of four strains of Bacillus megaterium [in each about 75% (w/w) meso-, 18% LL- and 7% DD-] and in walls of two strains of Bacillus cereus (about 85% meso-, 8% LL- and 7% DD-). One strain of B. cereus contained at least 95% meso-diaminopimelate, with only traces of LL- and DD-isomers. Peptidoglycan from Escherichia coli was assayed as containing at least 95% meso-isomer. The proportion of isomers in the wall of a strain of B. megaterium remained constant after growth in a variety of different media.
Project description:Isoleucyl-tRNA synthetase (IleRS) is an essential enzyme that covalently couples isoleucine to the corresponding tRNA. Bacterial IleRSs group in two clades, ileS1 and ileS2, the latter bringing resistance to the natural antibiotic mupirocin. Generally, bacteria rely on either ileS1 or ileS2 as a standalone housekeeping gene. However, we have found an exception in Bacillus species: in family Bacillaceae ileS1 appears mandatory, but some of the Bacilli posses additional, ileS2 gene. Taking Priestia (Bacillus) megaterium as a model organism, we showed that PmIleRS1 is constitutively expressed, while PmIleRS2 is stress-induced. As part of our investigation of ileS1/ileS2 function, we explored the expression of ileS1 and ileS2 under control conditions and upon mupirocin treatment.
Project description:The growth of material science and technology places high importance on creating better processes for synthesizing copper nanoparticles. Thus, an easy, ecological, and benign process for producing copper nanoparticles (CuNPs) has been developed using Priestia sp. bacteria utilizing a variety of low-cost agro-industrial wastes and byproducts. The biosynthesis of CuNPs was conducted using glucose medium and copper ions salt solution, then it was replaced by utilizing low-cost agro-industrial wastes. UV-visible spectroscopy, dynamic light scattering (DLS), X-ray diffraction (XRD), High-resolution transmission electron microscope (HR-TEM), Attenuated Total Reflectance and Fourier transform infrared (ATR-FTIR), and zeta potential were used to characterize the biosynthesized CuNPs. The cytotoxicity of CuNPs using Vero -CCL-81 cell lines, and antibacterial and antitumor properties using human colon epithelial colorectal adenocarcinoma Caco-2-HTB-37 cell lines were assessed. The UV-visible and DLS studies revealed CuNPs formation, with a maximum concentration of 6.19 ppm after 48 h, as indicated by a 0.58 Surface plasmon resonance (SPR) within 450 nm and 57.73 nm particle size. The 16S rRNA gene analysis revealed that Priestia sp. isolate is closely related to Priestia megaterium and has been deposited in the NCBI GenBank with accession number AMD 2024. The biosynthesis with various agro-industrial wastes indicated blackstrap sugar cane molasses being the most effective for reducing CuNPs size to 3.12 nm owing to various reducing and stabilizing active compounds. The CuNPs were free of contaminants, with a sphere-shaped structure and a cytotoxicity assessment with an IC50 of 367.27 μg/mL. The antibacterial activity exhibited by the most susceptible bacteria were Bacillus cereus ATCC 11788 and Staphylococcus aureus ATCC 6538 with inhibition zones of 26.0 mm and 28.0 mm, respectively. The antitumor effect showed an IC50 dose of 175.36 μg/mL. Based on the findings, the current work sought to lower product costs and provide a practical solution to the environmental contamination issues brought on by the buildup of agricultural wastes. In addition, the obtained CuNPs could be applied in many fields such as pharmaceuticals, water purification, and agricultural applications as future aspects.
Project description:Here, we report the high-quality complete genome sequences and plasmid arrays of Priestia megaterium ATCC 14581T and of two clinical strains (2008724129 and 2008724142) isolated from human samples in the United States.
Project description:We isolated an atmospheric contaminant, subsequently identified as a new strain of Bacillus mobilis, which showed a novel, robust, inducible filamentous sliding motility and completely colonized a bacterial culture plate in less than 48 h under some conditions. This flagella-independent sliding motility was characterized by long filamentous cells at the expanding edge, and was induced when cells were inoculated onto lawns of metabolically inactive Campylobacter jejuni cells, heat killed bacterial biomass, and milk or blood dried onto agar plates. Phosphatidylcholine (PC), bacterial membrane components, and sterile human fecal extracts were sufficient to induce filamentous expansion. Screening of eight other Bacillus spp. (five from the B. cereus group and three other Bacillus spp.) showed that filamentous motility was conserved amongst B. cereus group species to varying degrees. RNAseq of filamentously expanding cells collected from PC and milk lawn plates in comparison to rod-shaped cells from control plates revealed that genes related to metabolism, ion and amino acid transport were differently regulated, genes controlling sporulation were reduced, and some virulence genes (e.g., hblA/B/C/D and plcR) were increased. We hypothesize that the robust and conserved nature of filamentous motility in pathogenic B. cereus group species can enhance bacterial colonization during host colonization.