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Our results demonstrated that short ELP[I] n tags with low T t were useful in protein expression and purification, and poly-N linker played the key role in producing bioactive proteins without the need to remove the ELPs. © King Abdulaziz City for Science and Technology 2020.In this report, activity and stability of horseradish peroxidase (HRP) entrapped in polyacrylamide gel in the presence of proline (HEP) are compared with that of enzyme entrapped in absence of proline (HE). Within polyacrylamide (8%) beads, 80% entrapment yield for peroxidase was observed in the presence as well as absence of proline. The HEP (1.5 M proline) showed 170% higher activity compared to HE. HEP also showed significant increase in K M, V max and K cat. At 8th cycle of use, HEP retained 40% of its activity, whereas HE retained only 10% of activity. selleck compound In addition, in comparison with HE, HEP showed increased storage stability and thermo-stability. HEP showed higher activity compared to HE over an extensive range of pH (4-8), temperature (30-80 °C) and inhibitors such as NaN3, Cd2+ and Pb2+. Our results suggest that peroxidase entrapment in polyacrylamide gel in the presence of proline can be a useful approach for increasing activity and stability of horseradish peroxidase. © King Abdulaziz City for Science and Technology 2020.Biofilm formation of a nitrogen-fixing cyanobacterium Anabaena torulosa with a beneficial fungus Trichoderma viride (An-Tr) was examined under laboratory conditions. A gradual enhancement in growth over A. torulosa alone was recorded in the biofilm, with 15-20% higher values in nitrogen fixation, IAA and exopolysaccharide production illustrating the synergism among the partners in the biofilm. To investigate the role of such biofilms in priming seed attributes, mesocosm studies using primed seeds of two maize inbred lines (V6, V7) were undertaken. Beneficial effects of biofilm (An-Tr) were recorded, as compared to uninoculated treatment and cyanobacterial consortium (Anabaena-Nostoc; BF 1-4) at both stages (7 and 21 DAS, days after sowing) with a significant increase of more than 20% in seedling attributes, along with 5-15% increment in seed enzyme activities. More than three- to fivefold higher values in nitrogen fixation and C-N mobilizing enzyme activities, and significant increases in leaf chlorophyll, proteins and PEP carboxylase activity were observed with V7-An-Tr biofilm. Cyanobacterial inoculation brought about distinct changes in the soil phospholipid fatty acid profiles (PLFA); particularly, significant changes in those representing eukaryotes and anaerobic bacteria. Principal component analyses illustrated the significant role of dehydrogenase activity and microbial biomass carbon and distinct elicited effects on soil microbial communities, as evidenced by the PLFA. This investigation highlighted the promise of cyanobacteria as valuable priming options to improve mobilization of nutrients at seed stage, modulating the abundance and activities of various soil microbial communities, thereby, enhanced plant growth and vigour of maize plants. © King Abdulaziz City for Science and Technology 2020.The desulfurization of fuel is currently enforced to meet environmental legislation and prevent pollution. The use of specific biodesulfurizing microbes with a unique 4S pathway allows the desulfurization without compromising the quality of fuel. These specific microbes can be screened by the detection of 2-hydroxybiphenol (2-HBP) in desulfurizing mixture of dibenzothiophene (DBT). At present, colorimetric Gibb's assay is the most commonly employed screening method which requires a specific reagent, i.e., 2,6-dichloroquninone-4-chloramide. In the present study, a novel and simple spectrophotometric method was developed for the detection of 2-HBP for screening purpose based on dual wavelength method. The developed method facilitates the simultaneous analysis of DBT desulfurization and 2-HBP production in a sample by merely measuring the absorbance differences at two specified wavelengths, i.e., ΔA (λ 320-λ 247) for DBT and ΔA (λ 286-λ 324) for 2-HBP. The developed method was used to screen 57 microbes and two specific desulfurizing microbes Bacillus flexus MS-5 and Bacillus cereus BR-31 were selected based on 2-HBP production. The outcomes of developed method were validated by HPLC analysis. The strains MS-5 and BR-31 were employed in biodesulfurization and resulted in 54.88 ± 1.12% and 55.72 ± 1.32% desulfurization of 1.0 mM DBT, respectively. The developed method for screening of specific desulfurizing microbes does not require any specific reagent or sophisticated instrument in spite of being quick and reliable. The microbes selected by developed method exhibited excellent potential for biodesulfurization of fuel. © King Abdulaziz City for Science and Technology 2020.The anti-amyloidogenic potential and cyclooxygenase anti-inflammatory activity of Lasianthus trichophlebus extracts were evaluated. The MeOH extract (LTM) and chloroform extract (LTC) exhibited significant cytotoxic inhibition against the neuroblastoma SH-SY5Y cell with an IC50 of 17.52 μg/mL and 12.28 μg/mL, respectively. Thioflavin T assay indicated the LTC extract inhibition (70.56% at 50 μg/mL) to be statistically comparable (p less then 0.05) to the positive control. Cyclooxygenase inhibition against COX-1 and COX-2 enzymes gave IC50 values for the LTM extract to be 18.20 and 29.60 µg/mL, respectively; while, the LTC extract showed 4.11 and 2.78 µg/mL, respectively. LC-MS of the LTM extract identified 22 putative compounds, which may prove to be pharmacologically relevant. This study has provided potential insights into the utilization of L. trichophlebus to develop safer plant-based agents for anti-inflammatory or neurodegenerative diseases. © King Abdulaziz City for Science and Technology 2020.Streptomyces toxytricini produces bioactive metabolite recognized as lipstatin and its intermediate orlistat. The main focus of this study is to enhance lipstatin production by strain improvement and precursor feeding. In this study, strain improvement to enhance the production of lipstatin was carried out by different doses (50, 100, 150, 200, and 250 Gy) of gamma radiation and precursors (Linoleic acid, Oleic acid, and l-Leucine). Screening showed that the highest yield of lipstatin (4.58 mg/g) was produced by mutant designated as SRN 7. The production of lipstatin (5.011 mg/g) increased significantly when the medium was supplemented with ratio 11.5 (linoleic acid + oleic acid). The addition of 1.5% l-Leucine leads to further increment in the production of lipstatin (5.765 mg/g). The addition of 10% soy flour in the culture medium resulted in the maximum production of lipstatin to 5.886 mg/g. © King Abdulaziz City for Science and Technology 2020.Carotenoids are group of colored terpenoids with antioxidant properties and widespread in nature including in microorganisms. Lactobacillus plantarum subsp. plantarum KCCP11226 was previously isolated from kimchi, while exhibiting the production of 4,4'-diaponeurosporene as a C30 carotenoid. In this study, full genome sequencing of the strain KCCP11226 was performed. Genome analysis revealed that the dehydrosqualene synthase (crtM) and dehydrosqualene desaturase (crtN) genes, which are major genes for biosynthesis of 4,4'-diaponeurosporene, were shown to act as an operon in most L. plantarum strains, but they were uncommon in other Lactobacillus species. In vitro experiments revealed that the production of 4,4'-diaponeurosporene was greatly increased by oxidative stress. In this situation, mRNA expressions of crtN and crtM were also significantly increased. In conclusion, genome analysis of L. plantarum subsp. plantarum KCCP11226 suggested the presence of a well-conserved C30 carotenoid biosynthetic pathway that includes the crtM-crtN operon. link2 The genomic information on L. plantarum subsp. plantarum KCCP11226 could further elucidate the functions of genes involved in isoprenoid biosynthetic pathway, especially in C30 carotenoid biosynthesis. © King Abdulaziz City for Science and Technology 2020.Statins, the drugs for the treatment of dyslipidemia, have been suggested to impact insulin sensitivity, resulting in pancreatic β-cell dysfunction, and consequently, lead to new onset of diabetes. Taking this as a clue, the present study was designed to evaluate the protective effect of sesamol (a known antioxidant, antidiabetic and antidyslipidemic agent) against the diabetogenic potential of simvastatin. The toxic effects of simvastatin and sesamol on MIN6 insulinoma (Mouse pancreatic β cells) cells were evaluated separately by MTT assay. The protective effect of sesamol was evaluated at the IC50 value of simvastatin at doses ranging from 7.8 to 62.5 micromolar (µM). Further, the reversal of the impact of simvastatin on cell cycle and mitochondrial membrane potential by sesamol pretreatment was studied. The IC50 for simvastatin and sesamol were found to be 70.05 ± 2.34 μM and 2134 ± 8.41 μM, respectively, after 48 h and 72 h of incubation. Sesamol pretreatment protected the MIN6 cells from simvastatin toxicity (70 µM) in a dose-dependent manner from 7.8 to 31.25 µM. Simvastatin induced cell cycle arrest in G0/G1 phase. However, when cells were preincubated with sesamol for 24 h, a reversal in the cell cycle arrest was observed in simvastatin-treated cells (G0/G1). Pretreatment with sesamol also reduced the mitochondrial membrane potential loss compared to simvastatin treatment alone. These in vitro findings indicate that sesamol has a protective effect against simvastatin-induced toxicity on the pancreatic beta cells. © King Abdulaziz City for Science and Technology 2020.In the current study, the production of novel glutaminase free l-asparaginase from a new microbial source (Pseudomonas resinovorans IGS-131) is reported. Optimization of l-asparaginase production using conventional and statistical optimization techniques resulted in an enzyme yield of 37.63 IU/mL, which was 3.45-fold higher than the initial enzyme activity (i.e., 10.91 IU/mL). l-Asparaginase production from P. resinovorans IGS-131 was successfully carried out at the bioreactor level and investigations on the effect of agitation rates showed a maximum asparaginase yield of 38.88 IU/mL after 24 h fermentation at 400 rpm. The l-asparaginase gene from this source, showing 78% identity with a reported sequence in GenBank, was expressed in Escherichia coli rosetta DE3. The molecular weight of the recombinant protein was determined as 35.6 kDa. Downstream processing of recombinant l-asparaginase resulted in a purified protein concentration of 62.53 mg/L, which showed good free radical scavenging activity of 62%. The current findings provide promising results for a process of l-asparaginase production from P. resinovorans IGS-131. link3 Furthermore, the recombinant production of this enzyme could help in avoiding the complexity of down streaming processes associated with the purification of this enzyme from wild-type organisms. © King Abdulaziz City for Science and Technology 2020.Osteosarcoma (OS) is a rare aggressive bone, presenting low patient survival rate, high metastasis and relapse occurrence, mostly due to multi-drug resistant cells. To surpass that, the use of nanomedicine for the targeted delivery of genetic material, drugs or both have been extensively researched. In this review, we address the current situation of the disorder and some gene therapy options in the nanomedicine field that have been investigated. Among them, polymeric micelles (PM) are an advantageous therapeutic alternative highly explored for OS, as they allow for the targeted transportation of poorly water-soluble drugs to cancer cells. In addition, micelleplexes are PMs with cationic properties with promising features, such as the possibility for a dual therapy, which have made them an attractive research subject. The aim of this review article is to elucidate the application of a micelleplex formulation encapsulating the underexpressed miRNA145 to achieve an active targeting to OS cells and overcome multi-drug resistance, as a new and viable therapeutic strategy.
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