Search

Temperature is a key parameter that influences the uptake and subsequent utilization of nitrogen by plants. Both suboptimal and supraoptimal temperatures can impair nutrient uptake. The close relatives of bread wheat provide a possible source for breeders to increase stress tolerance. The effect of the increasing temperature (5, 10, 15, 20 and 30 °C) on nitrate uptake and metabolism in five modern spring cultivars of bread wheat (Triticum aestivum L.) and two cultivars of emmer wheat (Triticum dicoccon Schrank), was monitored. Wheat plants were grown under controlled conditions in hydroponics. The parameters of Michaelis-Menten kinetics, maximum uptake rate (Vmax), the Michaelis constant (Km) and selected characteristics of nitrate metabolism, the activity of nitrate reductase (NR) and contents of nitrate in leaves were observed. The effect of temperature was significant for all studied traits except Km, while the cultivar factor was significant for Vmax, Km, NR and root/shoot ratio (R/S). Emmer wheat cultivar Rudico had significantly higher Vmax at 5, 15, 20 and 30 °C than all bread wheat cultivars, on average 7.07, in comparison with 4.09-4.43 μmol NO3-/g FW/h in bread wheat cultivars. Emmer wheat Rudico and Tapiruz had significantly higher Km (on average, 41.59 and 47.22 μM NO3-) than bread wheat cultivars (27.59-33.44 μM NO3-). Differences in the studied kinetic parameters of nitrate uptake offer the possibility of using T. dicoccon genotypes in breeding for better nitrogen use efficiency.

The NHX1 gene encodes a Na+/H+ antiporter located in the tonoplast membrane, which plays critical role in regulating plant salt tolerance. It is also involved in the uptake and accumulation of K in plants; however, its precise mechanism is unknown. In this research, we elucidated the physiological basis underlying the increases in K content induced by NHX1. We evaluated main agronomic traits, leaf K content, K+ uptake kinetics, and root morphological and physiological characteristics from field-planted and hydroponic plants. We included a wild-type tobacco (Nicotiana tabacum) variety (K326) and three transgenic tobacco lines (NK7, NK9, NK10) that overexpress AtNHX1 from Arabidopsis thaliana. Results demonstrated that the agronomic performance of the AtNHX1 -overexpressing tobacco lines was similar to K326 in field and hydroponic settings. The three AtNHX1 -overexpressing tobacco lines had significantly higher leaf K contents than K326. Under hydroponic condition, enhanced K uptake capacity and a larger maximum K uptake rate were seen in AtNHX1 -overexpressing tobacco lines. AtNHX1 -overexpressing lines also exhibited significantly superior root morphological and physiological traits relative to K326, including root biomass, root volume, absorption area, root activity, cation exchange capacity, soluble protein content, and H+-ATPase activity. Overexpression of AtNHX1 in tobacco significantly improves the K uptake and accumulation. Therefore, leaf K content greatly increased in these transgenic lines in the end. Our findings strongly suggest that AtNHX1 overexpression increased leaf K content by boosting the capacity of enriching K in tobacco roots, thereby advancing the understanding of the function of AtNHX1. NHX1 is closely related to K uptake and accumulation in plants. AtNHX1 -overexpressing tobacco (Nicotiana tabacum) exhibited significantly higher leaf K content than the wild-type tobacco variety K326. The AtNHX1 -overexpressing tobacco has significantly improved root morphology and physiology compared to the wild-type variety K326. AtNHX1 -overexpressing tobacco showed higher V max levels in roots than the wild-type variety K326. Overexpression of AtNHX1 is able to increase leaf potassium content by boosting the ability of enriching potassium in tobacco roots. [ABSTRACT FROM AUTHOR]

Temperature is a key parameter that influences the uptake and subsequent utilization of nitrogen by plants. Both suboptimal and supraoptimal temperatures can impair nutrient uptake. The close relatives of bread wheat provide a possible source for breeders to increase stress tolerance. The effect of the increasing temperature (5, 10, 15, 20 and 30 °C) on nitrate uptake and metabolism in five modern spring cultivars of bread wheat (Triticum aestivum L.) and two cultivars of emmer wheat (Triticum dicoccon Schrank), was monitored. Wheat plants were grown under controlled conditions in hydroponics. The parameters of Michaelis-Menten kinetics, maximum uptake rate (Vmax), the Michaelis constant (Km) and selected characteristics of nitrate metabolism, the activity of nitrate reductase (NR) and contents of nitrate in leaves were observed. The effect of temperature was significant for all studied traits except Km, while the cultivar factor was significant for Vmax, Km, NR and root/shoot ratio (R/S). Emmer wheat cultivar Rudico had significantly higher Vmax at 5, 15, 20 and 30 °C than all bread wheat cultivars, on average 7.07, in comparison with 4.09-4.43 μmol NO3-/g FW/h in bread wheat cultivars. Emmer wheat Rudico and Tapiruz had significantly higher Km (on average, 41.59 and 47.22 μM NO3-) than bread wheat cultivars (27.59-33.44 μM NO3-). Differences in the studied kinetic parameters of nitrate uptake offer the possibility of using T. dicoccon genotypes in breeding for better nitrogen use efficiency. [ABSTRACT FROM AUTHOR]

The microsomal metabolism of phenol (11 °C) over an annual reproductive cycle from June to December was studied using fall spawning adult brook trout (Salvelinus fontinalis). Hepatic microsomes were isolated from three male and three female fish each month. Incubations were optimized for time, cofactor concentration, pH, and microsomal protein concentration. The formation of phase I ring-hydroxylation metabolites, i.e., hydroquinone (HQ) and catechol (CAT), was quantified by HPLC with dual-channel electrochemical detection. Sample preparation and chromatographic conditions were optimized to achieve the separation and sensitivity required for the analysis of these labile products. Biotransformation of phenol over a range of substrate concentrations (1 to 150 mM) was quantified for the calculation of Michaelis–Menten constants (Km and Vmax) for each month. Results indicate a nearly equal production of HQ and CAT among males and females in late June. At the peak of maturity in October, there was an approximate ten-fold greater production of ring-hydroxylation metabolites noted in females in comparison with males on a total liver basis. In vitro phase II biotransformation of phenol glucuronidation was assessed by determining the Michaelis–Menten constants (Km, Vmax) using brook trout hepatic microsomes over a range of substrate concentrations (1 to 60 mM). Initially, there were no significant differences in the glucuronide rate of formation (pmol/min/mg protein) or total capacity (nmol/min/liver) between females and males. At the peak of maturation, the maximum rate of glucuronide formation was 4-fold less in females; however, the total capacity was 2-fold less in females due to the increased liver size in the females. The alterations in biotransformation coincided with increases in the hepatic and gonadal somatic indices and with changes in plasma hormone concentrations. These experiments provide insight into the metabolic deactivation of xenobiotics and to provide data for the prediction of altered hepatic biotransformation rates and pathways during the reproductive cycle. [ABSTRACT FROM AUTHOR]

The medical field now needs more novel drugs to treat obesity and type-2 diabetes mellitus (T2D) than ever before. Obesity and T2D are both characterized by resistance to the hormones leptin and insulin. PTP-1B is a promising target for drug growth, as strong genetic, pharmacological, and biochemical evidence points to the possibility of treating diabetes and obesity by blocking the PTP-1B enzyme. Studies have also found that PTP-1B is overexpressed in patients with diabetes and obesity, suggesting that inhibiting PTP-1B may be a useful technique in their care. There are no clinically used PTP-1B inhibitors, despite the fact that numerous naturally occurring PTP-1B inhibitors have demonstrated great therapeutic promise. This is most likely due to their low activity or lack of selectivity. It is still important to look for more effective and focused PTP-1B inhibitors. A few organovanadium metal complexes were synthesized and characterized, and binding studies on vanadium complexes with PTP-B were also performed using fluorescence emission spectroscopy. Additionally, we theoretically (molecular modeling) and experimentally (enzyme kinetics) examined the PTP-1B inhibitory effects of these vanadium metal complexes and found that they have excellent PTP-1B inhibitory properties. [ABSTRACT FROM AUTHOR]

Alpha-amylase is one of the most widely used enzymes in the starch industry. However, industrial application of soluble alpha-amylase is hampered by changes in pH and temperature (adverse effects on enzyme stability) and activity loss, leading to higher costs. Immobilization of alpha-amylase is an efficient strategy to reduce the enzyme losing and subsequently reduces costs in this regard. Alpha-amylases are immobilized by adsorption, entrapment, covalent attachment, and cross-linking. A barrier in alpha-amylase immobilization is the large size of its substrate, namely amylose and amylopectin. Most of these immobilization methods decrease the affinity of the enzyme for its substrate as well as the maximum rate of reaction (Vmax). This review aims to study different aspects of alpha-amylase including enzyme activity, applications, structure, starch, immobilization methods, and immobilization’s obstacles to improve alpha-amylase efficiency in the industry and also lowering the costs related to providing this enzyme.

Abstract Background Complex enzymatic models are required for analyzing kinetic data derived under conditions that may not satisfy the assumptions associated with Michaelis–Menten kinetics. To analyze these data, several software packages have been developed. However, the complexity introduced by these programs is often dispensable when analyzing data conforming to the canonical Michaelis–Menten model. In these cases, the sophisticated routines of these packages become inefficient and unnecessarily intricated for the intended purpose, reason for which most users resort to general-purpose graphing programs. However, this approach, in addition of being time-consuming, is prone to human error, and can lead to misleading estimates of kinetic parameters, particularly when unweighted regression analyses of transformed kinetic data are performed. Results To fill the existing gap between highly specialized and general-purpose software, we have developed an easy-to-use R package, renz, designed for accurate and efficient estimation of enzyme kinetic parameters. The package provides different methods that can be clustered into four categories, depending on whether they are based on data fitting to a single progress curve (evolution of substrate concentration over time) or, alternatively, based on the dependency of initial rates on substrate concentration (differential rate equation). A second criterion to be considered is whether the experimental data need to be manipulated to obtain linear functions or, alternatively, data are directly fitted using non-linear regression analysis. The current program is a cross-platform, free and open-source software that can be obtained from the CRAN repository. The package is accompanied by five vignettes, which are intended to guide users to choose the appropriate method in each case, as well as providing the basic theoretical foundations of each method. These vignettes use real experimental data to illustrate the use of the package utilities. Conclusions renz is a rigorous and yet easy-to-use software devoted to the analysis of kinetic data. This application has been designed to meet the needs of users who are not practicing enzymologists, but who need to accurately estimate the kinetic parameters of enzymes. The current software saves time and minimizes the risk of making mistakes or introducing biases due to uncorrected error propagation effects.

Fungal metabolic rates could increase under climate warming but may be counteracted by mechanisms of physiological acclimation and evolutionary adaptation. We hypothesized that Vmax and Km parameters of Neurospora discreta extracellular enzymes would acclimate to warmer temperatures through compensatory mechanisms. We also predicted that evolution under warmer temperatures would alter enzyme parameters and fungal respiration through adaptive mechanisms. In contrast to these predictions, growth at higher temperature (22 °C versus 16 °C) increased the temperature-corrected Vmax of three enzymes. The carbon substrate used for fungal growth (lignin versus sucrose) had a much greater impact on enzyme Vmax than temperature. Following experimental evolution, the enzymatic parameters of Neurospora strains did not adapt to higher temperatures as hypothesized; rather, enzyme Vmax values were unaffected, and respiration rates increased. Together, these results suggest that physiological and evolutionary mechanisms are unlikely to counteract soil carbon losses driven by saprotrophic fungi under climate warming.

Fungal metabolic rates could increase under climate warming but may be counteracted by mechanisms of physiological acclimation and evolutionary adaptation. We hypothesized that Vmax and Km parameters of Neurospora discreta extracellular enzymes would acclimate to warmer temperatures through compensatory mechanisms. We also predicted that evolution under warmer temperatures would alter enzyme parameters and fungal respiration through adaptive mechanisms. In contrast to these predictions, growth at higher temperature (22 °C versus 16 °C) increased the temperature-corrected Vmax of three enzymes. The carbon substrate used for fungal growth (lignin versus sucrose) had a much greater impact on enzyme Vmax than temperature. Following experimental evolution, the enzymatic parameters of Neurospora strains did not adapt to higher temperatures as hypothesized; rather, enzyme Vmax values were unaffected, and respiration rates increased. Together, these results suggest that physiological and evolutionary mechanisms are unlikely to counteract soil carbon losses driven by saprotrophic fungi under climate warming.

The magnitude and direction of carbon cycle feedbacks under climate warming remain uncertain due to insufficient knowledge about the temperature sensitivities of soil microbial processes. Enzymatic rates could increase at higher temperatures, but this response could change over time if soil microbes adapt to warming. We used the Arrhenius relationship, biochemical transition state theory, and thermal physiology theory to predict the responses of extracellular enzyme Vmax and Km to temperature. Based on these concepts, we hypothesized that Vmax and Km would correlate positively with each other and show positive temperature sensitivities. For enzymes from warmer environments, we expected to find lower Vmax , Km , and Km temperature sensitivity but higher Vmax temperature sensitivity. We tested these hypotheses with isolates of the filamentous fungus Neurospora discreta collected from around the globe and with decomposing leaf litter from a warming experiment in Alaskan boreal forest. For Neurospora extracellular enzymes, Vmax Q10 ranged from 1.48 to 2.25, and Km Q10 ranged from 0.71 to 2.80. In agreement with theory, Vmax and Km were positively correlated for some enzymes, and Vmax declined under experimental warming in Alaskan litter. However, the temperature sensitivities of Vmax and Km did not vary as expected with warming. We also found no relationship between temperature sensitivity of Vmax or Km and mean annual temperature of the isolation site for Neurospora strains. Declining Vmax in the Alaskan warming treatment implies a short-term negative feedback to climate change, but the Neurospora results suggest that climate-driven changes in plant inputs and soil properties are important controls on enzyme kinetics in the long term. Our empirical data on enzyme Vmax , Km , and temperature sensitivities should be useful for parameterizing existing biogeochemical models, but they reveal a need to develop new theory on thermal adaptation mechanisms.

In this work, the electrodes of a microbial desalination cell (MDC) are investigated as the positive and negative electrodes of an internal supercapacitor. The resulting system has been named a supercapacitive microbial desalination cell (SC-MDC). The electrodes are self-polarized by the red-ox reactions and therefore the anode acts as a negative electrode and the cathode as a positive electrode of the internal supercapacitor. In order to overcome cathodic losses, an additional capacitive electrode (AdE) was added and short-circuited with the SC-MDC cathode (SC-MDC-AdE). A total of 7600 discharge/self-recharge cycles (equivalent to 44 h of operation) of SC-MDC-AdE with a desalination chamber filled with an aqueous solution of 30 g L-1 NaCl are reported. The same reactor system was operated with real seawater collected from Pacific Ocean for 88 h (15,100 cycles). Maximum power generated was 1.63 ± 0.04 W m-2 for SC-MDC and 3.01 ± 0.01 W m-2 for SC-MDC-AdE. Solution conductivity in the desalination reactor decreased by ∼50% after 23 h and by more than 60% after 44 h. There was no observable change in the pH during cell operation. Power/current pulses were generated without an external power supply.

ABSTRACT Understanding the kinetics of Zn absorption by roots and its effect on morphology of this organ is relevant for improving crop management, but still poorly studied for common beans. Therefore, an experiment was conducted in a hydroponic system with five initial concentrations of Zn (CZnI): 0.0; 1.0; 4.0; 16.0 and 48.0 µmol L-1. The experiment was installed with plants at V3 stage of development and aliquots of the solution collected over 24 h. The maximum absorption rate (Vmax), Michaelis-Menten constant (Km) and the absorption power (α) increased as a function of CZnI. The minimum concentration of Zn estimated for its absorption (Cmin) was at 0.0028 mg L-1. The influx of Zn (Imax) was higher in higher CZnI, 16,0 µmol L-1. Root length, root volume, root Zn content and Zn absorption efficiency increased with the increase of CZnI. Therefore, the increase of CZnI positively influenced kinetic parameters of root Zn absorption and common bean root morphology, characteristics that favor Zn absorption by roots and improves overall plant nutrition, favoring agronomical biofortification practices for Zn and other nutrients.

The volatile profile of ‘Redchief Delicious’ apple (Malus ×domestica Borkh.) fruit was evaluated at 18 time points from 3 weeks before to 8 weeks after onset of autocatalytic ethylene production to capture the dynamics associated with development from mature green to senescent fruit. Minor amounts of ester production began several days before the onset of ethylene production. Ester production rose rapidly as internal ethylene levels increased beyond 22 nmol·L−1 (0.5 µL·L−1). Peak ester production roughly coincided with maximum ethylene synthesis, declining thereafter. Ester production was further evaluated according to the acid- (alkanoate) and alcohol- (alkyl) derived portions of the ester. The maximum rate of production for a given ester tended to occur later in development as the chain length of the alcohol-derived portion declined. The production rate for many esters paralleled the rate of emanation of their respective alcohol substrates, suggesting that availability of the alcohols limits ester production more than availability of the acid substrates. Combining production rates with sensory descriptors and human sensitivity to individual volatiles permitted approximations of aroma sensations likely engendered by the fruit throughout ripening. Overripe and alcoholic sensations are predicted to increase 2 weeks after the initiation of ripening in response to an increase in the production of ethyl esters. Acetate esters predominated, comprising 50% to 80% of esters throughout maturation and ripening, indicating that the substrate acetyl-CoA may be at saturating levels for alcohol acyl transferase (AAT) at the final step of ester formation. Acetate feeding did not enhance ester production, although label from 13C-acetate was extensively incorporated into esters. The data are consistent with the action of multiple AAT isozymes differing in activity and substrate preference. Incorporation of labeled 13C-acetate into precursors of esters, alcohols, and acids, reflected ester biosynthesis via 1- and 2-carbon chain elongation pathways in ripening ‘Redchief Delicious’ apple fruit.

Background: Complex enzymatic models are required for analyzing kinetic data derived under conditions that may not satisfy the assumptions associated with Michaelis–Menten kinetics. To analyze these data, several software packages have been developed. However, the complexity introduced by these programs is often dispensable when analyzing data conforming to the canonical Michaelis–Menten model. In these cases, the sophisticated routines of these packages become inefficient and unnecessarily intricated for the intended purpose, reason for which most users resort to general-purpose graphing programs. However, this approach, in addition of being time-consuming, is prone to human error, and can lead to misleading estimates of kinetic parameters, particularly when unweighted regression analyses of transformed kinetic data are performed. Results: To fill the existing gap between highly specialized and general-purpose software, we have developed an easy-to-use R package, renz, designed for accurate and efficient estimation of enzyme kinetic parameters. The package provides different methods that can be clustered into four categories, depending on whether they are based on data fitting to a single progress curve (evolution of substrate concentration over time) or, alternatively, based on the dependency of initial rates on substrate concentration (differential rate equation). A second criterion to be considered is whether the experimental data need to be manipulated to obtain linear functions or, alternatively, data are directly fitted using non-linear regression analysis. The current program is a cross-platform, free and open-source software that can be obtained from the CRAN repository. The package is accompanied by five vignettes, which are intended to guide users to choose the appropriate method in each case, as well as providing the basic theoretical foundations of each method. These vignettes use real experimental data to illustrate the use of the package utilities. Conclusions: renz is a rigorous and yet easy-to-use software devoted to the analysis of kinetic data. This application has been designed to meet the needs of users who are not practicing enzymologists, but who need to accurately estimate the kinetic parameters of enzymes. The current software saves time and minimizes the risk of making mistakes or introducing biases due to uncorrected error propagation effects. [ABSTRACT FROM AUTHOR]

β-glucosidases (3.2.1.21) are present in all domains of living organisms, and their importance in a number of essential biological processes and industrial applications has been highlighted. They are interesting for biomass conversion because b-glucans are the world's largest source of biomass. For this reason, several fungal β-glucosidases have been investigated. The β-glucosidase gene of Aspergillus fumigatus, as well as its mutants D262E and W263F, were cloned and expressed in Pichia pastoris in this study. Their optimum temperature, pH, glucose tolerance, metal ion effect, and Vmax, km, and kcat were determined. The optimal temperature for recombinant β-glucosidase was 65 °C. For mutant D262E, there is an improvement in pH stability ranging from 4 to 6. As compared to the D262E mutant and recombinant β-glucosidase, mutant W263F showed a higher glucose tolerance and kcat. [ABSTRACT FROM AUTHOR]

Introduction: Children in families where there is substance misuse are at high risk of being removed from their parents' care. This study describes the characteristics of a community sample of parents who primarily smoke methamphetamine and their child or children's residential status. Design and Methods: Baseline data from a prospective study of methamphetamine smokers ('VMAX'). Participants were recruited via convenience, respondent‐driven and snowball sampling. Univariable and multivariable logistic regression analyses were used to estimate associations between parental status; fathers' or mothers' socio‐demographic, psychosocial, mental health, alcohol, methamphetamine use dependence, alcohol use and child or children's co‐residential status. Results: Of the 744 participants, 394 (53%) reported being parents; 76% (88% of fathers, 57% of mothers) reported no co‐resident children. Compared to parents without co‐resident children, parents with co‐resident children were more likely to have a higher income. Fathers with co‐resident children were more likely to be partnered and not to have experienced violence in the previous 6 months. Mothers with co‐resident children were less likely to have been homeless recently or to have accessed treatment for methamphetamine use. Discussion and Conclusions: The prevalence of non‐co‐resident children was much higher than previously reported in studies of parents who use methamphetamine; irrespective of whether in or out of treatment. There is a need for accessible support and services for parents who use methamphetamine; irrespective of their child or children's co‐residency status. Research is needed to determine the longitudinal impact of methamphetamine use on parents' and children's wellbeing and to identify how parents with co‐resident children (particularly mothers) can be supported. [ABSTRACT FROM AUTHOR]

Glucose Oxidase (GOX) is naturally produced by several microorganisms. In this article, the enzymes isolated from Pseudomonas and Actinomyces species were studied for enzyme characteristics, activity, stability and kinetic parameters. Enzyme extracted from Strain 1 (ES1), has shown optimum activity at 27°C and pH 5, half-life at 30°C. The enzyme was highly tolerant to AgNO3 (1.4mM) and less tolerant to NaCl; but was stable at 2.4mM of NaCl. 96% of activity was observed at 1.7mmol of Mg2+. 94% and 83% of activity were seen for Co and Cu when used as chelating agents. Denaturation of the enzyme occurred when DTAB was tested for its denaturing effect. 92% of enzyme activity was recorded by D-glucose when used as a substrate. The activation energy of 23.95 kJmol/l, 27°C, Vmax of 1.2U and Km of 6.91mM were recorded. Whereas Enzyme extracted from Strain 2 (ES2), reported optimum activity at 30°C and 5 pH, attained half-life after 30 minutes at 45°C. ES2 exhibited tolerance CoCl2 at 1.6mM and HgCl2 at 0.6mM. Stability of the enzyme observed at 3mM concentration for all salts used in experimentation. Enzyme activity of 98% for Mg2+ and 0% for Fe2+ were recorded among other metal ions. Enzyme activity of 87% for Co and 78% for Cu when used as chelating agents. Denaturation of the enzyme occurred when urea was used. 96% for D-glucose and 20% for sucrose were calculated as enzyme activity. Activation energy of 39.5 kJmol, 30°C, Vmax of 0.7U and Km of 72mM were recorded.

In order to study the role of agricultural exploitation in the activity of Alkaline phosphatase in the province of Diwaniyah, six sites that differ in some of their chemical, physical and biological properties were selected (Diwaniyah, Sunniya, Shamiya, Daghara, Afak and Al Budair). Three types of soils were chosen in each location, namely, orchard soil, field soil and jungle soil. The kinetic parameters of the enzymes maximum velocity (Vmax) and Michaelis constant (Km) were estimated in all study soils using increasing concentrations of the controlled substance. The results present that the highest efficacy of this enzyme is in the orchard soil of all sites except the location of the center Diwaniyah and Shamia. The average efficacy values ranged between (208.11 - 234.95) μg P-Nitro phenol. g-1 soil. 1 hour-1. The highest value of maximum velocity (Vmax) recorded at the field soil of Al-Shamia site (108.57) μg P-Nitro phenol. g-1 soil. 1 hour-1. The lowest value recorded at the soil of a jungle field at the Sunniya location (49.62) μg P-nitro phenol. g-1 soil. 1 hour-1. While the orchard soil of Al-Budair had the highest value of the Michaelis constant (Km) of (85.90) mM, the orchard soil of the Sunniya location had the lowest value (24.34) mM. .

Background: The lactate dehydrogenase C4 (LDH-C4) isoenzyme is an important enzyme involved in metabolic processes that are needed for spermatogenesis and sperm motility. Objectives: This study aims to assess the activity and kinetic parameters (maximum velocity, Vmax and Michaelis constant, Km) of LDH-C4 in fertile and infertile (azoospermia and oligospermia) men in Baghdad City, Iraq. Methods: A total of 120 participants (80 infertile and 40 healthy fertile men) were included in the current study. The patients were sub-grouped into: 40 infertile men with oligospermia, and 40 infertile men with azoospermia. The oligospermia patients were further subdivided into subgroups based on sperm count and motility. Semen samples were obtained by masturbation after 3-5 days abstain for seminal fluid analysis. The microscopic test included the assessment of the count, motility and morphology of the sperms. In addition, the coulometric assay was used for measuring the activity and kinetic parameters of LDH-C4 enzyme. Results: The activity of LDH-C4 is significantly higher in fertile men when compared with infertile subjects (fertile: 403.13±189.90, oligospermia: 110.01±58.13, azoospermia: 39.06±28.15; p≤0.01). Statistically significant differences in LDH-C4 activity were also noted among patients with oligospermia based on sperms count and motility. Based on sperms’ count in patients with oligospermia, a significantly higher LDH-C4 activity (p≤0.01) was noted in those with higher sperm count (10-15 million/ml) when compared to others who have lesser count. Significant elevation in enzyme activity (p≤0.01) was also observed in oligospermia patients with higher percentages of motile sperms when compared with others who have fewer percentages of motile sperms. Moreover, the highest Vmax value (0.483 mmol/L.min) and the lowest Km value (0.39 mmol/L.min) were recorded in fertile men. While, the lowest Vmax value (0.174 mmol/L.min) and the lowest Km value (0.75 mmol/L.min) were detected in azoospermia patients. Conclusions: Our results suggest that LDH-C4 is essential for the count and motility of sperm and may be considered as a therapeutic approach for infertility.

Nitrogen, the most demanded nutrient by coffee plants, has a rate of recovery from the soil of about 50%. Because of that high doses of nitrogenous fertilizers are used to reach high production, and consequently high amounts of N are lost to the environment. Knowing the kinetic parameters of nitrate (NO3-) absorption over the fruit development cycle is important as a mean of achieve more adjusted fertilizer doses and better recovery rates for the N applied as fertilizers. This study aimed determining the kinetic parameters of NO3- absorption in different development stages of fruits from adult coffee plants. The kinetic parameters Vmax and Km were determined in a low production year, at the pinhead (PH), rapid expansion (RE), grain filling (GF), and maturation (MT) stages. One month before each kinetics assay, lateral roots of eight plants were excavated and wrapped into non-woven fabrics grow cylinders filled in with vermiculite to produce absorbent roots. On the assay day, the roots were washed and immersed into a container with 1 L of 90 μmol L−1 NO3- solution. Sampling began one and a half hours after that, and was taken every hour over 7 h. Data on NO3- depletion were used to calculate the absorption kinetic parameters Vmax and Km. In a low production year the Vmax ranged from 0.14 to 0.72 μmol g−1 h−1 in a root fresh matter basis and Km from 6.47 to 50.31 μmol L−1. The Vmax values were highest at the PH and MT stages; the lowest absorption rate was recorded at GF and Km was lowest at RE. As at the RE stage of fruits Vmax shows a positive correlation with grain production, adequate nitrogen availability must be ensured before this phase to not to affect coming coffee production.

Understanding the kinetics of Zn absorption by roots and its effect on morphology of this organ is relevant for improving crop management, but still poorly studied for common beans. Therefore, an experiment was conducted in a hydroponic system with five initial concentrations of Zn (CZnI): 0.0; 1.0; 4.0; 16.0 and 48.0 µmol L-1. The experiment was installed with plants at V3 stage of development and aliquots of the solution collected over 24 h. The maximum absorption rate (Vmax), Michaelis-Menten constant (Km) and the absorption power (a) increased as a function of CZnI. The minimum concentration of Zn estimated for its absorption (Cmin) was at 0.0028 mg L-1. The influx of Zn (Imax) was higher in higher CZnI, 16,0 µmol L-1. Root length, root volume, root Zn content and Zn absorption efficiency increased with the increase of CZnI. Therefore, the increase of CZnI positively influenced kinetic parameters of root Zn absorption and common bean root morphology, characteristics that favor Zn absorption by roots and improves overall plant nutrition, favoring agronomical biofortification practices for Zn and other nutrients. [ABSTRACT FROM AUTHOR]

The objective of the present study was to assess if trypsin, a key enzyme involved in protein digestion, presents some kind of functional adaptations to seasonal changes in water temperature in freshwater fish. In order to test this hypothesis, individuals of two fish species Carassius gibelio (agastric) and Perca fluviatilis (gastric) were sampled in the basin of Chany Lake (Siberia, Russia) at two different seasons (spring and summer). Apparent kinetic parameters (Km and Vmax) were determined for both species and seasons at the actual pH values in fish guts, and at actual temperatures. Results showed a significant effect of both the species and sampling season on the apparent kinetic parameters of trypsin. In the case of Prussian carp, Km and Vmax were lower for each assayed temperature (for 5 and 15 °C the differences were significant) for fish sampled in summer when compared to those sampled in spring. In contrast, values of Km in perch tended to be lower in spring at 5 and 25 °C but these differences were not significant, while Vmax showed a significant decrease in summer samples. This suggests a sort of functional adaptation of the same trypsin enzymes to seasonal changes, oriented to maximize protein digestion under variable conditions. [ABSTRACT FROM AUTHOR]

Maximal velocity (Vmax) is a well established biomarker for the assessment of tissue redox status. There is scarce evidence, though, that it does not probably reflect sufficiently in vivo tissue redox profile. Instead, the Michaelis constant (Km) could more adequately image tissue oxidative stress and, thus, be a more physiologically relevant redox biomarker. Therefore, the aim of the present study was to side-by-side compare Vmax and Km of an antioxidant enzyme after implementing an in vivo set up that induces alterations in tissue redox status. Forty rats were divided into two groups including rats injected with blood plasma originating from rats that had previously swam until exhaustion and rats injected with blood plasma originating from sedentary rats. Tail-vein injections were performed daily for 21 days. Catalase Vmax and Km measured in gastrocnemius muscle were increased after administration of the exercise-conditioned plasma, denoting enhancement of the enzyme activity but impairment of its affinity for the substrate, respectively. These alterations are potential adaptations stimulated by the administered plasma pointing out that blood is an active fluid capable of regulating tissue homeostasis. Our findings suggest that Km adequately reflects in vivo modifications of skeletal muscle catalase and seems to surpass Vmax regarding its physiological relevance and biological interpretation. In conclusion, Km can be regarded as an in vivo-like biomarker that satisfactorily images the intracellular environment, as compared to Vmax that could be aptly parallelized with a biomarker that describes tissue oxidative stress in an in vitro manner.

Leaf CO2 uptake (A) in C4 photosynthesis is limited by the maximum apparent rate of PEPc carboxylation (Vpmax) at low intercellular [CO2] (ci) with a sharp transition to a ci‐saturated rate (Vmax) due to co‐limitation by ribulose‐1:5‐bisphosphate carboxylase/oxygenase (Rubisco) and regeneration of PEP. The response of A to ci has been widely used to determine these two parameters. Vmax and Vpmax depend on different enzymes but draw on a shared pool of leaf resources, such that resource distribution is optimized, and A maximized, when Vmax and Vpmax are co‐limiting. We collected published A/ci curves in 49 C4 species and assessed variation in photosynthetic traits between phylogenetic groups, and as a function of atmospheric [CO2]. The balance of Vmax‐Vpmax varied among evolutionary lineages and C4 subtypes. Operating A was strongly Vmax‐limited, such that re‐allocation of resources from Vpmax towards Vmax was predicted to improve A by 12% in C4 crops. This would not require additional inputs but rather altered partitioning of existing leaf nutrients, resulting in increased water and nutrient‐use efficiency. Optimal partitioning was achieved only in plants grown at pre‐industrial atmospheric [CO2], suggesting C4 crops have not adjusted to the rapid increase in atmospheric [CO2] of the past few decades. This study presents a data set of photosynthetic traits derived from published A‐ci curves measured in 49 C4 species. We determined that leaf photosynthetic resource allocation remains adapted to pre‐industrial atmospheric conditions and is not optimized for today's high [CO2] atmosphere. [ABSTRACT FROM AUTHOR]

Leaf CO2 uptake (A) in C4 photosynthesis is limited by the maximum apparent rate of PEPc carboxylation (Vpmax) at low intercellular [CO2] (ci) with a sharp transition to a ci‐saturated rate (Vmax) due to co‐limitation by ribulose‐1:5‐bisphosphate carboxylase/oxygenase (Rubisco) and regeneration of PEP. The response of A to ci has been widely used to determine these two parameters. Vmax and Vpmax depend on different enzymes but draw on a shared pool of leaf resources, such that resource distribution is optimized, and A maximized, when Vmax and Vpmax are co‐limiting. We collected published A/ci curves in 49 C4 species and assessed variation in photosynthetic traits between phylogenetic groups, and as a function of atmospheric [CO2]. The balance of Vmax‐Vpmax varied among evolutionary lineages and C4 subtypes. Operating A was strongly Vmax‐limited, such that re‐allocation of resources from Vpmax towards Vmax was predicted to improve A by 12% in C4 crops. This would not require additional inputs but rather altered partitioning of existing leaf nutrients, resulting in increased water and nutrient‐use efficiency. Optimal partitioning was achieved only in plants grown at pre‐industrial atmospheric [CO2], suggesting C4 crops have not adjusted to the rapid increase in atmospheric [CO2] of the past few decades. This study presents a data set of photosynthetic traits derived from published A‐ci curves measured in 49 C4 species. We determined that leaf photosynthetic resource allocation remains adapted to pre‐industrial atmospheric conditions and is not optimized for today's high [CO2] atmosphere. [ABSTRACT FROM AUTHOR]

Phenytoin follows Michaelis-Menten, a non-linear pharmacokinetics that occurs when drug molecules saturates the enzymes ability to metabolise the drug. When this occurs, steady state phenytoin serum concentration increases in a disproportionate manner after a dosage increase. General population data are usually used for the phenytoin dose calculation. However, many studies show that population pharmacokinetic parameters of phenytoin have high variations. Thus, use of specific local pharmacokinetic parameters for each population group in estimating individualised phenytoin dose can reduce phenytoin toxicity cases. This prospective, observational study was conducted to estimate a local Vmax and Km of phenytoin for adult epileptic patients in neurological ward and clinic at Hospital Pulau Pinang, Malaysia. All therapeutic drug monitoring of oral capsule phenytoin were studied in a three-month data collection period. Out of the 17 subjects in our study, there are 13 male subjects (76.47%) and 4 female subjects (23.53%). A total 11 Malay subjects (64.71%), 4 Chinese subjects (23.53%) and 2 Indian subjects (11.76%) were included. Median Vmax and Km were found to be 8.25 mg/kg/day and 3.80 mg/l. Male subjects have a higher Vmax (8.30 mg/kg/day) but a lower Km (3.3 mg/l). Chinese population has the highest Vmax (8.80 mg/kg/day). For Km, Indian population is the highest, with a value of 5.5 mg/l. From our study, gender does not correlate with Vmax and Km of phenytoin (p-value > 0.05). Ethnicity was also found to have no association with Vmax and Km (p-value > 0.05). Local Vmax (8.25 mg/kg/day) is higher and Km (3.8 mg/l) is lower when compared with standard Vmax (7 mg/kg/day) and Km (4 mg/l) obtained from Caucasian population. [ABSTRACT FROM AUTHOR]

The natural history of aortic stenosis (AS) progression, especially before severe AS development, is not well documented. We aimed to investigate the time course of peak aortic jet velocity (Vmax) and AS progression risk according to baseline Vmax, particularly whether there is a Vmax threshold. In a retrospective multicenter cohort study of patients on hemodialysis with aortic valve calcification, we investigated the time series of Vmax and the relationship between the baseline Vmax and progression to severe AS by analyzing longitudinal echocardiographic data. Among 758 included patients (mean age, 71 years; 65% male), patients with Vmax <1.5, 1.5–1.9, 2.0–2.4, 2.5–2.9, and 3.0–3.9 m/s were 395 (52%), 216 (29%), 85 (11%), 39 (5.1%), and 23 (3.0%), respectively. The Vmax slope was gradual (mean 0.05–0.07 m/s/year) at Vmax <2 m/s, but steeper (mean 0.13–0.21 m/s/year) at Vmax ≥2 m/s. During a median 3.2-year follow-up, 52 (6.9%) patients developed severe AS. While patients with Vmax <2 m/s rarely developed severe AS, the risk of those with Vmax ≥2 m/s increased remarkably with an increasing baseline Vmax; the adjusted incidence rates in patients with Vmax <1.5, 1.5–1.9, 2.0–2.4, 2.5–2.9, and 3.0–3.9 m/s were 0.59, 0.57, 4.25, 13.8, and 56.1 per 100 person-years, respectively; the adjusted hazard ratio per 0.2 m/s increase in the baseline Vmax was 1.49 (95% confidence interval: 1.32–1.68) when Vmax ≥2 m/s. The risk of progression to severe AS increased with the baseline Vmax primarily at ≥2 m/s; a Vmax threshold of 2 m/s was observed. [Display omitted] • The time course of Vmax and AS progression risk were investigated in hemodialysis patients with aortic valve calcification. • The slope of Vmax was gradual below 2 m/s but became steeper above 2 m/s. • The risk of progression to severe AS increased markedly with an increasing baseline Vmax at Vmax ≥2 m/s. • The risk increased 1.5-fold for every 0.2 m/s increase in Vmax at Vmax ≥2 m/s. [ABSTRACT FROM AUTHOR]

Phenytoin follows Michaelis-Menten, a non-linear pharmacokinetics that occurs when drug molecules saturates the enzymes ability to metabolise the drug. When this occurs, steady state phenytoin serum concentration increases in a disproportionate manner after a dosage increase. General population data are usually used for the phenytoin dose calculation. However, many studies show that population pharmacokinetic parameters of phenytoin have high variations. Thus, use of specific local pharmacokinetic parameters for each population group in estimating individualised phenytoin dose can reduce phenytoin toxicity cases. This prospective, observational study was conducted to estimate a local Vmax and Km of phenytoin for adult epileptic patients in neurological ward and clinic at Hospital Pulau Pinang, Malaysia. All therapeutic drug monitoring of oral capsule phenytoin were studied in a three-month data collection period. Out of the 17 subjects in our study, there are 13 male subjects (76.47%) and 4 female subjects (23.53%). A total 11 Malay subjects (64.71%), 4 Chinese subjects (23.53%) and 2 Indian subjects (11.76%) were included. Median Vmax and Km were found to be 8.25 mg/kg/day and 3.80 mg/l. Male subjects have a higher Vmax (8.30 mg/kg/day) but a lower Km (3.3 mg/l). Chinese population has the highest Vmax (8.80 mg/kg/day). For Km, Indian population is the highest, with a value of 5.5 mg/l. From our study, gender does not correlate with Vmax and Km of phenytoin (p-value > 0.05). Ethnicity was also found to have no association with Vmax and Km (p-value > 0.05). Local Vmax (8.25 mg/kg/day) is higher and Km (3.8 mg/l) is lower when compared with standard Vmax (7 mg/kg/day) and Km (4 mg/l) obtained from Caucasian population. [ABSTRACT FROM AUTHOR]

Glucose Oxidase (GOX) is naturally produced by several microorganisms. In this article, the enzymes isolated from Pseudomonas and Actinomyces species were studied for enzyme characteristics, activity, stability and kinetic parameters. Enzyme extracted from Strain 1 (ES1), has shown optimum activity at 27°C and pH 5, half-life at 30°C. The enzyme was highly tolerant to AgNO3 (1.4mM) and less tolerant to NaCl; but was stable at 2.4mM of NaCl. 96% of activity was observed at 1.7mmol of Mg2+. 94% and 83% of activity were seen for Co and Cu when used as chelating agents. Denaturation of the enzyme occurred when DTAB was tested for its denaturing effect. 92% of enzyme activity was recorded by D-glucose when used as a substrate. The activation energy of 23.95 kJmol/l, 27°C, Vmax of 1.2U and Km of 6.91mM were recorded. Whereas Enzyme extracted from Strain 2 (ES2), reported optimum activity at 30°C and 5 pH, attained half-life after 30 minutes at 45°C. ES2 exhibited tolerance CoCl2 at 1.6mM and HgCl2 at 0.6mM. Stability of the enzyme observed at 3mM concentration for all salts used in experimentation. Enzyme activity of 98% for Mg2+ and 0% for Fe2+ were recorded among other metal ions. Enzyme activity of 87% for Co and 78% for Cu when used as chelating agents. Denaturation of the enzyme occurred when urea was used. 96% for D-glucose and 20% for sucrose were calculated as enzyme activity. Activation energy of 39.5 kJmol, 30°C, Vmax of 0.7U and Km of 72mM were recorded. [ABSTRACT FROM AUTHOR]

Copyright of Environmental Science & Technology (10036504) is the property of Editorial Board of Environmental Science & Technology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

A comparison among VMIX, VMAX and the adapted step-down Sullivan et al. (SE) tests for covariance matrix under bivariate normal assumption is presented. The type I error and power estimates were obtained by using Monte Carlo simulation under different scenarios with respect to covariance and correlation structures. In general, VMIX was more powerful than VMAX being SE more powerful than both, with few exceptions. SE test is more general since it can be used for normal and non-normal data, with no restriction with respect to the pattern of the covariance matrix shifts, and for larger dimension than the bivariate case. [ABSTRACT FROM AUTHOR]

To study the characteristics of two malate dehydrogenases (MDHs), their coding genes MIMDH1 and MIMDH2 were cloned and expressed in Escherichiacoli BL21 cells. The molecular weights of both recombinant enzymes partially purified using Ni-NTA affinity chromatography were 32 kDa, and the specific activities of purified MIMDH1 and MIMDH2 were 329.3 and 241.3 U/mg protein, respectively. The optimal temperatures for MIMDH1 and MIMDH2 activities were 55 and 30oC, respectively, with 70% MIMDH1 activity and ≥70% MIMDH2 activity remaining after 30 min incubation at 45oC. Addition of 2 mM Zn2+ enhanced MIMDH1 activity, whereas the addition of other metal ions resulted in different degrees of inhibition. The inhibitory effect of Co2+ was most pronounced on MIMDH1, reaching 82.8% inhibition. Addition of 2 mM Ba2+ or Mn2+ increased MIMDH2 activity, the addition of other metal ions resulted in different degrees of inhibition. Furthermore, MIMDH1 was stable at pH range of 7.5–8.5, with optimal activity observed at pH of 8.0. MIMDH2 showed similar stability at the same pH range, but was optimal at pH of 7.5. The Vmax and KM values for recombinant MIMDH1 and MIMDH2 catalyzing the reduction of oxaloacetate to malate were 17.66 µmol mg–1min–1 and 0.541 mM, 15.59 µmol mg–1min–1 and 0.683 mM, respectively. [ABSTRACT FROM AUTHOR]

A novel temperature stable alkaline protease yielding bacteria was isolated from the soils of Dachigam National Park, which is known to be inhabited by a wide variety of endemic plant and animal species of Western Himalaya. This high-potential protease producing isolate was characterized and identified as Bacillus amyloliquefaciens strain HM48 by morphological, Gram’s staining and biochemical techniques followed by molecular characterization using 16S rRNA approach. The extracellular protease of B. amyloliquefaciens HM48 was purified by precipitating with ammonium sulfate (80%), followed by dialysis and Gel filtration chromatography increasing its purity by 5.8-fold. The SDS–PAGE analysis of the purified enzyme confirmed a molecular weight of about ≈25 kDa. The enzyme displayed exceptional activity in a broad temperature range (10–90 °C) at pH 8.0, retaining its maximum at 70 °C, being the highest reported for this proteolytic Bacillus sp., with KM and Vmax of 11.71 mg/mL and 357.14 µmol/mL/min, respectively. The enzyme exhibited remarkable activity and stability against various metal ions, surfactants, oxidizing agent (H2O2), organic solvents and displayed outstanding compatibility with widely used detergents. This protease showed effective wash performance by exemplifying complete blood and egg-yolk stains removal at 70 °C and efficiently disintegrated chicken feathers making it of vital importance for laundry purpose and waste management. For functional analysis, protease gene amplification of strain HM48 yielded a nucleotide sequence of about 700 bp, which, when checked against the available sequences in NCBI, displayed similarity with subtilisin-like serine protease of B. amyloliquefaciens. The structure of this protease and its highest-priority substrate β-casein was generated through protein modeling. These protein models were validated through futuristic algorithms following which protein–protein (protease from HM48 and β-casein) docking was performed. The interaction profile of these proteins in the docked state with each other was also generated, shedding light on their finer details. Such attributes make this thermally stable protease novel and suitable for high-temperature industrial and environmental applications.

Halogen bonds have recently gained attention in life sciences and drug discovery. However, it can be difficult to harness their full potential, when newly introducing them into an established hit or lead structure by molecular design. A possible solution to overcome this problem is the use of halogen-enriched fragment libraries (HEFLibs), which consist of chemical probes that provide the opportunity to identify halogen bonds as one of the main features of the binding mode. Initially, we have suggested the HEFLibs concept when constructing a focused library for finding p53 mutant stabilizers. Herein, we broaden and extent this concept aiming for a general HEFLib comprising a huge diversity of binding motifs and, thus, increasing the applicability to various targets. Using the construction principle of feature trees, we represent each halogenated fragment by treating all simple to complex substituents as modifiers of the central (hetero)arylhalide. This approach allows us to focus on the proximal binding interface around the halogen bond and, thus, its integration into a network of interactions based on the fragment's binding motif. As a first illustrative example, we generated a library of 198 fragments that unifies a two-fold strategy: Besides achieving a diversity-optimized basis of the library, we have extended this “core” by structurally similar “satellite compounds” that exhibit quite different halogen bonding interfaces. Tuning effects, i.e., increasing the magnitude of the σ-hole, can have an essential influence on the strength of the halogen bond. We were able to implement this key feature into the diversity selection, based on the rapid and efficient prediction of the highest positive electrostatic potential on the electron isodensity surface, representing the σ-hole, by VmaxPred.

Abstract Recent developments of enzyme-based decomposition models highlight the importance of enzyme kinetics with warming, but most modeling exercises are based on studies with a step-wise warming. This approach may mask the effect of temperature in controlling in-situ activities as in most ecosystems the rate of warming is more gradual than these step warming studies. We conducted an experiment to test the effects of contrasting warming rates on the kinetics of carbon (C), nitrogen (N), and phosphorus (P) degradation enzymes in subtropical peat soils. We also wanted to evaluate if the stoichiometry of enzyme kinetics shifts under contrasting warming rates and if so, how does it relate to the stoichiometry in microbial biomass. Contrasting warming rates altered microbial biomass stoichiometry leading to differing patterns of microbial demand for C vs. nutrient (N and P) and enzyme expression following the optimum foraging strategy. Activity (higher Vmax) and efficiency (lower Km) of C acquisition enzymes were greater in the step treatment; however, expressions of nutrient (N and P) acquiring enzymes were enhanced in the ramp treatment at the end of the experiment. In the step treatment, there was a typical pattern of an initial peak in the Vmax and drop in the Km for all enzyme groups followed by later adjustments. On the other hand, a consistent increase in Vmax and decline in Km of all enzyme groups were observed in the ramp treatment. These changes were sufficient to alter microbial identity (as indicated by enzyme Km and biomass stoichiometry) with two apparently different endpoints under contrasting warming rates. This observation resembles the concept of alternate stable states and highlights a need for improved representation of warming effects on enzymes in decomposition models. Using peat soils of Florida Everglades, here we have demonstrated that contrasting warming rates can influence the dynamics of microbial and enzymatic kinetics. Hence, we suggest that future laboratory and field warming studies could consider our approach to accurately represent microbial and enzymatic kinetics in biogeochemical models. Highlights • Contrasting warming rates altered microbial stoichiometry & enzyme kinetics. • Fast warming had higher Vmax and lower Km of C-related enzymes. • Slow warming had increased Vmax & lower Km of nutrient-related enzymes. • Enzyme Vmax & Km followed different patterns under contrasting warming rates. • Enzyme kinetics followed optimal foraging strategy and ecological stoichiometry theory. [ABSTRACT FROM AUTHOR]

Background: Enzyme activities can be utilized as good indicators for diagnostic purposes. Thus, determination of kinetic parameters of the target enzymes facilitates the interpretation of the effect that may be occurred on activities of the studied enzymes. The aim of this study was to determine of Vmax and Km of salivary lactate dehydrogenase (LDH) enzyme in presence and absence of the titanium dioxide nanoparticles (TiO2 NPs). Methods: The structural properties of (TiO2 NPs) have been determined using UV-Vis spectrophotometer and SEM. TiO2 NPs and sharp round peaks around 220nm with a spherical shape and average particle size ˂50 nm. Results: Km and Vmax values of LDH in absence and presence of TiO2 NPs were found to be 0.02mmol/L, 15.6U/L and 0.01mmol/L, 17.2U/L, respectively. Conclusion: our results confirmed the activating effects of TiO2 NPs on salivary LDH activity. The effects of TiO2 NPs on salivary LDH activity may be due to the activity of this NPs type and/or the conformational changes that can be occurred on the protein structure of the enzyme after interaction with these NPs. [ABSTRACT FROM AUTHOR]

This study looks at using partially purified peroxidase extracted from peels of sweet lime (Citrus limetta) for decolorizing textile industry effluent. The ideal pH and thermal conditions of the enzyme were 7 and 35°C. The Km and Vmax for guaiacol were 0.66 mM and 6666 μmol/mL/min, respectively. We found that sweet lime peroxidase was very effective in decolorizing textile industry effluent. Almost complete decolorization (>99 %) of effluent was attained at a pH of 5.0, temperature of 55°C, H2O2 concentration of 2 mM, and enzyme dose of 40 U/mL within 60 minutes of incubation. The effluent was also analysed in terms of physicochemical parameters before and after treatment with sweet lime peroxidase. The reduction in toxicity after the enzymatic treatment was evidenced by chemical oxygen demand (COD) and total suspended solids (TSS) values. [ABSTRACT FROM AUTHOR]

Abstract Fungal metabolic rates could increase under climate warming but may be counteracted by mechanisms of physiological acclimation and evolutionary adaptation. We hypothesized that Vmax and Km parameters of Neurospora discreta extracellular enzymes would acclimate to warmer temperatures through compensatory mechanisms. We also predicted that evolution under warmer temperatures would alter enzyme parameters and fungal respiration through adaptive mechanisms. In contrast to these predictions, growth at higher temperature (22 °C versus 16 °C) increased the temperature-corrected Vmax of three enzymes. The carbon substrate used for fungal growth (lignin versus sucrose) had a much greater impact on enzyme Vmax than temperature. Following experimental evolution, the enzymatic parameters of Neurospora strains did not adapt to higher temperatures as hypothesized; rather, enzyme Vmax values were unaffected, and respiration rates increased. Together, these results suggest that physiological and evolutionary mechanisms are unlikely to counteract soil carbon losses driven by saprotrophic fungi under climate warming. Highlights • We tested for physiological acclimation of extracellular enzyme activity. • There was little evidence for enzyme acclimation to increased growth temperature. • We tested for temperature adaptation using experimental evolution. • No evidence for temperature adaptation of enzyme activity. • Respiration rates increased in warm-evolved strains contrary to adaption hypothesis. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Harbin Institute of Technology. Social Sciences Edition / Haerbin Gongye Daxue Xuebao. Shehui Kexue Ban is the property of Harbin Institute of Technology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In this article, we propose the two control charts, i.e. the 'VMAX Group Runs' (VMAX-GR) and 'VMAX Modified Group Runs' (VMAX-MGR) control charts based on the bivariate normal processes, for monitoring the covariance matrix. The proposed charts give the faster detection of a process change and have better diagnostic feature. It is verified that the VMAX-GR and the VMAX-MGR charts give a significant reduction in the out-of-control 'Average Run Length' (ARL) in the zero state, as well as in the steady state, as compared to the synthetic control chart based on the VMAX statistic and the generalized variance ∣S∣ chart. [ABSTRACT FROM AUTHOR]

Marine microorganisms and their extracellular enzymes (ECEs) play an important role in the remineralization of organic material by hydrolyzing high-molecular-weight substrates to sizes sufficiently small to be transported through cell membrane, yet the diversity of the enzyme-producing bacteria and the types of ECEs involved in the degradation process are largely unknown. In this work, we investigated the diversity of cultivable bacteria and their ECEs and the potential activities of aminopeptidase in the water column at eight different depths of the New Britain Trench. There was a great diversity of cultivable bacteria and ECEs, and depth appears an important driver of the diversity. The 16S rRNA sequence analysis revealed that the cultivable bacteria were affiliated mostly with the phyla Proteobacteria and Actinobacteria, and the predominant genera were Pseudoalteromonas (62.7%) and Halomonas (17.3%). Moreover, 70.7% of the isolates were found to produce hydrolytic zone on casein and gelatin plates, in which Pseudoalteromonas was the predominant group, exhibiting relatively high protease production. Inhibitor analysis showed that the extracellular proteases from the isolated bacteria were serine proteases in the surface water and metalloproteases in the deep water. Meanwhile, the Vmax and Km of aminopeptidase exhibited a maximum in the surface water and low values in the deep bathy- and abyssopelagic water, indicating lower rates of hydrolysis and higher substrate affinity in the deeper waters. These results shed new insights into the diversity of the cultivable bacteria and bacterial ECEs and their likely biogeochemical functions in the trench environment.

Background: With emergence of multidrug-resistant strains of Staphylococcus aureus, there is an urgent need for the development of new antimicrobials which are narrow and pathogen specific. In this context, pyruvate kinase (PK) an important enzyme in the glycolysis, which catalyses the formation of pyruvate which is the key intersection in the network of metabolic pathways was isolated and purified from Staphylococcus aureus ATCC12600. Methods: Purification steps included 10%-20% ammonium sulphate fractionation, diethyl aminoethyl cellulose ion exchange chromatography followed by gel filtration on Sephadex G-100. The pure PK molecular weight was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Km and Vmax for the PK was demonstrated. Results: The pure PK obtained from Sephadex G-100 gel filtration column exhibited Km of 0.78+0.18 µM and Vmax 76.47+0.82 µM NADH/mg/min and molecular weight of 250 kDa in solution. However, in SDS-PAGE showed single band with a molecular weight of 63 kDa confirming the homotetramer nature. In all steps of purification the Km remained constant indicating presence of only one kind of enzyme. The PK gene searched in the genomic sequences of Staphylococcus aureus also confirmed the same. Interpretation and conclusions: In Staphylococcus aureus presence of only one kind of PK unlike in other Gram positive bacteria exhibiting distinct differences in enzyme kinetics. This enzyme also showed the functionality of PK is found to be different from its human host. Therefore, PK probably is regarded as an ideal drug target in the development of new potent antimicrobials.

Marine microorganisms and their extracellular enzymes (ECEs) play an important role in the remineralization of organic material by hydrolyzing high-molecular-weight substrates to sizes sufficiently small to be transported through cell membrane, yet the diversity of the enzyme-producing bacteria and the types of ECEs involved in the degradation process are largely unknown. In this work, we investigated the diversity of cultivable bacteria and their ECEs and the potential activities of aminopeptidase in the water column at eight different depths of the New Britain Trench. There was a great diversity of cultivable bacteria and ECEs, and depth appears an important driver of the diversity. The 16S rRNA sequence analysis revealed that the cultivable bacteria were affiliated mostly with the phyla Proteobacteria and Actinobacteria, and the predominant genera were Pseudoalteromonas (62.7%) and Halomonas (17.3%). Moreover, 70.7% of the isolates were found to produce hydrolytic zone on casein and gelatin plates, in which Pseudoalteromonas was the predominant group, exhibiting relatively high protease production. Inhibitor analysis showed that the extracellular proteases from the isolated bacteria were serine proteases in the surface water and metalloproteases in the deep water. Meanwhile, the Vmax and Km of aminopeptidase exhibited a maximum in the surface water and low values in the deep bathy- and abyssopelagic water, indicating lower rates of hydrolysis and higher substrate affinity in the deeper waters. These results shed new insights into the diversity of the cultivable bacteria and bacterial ECEs and their likely biogeochemical functions in the trench environment. [ABSTRACT FROM AUTHOR]

Copyright of Food Research & Development is the property of Food Research & Development Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)