Identification and characterization of a novel heparinase PCHepII from marine bacterium Puteibacter caeruleilacunae
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SUBMITTER: Lu D
PROVIDER: S-EPMC10656541 | biostudies-literature | 2023 Jan
REPOSITORIES: biostudies-literature
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Project description:An additive- and pollution-free method for the preparation of biogenic silver and silver chloride nanoparticles (Ag@AgCl NPs) was developed from the bacteria Shewanella sp. Arc9-LZ, which was isolated from the deep sea of the Arctic Ocean. The optimal synthesizing conditions were explored, including light, pH, Ag+ concentration and time. The nanoparticles were studied by means of ultraviolet-visible (UV-Vis) spectrophotometry, energy dispersive spectrometry (EDS), X-ray diffraction (XRD) and inductively coupled plasma optical emission spectrometers (ICP-OES). The transmission electron microscope (TEM) showed that the nanoparticles were spherical and well dispersed, with particle sizes less than 20.00 nm. With Ag@AgCl nanoparticles, the kinetic rate constants for congo red (CR) and rhodamine B (RhB) dye degradation were 2.74 × 10-1 min-1 and 7.78 × 10-1 min-1, respectively. The maximum decolourization efficiencies of CR and RhB were 93.36% and 99.52%, respectively. Ag@AgCl nanoparticles also showed high antibacterial activities against the Gram-positive and Gram-negative bacteria. The Fourier transform infrared spectroscopy (FTIR) spectrum indicated that the O-H, N-H and -COO- groups in the supernatant of Arc9-LZ might participate in the reduction, stabilization and capping of nanoparticles. We mapped the schematic diagram on possible mechanisms for synthesizing Ag@AgCl NPs.
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Project description:BackgroundThe identification of microorganisms with excellent flocculant-producing capability and optimization of the fermentation process are necessary for the wide-scale application of bioflocculants. Thus, we evaluated the flocculant-producing ability of a novel strain identified by the screening of marine bacteria, and we report for the first time the properties of the bioflocculant produced by Alteromonas sp. in the treatment of dye wastewater.ResultsA bioflocculant-producing bacterium was isolated from seawater and identified as Alteromonas sp. CGMCC 10612. The optimal carbon and nitrogen sources for the strain were 30 g/L glucose and 1.5 g/L wheat flour. In a 2-L fermenter, the flocculating activity and bioflocculant yield reached maximum values of 2575.4 U/mL and 11.18 g/L, respectively. The bioflocculant was separated and showed good heat and pH stability. The purified bioflocculant was a proteoglycan consisting of 69.61% carbohydrate and 21.56% protein (wt/wt). Infrared spectrometry further indicated the presence of hydroxyl, carboxyl and amino groups preferred for flocculation. The bioflocculant was a nanoparticle polymer with an average mass of 394,000 Da. The purified bioflocculant was able to remove Congo Red, Direct Black and Methylene Blue at efficiencies of 98.5%, 97.9% and 72.3% respectively.ConclusionsThe results of this study indicated that the marine strain Alteromonas sp. is a good candidate for the production of a novel bioflocculant and suggested its potential industrial utility for biotechnological processes.
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Project description:Naegleria fowleri is an opportunistic protozoan, belonging to the free-living amoeba group, that can be found in warm water bodies. It is causative agent the primary amoebic meningoencephalitis, a fulminant disease with a rapid progression that affects the central nervous system. However, no 100% effective treatments are available and those that are currently used involve the appearance of severe side effects, therefore, there is an urgent need to find novel antiamoebic compounds with low toxicity. In this study, the in vitro activity of six oxasqualenoids obtained from the red algae Laurencia viridis was evaluated against two different strains of N. fowleri (ATCC® 30808 and ATCC® 30215) as well as their cytotoxicity against murine macrophages. Yucatecone was the molecule with the highest selectivity index (>2.98 and 5.23 respectively) and it was selected to continue with the cell death type determination assays. Results showed that yucatone induced programmed cell death like responses in treated amoebae causing DNA condensation and cellular membrane damage among others. In this family of oxasqualenoids, it seems that the most significative structural feature to induce activity against N. fowleri is the presence of a ketone at C-18. This punctual oxidation transforms an inactive compound into a lead compound as the yucatecone and 18-ketodehydrotyrsiferol with IC50 values of 16.25 and 12.70 μM, respectively. The assessment of in silico ADME/Tox analysis revealed that the active compounds showed good Human Oral Absorption and demonstrate that are found to be within the limit of approved drug parameter range. Hence, the study highlights promising potential of yucatone to be tested for therapeutic use against primary amoebic meningoencephalitis.
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Project description:Chitooligosaccharides (COS), a high-value chitosan derivative, have many applications in food, pharmaceuticals, cosmetics and agriculture owing to their unique biological activities. Chitosanase, which catalyzes the hydrolysis of chitosan, can cleave β-1,4 linkages to produce COS. In this study, a chitosanase-producing Bacillus paramycoides BP-N07 was isolated from marine mud samples. The chitosanase enzyme (BpCSN) activity was 2648.66 ± 20.45 U/mL at 52 h and was able to effectively degrade chitosan. The molecular weight of purified BpCSN was approximately 37 kDa. The yield and enzyme activity of BpCSN were 0.41 mg/mL and 8133.17 ± 47.83 U/mg, respectively. The optimum temperature and pH of BpCSN were 50 °C and 6.0, respectively. The results of the high-performance liquid chromatography (HPLC) and thin-layer chromatography (TLC) of chitosan treated with BpCSN for 3 h showed that it is an endo-chitosanase, and the main degradation products were chitobiose, chitotriose and chitotetraose. BpCSN was used for the preparation of oligosaccharides: 1.0 mg enzyme converted 10.0 g chitosan with 2% acetic acid into oligosaccharides in 3 h at 50 °C. In summary, this paper reports that BpCSN has wide adaptability to temperature and pH and high activity for hydrolyzing chitosan substrates. Thus, BpCSN is a chitosan decomposer that can be used for producing chitooligosaccharides industrially.
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Project description:An esterase gene, estA1, was cloned from Alteromonas sp. 39-G1 isolated from the Beaufort Sea. The gene is composed of 1,140 nucleotides and codes for a 41,190 Da protein containing 379 amino acids. As a result of a BLAST search, the protein sequence of esterase EstA1 was found to be identical to Alteromonas sp. esterase (GenBank: PHS53692). As far as we know, no research on this enzyme has yet been conducted. Phylogenetic analysis showed that esterase EstA1 was a member of the bacterial lipolytic enzyme family IV (hormone sensitive lipases). Two deletion mutants (Δ20 and Δ54) of the esterase EstA1 were produced in Escherichia coli BL21 (DE3) cells with part of the N-terminal of the protein removed and His-tag attached to the C-terminal. These enzymes exhibited the highest activity toward p-nitrophenyl (pNP) acetate (C2) and had little or no activity towards pNP-esters with acyl chains longer than C6. Their optimum temperature and pH of the catalytic activity were 45°C and pH 8.0, respectively. As the NaCl concentration increased, their enzyme activities continued to increase and the highest enzyme activities were measured in 5 M NaCl. These enzymes were found to be stable for up to 8 h in the concentration of 3-5 M NaCl. Moreover, they have been found to be stable for various metal ions, detergents and organic solvents. These salt-tolerant and chemical-resistant properties suggest that the enzyme esterase EstA1 is both academically and industrially useful.
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