Partial Characterization Research Articles

Venom characterization of Venezuelan scorpion Tityus caripitensis.

Tityus caripitensis is an endemic scorpion species found in the northeastern region from Venezuela, being responsible for sting accidents in this area. This study describes for the first time a biological, biochemical and electrophysiological partial characterization of Tityus caripitensis scorpion venom. The venom is toxic to mice with a LD50 of 20.2 μg/gr mouse. Animals experimentally envenomed with Tityus caripitensis venom gradually manifested clinical signs in response to sublethal doses. SDS-PAGE of the venom resulted in 7 fractions ranging in size from ∼3.5 to ≥ 38 kDa. The 6-8 kDa proteins could correspond to neurotoxins. In addition, the components of Tityus caripitensis venom were similar to those obtained in the electrophoretic profile of Tityus discrepans. The commercial anti- Tityus discrepans IgG showed reactivity against Tityus caripitensis venom. Tityus caripitensis venom could induce hematological changes such as hyperamylasemia and hyperglycemia. The venom modified voltage dependent Na+v1.4 channels and blocked Kv+ channels. Although Tityus caripitensis venom is less toxic than Tityus discrepans, they share molecular and antigenic components. This aspect should be considered in the application of antivenom treatment.

Optimizing Exopolysaccharide Production by Bacillus subtilis Using Spoiled Fig and Grape.

The study aimed to enhance exopolysaccharides (EPSs) production by the bacterial strain Bacillus subtilis ES (OR501464) isolated from sugar cane juice. Spoiled grape and fig extract were utilized as cost-effective substrates for EPS synthesis by B. subtilis ES (OR501464), and the impact of nutritional factors on EPS synthesis was assessed. Among nineteen bacterial isolates evaluated for EPS production, the isolate with the highest EPS yield was identified through a combination of phenotypic and genotypic analyses. The optimization process revealed that the highest EPS yield of 4.7g/L was achieved in a production medium containing 4% sucrose, 0.1% NaNO3, 0.002% Na2SO4, and 2% NaCl at 30°C and pH 9. Additionally, the study explored EPS generation by B. subtilis ES (OR501464) using spoiled grape and fig extract as substrates. The addition of 2% NaCl to spoiled grape extract increased EPS production to 4.357mg/mL compared to 3.977mg/mL with grape alone. However, 2% NaCl did not enhance EPSs production in fig waste. Supplementing spoiled fig or grape extract with 0.2g/L Na2SO4 and 1g/L NaNO3 increased EPS production by B. subtilis ES (OR501464). The EPS was analyzed using GC-MS and FTIR spectroscopy for partial characterization. The study found that spoiled figs and grapes can be used as effective substrates for EPS production. The highest yield was achieved by adding 0.2g/L Na2SO4 and 1g/L NaNO3. This study highlights the use of spoiled figs and grapes to produce valuable biopolymers, promoting sustainable and eco-friendly bioprocessing technologies.

Partial Near-Field Antenna Characterization

Partial Near-Field Antenna Characterization

Expression, Purification and Partial Characterization of Recombinant S-Adenosyl-L-Homocysteine Hydrolase from Banana

Expression, Purification and Partial Characterization of Recombinant S-Adenosyl-L-Homocysteine Hydrolase from Banana

Purification and partial characterization of a new melibiose-specific lectin from Bauhinia catingae Harms

Lectins are ubiquitous proteins that selectively bind to carbohydrates, serving as vital models for understanding protein-carbohydrate interactions. While extensively distributed across various life forms, plant lectins, especially from the Leguminosae family, have garnered significant attention. However, limited research exists on lectins from the Caesalpinioideae subfamily, suggesting a source of untapped biotechnological potential. This underscores the imperative for further exploration, particularly in isolating lectins from the Bauhinia genus, which remains relatively understudied, despite harboring lectins with diverse characteristics and promising biotechnological activities. In this study, a novel lectin extracted from Bauhinia catingae Harms seeds (BCL) was isolated in three chromatographic steps. BCL exhibited affinity for galactose and derivatives, akin to other Bauhinia lectins, with SDS-PAGE confirming its molecular weight around 30 kDa. Notably, BCL demonstrated stability across temperature and pH ranges and lacked metalloprotein characteristics. Electrospray ionization mass spectrometry revealed a partial sequence covering 81 % of the total protein sequence with nearly 80 % identity to Bauhinia forficata. Structural analysis suggested a β-sheet-rich secondary structure similar to that of other lectins. Further structural elucidation of BCL is essential to unveil its full potential and applications.

Partial Characterisation of Three Isolates of Potato virus Y

Partial Characterisation of Three Isolates of Potato virus Y

Optimization of the production of a new milk clotting enzyme by Aspergillus flavus thermophile on agro-industrial residues medium and characterization of the enzyme

This study focused on the optimization of the production of a milk clotting protease by Aspergillus flavus, isolated from a thermal station. The fermentation was carried out on wheat bran as a medium, followed by a partial characterization of the enzyme. The fungal isolate was screened for protease production based on clear zone hydrolysis on skim milk agar, and then it was identified as belonging to the genus Aspergillus based on its morphological characteristics. This identification was confirmed by molecular analyzes with 100% similarity with the A. flavus strain. After 7 days of fermentation, the crude extract is recovered by simple filtration. The production and optimization of the milk coagulant protease using wheat bran and agro-industrial waste as a medium was carried out using a statistical method of experimental design based on the Plackett and Burman matrices. The results showed that the optimal fermentation medium consisted of wheat bran (4% w/w), incubated at a temperature of 30 °C, with a pH of 5.0 after 6 days of fermentation. The enzyme was partially purified by ammonium sulfate precipitation, yielding an estimated proteolytic activity of 2,985 U. The partially purified protease exhibits optimal activity at pH 5 with hemoglobin as a substrate and an optimal temperature of 50 °C. The Lineweaver-Burk plot showed a KM value of 4.83 g/L and a Vm of 1,560 U. This enzyme causes very rapid coagulation of fresh cow’s milk with an optimum at 45 °C, a pH 7 and a concentration of 50 mM of calcium.

Exopolysaccharide from Leuconostoc mesenteroides XR1: Yield optimization, partial characterization and properties

The production conditions of exopolysaccharide (EPS) from Leuconostoc mesenteroides XR1 were optimized by response surface methodology (RSM). Maximum EPS yield was 56.59 ± 0.51 g/L under fermentation conditions with 2.6 g/L ammonium citrate, initial pH 6.5 and temperature 23 °C, which was 6.21-fold greater than the EPS yield before optimization. Characterization of the chain conformation using Congo red test and circular dichroism (CD) showed that EPS exhibited a random coil structure in aqueous solution. The CD results revealed that the EPS concentration altered its hydrogen-bond interactions and chirality, but did not change its chain conformation. The average polydispersity index (PDI) of the EPS solution was only 27.16 %, indicating that it was uniformly distributed in the aqueous solution with high stability. The degradation temperature of EPS was 253.11 °C, indicating high thermal stability. EPS possessed the ability to scavenge activities of free radicals and was protective against oxidative stress-induced plasmid DNA damage. In addition, stable hydrogels could be formed at EPS concentrations above 5 % (w/v). These results collectively showed that EPS can be used commercially as an antioxidant and drug delivery carrier.

Isolation and Partial Characterization of 11-Octadecenoic Acid Methyl Ester and Bis (2-ethylhexyl) Phthalate from Celtis integrifolia Lam

Introduction: Celtis integrifolia Lam is a traditional medicinal plant used as a remedy for the cure of diarrhea, measles, bleeding, ecbolic, sore throat and as an antinociceptive agent. Aim: The study was aimed at the isolation and partial characterization of phytocompounds from Celtis integrifolia crude extract. Methodology: The stembark sample was powdered and extracted using maceration technique with methanol. The extract was filtered using cotton wool and clarified using Whatmann No.1 filter paper. The filtrate was concentrated on a rotavapor at 45oC. The crude extract was then reconstituted in water and partitioned successively with n-hexane, ethyl acetate, and n-butanol. The ethyl acetate and the n-hexane fractions were concentrated and subjected to purification on column chromatography. Results: Gradient elution of extract fractions on open-column chromatography yielded two compounds coded CiL1 and CiL2 from the n-hexane and ethyl acetate fractions respectively. The isolated compounds were characterized using spectroscopic data from Fourier transform infrared (FT-IR) spectroscopy, proton nuclear magnetic resonance (1H NMR) spectroscopy, gas chromatography-mass spectrometry (GC-MS) and literature database. Compound CiL1 with Rf = 0.43 in hexane: Ethyl acetate (9:1) was identified as 11-Octadecenoic acid methyl ester while Compound CiL2 isolated from ethyl acetate fraction as a yellow substance with Rf = 0.38 in hexane: ethyl acetate (8:2) was identified as bis (2-ethylhexyl) phthalate. Conclusion: The study showed the presence of 11-octadecenoic acid methyl ester and bis (2-ethylhexyl) phthalate in the stembark extract of Celtis integrifolia Lam. These two phytocompounds are reported for the first time from Celtis integrifolia Lam based on available literature. Since the presence of phytochemical substances is believed to be the basis of any observed pharmacological activity, the study therefore lends credence to the traditional medicinal uses of Celtis integrifolia Lam.

Molecular characterization and gene expression of pattern recognition receptors in brown-marbled grouper (Epinephelus fuscoguttatus) fingerlings responding to vibriosis infection

The pathogen recognition system involves receptors and genes that play a crucial role in activating innate immune response in brown-marbled grouper (Epinephelus fuscoguttatus) as a control agent against various infections including vibriosis. Here, we report the molecular cloning of partial open reading frames, sequences characterization, and expression profiles of Pattern Recognition Receptors (PRRs) in brown-marbled grouper. The PRRs, namely pglyrp5, tlr5, ctlD, and ctlE in brown-marbled grouper, possess conserved domains and showed shared evolutionary relationships with other fishes, humans, mammals, birds, reptilians, amphibians, and insects. In infection experiments, up to 50% mortality was found in brown-marbled grouper fingerlings infected with Vibrio alginolyticus compared to 27% mortality infected Vibrio parahaemolyticus and 100% survival of control groups. It is also demonstrated that all four PRRs had higher expression in samples infected with V. alginolyticus compared to V. parahaemolyticus. This PRRs gene expression analysis revealed that all four PRRs expressed rapidly at 4-h post-inoculation even though the Vibrio count was only detected earliest at 12-h post-inoculation in samples. The highest expression recorded was from V. alginolyticus inoculated fish spleen with up to 73-fold change for pglyrp5 gene, followed by 14 to 38-fold expression for the same treatment in spleen, head kidney, and blood samples for other PRRs, namely tlr5, ctlD, and ctlE genes. Meanwhile less than a 10% increase in expression of all four genes was detected in spleen, head kidney, and blood samples inoculated with V. parahaemolyticus. These findings indicated that pglyrp5, tlr5, ctlD, and ctlE play important roles in the early immune response to vibriosis infected, brown-marbled grouper fingerlings.

Partial Discharge Characterization in a Defect Subjected to HVDC Cable Operating Conditions

Partial Discharge Characterization in a Defect Subjected to HVDC Cable Operating Conditions

Optimization and Characterization of an Ultra-Thermostable, Acidophilic, Cellulase-Free Xylanase from a New Obligate Thermophilic Geobacillus thermoleovorans AKNT10 and its Application in Saccharification of Wheat Bran.

Microbial xylanases are enzymes of great importance due to their wide industrial applications, especially in the degradation of lignocellulosic biomass into fermentable sugars. This study aimed to describe the production optimization and partial characterization of an ultra-thermostable, acidophilic, cellulase-free xylanase from an obligate thermophilic eubacterium Geobacillus thermoleovorans strain-AKNT10 (Ac.No. LT158229) isolated from a hot-spring of Puga Valley located at an altitude of 4419m in Ladakh, India. The optimization of cultural conditions improved enzyme yield by 10.49-fold under submerged fermentation. The addition of 1% (w/v) xylose induced the enzyme synthesis by ~ 165 and 371% when supplemented in the fermentation medium containing wheat bran (WB) 1 and 3%, respectively. The supplementation of sucrose reduced the xylanase production by ~ 25%. Results of partial characterization exhibited that xylanase was optimally active at pH 6.0 and 100°C. Enzyme retained > 75%, > 83%, and > 84% of activity at 4°C for 28days, 100°C for 60min, and pHs 3-8 for 60min, respectively. An outstanding property of AKNT10-xylanase, was the retention of > 71% residual activity at extreme conditions (121°C and 15 psi pressure) for 15min. Enzymatic saccharification showed that enzyme was also capable to liberate maximum reducing sugars within 4-8h under optimized conditions thus it could be a potential candidate for the bioconversion of lignocellulosic biomass as well as other industrial purposes. To the best of our knowledge, this is the first report on such an ultra-thermo-pressure-tolerant xylanase optimally active at pH6 and 100°C from the genus Geobacillus.

Unveiling the genomic landscape: Whole genome draft and phenotypic profiling of Oceanobacillus sp. SE10311 isolated from Mount Makiling, Philippines

The genus Oceanobacillus remains one of the least understood groups of Bacillaceae due to limited information and a scarcity of accessions. With the full genome sequencing and partial phenotypic characterization of the cellulase and brown-pigment-producing Oceanobacillus sp. SE10311, an isolate from Mount Makiling, this study provided taxonomic, genomic, and functional insights into this bacterial discovery. The results showed that Oceanobacillus sp. SE10311 draft genome contained 3.946 million base pairs assembled from 137 contigs and 39.30 percent G+C. The whole genome phylogeny and pangenome analyses revealed the identity of Oceanobacillus sp. SE10311 as an isolate of Oceanobacillus indicireducens with greater than 98% genome similarity to the type strain of the species. The confirmed presence of the associated genes from the genome annotation rationalized the pigmentation and cellulose degradation of Oceanobacillus sp. SE10311. This research milestone presents Oceanobacillus sp. SE10311, with distinct chromogenic and cellulolytic properties as a completed whole genome-sequenced bacterial accession from the Philippines.

Partial purification and characterization of bacteriocin from novel Brevibacillus borstelensis sp. isolated from Donkey’s lactation

Partial purification and characterization of bacteriocin from novel Brevibacillus borstelensis sp. isolated from Donkey’s lactation

Isolation and Partial Characterization of Novel, Structurally Uniform (Hfq6)n≥8 Assemblies Carrying Accessory Transcription and Translation Factors.

In growing E. coli cells, the transcription-translation complexes (TTCs) form characteristic foci; however, the exact molecular composition of these superstructures is not known with certainty. Herein, we report that, during our recently developed "fast" procedures for purification of E. coli RNA polymerase (RP), a fraction of the RP's α/RpoA subunits is displaced from the core RP complexes and copurifies with multiprotein superstructures carrying the nucleic acid-binding protein Hfq and the ribosomal protein S6. We show that the main components of these large multiprotein assemblies are fixed protein copy-number (Hfq6)n≥8 complexes; these complexes have a high level of structural uniformity and are distinctly unlike the previously described (Hfq6)n "head-to-tail" polymers. We describe purification of these novel, structurally uniform (Hfq6)n≥8 complexes to near homogeneity and show that they also contain small nonprotein molecules and accessory S6. We demonstrate that Hfq, S6, and RP have similar solubility profiles and present evidence pointing to a role of the Hfq C-termini in superstructure formation. Taken together, our data offer new insights into the composition of the macromolecular assemblies likely acting as scaffolds for transcription complexes and ribosomes during bacterial cells' active growth.

Production and partial characterization of endoglucanase by Thermothelomyces heterothallicus PA2S4T and its application in biopolishing of denim jeans

Production and partial characterization of endoglucanase by Thermothelomyces heterothallicus PA2S4T and its application in biopolishing of denim jeans

Antibacterial activity and partial characterization of antibacterial agent from combretum glutinosum leaf extracts against microbes isolated from dental caries

Antibacterial activity and partial characterization of antibacterial agent from combretum glutinosum leaf extracts against microbes isolated from dental caries

Production, Extraction and Partial Characterization of Natural Pigments from Chryseobacterium sp. kr6 Growing on Feather Meal Biomass

Obtaining natural pigments from microorganisms is an alternative with high potential for biotechnological application. The use of agro-industrial wastes as substrate for cultivations enables a reduction of the production cost and may add value to potentially polluting byproducts. In this work, the extraction of pigments produced by the bacterium Chryseobacterium sp. strain kr6 was evaluated, employing feather meal as the sole carbon source for bacterial growth. The maximum production of the yellow pigments was observed for cultivation at 30 °C, during 48 h, with 5 g/L feather meal. The pigment extraction from the bacterial biomass was performed with the aid of physical methods and the testing of different organic solvents. The conditions that provided better extraction were using ultrasound with acetone as the solvent, reaching a yield of 180 μg/g biomass after optimization. The pigment was partially characterized via UV-visible, FTIR and mass spectroscopy and CIELAB color parameters, suggesting the presence of molecules belonging to the flexirubin group (aryl polyenes). The antioxidant capacity of the pigment was confirmed via the scavenging of DPPH radical and thiobarbituric acid reactive substances (TBARS) methodologies. Moreover, the pigment extract showed antimicrobial activity against Staphylococcus aureus and Enterococcus faecalis.

Establishment and partial characterisation of a new cell line derived from adult tissues of the tsetse fly Glossina morsitans morsitans

BackgroundInsect cell lines play a vital role in many aspects of research on disease vectors and agricultural pests. The tsetse fly Glossina morsitans morsitans is an important vector of salivarian trypanosomes in sub-Saharan Africa and, as such, is a major constraint on human health and agricultural development in the region.MethodsHere, we report establishment and partial characterisation of a cell line, GMA/LULS61, derived from tissues of adult female G. m. morsitans. GMA/LULS61 cells, grown at 28 °C in L-15 (Leibovitz) medium supplemented with foetal bovine serum and tryptose phosphate broth, have been taken through 23 passages to date and can be split 1:1 at 2-week intervals. Karyotyping at passage 17 revealed a predominantly haploid chromosome complement. Species origin and absence of contaminating bacteria were confirmed by PCR amplification and sequencing of fragments of the COI gene and pan-bacterial 16S rRNA gene respectively. However, PCR screening of RNA extracted from GMA/LULS61 cells confirmed presence of the recently described Glossina morsitans morsitans iflavirus and Glossina morsitans morsitans negevirus, but absence of Glossina pallipides salivary gland hypertrophy virus. GMA/LULS61 cells supported infection and growth of 6/7 different insect-derived strains of the intracellular bacterial symbiont Wolbachia.ConclusionsThe GMA/LULS61 cell line has potential for application in a variety of studies investigating the biology of G. m. morsitans and its associated pathogenic and symbiotic microorganisms.Graphical

Preliminary Purification and Partial Characterization of a Functional Bacteriocin of Lacticaseibacillus paracasei Zhang and Mining for its Gene Cluster.

Bacteriocins produced by lactic acid bacteria (LAB) have good potential for use as food biopreservatives. Lacticaseibacillus paracasei Zhang (L. paracasei Zhang) is both a food use and a probiotic bacterium. This study aimed to purify and preliminary characterize the active antibacterial metabolite of L. paracasei Zhang. The cell-free supernatant of L. paracasei Zhang was collected and purified by ultrafiltration and gel filtration chromatography. The 1-3kDa active fraction could inhibit the growth of Staphylococcus aureus but not Escherichia coli. Further antibacterial activity assays revealed its capacity to suppress various foodborne and human opportunistic pathogens (including Staphylococcus aureus, Pseudomonas fluorescens, Pseudomonas aeruginosa, Listeria monocytogenes, and Bacillus cereus), but not fungi. The antibacterial activity showed good tolerance to heat (40 to 100°C), acid-base (pH 2-3 and pH 6-10), and digestions by a number of industrial and animal/human enzymes (such as trypsin, pepsin, α-amylase, and protease K, except papain); these desired properties make it a suitable biopreservative to be used in harsh and complex industrial production processes. The high papain sensitivity suggested a proteinaceous/peptide nature of the bioactivity. Moreover, our genomic data mining for bacteriocin through BAGEL4 revealed an area of interest encoding a complete set of putative genes required for bacteriocin production. In conclusion, our study showed that L. paracasei Zhang can produce extracellular functional antibacterial metabolite, likely a class II bacteriocin. Our preliminary extraction and characterization of the active metabolite demonstrated that it has good potential to be used as a biopreservative or an agent for suppressing gastrointestinal infections.