Hydrolysis Test Research Articles

The Gut Bacteria of Gampsocleis gratiosa (Orthoptera: Tettigoniidae) by Culturomics

Gampsocleis gratiosa Brunner von Wattenwyl, 1862, is a type of omnivorous chirping insect with a long history of artificial breeding. It has high economic value and is also an excellent orthopteran model organism. In this study, 12 types of culture media combined with 16S rRNA sequencing were employed to isolate 838 bacterial strains from the gut of G. gratiosa. After sequence comparison, a total of 98 species of bacteria were identified, belonging to 3 phyla, 5 classes, 11 orders, 20 families, and 45 genera. Firmicutes and Proteobacteria accounted for the majority (92.86%). At the order level, Enterobacteriaceae, Bacillales, and Lactobacillales predominated (79.59%). At the genus level, Klebsiella (11.22%) and Enterococcus (7.14%) predominated. This study also enumerated the strain morphological, physiological and biochemical properties of 98 species of bacteria, including colony morphology, Gram staining, bacterial motility test, temperature gradient growth, pH gradient growth, citrate utilization test, temperature oxidase test, contact enzyme test, methyl red test, V-P test, indole test, gelatin liquefaction test, nitrate reduction test, hydrogen sulfide test, starch hydrolysis test, cellulose decomposition test, esterase (corn oil) test and antibiotic susceptibility testing. Additionally, 16 antibiotics were utilized to test the bacterial susceptibility of the strains. This study explored the types and community structure of some culturable microorganisms in the intestinal tract of G. gratiosa and recorded their physiological characteristics. These data reflect the physiological functions of the intestinal microorganisms of G. gratiosa and provide support for subsequent research on the interaction mechanism between microorganisms and their hosts.

Isolation and Characterization of Lactic Acid Bacteria: A Strategy to Produce Inoculum for Forage Ensiling

Purpose: Sri Lankan dairy sector faces challenges due to seasonal variability and suboptimal quality of forage for dairy cattle feeding. While silage offers year-round availability of forage, concerns arise regarding potential quality degradation during forage ensiling. Microbial inoculants are utilized to expedite the ensiling and minimize quality deterioration. This study involves the isolation and characterization of lactic acid bacteria (LAB) from silage, with the aim of identifying potential candidates to be used as silage inoculants.Research Method: Lactobacillus were isolated from 10 silage samples of each forage type: sorghum, maize, and guinea grass. They were characterized using the 16S rRNA gene sequencing procedure. The anaerobic sugar fermentation ability of the isolated LAB was evaluated by inoculating them into sterilized sugar solutions, and measuring the titratable acidity (TA) and pH value over time.Findings: The isolates were identified to be non-motile, non-spore-forming and Gram-positive rods. Further, they tested negative for catalase, triple sugar iron, citrate, urease, indole, gelatine hydrolysis, motility, and oxidase tests. The LAB isolated from guinea grass, sorghum, and maize corresponded to Lactobacillus oris, Lactobacillus rhamnosus, and Lactobacillus plantarum, respectively. The TA in all inoculated solutions was higher compared to the control (0.55-0.66% vs. 0.48%), with L. plantarum showing the highest TA (0.66%) at 30 hours incubation. Additionally, all sugar solutions treated with LAB isolates exhibited a lower pH value at 30 hours compared to the control (4.70-5.07 vs. 5.75). Notably, L. plantarum demonstrated the most significant pH value reduction at 18 hours (5.85 vs. 6.29-6.64).Value: Lactobacillus oris, L. rhamnosus, and L. plantarum, isolated from guinea grass, sorghum, and maize silage, respectively, have the potential to be developed as inoculants for ensiling forage.

Enhanced hydrolysis of MgH₂ by transition metal oxides and NaH: Performance, mechanisms, and optimization

MgH₂ is recognized as a promising material for hydrogen production due to its high hydrogen density, abundant availability, and non-polluting hydrolysis by-products, making it a viable hydrogen source for fuel cells. This study, for the first time, investigates the hydrogen production performance of the MgH₂-MOx-NaH (M = Ti, Zr, Mo, Fe, V, Bi) system, which exhibits remarkable hydrolysis properties in aqueous solutions. A series of MgH₂-MOx-NaH composite powders were synthesized via mechanical ball milling. The results revealed that V₂O₅-NaH and Bi₂O₃-NaH significantly enhanced the hydrolysis performance of MgH₂ for hydrogen production. Specifically, Bi₂O₃-NaH and V₂O₅-NaH effectively refined MgH₂ particles and mitigated agglomeration during the milling process. Hydrolysis tests demonstrated that the conversion rate and the hydrogen generation rate (mHGR) of composites initially increased with ball milling time and then decreased. The maximum hydrolysis hydrogen production yield and conversion rate for MgH₂-Bi₂O₃-NaH and MgH₂-V₂O₅-NaH composites occurred at a ball milling duration of 10 hours, and reached up to 1354.1 mL g⁻¹ (88.5 %) and 1273.5 mL g⁻¹ (81.8 %), respectively, within 8 minutes. Kinetic tests of hydrolysis hydrogen production showed that the reaction rate and hydrogen conversion increased with elevated hydrolysis temperatures. The activation energies for the hydrolysis of MgH₂-MOx-NaH (M = Ti, Zr, Mo, Fe, V, Bi) were calculated from the hydrolysis-hydrogen production curves using the Arrhenius formula, yielding values of 41.32, 37.66, 36.40, 34.60, 30.80, and 27.17 kJ mol⁻¹, respectively. The lower activation energies for hydrolysis indicate improved reaction kinetics, suggesting that the incorporation of transition metal oxides and sodium hydride effectively enhances both the hydrolytic conversion and reaction rate of MgH₂.

Characterisation of ACE2-like Enzyme from Bacillus cereus sp. as an Alternative Treatment for COVID-19 Patients

SARS-CoV-2 infection can lead to downregulation of ACE2, raising pro-inflammatory Angiotensin II levels. Recombinant human ACE2 protein therapy can restore homeostasis but is costly. An alternative is an ACE2-like enzyme with similar effects. The previous research identified a carboxypeptidase protein from Bacillus cereus sp. (rBceCP) as a potential ACE2-like enzyme, but it has not been characterized well. This study characterizes, expresses, and tests the activity of the rBceCP). rBceCP structure and properties were predicted using Prabi and Expasy. The in vitro approach included protein expression optimization, hydrolysis tests, and inhibition tests. In silico analysis revealed the protein is a homodimer with 53.66% α-helix and a molecular weight of 58.99 kDa. The protein is stable, hydrophilic, and has an isoelectric point at pH 4.93, indicating it can be expressed using the E. coli system. Expression of rBceCP showed no significant differences across IPTG concentrations (p-value >0.05). The protein hydrolysis activity of rBceCP was similar to the control, though purified samples had lower activity than crude samples. Inhibition activity in crude and purified samples showed no significant differences (p-value >0.05) and was higher than the control. Thus, rBceCP has potential as an ACE2-like enzyme and a therapeutic candidate for COVID-19.

Evaluation of the CARBA PAcE test, a colorimetric imipenem hydrolysis test for rapid detection of carbapenemase activity.

There is an urgent need for accurate and fast diagnostic tests to identify carbapenemase-producing bacteria. Here, we evaluated a colorimetric imipenem hydrolysis test, called the CARBA PAcE test, to detect carbapenemase-producing Gram-negative bacteria (GNB). We tested a collection of 270 GNB isolates with a characterized carbapenemase content. Our testing set included 205 carbapenemase-producing, carbapenemase-resistant Enterobacterales (CP CRE) with 40 Klebsiella pneumoniae carbapenemase (KPC), 49 New Delhi metallo beta lactamase (NDM), 49 OXA-48-like, 15 Verona integron-mediated metallo-β-lactamase (VIM), three IMP, 43 IMI, six isolates producing more than one carbapenemase, and 65 non-carbapenemase-producing Enterobacterales (20 ESBL producers, 35 non-carbapenemase-producing, carbapenem-resistant [non-CP CRE], and 10 carbapenem-susceptible Enterobacterales [non-CP, non-CSE, third-generation cephalosporin and carbapenem susceptible]). We compared the performances of the CARBA PAcE test, the qualitative colorimetric β-CARBA test, and the modified CarbaNP test to a gold standard of carbapenemase gene detection by PCR. Specificities of all tests were high: 95.4% (62/65) for CARBA PAcE test, 98.5% (64/65) for β-CARBA test, and 100% (65/65) for the modified CarbaNP test. Sensitivity varied by carbapenemase: all three tests had a sensitivity of 100% for NDM, VIM, and IMP and 97.5% for KPC. Sensitivity to detect IMI was 0% for the CARBA PAcE and β-CARBA tests and 11.6% for the modified CarbaNP test. Sensitivity to detect OXA-48-like was 89.7% for the CARBA PAcE test, 87.7% for the β-CARBA test, and 14.2% for the modified CarbaNP test. Reading the results of the CARBA PAcE assay was difficult. The CARBA PAcE assay is highly sensitive for detecting NDM, VIM, IMP, and KPC, but slightly less sensitive for OXA-48-like. It does not detect IMI. It is highly specific, and its overall diagnostic accuracy is similar to that of β-CARBA. Its operational advantages are rapid turnaround time, ease of use, and long shelf life, but reading of results is subjective.IMPORTANCEWe evaluated the ability of the CARBA PAcE test to detect carbapenemases in 274 Gram-negative isolates with a known carbapenemase content. Specificity was high for all carbapenemases tested (96.9%). Sensitivity was high for KPC, NDM, VIM, and IMP (97.5-100%); but lower for OXA-48-like (89.7%). Activity of IMI could not be detected. Taken together, our results indicate that CARBA PAcE is a useful alternative in regions where NDM and KPC are predominant. The limitations of the test are difficulty in reading results and incompatibility with mSuperCARBA.

Enhanced saccharification levels of corn starch using as a strategy a novel amylolytic complex (AmyHb) from the thermophilic fungus Humicola brevis var. thermoidea in association with commercial enzyme.

Amylases represent a versatile group of catalysts that are used for the saccharification of starch because they can hydrolyze the glycosidic bonds of starch molecules to release glucose, maltose, and short-chain oligosaccharides. The amylolytic complex of the thermophilic filamentous fungus Humicola brevis var. thermoidea (AmyHb) was produced, biochemically characterized, and compared with the commercial amylase Termamyl. In addition, the biotechnological application of AmyHb in starch saccharification was investigated. The highest production was achieved using a wheat bran medium at 50°C for 5-6days in solid-state fermentation (849.6 ± 18.2U·g-1) without the addition of inducers. Optimum amylolytic activity occurred at pH 5.0 at 60°C, and stability was maintained between pH 5.0 and 6.0, with thermal stability at 50-60°C, especially in the presence of Ca2+. These results were superior to those found with Termamyl. Both enzymes were strongly inhibited by Hg2+, Cu2+, and Ag+; however, AmyHb displayed increased activity in the presence of Mn2+ and Na+. In addition, AmyHb showed greater tolerance to a wide range of ethanol concentrations. AmyHb appears to be a complex consisting of glucoamylase and α-amylase, based on its substrate specificity and TLC. The hydrolysis tests on cornstarch flour showed that the cocktail of AmyHb50% + Termamyl50% significantly increased the release of glucose and total reducing sugars (36.6%) when compared to the enzymes alone. AmyHb exhibited promising physicochemical properties and good performance with commercial amylase; therefore, this complex is a biotechnological alternative candidate for the bioprocessing of starch sources.

HPTLC – densitometric method of analysis for estimation of efonidipine hydrochloride and telmisartan used in treatment of hypertension

Precise, accurate, and sensitive high performance thin layer chromatographic method has been developed and validated for simultaneous determination of efonidipine hydrochloride (EFD) and telmisartan (TEL) in a tablet dosage form which is used in treatment of hypertension. The proposed method used aluminum plates pre-coated with silica gel 60 F254 as a stationary phase and n-butanol: toluene: acetic acid (3:7:0.2, v/v/v) as a mobile phase for separation purpose. Compact spots of TEL and EFD were obtained at Rf 0.73 and 0.37, respectively. Densitometric detection was carried out at 299 nm in UV. Linear responses were shown by the detector over the range of 200–1200 ng/band for both drugs. The proposed method was validated as per ICH guidelines. The method was successfully applied for estimation of both drugs in tablet formulations. Forced degradation study was carried and efonidipine was found to be susceptible to base hydrolysis and oxidative stress degradation while TEL was stable in acid-base hydrolysis, oxidative stress, dry heat and photo stability testing conditions. The developed method can be used for the analysis of stability samples. Greenness assessment of developed method was performed using AGREE software. The method was found to be greener having greenness score of 0.69.

Production of high-grade antimony oxide from smelter slag via leaching and hydrolysis process

This study aimed to investigate the recovery of antimony (Sb) from slag generated in an antimony smelting plant using leaching followed by hydrolysis processes. The leaching behaviors of rare earth elements (REEs) were also examined. The physicochemical properties of the slag were determined using various analytical techniques. The slag (4.12 % Sb) was mainly composed of quartz and minor minerals, including microline, magnetite, hedenbergite, and stibiconite. The Sb types in the slag determined by XPS were found to be in the oxide form. The concentrations of REEs (La, Y, Ce, and Nd) in the slag were 169.21 g/t. Preliminary leaching experiment results indicate that (i) HCl was selected rather than other acids due to its high extraction ability on the Sb from the slag, (ii) a sample with a d50 of < 25 µm should be used, (iii) the slurry should be mixed at 300 rpm. In the following leaching tests, the effects of leaching parameters (HCl acid concentration, amount of tartaric acid, solid-to-liquid ratio, reaction temperature, and time) on the extraction rates of Sb, impurities, and REEs were investigated. At the best leaching conditions (HCl: 8 M, amount of tartaric acid: 1 g/L, stirring speed: 300 rpm, reaction temperature: 75 °C, and time: 180 min), the extraction rates of Sb from the slag were determined to be 91.19 %, but the extraction rates of REEs were measured to be ≤ 50 %. The activation energy (Ea) for Sb leaching was found to be 46.75 kJ/mol, indicating that the reaction was governed by the chemically controlled mechanism. In particular, it was understood from the additional experimental results that the leaching procedure should be carried out for 20 h to extract La with an extraction rate of > 90 %. However, the extraction rate of Sb was negligible in extended times. It was determined that using tartaric acid positively affected La’s leaching mechanism, and the required leaching time for La decreased to 180 min from 20 h with the increase of tartaric acid from 1 g/L to 6 g/L. Hydrolysis tests were conducted using the Taguchi approach (L32, 2^1 4^3). The effects of the alkaline type (NH4OH and NaOH), stirring speed (100, 200, 300, and 400 rpm), temperature (50, 60, 70, and 80 °C), and pH (1.5, 2, 2.5, and 3) on the precipitation of Sb from the PLS were investigated. NH4OH was suggested for use in the hydrolysis test to obtain precipitates with higher purities. The product obtained under the optimal conditions comprised 81.43 % Sb, 16.23 % O, and 2.34 % Fe. The product was identified as antimony oxide by XRD.

Novel Biobased Copolymers Based on Poly(butylene succinate) and Cutin: In Situ Synthesis and Structure Properties Investigations.

The present work describes the synthesis of poly(butylene succinate) (PBSu)-cutin copolymers by the two-stage melt polycondensation method, esterification and polycondensation. Cutin was added in four different concentrations, 2.5, 5, 10, and 20 wt%, in respect to succinic acid. The obtained copolymers were studied using a variety of techniques such as Fourier transform infrared spectroscopy (FTIR), X-ray diffraction analysis (XRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), polarized light microscopy (PLM), as well as diffuse reflectance spectroscopy (DRS). A series of results, in agreement between different techniques, revealed the formation of PBSu-cutin interactions, confirming indirectly the successful in situ synthetic route of copolymers. DSC and XRD combined with PLM results provided indications that the crystallization temperature increases with the addition of small amounts of cutin and gradually decreases with increasing concentration. The crystallization process was easier and faster at 2.5%, 5%, and 10% concentrations, whereas at 20%, it was comparable to neat PBSu. The presence of cutin, in general, leads to the facilitated crystallizability of PBSu (direct effect), whereas a moderate drop in the glass transition temperature is recorded, the latter being an indirect effect of cutin via crystallization. The thermal stability improved in the copolymers compared to neat PBSu. Water contact angle measurements confirmed that the addition of cutin decreased the hydrophilicity. The local and segmental relaxation mapping is demonstrated for PBSu/cutin here for the first time. Enzymatic hydrolysis and soil degradation tests showed that, overall, cutin accelerated the decomposition of the polymers. The copolymers may be proven useful in several applications.

Isolation and Molecular Identification of Novel Chicken Feather Degrading Bacteria Bacillus tropicus KRKJVR 5

The isolation and molecular identification of a novel chicken feather-degrading bacterium, Bacillus tropicus KRKJVR 5, were conducted to explore its potential in biotechnological applications. Soil samples collected from various sites in Suryapet district, Telangana, were screened using the enrichment culture method to identify potential keratinolytic producers. A total of ten bacterial isolates were initially identified, out of which five isolates demonstrated significant feather degradation on feather-containing agar media. Among these, Isolate 5 exhibited the highest keratinase activity, completely degrading feathers within 48 hours and producing 8.66 U/ml of keratinase, as determined by the Azocasein assay. Morphological and biochemical tests characterized Isolate 5 as a rod-shaped, Gram-positive Bacillus species with positive results for spore staining, Voges-Proskauer, catalase, starch hydrolysis, and carbohydrate fermentation tests, and negative results for indole, methyl red, citrate, H2S, and urease tests. Definitive genetic identification through 16S rRNA sequencing and comparison with the NCBI database confirmed Isolate 5 as Bacillus tropicus KRKJVR 5, with a 92% similarity to other Bacillus tropicus strains. The sequence data was submitted to NCBI and assigned the accession number PP859287. These findings highlight the potential of Bacillus tropicus KRKJVR 5 for efficient keratin degradation, making it a promising candidate for various industrial and environmental applications.

A comprehensive investigation into prebiotic influence of orange pectin on synergistic modulation of probiotic viability

Synbiotics are well accepted for their nutraceutical value and also in therapeutics for lifestyle disorders like diabetes, cardiovascular, neurological diseases and PCOS. Current manuscript presents the study of prebiotic potential of Pectin isolated from Nagpuri orange on the strains of probiotic bacteria like L. acidophilus, L. plantarum, B. bifidum, and B. lactis. In vitro and in vivo evaluation shows a significant increase in the L. acidophilus colonies as 2.12 ± 0.04 cfu/g followed by L. plantarum as 1.38 ± 0.03 cfu/g, B. lactis as 0.68 ± 0.04 cfu/g and B. bifidum as 0.48 ± 0.05 cfu/g using a modified MRS media. The highest cfu/g complemented the declined pH of cultured media to 6.05 ± 1.32 from near neutral pH. In vivo study using Sprague Dawley rats presented enhanced fecal colony count for L. acidophilus as 8.92 ± 0.04 cfu/g along with confirmation for microscopical characteristics of Gram-positive, rod-shaped, non-motile, non-sporulating bacteria showing catalase-negative, carbohydrate fermentation, lactic acid production, and positive arginine hydrolysis test. All blends of probiotic strains with Pectin as prebiotic material shown significant increase of biomass as compared to single probiotic administration. All results suggest the co-administration of Pectin with the L. acidophilus L. plantarum, B. bifidum, and B. lactis probiotic strains enhances in vitro and in vivo viability of these bacteria. More thorough quantitative analysis of the extract for in vivo parameters can help utilising it in the nutraceutical formulations as a prebiotic compound or as a synbiotic composition along with selected probiotic strains.

Morphogenetic characterization of Stenotrophomonas maltophilia infecting white stripe disease of rice (Oryza sativa L.)

Rice is a major cereal crop which ensure food security to more than half of the global population. Several biotic factors impact rice grain quality and its final production. White stripe disease, caused by pathogen Stenotrophomonas maltophilia is considered among the major limiting factor for reducing rice yields and quality. Present study was performed to understand the white stripe disease, which has been frequently misdiagnosed as bacterial leaf blight (BLB) due to similar symptoms. A survey was carried out based on accessibility and farmer participation to monitor incidence and sample collection. The survey was conducted in districts Faisalabad, Gujranwala, Sialkot, Sheikhupura, and Hafizabad, these districts were selected for their importance for rice cultivation in Pakistan. The total sample size was around 500 leaves distributed evenly throughout each study area. The results of study indicated presence of new pathogen of rice. These isolates were biochemically identified and confirmed by gram staining negative, 3% KOH positive, 5% salt tolerance positive, oxidase test negative, catalase test positive, starch hydrolysis test negative, nitrate reductase test positive, indole test negative, lactose test positive, maltose test positive, methyl red test negative, Voges-prokauer test negative and urea hydrolysis test negative. The pathogenicity test was confirmed that pathogen and the Sialkot isolate were the most aggressive isolate among the five isolates collected form the studied areas. The molecular characterization was accomplished by PCR and sequencing. The results of the phylogenetic study indicate that this pathogen belongs to a distinct group, as it is distantly related to Xanthomonas oryzae pv. oryzae (Xoo). This study provides important findings into a novel clade of pathogen causing white stripe disease in rice.

Isolation and characterization of plant growth promoting rhizobacteria (PGPR) from rhizosphere of Helianthus annuus L.

Plant growth-promoting rhizobacteria (PGPR) support plant growth through direct and indirect mechanisms. To investigate PGPR strains that support plant growth, 21 bacterial isolates, mostly Bacillus ssp. and Pseudomonas ssp., were isolated from different rhizospheric soils of sunflowers in Kırşehir districts in 2020. All isolates were characterized morphologically, biochemically by screening under in vitro conditions for plant growth-promoting properties such as nitrogen fixation, IAA (indoleacetic acid) production, siderophore production, HCN (hydrogen cyanide) production, inorganic phosphate solubility. It was also screened for extracellular enzyme production and antifungal activity against Fusarium oxysporum. Among the 21 isolates, 3 isolates (MH-35-4, MH-49-4, MH-64-3) fixed nitrogen, 2 isolates (MH-59-6, MH-64-3), produced siderophores, 8 isolates (MH-35-4, MH-35-6, MH-54-3, MH-54-4, MH-59-1, MH-59-2, MH-59-4, MH-59-8) produced HCN, 6 isolates (MH-35-6, MH-54-4, MH-59-1, MH-59-2, MH-59-4, MH-59-8) produced IAA, and 7 isolates (MH-35-4, MH-35-6, MH-59-1, MH-59-2, MH-59-4, MH-59-8, MH-64-3) solubilized inorganic phosphate. Additionally, only 2 isolates (MH-54-3, MH-54-4) were positive amylase tests, 8 isolates (MH-35-6, MH-54-4, MH-59-1, MH-59-2, MH-59-4, MH-59-6, MH-59-7, MH-59-8) were positive citrate tests, 8 isolates (MH-35-1, MH-35-4, MH-35-7, MH-49-4, MH-54-4, MH-59-6, MH-59-7, MH-64-3) were positive protease tests, and 6 isolates (MH-35-1, MH-35-3, MH-35-7, MH-54-3, MH-54-4, MH-59-7) were positive gelatin hydrolysis tests. Among 21 isolates, 38% were determined as hydrogen cyanide producers, 10% as siderophore producers, 29% IAA producers, 33% as phosphate solubilizers and 14% as nitrogen fixers. Isolate MH-35-6 showed the highest antifungal activity against Fusarium oxysporum with an inhibition rate of 53.57%. This was followed by isolates MH-54-1 (51.19%), MH-54-3 (47.61%) and MH-59-2 (38.09%), respectively. Therefore, our study reveals that bacteria that promote plant growth in sunflowers can be used to increase crop yield and as a biocontrol agent.

Augmenting corn starch gel printability for architectural 3D modeling for customized food

The advent of direct-ink-writing 3D printing in food processing highlights potential for innovation but underscores challenges with food-grade inks, notably their inadequate self-supporting properties post-extrusion that impede maintaining structural integrity and crating complex 3D forms. This challenge is particularly pronounced with starch—a key food ingredient. This study aims to bolster the printability of normal corn starch (NCS) through integration with pregelatinized (PG) high-amylose starch (G50 and G70, with 55% and 68% amylose contents, respectively) and proteins (soy, wheat, pea protein isolates, and whey protein). The PG starch was prepared by disorganizing the high-amylose starches in 33% CaCl2 solution and then precipitating them with ethanol. The formulation featuring an NCS/PG-G70/soy protein isolate ratio of 5:5:3 emerged superior, yielding enhanced formability, precise line printing, and robust self-support. This adapted starch-based gel facilitated the 3D printing of sophisticated structures, such as hollow and overhanging architectural forms, without necessitating chemical modification or a support bath. In vitro enzymatic hydrolysis tests on the printed constructs manifested approximately 50% resistant starch and 15% slowly digestible starch. These results suggest that the composite biopolymer ink developed in this study showcases not only superior printability but also boasts improved digestion-resistance. Thus, the findings from this research provide a foundation for developing food-grade inks capable of crafting customizable, intricately structured food products while conferring health advantages.

Impact of aromatic sulfonate derivative on the crystallization, thermal resistance, and hydrolytic properties of poly(lactic acid)

AbstractThe thermal, crystallization kinetics and hydrolysis properties of poly(lactic acid)‐based blends containing different concentrations of 5‐dimethylthioisosulfate salts (LAK) were systematically investigated and compared with those of poly(lactic acid) (PLA). PLA/LAK blends were prepared by melt blending method‐using LAK as a nucleating agent of PLA. During the isothermal crystallization process, the morphology of PLA crystal in the neat PLA and PLA/LAK blends showed spherical crystals. With an increase in the concentration of LAK, the nucleating density of the PLA spherulites increased and the size of the PLA spherulites decreased. Differential scanning calorimetry indicates that after the addition of LAK, both the crystallinity and crystallization rate of PLA have been enhanced. In situ X‐ray diffraction analysis also shows that the sample exhibits the same crystal structure as neat PLA and continues to strengthen, indicating that LAK enhances the crystallization properties of PLA. Moreover, the crystallinity of PLA increased and crystal size of PLA decreased with the increase in the LAK content. The Vicat softening temperatures of PLA/LAK blends increased, so the heat resistance improved. Through hydrolysis testing under identical conditions, the PLA/LAK blends exhibits better hydrolytic stability.

Isolation and Characterization of Thermophilic Bacteria from a Hot Spring in the State of Hidalgo, Mexico, and Geochemical Analysis of the Thermal Water.

Hot springs worldwide can be a source of extremophilic microorganisms of biotechnological interest. In this study, samplings of a hot spring in Hidalgo, Mexico, were conducted to isolate, identify, and characterize morphologically, biochemically, and molecularly those bacterial strains with potential industrial applications. In addition, a physicochemical and geochemical examination of the hot spring was conducted to fully understand the study region and its potential connection to the strains discovered. The hot spring was classified as sulfate-calcic according to the Piper Diagram; the hydrogeochemical analysis showed the possible interactions between minerals and water. Eighteen bacterial strains were isolated with optimal growth temperatures from 50 to 55 °C. All strains are Gram-positive, the majority having a rod shape, and one a round shape, and 17 produce endospores. Hydrolysis tests on cellulose, pectin, and xylan agar plates demonstrated enzymatic activity in some of the strains. Molecular identification through the 16S rDNA gene allowed classification of 17 strains within the Phylum Firmicutes and one within Deinococcus-Thermus. The bacterial strains were associated with the genera Anoxybacillus, Bacillus, Anerunibacillus, Paenibacillus, and Deinococcus, indicating a diversity of bacterial strains with potential industrial applications.

Isolation and Identification of Soil-Derived Amylase-Producing Bacteria

Soil is a vital ecosystem for various life forms, including bacteria. Bacteria are the main group of soil microbes, mostly because of their ability to produce various extracellular enzymes. Amylases are the enzymes that break down starch, which are essential in biotechnology industries such as food, fermentation, textiles, and paper manufacturing. The demand for amylase is often high since many enterprises do not have access to local supplies of the enzyme. The objective of the current investigation is to isolate and identify bacterial isolates that can produce the enzyme amylase from soil samples. Soil samples were collected from the unexplored Biligiriranga hills (11.9988° N, 77.1398° E) in south-western Karnataka Bacterial isolates were isolated from soil samples and were primarily screened on starch agar medium to determine the amylase production. Three out of fifteen bacterial isolates that produced amylase were selected based on the highest clear zone around the bacterial colony. These three bacterial isolates were identified both microscopically and biochemically. Gram staining demonstrated that the isolates are rod-shaped and gram-positive organisms. Biochemical studies indicated that the isolates were positive for the catalase and starch hydrolysis tests, whereas negative for the potassium hydroxide and methyl red tests. Besides, 16S rRNA gene amplification and sequencing confirmed the isolate as Bacillus subtilis. The 16S rRNA sequence of B. subtilis strain B has been deposited in the NCBI GenBank. The optimal growth conditions observed in B. Subtilis. The optimal growth conditions observed in B. Subtilis were 37° C temperature at pH 7 and incubation period for 37h. Thus, the isolation and identification of soil-derived amylase-producing bacteria are crucial for understanding their potential applications in various industrial processes, such as bioremediation, waste management, and the production of enzymes for food, pharmaceuticals, and biofuel industries.

Identification and structural characterization of potential degradation products of baricitinib using liquid chromatography combined with quadrupole time-of-flight mass spectrometry

AbstractBaricitinib (BRT) is a drug substance with potent anti-inflammatory activity indicated in rheumatoid arthritis, atopic dermatitis, severe alopecia areata and recently for the treatment of Covid-19. Process impurities of the drug during its formulation are quite known, however studies regarding its degradation products (DPs) under stress conditions are limited. In this study, the drug was subjected to forced degradation under various degradation conditions, including acidic hydrolysis, alkaline hydrolysis, oxidative and thermal, to determine its inherent stability. To this purpose, a novel HPLC method was developed for the determination of degradation impurities of BRT. Alkaline hydrolysis test showed a selectivity towards breaking C–C bonds. This resulted to five DPs formed by chain scission reactions occurred at the pyrrolo-pyrimidine group between C6–C10 and C8–C9. Also, the ethylsulfonyl-azetidin-ylidene group was subjected to C–C bond cleavage at C12–C15 and C16–C18. Degradation products were further characterized with the use of liquid chromatography quadrupole time of flight tandem mass spectrometry (LC-Q-TOF-MS/MS).

Characterization and Evaluation of Native Rhizobium of Groundnut (Arachis hypogaea L.) and Soybean (Glycine max L.)

The present investigation was conducted to characterize and evaluate native Rhizobium isolates of groundnut and soybean to enhance the Legume-Rhizobium symbiosis as well as increase sustainable crop production. In this connection, all the native isolates under study were characterized biochemically and tested with N-free sand culture in the glasshouse to select effective native Rhizobium isolates through proper screening. 10 isolates of each crop were collected from the culture collection bank of the Department of Agricultural Microbiology, Indira Gandhi Krishi Vishwavidyalaya Raipur (Chhattisgarh). Out of 10 isolates, 3 isolates of each crop were selected on the basis of their growth performance and the purity of the culture. The biochemical studies of selected isolates include starch hydrolysis, catalase, urease, citrate utilization, indole production, gelatine iron, and methyl red tests, which were further evaluated for nodulation and biological nitrogen fixation in groundnut and soybean under controlled conditions with four treatments for each crop and five replications. The experiment result revealed that the evaluation of native Rhizobium isolate no. 1185 of groundnut and isolate no. 1076 of soybean was found to be superior among all the rhizobial isolates taken for study. These isolates accumulated 1.24, 1.44 mg/plant extra amount of atmospheric nitrogen over un-inoculated control plants, respectively. Further, it can be concluded that these native isolates may be the most effective nitrogen fixers for groundnut and soybean over the control.

Oriental beech decline caused by Brenneria alni

AbstractHyrcanian forests comprise a long strip along the southern coast of the Caspian Sea in northern Iran. Oriental beech (Fagus orientalis) is one of the most valuable forest tree species in the Hyrcanian forests. Bark necrosis and bleeding spots of oriental beech were observed in Savadkuh forests in Mazandaran province during summer 2021. Ten samples were collected from symptomatic oriental beech trees. The bark canker symptoms included stem tissue necrosis, dark brown to black spots on the trunk, bleeding with dark exudates and dark lesions in the inner bark. Twenty‐six strains of culturable bacteria were isolated from the samples which in culture were predominantly round, convex, smooth with entire margins on nutrient agar and a metallic green sheen pigment on eosin‐methylene blue (EMB) agar. Fourteen strains induced necrosis and browning of the tissues around the inoculation sites on acorns after 2 weeks. Five representative strains also caused dieback in twigs of oriental beech 3 weeks after inoculation. These bacteria were Gram‐negative, catalase positive and oxidase negative. All strains produced colonies with a green metallic sheen on EMB agar. Strains were negative in tests for starch and gelatine hydrolysis or production of levan. A phylogenetic tree based on concatenated sequences of gyrB and infB genes, of representative strains well as in each of the single gene‐based phylogenetic trees, indicated that these strains clustered with Brenneria alni NCPPB 3934T with high bootstrap values. To the best of our knowledge, this is the first report of B. alni associated with bark necrosis and bleeding spots on oriental beech.