Non-competitive Inhibition Model Research Articles

Optimizing pectin lyase production using the one-factor-at-a-time method and response surface methodology.

Pectinases are commonly industrially synthesized by molds. This study aimed to optimize pectin lyase synthesis by a bacterium, Pseudomonas fluorescens, using both the one-factor-at-a-time (OFAT) method and response surface methodology. First, on optimization of pectin lyase fermentation by the OFAT method, the effects of pectin, peptone, yeast extract, (NH4)2SO4, pH, and salts were investigated. The highest pectin lyase activity was determined to be 28.63U/mL at pH 8, 30°C, with 1% (w/v) pectin and 0.14% (w/v) (NH4)2SO4 concentration at the 90th hour. The effect of substrate inhibition on the microbial growth was also investigated, and the results showed that the process can be described by noncompetitive inhibition model. The values of kinetic constants were determined as µm=0.175h-1, KS=6.931g/L, and, KI=6.932g/L by nonlinear regression analysis. It was reported that pectin lyase enzymes exhibited peak activity at 50°C and pH 8. Finally, response surface methodology (RSM) was utilized to optimize pH, concentrations of ammonium sulfate, and pectin, which were chosen as independent variables. The interactions between these variables were also examined. According to RSM, the optimum values of the parameters to achieve a maximum pectin lyase activity of 35.62U/mL were determined to be pH 7.97, 1.25% (w/v) pectin concentration, and 0.25% (w/v) (NH4)2SO4 concentration.

Performance evaluation of Trichoderma reseei in tolerance and biodegradation of diuron herbicide in agar plate, liquid culture and solid-state fermentation.

The present study evaluated the performance of the fungus Trichoderma reesei to tolerate and biodegrade the herbicide diuron in its agrochemical presentation in agar plates, liquid culture, and solid-state fermentation. The tolerance of T. reesei to diuron was characterized through a non-competitive inhibition model of the fungal radial growth on the PDA agar plate and growth in liquid culture with glucose and ammonium nitrate, showing a higher tolerance to diuron on the PDA agar plate (inhibition constant 98.63 mg L-1) than in liquid culture (inhibition constant 39.4 mg L-1). Diuron biodegradation by T. reesei was characterized through model inhibition by the substrate on agar plate and liquid culture. In liquid culture, the fungus biotransformed diuron into 3,4-dichloroaniline using the amide group from the diuron structure as a carbon and nitrogen source, yielding 0.154mg of biomass per mg of diuron. A mixture of barley straw and agrolite was used as the support and substrate for solid-state fermentation. The diuron removal percentage in solid-state fermentation was fitted by non-multiple linear regression to a parabolic surface response model and reached the higher removal (97.26%) with a specific aeration rate of 1.0 vkgm and inoculum of 2.6 × 108 spores g-1. The diuron removal in solid-state fermentation by sorption on barley straw and agrolite was discarded compared to the removal magnitude of the biosorption and biodegradation mechanisms of Trichoderma reesei. The findings in this work about the tolerance and capability of Trichoderma reesei to remove diuron in liquid and solid culture media demonstrate the potential of the fungus to be implemented in bioremediation technologies of herbicide-polluted sites.

Rehmannioside A inhibits the activity of CYP3A4, 2C9 and 2D6 in vitro

To assess the effect of Rehmannioside A on CYP450s activity and to estimate its inhibitory properties. The effect of Rehmannioside A on the activity of major CYP450s in human liver microsomes (HLMs) was assessed with the corresponding substrates and marker reactions, and compared with a blank control and the respective inhibitors. Suppression of CYP3A4, 2C9 and 2D6 was assessed by the dose-dependent assay and fitted with non-competitive or competitive inhibition models. The inhibition of CYP3A4 was determined in a time-dependent manner. Rehmannioside A suppressed the activity of CYP3A4, 2C9, and 2D6 with IC50 values of 10.08, 12.62, and 16.43 μM, respectively. Suppression of CYP3A4 was fitted to a non-competitive model with Ki value of 5.08 μM, whereas CYP2C9 and 2D6 were fitted to a competitive model with Ki values of 6.25 and 8.14 μM. Additionally, the inhibitory effect on CYP3A4 was time-dependent with KI value of 8.47 μM−1 and a Kinact of 0.048 min−1. In vitro suppression of CYP3A, 2C9 and 2D6 by Rehmannioside A indicated that Rehmannioside A or its source herbs may interact with drugs metabolised by these CYP450s, which could guide the clinical application.

Acute responses of bio-denitrification to short-term clopyralid exposure: Kinetic analysis and biological mechanisms

The mass production and extensive application of clopyralid (CLP) have attracted much attention regarding its environmental effect in recent decades. However, studies on the acute responses and the underlying inhibition mechanisms of denitrifiers to CLP exposure are scarce. Herein, the crucial factors in denitrification were quantified through kinetics, metabolic activity analysis, and structural equation modeling (SEM). The acute response mechanisms of denitrifiers under short-term CLP exposure were further proposed according to the quantitative correlation of denitrification performance with C and N metabolism, ETS activity, and oxidative stress. The modified non-competitive inhibition model fitted well to describe the inhibitory characteristics of CLP on the nitrate reduction (R2 = 0.99), which was significantly inhibited with increasing CLP exposure concentrations. Short-term exposure to CLP could inhibit C and N metabolism and electron transport by inducing oxidative stress during denitrification. Quantitative PCR (qPCR) analysis demonstrated that CLP restrained the abundance of narG and nirS, suggesting that down-regulation of denitrification enzyme expression was highly related to the reduction in nitrogen conversion. Moreover, SEM indicated that ETS activity and NAR were the major factors in predicting NO3− reduction rate (DNR) and were mainly influenced by NADH and reactive oxygen species (ROS). Consistently, NADH and NIR were closely correlated with total nitrite accumulation (NIA). Short-term CLP exposure could affect denitrification performance by directly affecting microbial physicochemical properties or indirectly interfering with the abundance of denitrifying functional genes and metabolic activity through oxidative stress. This study provided new insights into acute responses of denitrifiers to short-term CLP exposure.

Effects of Simvastatin on the Metabolism of Vonoprazan in Rats Both in vitro and in vivo.

PurposeTo study the potential drug–drug interactions between simvastatin and vonoprazan and to provide the scientific basis for rational use of them in clinical practice.MethodsAn incubation system was established with rat liver microsomes, and the main metabolite of vonoprazan M-I was detected by UPLC-MS/MS. The IC50 value of simvastatin was then calculated and its inhibitory mechanism against vonoprazan was also analyzed. Twelve SD rats were randomly divided into 2 groups, then they were given simvastatin or saline for 2 weeks continuously. On the day of the experiment, both groups were intragastrically administered with vonoprazan once, followed by the collection of blood at different time points. Then the plasma concentration of vonoprazan and M-I in rats were detected by UPLC-MS/MS.ResultsIn vitro experiments revealed that simvastatin could inhibit the metabolism of vonoprazan, and its inhibition type belonged to the mixed non-competitive and competitive inhibition model. In vivo experiments in rats demonstrated that the area under concentration time curve (AUC) of vonoprazan was decreased but the clearance (CLz/F) of it was increased in the simvastatin administrated group, as compared to those of the control group. However, M-I in simvastatin treated group exhibited the higher AUC and lower CLz/F values compared to those in the control group. These data indicated that multiple doses of simvastatin administration could reduce the plasma concentration of vonoprazan and accelerate its metabolic rate in rats.ConclusionSimvastatin could inhibit the metabolism of vonoprazan in vitro but multiple doses of simvastatin exhibited the opposite effect In vivo. Altogether, our data indicated that an interaction existed between simvastatin and vonoprazan and additional cares might be taken when they were co-administrated in clinic.

The Inhibitory Effect of Free Nitrous Acid and Free Ammonia on the Anoxic Phosphorus Uptake Rate of Polyphosphate-Accumulating Organisms

The purpose of this study is to investigate the effect of free nitrous acid (FNA) and free ammonia (FA) on the anoxic phosphorus uptake rate (PUR) of polyphosphate-accumulating organisms (PAOs) via the utilization of nitrite. With this goal, upon developing a PAO-enriched culture in a sequential batch reactor, a series of batch experiments were conducted to examine the effects of nitrite and ammonium on the anoxic phosphorus uptake rate at different pH levels. According to the results, both free nitrous acid and free ammonia were found to inhibit anoxic PUR to a degree similar to their respective effects on aerobic PUR reported in previous studies, suggesting that phosphorus removal via the anoxic pathway may be just as susceptible as that via the aerobic pathway. The effect of FNA on anoxic PUR is optimally described by a non-competitive inhibition model with a KiFNA value of 1.6 μg N L−1, while the Levenspiel model with an SFA* value of 37 mg N L−1 provided the best fit for the FA effect on PAOs anoxic activities. The results of this study provide new insights regarding the viability of EBPR under high nitrogen loading conditions.

In vitro evaluation of the inhibition potential of echinacoside on human cytochrome P450 isozymes

BackgroundEchinacoside (ECH) possesses a wide range of biological activity. This present study analyzes the effect of ECH on cytochrome P450 isozymes (CYPs) activities of human liver microsomes.MethodsThe effect of ECH on CYPs enzyme activities were studied using the enzyme-selective substrates phenacetin (1A2), chlorzoxazone (2E1), S-mephenytoin (2C19), testosterone (3A4), coumarin (2A6), diclofenac (2C9), paclitaxel (2C8), and dextromethorphan (2D6). The IC50 values for CYP1A2, CYP2E1, CYP2C19, and CYP3A4 isoforms were examined to express the strength of inhibition. Further, the inhibition of CYPs was checked for time-dependent or not, and then fitted with competitive or non-competitive inhibition models. The corresponding parameters were also obtained.ResultsECH caused inhibitions on CYP1A2, CYP2E1, CYP2C19 and CYP3A4 enzyme activities in HLMs with IC50 of 21.23, 19.15, 8.70 and 55.42 μM, respectively. The obtained results showed that the inhibition of ECH on CYP3A4 was time-dependent with the KI/Kinact value of 6.63/0.066 min− 1·μM− 1. Moreover, ECH inhibited the activity of CYP1A2 and CYP2E1 via non-competitive manners (Ki = 10.90 μM and Ki = 14.40 μM, respectively), while ECH attenuated the CYP2C19 activity via a competitive manner (Ki = 4.41 μM).ConclusionsThe results of this study indicate that ECH inhibits CYP1A2, CYP2E1, CYP2C19 and CYP3A4 activities in vitro. In vivo and clinical studies are warranted to verify the relevance of these interactions.

Inhibitory effects of Thai herbal extracts on the cytochrome P450 3A-mediated the metabolism of gefitinib, lapatinib and sorafenib

Herbal products are widely used in cancer patients via co−administration with chemotherapy. Previous studies have demonstrated that pharmacokinetic interactions between herbs and anticancer drugs exist due to inhibition of drug−metabolizing enzymes, particularly cytochrome P450s (CYPs). The aim of this study was to determine the inhibitory effects of Andrographis paniculata, Curcuma zedoaria, Ganoderma lucidum, Murdannia loriformis and Ventilago denticulata extracts on the metabolism of gefitinib, lapatinib and sorafenib. The activities of CYP3A in human liver microsome on the metabolism of gefitinib, lapatinib and sorafenib in the absence and presence of Thai herbal extracts were assayed using high-performance liquid chromatography analysis. Curcuma zedoaria extract potently inhibited CYP3A−mediated lapatinib and sorafenib metabolism with IC50 values of 4.18 ± 3.20 and 7.59 ± 1.23 μg/mL, respectively, while the metabolism of gefitinib was strongly inhibited by Murdannia loriformis and Ventilago denticulata extracts with IC50 values of 7.53 ± 2.87 and 7.06 ± 1.23 μg/mL, respectively. Andrographis paniculata and Ganoderma lucidum extracts had less effect on the metabolism of the tested anticancers (IC50 values >10 μg/mL). In addition, kinetic analysis of the ability of Curcuma zedoaria extract to inhibit CYP3A-mediated metabolism of anticancer drugs was best described by the noncompetitive and competitive inhibition models with Ki values of 20.08 and 11.55 μg/mL for the metabolism of gefitinib and sorafenib, respectively. The present study demonstrated that there were potential pharmacokinetic interactions between tyrosine kinase inhibitors and herbal extracts.

Incremental Model Identification of Bio-processes from Data: Application to Microbial Production of Hyaluronic Acid

The development of reliable kinetic models of bioprocesses from data is a challenging task. In this work, a systematic approach to develop a kinetic model of bioprocess involving single biomass from concentration data is proposed without imposing any kinetic model a priori. The proposed incremental model identification approach decomposes the model-building task into a set of sub-tasks such as determining the yield coefficients and maintenance coefficient, specific growth rate structure identification, and parameter estimation. It is shown that the proposed approach allows identifying the mechanism of product formation. The proposed approach is applied to the microbial production of Hyaluronic acid (HA), an important biopolymer, using a recombinant Lactococcus lactis MKG6. An unstructured kinetic model is developed for the HA production from data. It is shown that HA production is a growth-associated process. Further, the specific growth rate of HA production is identified from a set of rate candidates. It is revealed that the specific growth rate in the HA production follows the non-competitive HA inhibition model. The parameters obtained by the incremental identification are further refined to obtain statistically optimal estimates using the simultaneous model identification. Validation of the identified kinetic model of HA production on new experimental data shows that the proposed approach leads to a reliable kinetic model with the optimal parameter estimates.

Assessing and modeling nitrite inhibition in microalgae-bacteria consortia for wastewater treatment by means of photo-respirometric and chlorophyll fluorescence techniques

Total nitrite (TNO2 = HNO2 + NO−2) accumulation due to the activity of ammonia-oxidizing bacteria (AOB) was monitored in microalgae-bacteria consortia, and the inhibitory effect of nitrite/free nitrous acid (NO2-N/FNA) on microalgae photosynthesis and inhibition mechanism was studied. A culture of Scenedesmus was used to run two sets of batch reactors at different pH and TNO2 concentrations to evaluate the toxic potential of NO2-N and FNA. Photo-respirometric tests showed that NO2-N accumulation has a negative impact on net oxygen production rate (OPRNET). Chlorophyll a fluorescence analysis was used to examine the biochemical effects of NO2-N stress and the mechanism of NO2-N inhibition. The electron transport rate (ETR), non-photochemical quenching (NPQ), and JIP-test revealed that the electron transport chain between Photosystems II and I (PS II and PS I) was hindered at NO2-N concentrations above 25 g N m−3. Electron acceptor QA was not able to reoxidize and could not transfer electrons to the next electron acceptor, QB, accumulating P680+ (excited PS II reaction center) and limiting oxygen production.A semi-continuous reactor containing a Scenedesmus culture was monitored by photo-respirometry tests and Chlorophyll a fluorescence to calibrate NO2-N inhibition (5–35 g N m−3). Non-competitive inhibition and Hill-type models were compared to select the best-fitting inhibition equations. Inhibition was correctly modeled by the Hill-type model and a half inhibition constant (KI) for OPRNET, NPQ, maximum photosynthetic rate (ETRMAX) and the performance index PIABS was 23.7 ± 1.2, 26.36 ± 1.10, 39 ± 2 and 26.5 ± 0.4, respectively.

In vitro inhibitory effect of obtusofolin on the activity of CYP3A4, 2C9, and 2E1

BackgroundObtusofolin is the major active ingredient of Catsia tora L., which possesses the activity of improving eyesight and protecting the optic nerve. Investigation on the interaction of obtusofolin with cytochrome P450 enzymes (CYP450s) could provide a reference for the clinical application of obtusofolin.MethodsThe effect of obtusofolin on the activity of CYP450s was investigated in the presence of 100 μM obtusofolin in pooled human liver microsomes (HLMs) and fitted with the Lineweaver–Burk plots to characterize the specific inhibition model and kinetic parameters.ResultsObtusofolin was found to significantly inhibited the activity of CYP3A4, 2C9, and 2E1. In the presence of 0, 2.5, 5, 10, 25, 50, and 100 μM obtusofolin, the inhibition of these CYP450s showed a dose-dependent manner with the IC50 values of 17.1 ± 0.25, 10.8 ± 0.13, and 15.5 ± 0.16 μM, respectively. The inhibition of CYP3A4 was best fitted with the non-competitive inhibition model with the Ki value of 8.82 μM. While the inhibition of CYP2C9 and 2E1 was competitive with the Ki values of 5.54 and 7.79 μM, respectively. After incubating for 0, 5, 10, 15, and 30 min, the inhibition of CYP3A4 was revealed to be time-dependent with the KI value of 4.87 μM− 1 and the Kinact value of 0.0515 min− 1.ConclusionsThe in vitro inhibitory effect of obtusofolin implying the potential drug-drug interaction between obtusofolin and corresponding substrates, which needs further in vivo validations.

In vitro study on the effect of leonurine hydrochloride on the enzyme activity of cytochrome P450 enzymes in human liver microsomes

Leonurine hydrochloride (LH) is derived from an ingredient of Leonurus japonicus Houtt which is widely used for diseases in women. The influence of LH on the activity of cytochrome P450 (CYPs) enzymes was investigated in this study. The effect of LH on CYPs enzyme activities were studied using the enzyme-selective substrates phenacetin (1A2), coumarin (2A6), diclofenac (2C9), S-mephenytoin (2C19), paclitaxel (2C8), dextromethorphan (2D6), chlorzoxazone (2E1) and testosterone (3A4). The IC50 value was calculated to express the strength of inhibition. The inhibition of CYPs was fitted with competitive or non-competitive inhibition models and corresponding parameters were also obtained. LH exerted inhibitory effects on the activity of CYP1A2, 2D6, and 3A4 with the IC50 values of 18.05, 15.13, and 20.09 μM, respectively. The obtained results showed that LH inhibited the activity of CYP1A2 and CYP2D6 via competitive manners (Ki = 8.667 μM and Ki = 7.805 μM, respectively), while LH attenuated the CYP3A4 activity via a non-competitive manner (Ki = 9.507 μM). Moreover, LH showed time-dependent inhibition on CYP3A4 with the KI/Kinact value of 4.31/0.044 min−1·μM−1. The inhibition of CYP1A2, CYP2D6, and CYP3A4 by LH, demonstrated in vitro, indicated the potential herb–drug interaction. Therefore, pharmacokinetic interactions involving LH and CYP1A2 or CYP2D6 or CYP1A2 substrates are likely to occur.

Effect of the tetracycline antibiotics on performance and microbial community of microbial fuel cell.

The adverse effect of tetracycline antibiotics on microbial activity is one of the serious risks for the biologic wastewater treatment process. The microbial fuel cells (MFCs) are a promising technology for wastewater treatment and renewable power generation process. For this reason, the investigation of the inhibition effect of the tetracyclines on the MFCs is essential for reducing damage on the environment. This paper focused on the performance of MFCs under different antibiotic concentrations at the range of 0.25-50mg/L. The power generation performance, microbial community and biofilm characteristics (morphology, resistance and viability) of MFCs were investigated in detail. The results indicated that the increase in the antibiotic concentration significantly affected the MFC performance and microbial community. A modified non-competitive inhibition model was used to predict the inhibition effect of tetracycline antibiotics on the MFCs.

A rapid assay to screen adenosine deaminase inhibitors from Ligustri Lucidi Fructus against metabolism of cordycepin utilizing ultra-high-performance liquid chromatography-tandem mass spectrometry.

Cordycepin has recently received increased attention owing to its extensive pharmacological activity. Adenosine deaminase (ADA) is widely distributed in mammalian blood and tissues; as a result, cordycepin is quickly metabolized upon entering into the body and converted into the inactive metabolite 3'-deoxyinosine, thus limiting its activity when administered alone. We herein present a novel ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for screening ADA inhibitors against the metabolism of cordycepin. Cordycepin and 3'-deoxyinosine were chosen as substrate and product, respectively. A proper separation was achieved for all analytes within 3 min. 3'-Deoxyinosine was quantified in the presence or absence of potential ADA inhibitors to evaluate ADA activity. The assay can simultaneously determine substrate and product, with the endogenous substance and ADA inhibitors added not interfering in its activity. After optimizing the enzymatic incubation and UHPLC-MS/MS conditions, Km and Vmax values for ADA deamination of cordycepin were 95.18 ± 7.85 μm and 363.90 ± 12.16 μmol/min/unit, respectively. Oleanolic acid and ursolic acid from Ligustri Lucidi Fructus were chosen as ADA inhibitors with half maximal inhibitory concentration values of 21.82 ± 0.39 and 18.41 ± 0.14 μm, respectively. A non-competitive inhibition model was constructed and this assay can be used to screen other potential ADA inhibitors quickly and accurately.

Succinic acid inhibits the activity of cytochrome P450 (CYP450) enzymes

Context Succinic acid, extracted from amber, is widely used in cardiovascular therapy. Objective The effect of succinic acid on the activity of cytochrome P450 (CYP450) enzymes was investigated in this study. Materials and methods The effect of succinic acid (100 μM) on the activity of eight isoforms of CYP450 (i.e., 1A2, 3A4, 2A6, 2E1, 2D6, 2C9, 2C19 and 2C8) was investigated compared to the specific inhibitor and blank controls in pooled human liver microsomes in vitro. The inhibition of CYPs was fitted with competitive or non-competitive inhibition models and corresponding parameters were also obtained. Results Succinic acid exerted inhibitory effect on the activity of CYP3A4, 2D6, and 2C9 with the IC50 values of 12.82, 14.53, and 19.60 μM, respectively. Succinic acid inhibited the activity of CYP3A4 in a non-competitive manner with the Ki value of 6.18 μM, and inhibited CYP2D6 and 2C9 competitively with Ki values of 7.40 and 9.48 μM, respectively. Furthermore, the inhibition of CYP3A4 was found to be time-dependent with the KI/Kinact value of 6.52/0.051 min−1·μM−1. Discussion and conclusions Succinic acid showed in vitro inhibitory effects on the activity of CYP3A4, 2D6, and 2C9, which indicated the potential drug-drug interactions. Succinic acid should be carefully co-administrated with the drugs metabolized by CYP3A4, 2D6, and 2C9.

Identification of the Methanogenesis Inhibition Mechanism Using Comparative Analysis of Mathematical Models.

The application of cationic polymers to enhance membrane fluxes in anaerobic membrane bioreactors has been proposed by several authors. However, literature shows contradictory results on the influence of those chemicals on the biological activity. In this research, we studied the effect of a cationic polymer on the production of methane from acetate by acetoclastic methanogens. We assessed the effect of polymer concentration on the accumulated methane production (AMP) and the specific methanogenic activity (SMA) in batch tests. Batch tests results showed lower SMA values at higher concentrations of polymer and no effect on the final AMP. Different inhibition models were calibrated and compared to find the best fit and to hypothesize the prevailing inhibition mechanisms. The assessed inhibition models were: competitive (M1a), non-competitive (M2a), un-competitive (M3a), biocide-linear (M4a), and biocide-exponential (M5a). The parameters in the model related to the polymer characteristics were adjusted to fit the experimental data. M2a and M3a were the only models that fitted both experimental SMA and AMP. Although M1a and M4a adequately fitted the experimental SMA, M1a simulations slightly deviated from the experimental AMP, and M4a considerably underpredicted the AMP at concentrations of polymer above 0.23 gCOD L−1. M5a did not adequately fit either experimental SMA and AMP results. We compared models a (M1a to M5a), which consider the inhibition by the concentration of polymer in the bulk liquid, with models b (M1b to M5b) considering the inhibition being caused by the total concentration of polymer in the reactor. Results showed that the difference between a and b models' simulations were negligible for all kinetic models considered (M1, M2, M3, M4, and M5). Therefore, the models that better predicted the experimental data were the non-competitive (M2a and M2b) and un-competitive (M3a and M3b) inhibition models, which are biostatic inhibition models. Consequently, the decreased methanogenic activity caused by polymer additions is presumably a reversible process

Production of a Thermostable Chitosanase from Shrimp Heads via Paenibacillus mucilaginosus TKU032 Conversion and its Application in the Preparation of Bioactive Chitosan Oligosaccharides.

Chitosanase has attracted great attention due to its potential applications in medicine, agriculture, and nutraceuticals. In this study, P. mucilaginosus TKU032, a bacterial strain isolated from Taiwanese soil, exhibited the highest chitosanase activity (0.53 U/mL) on medium containing shrimp heads as the sole carbon and nitrogen (C/N) source. Using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, a chitosanase isolated from P. mucilaginosus TKU032 cultured on shrimp head medium was determined at approximately 59 kDa. The characterized chitosanase showed interesting properties with optimal temperature and thermal stability up to 70 °C. Three chitosan oligosaccharide (COS) fractions were isolated from hydrolyzed colloidal chitosan that was catalyzed by TKU032 chitosanase. Of these, fraction I showed the highest α-glucosidase inhibitor (aGI) activity (65.86% at 20 mg/mL); its inhibitory mechanism followed the mixed noncompetitive inhibition model. Fractions II and III exhibited strong 2,2-diphenyl1-picrylhydrazyl (DPPH) radical scavenging activity (79.00% at 12 mg/mL and 73.29% at 16 mg/mL, respectively). In summary, the COS fractions obtained by hydrolyzing colloidal chitosan with TKU032 chitosanase may have potential use in medical or nutraceutical fields due to their aGI and antioxidant activities.

Nitrous Oxide Is a Potent Inhibitor of Bacterial Reductive Dechlorination

Organohalide-respiring bacteria are key players for the turnover of organohalogens. At sites impacted with chlorinated ethenes, bioremediation promotes reductive dechlorination; however, stoichiometric conversion to environmentally benign ethene is not always achieved. We demonstrate that nitrous oxide (N2O), a compound commonly present in groundwater, inhibits organohalide respiration. N2O concentrations in the low micromolar range decreased dechlorination rates and resulted in incomplete dechlorination of tetrachloroethene (PCE) in Geobacter lovleyi strain SZ and of cis-1,2-dichloroethene ( cDCE) and vinyl chloride (VC) in Dehalococcoides mccartyi strain BAV1 axenic cultures. Presumably, N2O interferes with reductive dechlorination by reacting with super-reduced Co(I)-corrinoids of reductive dehalogenases, which is supported by the finding that N2O did not inhibit corrinoid-independent fumarate-to-succinate reduction in strain SZ. Kinetic analyses revealed a best fit to the noncompetitive Michaelis-Menten inhibition model and determined N2O inhibitory constants, KI, for PCE and cDCE dechlorination of 40.8 ± 3.8 and 21.2 ± 3.5 μM in strain SZ and strain BAV1, respectively. The lowest KI value of 9.6 ± 0.4 μM was determined for VC to ethene reductive dechlorination in strain BAV1, suggesting that this crucial dechlorination step for achieving detoxification is most susceptible to N2O inhibition. Groundwater N2O concentrations exceeding 100 μM are not uncommon, especially in watersheds impacted by nitrate runoff from agricultural sources. Thus, dissolved N2O measurements can inform about cDCE and VC stalls at sites impacted with chlorinated ethenes.

Investigating the effect of copper and magnesium ions on nitrogen removal capacity of pure cultures by modified non-competitive inhibition model

Copper, a common heavy metal, may be beneficial for or poisonous to microbial activity. The objective of this study was to determine the effect of different copper ion concentrations on the nitrogen removal performance of Arthrobacter arilaitensis strain Y-10 and Pseudomonas taiwanensis strain J488. The non-competitive inhibition model was employed to evaluate the 50% inhibition concentrations (IC50 values) of copper ions toward the pure strains. In the absence of magnesium ions, a low concentration of copper (0.1 mg/L) significantly enhanced the ammonium removal ability of strain Y-10 and its removal efficiency increased by 10.88% compared with the control treatment. Copper ranging from 0 to 0.1 mg/L had no significant effect on the ammonium removal capacity of strain J488. After adding 9.90 mg/L of magnesium to the basal medium, the effects of copper on nitrification of ammonium or denitrification of nitrate or nitrite were also assessed. In these conditions, 0.25 mg/L copper ions could strongly inhibit the ammonium, nitrate and nitrite removal activities for strain Y-10. For strain J488, no clear deterioration in ammonium removal efficiency was observed at copper ion concentrations below 0.5 mg/L, but 0.25 mg/L copper ions significantly inhibited nitrate and nitrite removal efficiencies, which were only 45.88% and 6.35%, respectively. The IC50 values of copper ions for nitrate and nitrite removal by strain Y-10 were 0.195 and 0.090 mg/L respectively; for strain J488, the IC50 values were 0.175 and 0.196 mg/L. The magnesium ions could improve the cell growth, nitrogen removal efficiency and copper ion resistance of bacteria.

Insight into the short- and long-term effects of quinoline on anammox granules: Inhibition and acclimatization

The short- and long-term influence of quinoline on the properties of anaerobic ammonium oxidation (anammox) biogranules was evaluated. During batch tests, the bioactivity of anammox granules in the presence of different quinoline concentrations was monitored, and the IC50 of quinoline was calculated to be 13.1 mg L−1 using a non-competitive inhibition model. The response of anammox granules to pre-exposure to quinoline was dependent on metabolic status, and the presence of both quinoline and NO2−-N had a rapid detrimental effect, resulting in a 64.5% decrease within 12 h. During continuous-flow experiments, the nitrogen removal rate (NRR) of the reactor decreased sharply within 3 days in the presence of 10 mg L−1 quinoline and then was restored to 2.6 kg N m−3 d−1. In the presence of quinoline-induced stress, the specific anammox activity and levels of extracellular polymeric substance and heme c were decreased, while settling velocity persistently increased. After cultivation and acclimation obtained by adding a medium level of quinoline to the influent, the anammox granule sludge was able to tolerate 10 mg L−1 quinoline in 178 days.