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1.
Sci Total Environ ; 935: 173299, 2024 Jul 20.
Article in English | MEDLINE | ID: mdl-38761954

ABSTRACT

As humanity embarks on the journey to establish permanent colonies on Mars, ensuring a reliable source of sustenance will be crucial. Therefore, detailed studies regarding crop cultivation using Martian simulants are of great importance. This study aimed to grow wheat on substrates based on soil and Martian simulants, with the addition of vermicompost, to investigate the differences in wheat development. Basic physical and chemical properties of substrates were examined, including determination of macro- and microelements as well as their microbiological properties. Plant growth parameters were also determined. The addition of vermicompost positively affected wheat grown on soil, but the effect on plants grown on substrate with Martian simulants was negligible. Comparing the microbiological and chemical components, it was observed that plants can defend themselves against the negative effects of growth on the Martian simulants, but their success depends on having the PGPR (Plant growth-promoting rhizobacteria) present, which can provide the plant with additional nitrogen. The presence of beneficial symbiotic microbiota will allow the wheat to wait out the negative growth time rather than adapt to the regolith environment.


Subject(s)
Soil , Triticum , Triticum/growth & development , Soil/chemistry , Mars , Soil Microbiology , Microbiota/drug effects , Composting/methods
2.
Article in English | MEDLINE | ID: mdl-38646860

ABSTRACT

The ß-glucosidase enzyme was obtained from Trichoderma koningii Oudem. NRRL 54330 under optimal conditions by solid substrate fermentation (SSF) using corn cobs as substrate. The enzyme was purified by two-step procedures, ammonium sulphate precipitation and cefarose-4B-l-tyrosine-1-naphthylamine hydrophobic interaction chromatography, followed by biochemical and kinetic characterisation. The ß-glucosidase was obtained from T. koningii using ground corn cob as substrate and Na2HPO4, pH 9, as humidification medium. The optimum conditions for enzyme production by SSF were 30 °C and 6 days. The purification efficiency of the obtained ß-glucosidase was calculated to be 22.56-fold with a yield of 73.51 %. In the determination of ß-glucosidase activity, p-nitrophenyl-ß-d-glucopyranoside (pNPG) substrate was used, and the optimum pH and temperature values at which ß-glucosidase showed high activity were determined to be pH 3.0 and 75 °C. The purity of the enzyme and the presence/number of subunits were checked using two different electrophoretic methods, SDS-PAGE and NATIVE-PAGE electrophoretic methods. The K m and V max values of the purified enzyme were determined to be 0.16 mM and 2000 EU respectively. It was also found that d-(+)-glucose and δ-gluconolactone inhibitors exhibited competitive inhibition of ß-glucosidase in the presence of pNPG.

3.
Heliyon ; 10(1): e23859, 2024 Jan 15.
Article in English | MEDLINE | ID: mdl-38234904

ABSTRACT

To amend physical properties of coastal saline soil for rice production, six biochar treatments (0, 0.5, 1, 2, 4 and 6 kg biochar per m2 soil) were set up as CK, T1, T2, T3, T4 and T5, respectively and their effect on the biochemical properties of coastal saline soil and rice growth characteristics were evaluated in a barrel planting experiment. The results showed that compared with CK (with no biochar added), the soil EC of T1 and T2-T5 was reduced by 11.5 %, but increased by 8.8-62.9 %, respectively. The available potassium and organic matter contents of T1-T5 increased ranging from 3.7-10.2 % to 8.0-46.8 %, respectively. With the increase of the biochar amount, the urease activity of soil in the 0-10 cm deep soil showed an increasing trend by 194.8-744.6 % with T1-T5, compared with that of the CK treatment. The activity of alkaline phosphatase in soil increased first and then decreased, and its increment with T1-T5 was between 28.2 and 64.8 % in comparison with that of CK. With more biochar added to soil, the leaf dry weight, root dry weight, total dry matter mass, total root length, single panicle quality and weight per 100 grains showed a trend of increase first and then decrease. The highest incremental values of all measurements were obtained with T1 by 21.8 %, 23.9 %, 13.8 %, 33.9 %, 30.8 % and 11.6 % respectively, compared with those with CK. However, adding biochar in soil demonstrated insignificant effect on the weight of single panicle, panicle length, stem thickness, tillers, setting rate, soil hydrolyzable nitrogen, available phosphorus content, rice protein, amylose, and taste quality among all treatments. In summary, the application of 0.5 kg m-2 biochar can improve the biochemical properties of saline soil and therefore increase rice yield.

4.
Prep Biochem Biotechnol ; 54(4): 455-469, 2024 Apr.
Article in English | MEDLINE | ID: mdl-37587838

ABSTRACT

Strain improvement via chemical mutagen could impart traits with better enzyme production or improved characteristics. The present study sought to investigate the physicochemical properties of pullulanase produced from the wild Bacillus sp and the mutant. The pullulanases produced from the wild and the mutant Bacillus sp. (obtained via induction with ethyl methyl sulfonate) were purified in a-three step purification procedure and were also characterized. The wild and mutant pullulanases, which have molecular masses of 40 and 43.23 kDa, showed yields of 2.3% with 6.0-fold purification and 2.0% with 5.0-fold purification, respectively, and were most active at 50 and 40 °C and pH 7 and 8, respectively. The highest stability of the wild and mutant was between 40 and 50 °C after 1 h, although the mutant retained greater enzymatic activity between pH 6 and 9 than the wild. The mutant had a decreased Km of 0.03 mM as opposed to the wild type of 1.6 mM. In comparison to the wild, the mutant demonstrated a better capacity for tolerating metal ions and chelating agents. These exceptional characteristics of the mutant pullulanase may have been caused by a single mutation, which could improve its utility in industrial and commercial applications.


Subject(s)
Bacillus , Bacillus/metabolism , Glycoside Hydrolases/metabolism , Hydrogen-Ion Concentration , Enzyme Stability , Temperature
5.
Int J Biol Macromol ; 254(Pt 2): 127859, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37924916

ABSTRACT

D-Allose and D-allulose are two important rare natural monosaccharides found in meager amounts. They are considered to be the ideal substitutes for table sugar (sucrose) for, their significantly lower calorie content with around 80 % and 70 % of the sweetness of sucrose, respectively. Additionally, both monosaccharides have gained much attention due to their remarkable physiological properties and excellent health benefits. Nevertheless, D-allose and D-allulose are rare in nature and difficult to produce by chemical methods. Consequently, scientists are exploring bioconversion methods to convert D-allulose into D-allose, with a key enzyme, L-rhamnose isomerase (L-RhIse), playing a remarkable role in this process. This review provides an in-depth analysis of the extractions, physiological functions and applications of D-allose from D-allulose. Specifically, it provides a detailed description of all documented L-RhIse, encompassing their biochemical properties including, pH, temperature, stabilities, half-lives, metal ion dependence, molecular weight, kinetic parameters, specific activities and specificities of the substrates, conversion ratio, crystal structure, catalytic mechanism as well as their wide-ranging applications across diverse fields. So far, L-RhIses have been discovered and characterized experimentally by numerous mesophilic and thermophilic bacteria. Furthermore, the crystal forms of L-RhIses from E. coli and Stutzerimonas/Pseudomonas stutzeri have been previously cracked, together with their catalytic mechanism. However, there is room for further exploration, particularly the molecular modification of L-RhIse for enhancing its catalytic performance and thermostability through the directed evolution or site-directed mutagenesis.


Subject(s)
Escherichia coli , Fructose , Escherichia coli/metabolism , Fructose/chemistry , Monosaccharides/metabolism , Sucrose/metabolism
6.
Biochemistry (Mosc) ; 88(11): 1956-1969, 2023 Nov.
Article in English | MEDLINE | ID: mdl-38105212

ABSTRACT

Euphorbiaceae, also known as the spurge family, is a large group of flowering plants. Despite being tropical natives, they are now widespread. Due to its medicinal and commercial importance, this family of plants attracted a lot of attention in the scientific community. The distinctive characteristic of the family is production of milky latex, which is a rich source of several lectins, the proteins that bind carbohydrates. Although their function is unclear, they are believed to defend plants against damaging phytopathogenic microorganisms, insects, and predatory animals. Additionally, they serve as crucial metabolic regulators under a variety of stressors. Detection, separation, purification, and characterization of lectins from the Euphorbiaceae family - mostly from the latex of plants - began over 40 years ago. This effort produced over 35 original research papers that were published. However, no systematic review that compiles these published data has been presented yet. This review summarizes and describes several procedures and protocols employed for extraction and purification of lectins belonging to this family. Physicochemical properties and biological activities of the lectins, along with their medicinal and pharmacological properties, have also been analyzed. Additionally, using examples of ricin and ricin agglutinin, we have structurally analyzed characteristics of the lectin known as Ribosome Inactivating Protein Type II (RIP-Type II) that belongs to this family. We anticipate that this review article will offer a useful compendium of information on this important family of lectins, show the scientists involved in lectin research the gaps in our knowledge, and offer insights for future research.


Subject(s)
Euphorbiaceae , Ricin , Animals , Ricin/chemistry , Plant Lectins/pharmacology , Latex/chemistry , Plants
7.
J Enzyme Inhib Med Chem ; 38(1): 2282379, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37985663

ABSTRACT

Acid phosphatases (EC 3.1.3.2) are the enzymes that catalyse transphosphorylation reactions and promotes the hydrolysis of numerous orthophosphate esters in acidic media, as a crucial element for the metabolism of phosphate in tissues. Inorganic phosphate (Pi) utilisation and scavenging, as well as the turnover of Pi-rich sources found in plant vacuoles, are major processes in which intracellular and secretory acid phosphatases function. Therefore, a thorough understanding of these enzymes' structural characteristics, specificity, and physiochemical properties is required to comprehend the function of acid phosphatases in plant energy metabolism. Furthermore, acid phosphatases are gaining increasing importance in industrial biotechnology due to their involvement in transphosphorylation processes and their ability to reduce phosphate levels in food products. Hence, this review aims to provide a comprehensive overview of the purification methods employed for isolating acid phosphatases from diverse plant sources, as well as their structural and functional properties. Additionally, the review explores the potential applications of these enzymes in various fields.


Subject(s)
Acid Phosphatase , Plants , Acid Phosphatase/chemistry , Acid Phosphatase/metabolism , Hydrolysis , Phosphates
8.
Diseases ; 11(4)2023 Sep 28.
Article in English | MEDLINE | ID: mdl-37873776

ABSTRACT

The current literature provides a body of evidence on C-Reactive Protein (CRP) and its potential role in inflammation. However, most pieces of evidence are sparse and controversial. This critical state-of-the-art monography provides all the crucial data on the potential biochemical properties of the protein, along with further evidence on its potential pathobiology, both for its pentameric and monomeric forms, including information for its ligands as well as the possible function of autoantibodies against the protein. Furthermore, the current evidence on its potential utility as a biomarker of various diseases is presented, of all cardiovascular, respiratory, hepatobiliary, gastrointestinal, pancreatic, renal, gynecological, andrological, dental, oral, otorhinolaryngological, ophthalmological, dermatological, musculoskeletal, neurological, mental, splenic, thyroid conditions, as well as infections, autoimmune-supposed conditions and neoplasms, including other possible factors that have been linked with elevated concentrations of that protein. Moreover, data on molecular diagnostics on CRP are discussed, and possible etiologies of false test results are highlighted. Additionally, this review evaluates all current pieces of evidence on CRP and systemic inflammation, and highlights future goals. Finally, a novel diagnostic algorithm to carefully assess the CRP level for a precise diagnosis of a medical condition is illustrated.

9.
Front Plant Sci ; 14: 1250669, 2023.
Article in English | MEDLINE | ID: mdl-37790782

ABSTRACT

Background and aims: In agriculture, biochar (BC) and nitrogen (N) fertilizers are commonly used for improving soil fertility and crop productivity. However, it remains unclear how different levels of BC and N fertilizer affect soil fertility and crop productivity. Methods: This study elucidates the impact of different application rates of BC (0, 600, and 1200 kg/ha) and N fertilizer (105 and 126 kg/ha) on biomass accumulation, soil microbial biomass of carbon (SMC) and nitrogen (SMN), and soil biochemical properties, including soil organic carbon (SOC), total nitrogen (TN), soil nitrate nitrogen (NO3--N), ammonium nitrogen (NH4+-N), urease (UE), acid phosphatase (ACP), catalase (CAT), and sucrase (SC) of tobacco plants. In addition, a high throughput amplicon sequencing technique was adopted to investigate the effect of different application rates of BC/N on rhizosphere bacterial communities of tobacco plants. Results: The results confirm that high dosages of BC and N fertilizer (B1200N126) significantly enhance dry matter accumulation by 31.56% and 23.97% compared with control B0N105 and B0N126 under field conditions and 23.94% and 24.52% under pot experiment, respectively. The soil biochemical properties, SMC, and SMN significantly improved under the high application rate of BC and N fertilizer (B1200N126), while it negatively influenced the soil carbon/nitrogen ratio. Analysis of rhizosphere bacteriome through amplicon sequencing of 16S rRNA revealed that the structure, diversity, and composition of rhizosphere bacterial communities dramatically changed under different BC/N ratios. Proteobacteria, Bacteroidetes, Actinobacteria, Firmicutes, and Acidobacteria were highly abundant bacterial phyla in the rhizosphere of tobacco plants under different treatments. Co-occurrence network analysis displayed fewer negative correlations among rhizosphere bacterial communities under high dosages of biochar and nitrogen (B1200N126) than other treatments, which showed less competition for resources among microbes. In addition, a redundancy analysis further proved a significant positive correlation among SMC, SMN, soil biochemical properties, and high dosage of biochar and nitrogen (B1200N126). Conclusions: Thus, we conclude that a high dosage of BC (1200 kg/ha) under a high application rate of N fertilizer (126 kg/ha) enhances the biomass accumulation of tobacco plants by improving the soil biochemical properties and activities of rhizosphere bacterial communities.

10.
Life (Basel) ; 13(10)2023 Oct 22.
Article in English | MEDLINE | ID: mdl-37895479

ABSTRACT

Biochar is an effective soil amendment with capabilities of boosting carbon sequestration and enhancing soil fertility, thus enhancing plant growth and productivity. While numerous studies have documented the positive effects of biochar on improving soil properties, a number of studies have reported conflicting results. Therefore, the current study was conducted to evaluate the impact of Prosopis juliflora biochar (0, 2.5, 5.0, and 7.5 t ha-1) on soil biochemical properties in Coastal Kenya to ascertain biochar's potential for soil fertility improvement. A randomized complete block design was used for setting up the experiment with three replicates, while Casuarina equisetifolia L. was planted as the test crop. Soil sampling for nutrient analysis was conducted quarterly for 12 months to assess nutrient dynamics under different biochar rates in the current study. Compared to soil untreated with Prosopis juliflora biochar, the results showed that there was a significant increase in soil pH by 21% following biochar utilization at the rate of 7.5 t ha-1. Total nitrogen was increased by 32% after the biochar application, whereas the total organic carbon was increased by four folds in comparison to biochar-untreated soil. Available phosphorus was increased by 264% following biochar application in comparison to the control treatment. In addition, the application of biochar resulted in an increment in the soil exchangeable cations (Ca2+, K+, Mg2+) across the assessment periods. Soil cation exchange capacity (CEC), bacteria and fungi were enhanced by 95, 33 and 48%, respectively, following biochar application at 7.5 t ha-1 in comparison to untreated soil. In conclusion, these results strongly suggest improvement of soil biochemical properties following Prosopis juliflora biochar application, thus providing potential for soil fertility improvement in regions such as the one in the study.

11.
Huan Jing Ke Xue ; 44(9): 5204-5213, 2023 Sep 08.
Article in Chinese | MEDLINE | ID: mdl-37699838

ABSTRACT

To investigate the effects of nano-copper oxide (CuO NPs) on plant growth, physio-biochemical characteristics, and heavy metal content under cadmium stress, a hydroponics experiment was conducted on the effects of single and combined treatments of CuO NPs (0, 10, 20, and 50 mg·L-1) and Cd (0, 1, and 5 µmol·L-1) on the fresh weight, photosynthetic pigment content, MDA content, antioxidant enzyme activity (CAT, POD, SOD, and GR), and Cu and Cd contents in Brassica chinensis L. The results showed that under the single addition of CuO NPs, the fresh weight and activities of CAT, POD, and GR were inhibited as a whole. Photosynthetic pigment content and SOD activity increased first and then decreased with the increase in CuO NPs concentration, whereas MDA content in leaves and roots, and Cu content in subcells of B. chinensis L. increased with the increasing of CuO NPs. As compared with that in the control, CuO NPs promoted the growth of B. chinensis L., and the fresh weight increased by 8.70%-44.87% at 1 µmol·L-1 Cd. When the content of Cd was up to 5 µmol·L-1, a low content (10 mg·L-1) of CuO NPs promoted the growth of B. chinensis L., whereas a high concentration (50 mg·L-1) showed an inhibitory effect. The addition of CuO NPs could increase photosynthetic pigment and MDA contents under different Cd stress, and MDA content in leaves and roots of B. chinensis L. increased by 4.34%-36.27% and 13.43%-131.04%, respectively, than that in the control groups. Under the same concentration of 1 µmol·L-1 Cd, the addition of CuO NPs decreased the activities of CAT and GR, whereas the activity of POD increased. When the content of Cd was up to 5 µmol·L-1, CuO NPs increased the POD activity and inhibited the activity of SOD and GR. The activities of CAT and CAT in the leaves of B. chinensis L. initially showed an increasing and then decreasing trend. CuO NPs and Cd showed antagonistic effects, the maximum reduction of Cd content in leaves and roots of Brassica chinensis L. under 1 µmol·L-1 Cd treatment was 45.64% and 33.39%, and that under 5 µmol·L-1 Cd treatment was 18.25% and 25.35%, respectively. The content of Cu and Cd in subcellular organs of the plants decreased, but the proportion of soluble components increased. These results indicated that CuO NPs at low concentrations promoted plant growth under Cd stress and further inhibited the absorption of Cd but increased the oxidative damage to B. chinensis L.


Subject(s)
Brassica , Metals, Heavy , Copper , Cadmium/toxicity , Metals, Heavy/toxicity , Antioxidants , Oxides , Superoxide Dismutase
12.
Anim Sci J ; 94(1): e13867, 2023.
Article in English | MEDLINE | ID: mdl-37642265

ABSTRACT

The effects of crude protein (CP) and neutral detergent fiber (NDF) percentages in the diet of Japanese Black steers on rumen fluid properties, blood biochemical properties, and carcass characteristics were examined. Twelve 13-month-old Japanese Black steers were used for this study and slaughtered at 30 months of age. Steers were assigned to a control group (n = 6) and test group (n = 6) and were fed a concentrate containing 12.9%-13.9% CP and 26.5%-29.8% NDF or 9.1%-9.6% CP and 29.9%-31.2% NDF, respectively. Lipopolysaccharide activity levels in rumen fluid were lower in the test group than in the control group. Plasma urea nitrogen concentration and activities of aspartate aminotransferase and γ-glutamyltransferase remained lower in the test group than in the control group. In contrast, plasma vitamin A concentrations remained higher in the test group than in the control group. Carcass characteristics did not significantly differ between the two groups. These results suggest that dietary CP and NDF percentages in feed for Japanese Black steers older than 13 months of age affected rumen fluid properties and blood biochemical properties, indicating a reduced load on the liver with a small effect on carcass characteristics.


Subject(s)
Body Fluids , Detergents , Animals , Detergents/metabolism , Rumen/metabolism , Animal Feed/analysis , Diet/veterinary , Body Fluids/metabolism , Dietary Fiber/metabolism , Digestion
13.
Rice (N Y) ; 16(1): 34, 2023 Aug 01.
Article in English | MEDLINE | ID: mdl-37526797

ABSTRACT

Soil fungal microbiomes facilitate a range of beneficial functions for their host plants, and rhizosphere fungal community composition, richness, and diversity affect plant growth and development, and crop yield. Therefore, exploring the community structure and assembly of the rhizosphere fungal microbiome and its relationship with soil biochemical properties are fundamental to elucidating how rice plants benefit from their fungal symbionts. In this study, soil samples were collected at seedling, tillering, heading, and ripening stages of rice subjected to three levels of nitrogen fertilization. Plant growth demonstrates a substantial influence on fungal community composition and diversity. From the tillering to the ripening stage, the fungal communities were governed by homogenizing dispersal and dispersal limitation. The prevalence of Glomeromycota, the beneficial fungi, was considerably higher during the heading stage compared to the three other growth stages. This increase in abundance was strongly associated with increased levels of soil nutrients and enhanced activity of nitrogen acquisition enzymes. This may be a strategy developed by rice grown in flooded soil to recruit beneficial fungi in the rhizosphere to meet high nitrogen demands. Our study findings contribute to elucidating the influence of plant development and nitrogen fertilization on the structure and composition of the fungal community as well as its relationship with soil key soil nutrient content and nitrogen-related enzyme activities. They also illustrate how a shift in the fungal community mediates and reflects the effects of nitrogen fertilization input in rice agroecosystems. These findings provide new insights into the effects of changes in nitrogen application in rice rhizosphere at different growth stages on fungal communities and soil biochemical characteristics.

14.
Plants (Basel) ; 12(15)2023 07 31.
Article in English | MEDLINE | ID: mdl-37570992

ABSTRACT

Drought stress in arid regions is a serious factor affecting yield quantity and quality of economic crops. Under drought conditions, the application of nano-elements and nano-agents of water retention improved the water use efficiency, growth performance, and yield quantity of drought-stressed plants. For this objective, two field experiments were performed and organized as randomized complete block designs with six replications. The treatments included kaolin (5 t. ha-1) bentonite (12.5 t. ha-1), perlite (1.25 t.ha-1), N-zeolite (1.3 L.ha-1), N-silicon (2.5 L.ha-1), and N-zinc (2.5 L.ha-1). The current study showed that the application of silicon, zinc, and zeolite nanoparticles only positively influenced the morphological, physiological, and biochemical properties of the drought-stressed coriander plant. Exogenous application of N-silicon, N-zinc, and N-zeolite recorded the higher growth parameters of drought-stressed plants; namely, plant fresh weight, plant dry weight, leaf area, and root length than all the other treatments in both seasons. The improvement ratio, on average for both seasons, reached 17.93, 17.93, and 18.85% for plant fresh weight, 73.46, 73.46, and 75.81% for plant dry weight, 3.65, 3.65, and 3.87% for leaf area, and 17.46, 17.46, and 17.16% for root length of drought-stressed plants treated with N-silicon, N-zinc, and N-zeolite, respectively. For physiological responses, the application of N-zeolite, N-silicon, and N-zinc significantly increased leaf chlorophyll content, photosynthetic rate, water use efficiency, chlorophyll fluorescence, and photosystem II efficiency compared with the control in both seasons, respectively. Similar results were observed in antioxidant compounds, nutrient accumulation, and phytohormones. In contrast, those treatments markedly reduced the value of transpiration rate, nonphotochemical quenching, MDA, ABA, and CAT compared to control plants. Regarding the seed and oil yield, higher seed and oil yields were recorded in drought-stressed plants treated with N-zeolite followed by N-silicon and N-zinc than all the other treatments. Application of N-zeolite, N-silicon and N-zinc could be a promising approach to improve plant growth and productivity as well as to alleviate the adverse impacts of drought stress on coriander plants in arid and semi-arid areas.

15.
Heliyon ; 9(6): e17365, 2023 Jun.
Article in English | MEDLINE | ID: mdl-37383213

ABSTRACT

Polyphenol oxidase which is responsible for oxidative conversion of phenolic compounds to polymers, has continued to attract the attention of scientists. Here, we report the extraction, purification and biochemical properties of polyphenol oxidase (PPO) from bitter leaf (Vernonia amygdalina). The enzyme was purified and concentrated using a non-conventional approach, aqueous two-phase partitioning (ATPS) and the biochemical properties of the purified enzyme were investigated. Substrate specificity studies revealed that the enzyme predominantly exhibits diphenolase activity. The order of substrate preference was catechol > L-DOPA > caffeic acid > L-tyrosine > resorcinol>2-naphthol > phenol. The optimum pH and temperature obtained for the enzyme using catechol as substrate were 5.5 and 50 °C respectively. The estimated Michaelis constant (Km) and maximum velocity (Vmax) for the purified vaPPO using catechol as substrate were 183 ± 5.0 mM and 2000 ± 15 units/mg protein respectively. The catalytic efficiency (Vmax/Km) of the purified vaPPO was 10.9 ± 0.03 min/mg. Na+, K+ and Ba2+ remarkably activated the enzyme and the level of activation was proportional to the concentration. The vaPPO presented stability in the presence of up to 50 mM of the different metal ions tested. In contrast, Cu2+ and NH4+ inhibited the enzyme even 10 mM concentrations. The enzyme was stable in chloroform retaining up to 60% relative activity at 50% (v/v) concentration. There was an increase in the activity (143%) of the enzyme at 30% (v/v) chloroform., revealing that vaPPO could catalyze the substrate more efficiently in 30% (v/v) chloroform. Total loss of enzyme activity was observed at 20% (v/v) concentrations of acetone, ethanol and methanol. In conclusion, the properties of the vaPPO such as its catalysis in the presence of organic solvents, metals and high temperature would be of interest in many biotechnological applications.

16.
Saudi J Biol Sci ; 30(7): 103682, 2023 Jul.
Article in English | MEDLINE | ID: mdl-37305655

ABSTRACT

A soil inhabiting Pseudomonas sp. has been examined for producing L- methionine gamma-lyase enzyme. The identity of the tested bacteria was verified by VITEK2, and MALDI-TOF analysis in addition to molecular confirmation by 16S rDNA sequence and submitted in Genbank under accession number ON993898.1. Production of the targeted enzyme was done using a commercial medium including L-methionine, as the main substrate. This obtained enzyme was precipitated using acetone (1:1v/v) followed by purification with Sephadex G100 and sepharose columns. The specific activity of the purified enzyme (105.8 µmol/ mg/min) increased by 1.89 folds after the purification steps. The peptide fingerprint of the native MGL was verified from the proteomics analysis, with identical conserved active site domains with database-deposited MGLs. The molecular mass of the pure MGL denatured subunit was (>40 kDa) and that of the native enzyme was (>150 kDa) ensuring their homotetrameric identity. The purified enzyme showed absorption spectra at 280 nm and 420 nm for the apo-MGL and PLP coenzyme, respectively. Amino acids suicide analogues analysis by DTNB, hydroxylamine, iodoacetate, MBTH, mercaptoethanol and guanidine thiocyanate reduced the relative activity of purified MGL. From the kinetic properties, the catalytic effectiveness (Kcat/km) of Pseudomonas sp. MGL was 10.8 mM -1 S-1 for methionine and 5.51 mM -1 S-1 for cysteine, respectively. The purified MGL showed highly significant antiproliferative activity towards the liver carcinoma cell line (HEPG-2) and breast carcinoma cell line (MCF-7) with half inhibitory concentration values (IC50) 7.23 U/ml and 21.14 U/ml, respectively. No obvious signs of toxicity on liver and kidney functions in the examined animal models were observed.

17.
Molecules ; 28(12)2023 Jun 10.
Article in English | MEDLINE | ID: mdl-37375248

ABSTRACT

Chromium (Cr) can exist in several oxidation states, but the two most stable forms-Cr(III) and Cr(VI)-have completely different biochemical characteristics. The aim of the present study was to evaluate how soil contamination with Cr(III) and Cr(VI) in the presence of Na2EDTA affects Avena sativa L. biomass; assess the remediation capacity of Avena sativa L. based on its tolerance index, translocation factor, and chromium accumulation; and investigate how these chromium species affect the soil enzyme activity and physicochemical properties of soil. This study consisted of a pot experiment divided into two groups: non-amended and amended with Na2EDTA. The Cr(III)- and Cr(VI)-contaminated soil samples were prepared in doses of 0, 5, 10, 20, and 40 mg Cr kg-1 d.m. soil. The negative effect of chromium manifested as a decreased biomass of Avena sativa L. (aboveground parts and roots). Cr(VI) proved to be more toxic than Cr(III). The tolerance indices (TI) showed that Avena sativa L. tolerates Cr(III) contamination better than Cr(VI) contamination. The translocation values for Cr(III) were much lower than for Cr(VI). Avena sativa L. proved to be of little use for the phytoextraction of chromium from soil. Dehydrogenases were the enzymes which were the most sensitive to soil contamination with Cr(III) and Cr(VI). Conversely, the catalase level was observed to be the least sensitive. Na2EDTA exacerbated the negative effects of Cr(III) and Cr(VI) on the growth and development of Avena sativa L. and soil enzyme activity.


Subject(s)
Avena , Soil Pollutants , Soil/chemistry , Edetic Acid , Soil Pollutants/analysis , Chromium/chemistry
18.
Biomolecules ; 13(5)2023 05 12.
Article in English | MEDLINE | ID: mdl-37238695

ABSTRACT

Acute Respiratory Distress Syndrome (ARDS) is an illness that typically develops in people who are significantly ill or have serious injuries. ARDS is characterized by fluid build-up that occurs in the alveoli. T-cells are implicated as playing a role in the modulation of the aberrant response leading to excessive tissue damage and, eventually, ARDS. Complementarity Determining Region 3 (CDR3) sequences derived from T-cells are key players in the adaptive immune response. This response is governed by an elaborate specificity for distinct molecules and the ability to recognize and vigorously respond to repeated exposures to the same molecules. Most of the diversity in T-cell receptors (TCRs) is contained in the CDR3 regions of the heterodimeric cell-surface receptors. For this study, we employed the novel technology of immune sequencing to assess lung edema fluid. Our goal was to explore the landscape of CDR3 clonal sequences found within these samples. We obtained more than 3615 CDR3 sequences across samples in the study. Our data demonstrate that: (1) CDR3 sequences from lung edema fluid exhibit distinct clonal populations, and (2) CDR3 sequences can be further characterized based on biochemical features. Analysis of these CDR3 sequences offers insight into the CDR3-driven T-cell repertoire of ARDS. These findings represent the first step towards applications of this technology with these types of biological samples in the context of ARDS.


Subject(s)
Complementarity Determining Regions , Respiratory Distress Syndrome , Humans , Complementarity Determining Regions/genetics , Receptors, Antigen, T-Cell, alpha-beta/genetics , Receptors, Antigen, T-Cell , Respiratory Distress Syndrome/genetics , Edema
19.
Molecules ; 28(8)2023 Apr 10.
Article in English | MEDLINE | ID: mdl-37110566

ABSTRACT

Heat stress is one of the most stressful events in livestock life, negatively impacting animal health, productivity, and product quality. Moreover, the negative impact of heat stress on animal product quality has recently attracted increasing public awareness and concern. The purpose of this review is to discuss the effects of heat stress on the quality and the physicochemical component of meat in ruminants, pigs, rabbits, and poultry. Based on PRISMA guidelines, research articles were identified, screened, and summarized based on inclusion criteria for heat stress on meat safety and quality. Data were obtained from the Web of Science. Many studies reported the increased incidences of heat stress on animal welfare and meat quality. Although heat stress impacts can be variable depending on the severity and duration, the exposure of animals to heat stress (HS) can affect meat quality. Recent studies have shown that HS not only causes physiological and metabolic disturbances in living animals but also alters the rate and extent of glycolysis in postmortem muscles, resulting in changes in pH values that affect carcasses and meat. It has been shown to have a plausible effect on quality and antioxidant activity. Acute heat stress just before slaughter stimulates muscle glycogenolysis and can result in pale, tender, and exudative (PSE) meat characterized by low water-holding capacity (WHC). The enzymatic antioxidants such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) act by scavenging both intracellular and extracellular superoxide radicals and preventing the lipid peroxidation of the plasma membrane. Therefore, understanding and controlling environmental conditions is crucial to successful animal production and product safety. The objective of this review was to investigate the effects of HS on meat quality and antioxidant status.


Subject(s)
Antioxidants , Chickens , Animals , Rabbits , Swine , Antioxidants/metabolism , Chickens/metabolism , Meat/analysis , Heat-Shock Response , Poultry
20.
Braz J Microbiol ; 54(2): 715-723, 2023 Jun.
Article in English | MEDLINE | ID: mdl-36917331

ABSTRACT

L-Asparaginase (L-ASNase) is a potent chemotherapeutic drug employed to treat leukemia and lymphoma. Currently, L-ASNases for therapeutic use are obtained from Escherichia coli and Dickeya chrysanthemi (Erwinia chrysanthemi). Despite their therapeutic potential, enzymes from bacteria are subject to inducing immune responses, resulting in a higher number of side effects. Eukaryote producers, such as fungi, may provide therapeutic alternatives through enzymes that induce relatively less toxicity and immune responses. Additional expected benefits from yeast-derived enzymes include higher activity and stability in physiological conditions. This work describes the new potential therapeutic candidate L-ASNase from the yeast Meyerozyma guilliermondii. A statistical approach (full factorial central composite design) was used to optimize L-ASNase production, considering L-asparagine and glucose concentration, pH of the medium, and cultivation time as independent factors. In addition, the crude enzymes were biochemically characterized, in terms of temperature and optimal pH, thermostability, pH stability, and associated glutaminase or urease activities. Our results showed that enzyme production increased after supplementing a pH 4.0 medium with 1.0% L-asparagine and 0.5% glucose during 75 h of cultivation. Under these optimized conditions, L-ASNase production reached 26.01 U mL-1, which is suitable for scale-up studies. The produced L-ASNase exhibits maximal activity at 37 °C and pH 7.0 and is highly stable under physiological conditions. In addition, M. guilliermondii L-ASNase has no associated glutaminase or urease activities, demonstrating its potential as a promising antineoplastic agent.


Subject(s)
Antineoplastic Agents , Asparaginase , Asparaginase/genetics , Asparagine , Urease , Glutaminase , Escherichia coli/genetics , Glucose
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