Inorganic Pyrophosphatase Activity Research Articles

Supplementation of Nitrogen and its Influence on Free Sugars, Amino Acid and Protein Metabolism in Roots and Internodes of Wheat

Effect of different doses of nitrogen (N) (90, 120, 150 and 180 kg Nha–1) on the activities of aminotransferases and alkaline inorganic pyrophosphatase (AIP) in relation to the accumulation of proteins, amino acids and sugars in roots and internodes at 15 and 40 days post anthesis (DPA) stages was studied in six wheat genotypes namely HD 2967, GLU 1101, PBW 343, BW 9022, PH-132–4840 and PBW 550. Supra-optimal N doses (150 kg Nha–1 and 180 kg Nha–1) accentuated glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and alkaline inorganic pyrophosphatase activities in correspondence with an increase in amino acid, protein and sugar content in both roots and internodes in all the six genotypes. Activities of analyzed enzymes were significantly high at 15 days post anthesis (DPA) stage and thereafter declined at maturity (40 DPA) in parallel with decrease in amino acid contents. Maximum activity of GOT, GPT and AIP was observed in HD 2967 and GLU 1101 genotypes along with higher build up of proteins and amino acids which resulted in higher grain yield. Activity of GPT was comparatively high over GOT, indicating its major role towards protein synthesis. Grain filling processes in terms of proteins and amino acids were positively correlated with GOT and GPT activities while sugars were correlated to AIP. Thus, nitrogen acquisition and assimilation resulted in favoured utilization of N in form of amino acid and proteins accumulation while sugar content was also stimulated. Due to immense activities of aminotransferases and higher contents of amino acids and proteins in GLU 1101 and HD 2967 genotypes at optimal dose and higher dose of N, these genotypes hold future potential for developing new cultivars with better grain quality characteristics.

Self-incompatibility in Papaver pollen: programmed cell death in an acidic environment.

Self-incompatibility (SI) is a genetically controlled mechanism that prevents self-fertilization and thus encourages outbreeding and genetic diversity. During pollination, most SI systems utilize cell-cell recognition to reject incompatible pollen. Mechanistically, one of the best-studied SI systems is that of Papaver rhoeas (poppy), which involves the interaction between the two S-determinants, a stigma-expressed secreted protein (PrsS) and a pollen-expressed plasma membrane-localized protein (PrpS). This interaction is the critical step in determining acceptance of compatible pollen or rejection of incompatible pollen. Cognate PrpS-PrsS interaction triggers a signalling network causing rapid growth arrest and eventually programmed cell death (PCD) in incompatible pollen. In this review, we provide an overview of recent advances in our understanding of the major components involved in the SI-induced PCD (SI-PCD). In particular, we focus on the importance of SI-induced intracellular acidification and consequences for protein function, and the regulation of soluble inorganic pyrophosphatase (Pr-p26.1) activity by post-translational modification. We also discuss attempts to identify protease(s) involved in the SI-PCD process. Finally, we outline future opportunities made possible by the functional transfer of the P. rhoeas SI system to Arabidopsis.

The role of anion in the impact of tetraethylammonium salts on soil phosphatase activities

Quaternary ammonium salts (QAS) are increasingly commonly used in the chemical industry. The diverse usage of these compounds results in high possibility of their penetration to the environment. The aim of this study was to compare the effect of three salts with tetraethylammonium cation with different anions such as chloride [TEA][Cl], bromide [TEA][Br], and iodine [TEA][I] on the activity of phosphatases in loamy sand. Organic carbon and total nitrogen content in the soil were 8.71 and 0.97 g·kg-1, respectively. The salts were applied to soil at doses of 0, 0.05, 0.50, and 5.00 mmol·kg-1. The samples were adjusted to 60% maximum water holding capacity, and they were incubated at a temperature of 20°C. Activities of acid phosphomonoesterase, alkaline phopshomonoesterase, phosphotriesterase, and inorganic pyrophosphatase were assayed on day 1, 7, 14, 28, 56, and 112. The obtained results demonstrated that the presence of [TEA] in the soil at the low dose (0.05 mmol·kg-1) may stimulate the activity of soil phosphatases. Higher doses of the soil had mostly inhibitory effect on phosphatases activity. Among the determined enzymes, phosphomonoesterases were the most sensitive to salts containing [TEA] cation. A considerable contribution of the anion type on the formation of activity of the determined enzymes of phosphorus compound changes. The strongest effect characterized tetraethylammonium bromide [TEA][Br].

Combined effect of low-molecular-weight organic acids and creosote on phosphatase activities in sandy soil

This paper assesses the impact of creosote and low-molecular-weight organic acids (LMWOAs) on the activity of acid phosphomonoesterase, alkaline phosphomonoesterase, phosphotriesterase, and inorganic pyrophosphatase in soil. The experiment was carried out on loamy sand samples with organic carbon content of 8.71 g · kg -1 , with the following variable factors: dosages of creosote: 0, 0.5%, and 2.5%; type of LMWOAs: oxalic acid, tartaric acid, and citric acid in the amount of 50 mmol · kg -1 of soil; days of experiment: 1, 7, 14, 28, 56, 112. Obtained results showed that contamination with creosote caused decrease in the activity of soil phosphatases. The observed effect did not always increase with increase in the dosage of the pollutant. Among the assayed phosphatases, the biggest changes were noted in the activity of phosphomonoesterases. Application of LMWOAs to contaminated soil mainly effected the inhibition of phosphatase, especially the activity of acid phosphomonoesterase. Comparison of the effects of LMWOAs showed that the citric acid was the least toxic to soil phosphatases.

Silencing PPA1 inhibits human epithelial ovarian cancer metastasis by suppressing the Wnt/β-catenin signaling pathway.

Inorganic pyrophosphatase (PPA1) activity is a key determinant of cellular inorganic pyrophosphate levels, and its expression is correlated with growth of several solid tumors. To investigate this relationship, we first examined PPA1 expression in human epithelial ovarian cancer (EOC) samples, and found that PPA1 was overexpressed in tumors from EOC patients. Higher PPA1 levels correlated with advanced grades, stages, and poor survival in EOC patients. Examination of PPA1 function in EOC revealed that silencing PPA1 inhibited EOC migration, epithelial-mesenchymal transition (EMT), and metastasis in vitro and in vivo. In addition, PPA1 may promote the dephosphorylation and translocation of β-catenin. These results demonstrate that silencing PPA1 inhibits EOC metastasis by suppressing the Wnt/β-catenin signaling pathway. Strategies for downregulating PPA1 may have therapeutic potential for the prevention and treatment of EOC.

Identification of Phosphorylation Sites Altering Pollen Soluble Inorganic Pyrophosphatase Activity.

Protein phosphorylation regulates numerous cellular processes. Identifying the substrates and protein kinases involved is vital to understand how these important posttranslational modifications modulate biological function in eukaryotic cells. Pyrophosphatases catalyze the hydrolysis of inorganic phosphate (PPi) to inorganic phosphate Pi, driving biosynthetic reactions; they are essential for low cytosolic inorganic phosphate. It was suggested recently that posttranslational regulation of Family I soluble inorganic pyrophosphatases (sPPases) may affect their activity. We previously demonstrated that two pollen-expressed sPPases, Pr-p26.1a and Pr-p26.1b, from the flowering plant Papaver rhoeas were inhibited by phosphorylation. Despite the potential significance, there is a paucity of data on sPPase phosphorylation and regulation. Here, we used liquid chromatographic tandem mass spectrometry to map phosphorylation sites to the otherwise divergent amino-terminal extensions on these pollen sPPases. Despite the absence of reports in the literature on mapping phosphorylation sites on sPPases, a database survey of various proteomes identified a number of examples, suggesting that phosphorylation may be a more widely used mechanism to regulate these enzymes. Phosphomimetic mutants of Pr-p26.1a/b significantly and differentially reduced PPase activities by up to 2.5-fold at pH 6.8 and 52% in the presence of Ca2+ and hydrogen peroxide over unmodified proteins. This indicates that phosphoregulation of key sites can inhibit the catalytic responsiveness of these proteins in concert with key intracellular events. As sPPases are essential for many metabolic pathways in eukaryotic cells, our findings identify the phosphorylation of sPPases as a potential master regulatory mechanism that could be used to attenuate metabolism.

Real-time assay of inorganic pyrophosphatase activity in the red region based on Li+-doped NaYF4:Yb,Er upconversion luminescent nanoparticles

Based on a competition assay approach, a new highly sensitive method for real-time detection of PPase activity using Li+-doped NaYF4:Yb,Er upconversion red luminescent nanoparticles was developed.

A novel dual-functional biosensor for fluorometric detection of inorganic pyrophosphate and pyrophosphatase activity based on globulin stabilized gold nanoclusters

A novel ultrasensitive dual-functional biosensor for highly sensitive detection of inorganic pyrophosphate (PPi) and pyrophosphatase (PPase) activity was developed based on the fluorescent variation of globulin protected gold nanoclusters (Glo@Au NCs) with the assistance of Cu2+. Glo@Au NCs and PPi were used as the fluorescent indicator and substrate for PPase activity evaluation, respectively. In the presence of Cu2+, the fluorescence of the Glo@Au NCs will be quenched owing to the formation of Cu2+-Glo@Au NCs complex, while PPi can restore the fluorescence of the Cu2+-Glo@Au NCs complex because of its higher binding affinity with Cu2+. As PPase can catalyze the hydrolysis of PPi, it will lead to the release of Cu2+ and re-quench the fluorescence of the Glo@Au NCs. Based on this mechanism, quantitative evaluation of the PPi and PPase activity can be achieved ranging from 0.05 μM to 218.125 μM for PPi and from 0.1 to 8 mU for PPase, with detection limits of 0.02 μM and 0.04 mU, respectively, which is much lower than that of other PPi and PPase assay methods. More importantly, this ultrasensitive dual-functional biosensor can also be successfully applied to evaluate the PPase activity in human serum, showing great promise for practical diagnostic applications.

Fluorescence biosensor for inorganic pyrophosphatase activity.

A highly sensitive and selective fluorescence biosensor for inorganic pyrophosphatase (PPase) activity has been developed based on special click ligation trigger hyperbranched rolling circle amplification (CLT-HRCA). Pyrophosphate ion (PPi) can coordinate with Cu2+ to form stable PPi/Cu2+ complex and Cu2+ in the complex cannot be reduced to Cu+. The addition of PPase causes the hydrolysis of PPi into orthophosphate (Pi) and therefore induces the releasing of Cu2+ from the stable PPi/Cu2+ complex, and the free Cu2+ is easily reduced to Cu+ by sodium ascorbate. Then Cu+ catalyzes the cyclization reaction between the specially designed 5'-azide and 3'-alkyne tagged padlock probes through Cu+ catalyzed azide-alkyne cycloaddition (CuAAC), which in turn initiates the hyperbranched rolling circle amplification (HRCA). Given that the CLT-HRCA products contain large amounts of double-stranded DNAs (dsDNAs), the addition of SYBR Green I resulted in the enhanced fluorescence signal. There was a linear relationship between the enhanced fluorescence intensity and the logarithm PPase activity ranging from 0.05 to 25mU with a detection limit of 0.02mU. Such proposed biosensor has been successfully applied to screen the potential PPase inhibitors and has accessed the related inhibit ability with high efficiency.

Sudden Cardiac Death Due to Deficiency of the Mitochondrial Inorganic Pyrophosphatase PPA2

We have used whole-exome sequencing in ten individuals from four unrelated pedigrees to identify biallelic missense mutations in the nuclear-encoded mitochondrial inorganic pyrophosphatase (PPA2) that are associated with mitochondrial disease. These individuals show a range of severity, indicating that PPA2 mutations may cause a spectrum of mitochondrial disease phenotypes. Severe symptoms include seizures, lactic acidosis, cardiac arrhythmia, and death within days of birth. In the index family, presentation was milder and manifested as cardiac fibrosis and an exquisite sensitivity to alcohol, leading to sudden arrhythmic cardiac death in the second decade of life. Comparison of normal and mutant PPA2-containing mitochondria from fibroblasts showed that the activity of inorganic pyrophosphatase was significantly reduced in affected individuals. Recombinant PPA2 enzymes modeling hypomorphic missense mutations had decreased activity that correlated with disease severity. These findings confirm the pathogenicity of PPA2 mutations and suggest that PPA2 is a cardiomyopathy-associated protein, which has a greater physiological importance in mitochondrial function than previously recognized.

Facile Synthesis of Enhanced Fluorescent Gold-Silver Bimetallic Nanocluster and Its Application for Highly Sensitive Detection of Inorganic Pyrophosphatase Activity.

Herein, gold-silver bimetallic nanoclusters (Au-Ag NCs) with the high fluorescent intensity were first synthesized successfully and utilized for the fabrication of sensitive and specific sensing probes toward inorganic pyrophosphatase (PPase) activity with the help of copper ion (Cu(2+)) and inorganic pyrophosphate ion (PPi). Cu(2+) was used as the quencher of fluorescent Au-Ag NC, while PPi was employed as the hydrolytic substrate of PPase. The system consisted of PPi, Cu(2+) ion, and bovine serum albumin (BSA)-stabilized Au-Ag NC. The detection was carried out by enzyme-induced hydrolysis of PPi to liberate copper ion from the Cu(2+)-PPi complex. In the absence of target PPase, free copper ions were initially chelated with inorganic pyrophosphate ions to form the Cu(2+)-PPi complexes via the coordination chemistry, thus preserving the natural fluorescent intensity of the Au-Ag NCs. Upon addition of target PPase into the detection system, the analyte hydrolyzed PPi into phosphate ions and released Cu(2+) ion from the Cu(2+)-PPi complex. The dissociated copper ions readily quenched the fluorescent signal of Au-Ag NCs, thereby resulting in the decrease of fluorescent intensity. Under optimal conditions, the detectable fluorescent intensity of the as-prepared Au-Ag NCs was linearly dependent on the activity of PPase within a dynamic linear range of 0.1-30 mU/mL and allowed the detection at a concentration as low as 0.03 mU/mL at the 3sblank criterion. Good reproducibility (CV < 8.5% for the intra-assay and interassay), high specificity, and long-term stability (90.1% of the initial signal after a storage period of 48 days) were also received by using our system toward target PPase activity. In addition, good results with the inhibition efficiency of sodium fluoride were obtained in the inhibitor screening research of pyrophosphatase. Importantly, this system based on highly enhanced fluorescent Au-Ag NCs offer promise for simple and cost-effective screening of target PPase activity without the needs of sample separation and multiple washing steps.

Water-soluble gold nanoclusters prepared by protein-ligand interaction as fluorescent probe for real-time assay of pyrophosphatase activity

This paper reports a new and facile method for the synthesis of water-soluble thiolate-protected AuNCs via protein-ligand interaction. Using 3-mercaptopropionic acid (MPA) as a model ligand and bovine serum albumin (BSA) as a model protein, water-soluble AuNCs (BSA/MPA-AuNCs) with intense orange-yellow fluorescent emission (quantum yield=16%) are obtained. Results show that AuNCs produced with this method have hydrophobic interactions with BSA. The synthetic strategy is then successfully extended to produce water-soluble AuNCs protected by other thiolates. Moreover, a sensitive and eco-friendly sensing system is established for detection of the activity of inorganic pyrophosphatase (PPase), which relies on the selective coordination of Fe3+with BSA/MPA-AuNCs, the higher affinity between pyrophosphate (PPi) and Fe3+, and the hydrolysis of PPi by PPase. A good linearity between the fluorescence intensity and PPase activity within the range from 0.1 to 3U/L is found, with a detection limit down to 0.07U/L. Additionally, the fluorescent assay developed here is utilized to assay the PPase activity in real biological samples and as well as to evaluate PPase inhibitor, illustrating the great potential for biological analysis.

Comparison of glyphosate and Roundup preparations influence on inorganic pyrophosphatase activity and available phosphorus content in sandy loam

Comparison of glyphosate and Roundup preparations influence on inorganic pyrophosphatase activity and available phosphorus content in sandy loam

Fluorescent and Colorimetric Dual-Readout Assay for Inorganic Pyrophosphatase with Cu2+-Triggered Oxidation of o-Phenylenediamine

We demonstrate a rationally designed fluorescent and colorimetric dual-readout strategy for the highly sensitive, quantitative determination of inorganic pyrophosphatase (PPase) activity, a key hydrolytic enzyme involved in a variety of metabolic processes. Inspired by the selective oxidative and chromogenic reaction of o-phenylenediamine (OPD) with Cu(2+), the special inhibitory effects of pyrophosphate (PPi) on the oxidative ability of Cu(2+), and the specific hydrolysis of PPi into orthophosphate by PPase, a convenient small molecule OPD-based analytical system was developed for Cu(2+)/PPi recognition and PPase activity assay. We have confirmed that Cu(2+) acts as the oxidant in the reaction and the main chromogenic product of OPD is 2,3-diaminophenazine (usually called OPDox) in the assay by combining the ESI-MS, (1)H NMR, and XPS spectra analysis. Direct electrochemical insights into the Cu(2+)-triggered and PPi-inhibited mechanism were performed by cyclic voltammetry characterizations of the Cu(2+) in the absence and presence of PPi for the first time. Furthermore, the proposed analytical system with clear response mechanism exhibits a promising outlook for the PPase activity assay in real biological samples and inhibitor screening.

A novel fluorescence assay for inorganic pyrophosphatase based on modulated aggregation of graphene quantum dots.

A simple and highly sensitive fluorometric method has been developed for inorganic pyrophosphatase (PPase) activity detection based on the disaggregation and aggregation of graphene quantum dots (GQDs). Copper ions can trigger the severe aggregation of GQDs with rich carboxyl groups, which results in effective fluorescence quenching. While, with the addition of pyrophosphate (PPi), the quenched fluorescence is effectively recovered owing to the strong interaction between PPi and Cu(2+). Furthermore, under the catalytic hydrolysis of PPase, the complex of PPi-Cu(2+)-PPi is rapidly disassembled, and the fluorescence is re-quenched. This method is highly sensitive and selective for PPase detection, with a linear correlation between the fluorescence intensity and the PPase concentration in the range from 1 to 200 mU mL(-1) with a detection limit down to 1 mU mL(-1) (S/N = 3). Additionally, the inhibition effect of NaF on the PPase activity is also studied. Thus, the proposed method may hold a potential application in the diagnosis of PPase-related diseases and screening of PPase inhibitors, to evaluate the function and inhibition of PPase in biological systems.

Self-incompatibility-induced programmed cell death in field poppy pollen involves dramatic acidification of the incompatible pollen tube cytosol.

Self-incompatibility (SI) is an important genetically controlled mechanism to prevent inbreeding in higher plants. SI involves highly specific interactions during pollination, resulting in the rejection of incompatible (self) pollen. Programmed cell death (PCD) is an important mechanism for destroying cells in a precisely regulated manner. SI in field poppy (Papaver rhoeas) triggers PCD in incompatible pollen. During SI-induced PCD, we previously observed a major acidification of the pollen cytosol. Here, we present measurements of temporal alterations in cytosolic pH ([pH]cyt); they were surprisingly rapid, reaching pH 6.4 within 10 min of SI induction and stabilizing by 60 min at pH 5.5. By manipulating the [pH]cyt of the pollen tubes in vivo, we show that [pH]cyt acidification is an integral and essential event for SI-induced PCD. Here, we provide evidence showing the physiological relevance of the cytosolic acidification and identify key targets of this major physiological alteration. A small drop in [pH]cyt inhibits the activity of a soluble inorganic pyrophosphatase required for pollen tube growth. We also show that [pH]cyt acidification is necessary and sufficient for triggering several key hallmark features of the SI PCD signaling pathway, notably activation of a DEVDase/caspase-3-like activity and formation of SI-induced punctate actin foci. Importantly, the actin binding proteins Cyclase-Associated Protein and Actin-Depolymerizing Factor are identified as key downstream targets. Thus, we have shown the biological relevance of an extreme but physiologically relevant alteration in [pH]cyt and its effect on several components in the context of SI-induced events and PCD.

Fluorometric method for inorganic pyrophosphatase activity detection and inhibitor screening based on click chemistry.

A fluorometric method for pyrophosphatase (PPase) activity detection was developed based on click chemistry. Cu(II) can coordinate with pyrophosphate (PPi), the addition of pyrophosphatase (PPase) into the above system can destroy the coordinate compound because PPase catalyzes the hydrolysis of PPi into inorganic phosphate and produces free Cu(II), and free Cu(II) can be reduced by sodium ascorbate (SA) to form Cu(I), which in turn initiates the ligating reaction between nonfluorescent 3-azidocoumarins and terminal alkynes to produce a highly fluorescent triazole complex, based on which, a simple and sensitive turn on fluorometric method for PPase can be developed. The fluorescence intensity of the system has a linear relationship with the logarithm of the PPase concentration in the range of 0.5 and 10 mU with a detection limit down to 0.2 mU (S/N = 3). This method is cost-effective and convenient without any labels or complicated operations. The proposed system was applied to screen the potential PPase inhibitor with high efficiency. The proposed method can be applied to diagnosis of PPase-related diseases.

Changes in the inorganic status and enzyme activities in senescent leaves of chickpea, Cicer arietinum L.

The changes in the level of some inorganic constituents and the activities of some important enzyme systems in senescent leaves of chickpea (&lt;em&gt;Cicer arietinum&lt;/em&gt; L.) have been studied. In senescent leaves, a marked decline in the potassium and phosphorus contents was evident which was accompanied by the accumulation of calcium, silicon, chloride and manganese. Leaf senescence was accompanied by a great increase in hydrolytic processes, as revealed by the increase in the activities of acid phosphatase, alkaline phosphatase, ATPase, inorganic pyrophosphatase and 3-phosphoglycerate phosphatase. The activities of nitrogen metabolism enzymes, namely nitrate reductase, nitrite reductase, glutamine synthetase and alanine aminotransferase, and of photorespiratory enzymes -- phosphoglycolate phosphatase, glycolate oxidase and catalase, were lower in senescent leaves. Leaf senescence was further associated with an increase in the activities of peroxidase and polyphenol oxidase, a considerable depression in pyruvate kinase activity, and a slight elevation in aldolase activity.

Age-dependent increased expression and activity of inorganic pyrophosphatase in the liver of male mice and its further enhancement with short- and long-term dietary restriction

Intracellular orthophosphate and inorganic pyrophosphate (PPi) are by-products of multiple biosynthetic reactions. PPi hydrolysis by soluble inorganic pyrophosphatase (iPPase) has been considered as an important homeostatic mechanism. We investigated the expression and activities (U/mg protein) of iPPase in the liver of young and old mice subjected to short- and long-term dietary restriction. The expression level of iPPase was ascertained by the Western blot analysis using anti-iPPase and differential polymerase chain reaction using iPPase specific primer. Older mice showed a significant increase in the expression and activity of iPPase as compared to younger ones. Short-term fasting of 24h increased the expression and activity of iPPase in the liver of both young and old mice which were reversed upon 24h of re-feeding them. However, both young and old mice on long-term dietary restriction showed a cumulative increase in the expression and activity of iPPase when compared with their age-matched controls. This might be due to accumulative adaptation to refill energy deficiency of long-term dietary restricted mice for ATP generation via oxidative phosphorylation, where fatty acid activation could be driven by elevated iPPase.

Real-Time Colorimetric Assay of Inorganic Pyrophosphatase Activity Based on Reversibly Competitive Coordination of Cu2+ between Cysteine and Pyrophosphate Ion

In this study we demonstrate a new colorimetric method for real-time pyrophosphatase (PPase) activity assay based on reversible tuning of the dispersion/aggregation states of gold nanoparticles (Au-NPs) by controlling the coordination of Cu(2+) between cysteine and pyrophosphate ion (PPi) with PPase. The addition of Cu(2+) to the cysteine-stabilized Au-NP dispersion results in the aggregation of Au-NPs, while the further addition of PPi to this aggregation turns the aggregated Au-NPs into their dispersed state because of the higher coordination reactivity between Cu(2+) and PPi than that between Cu(2+) and cysteine. The subsequent addition of PPase to the PPi-triggered dispersed Au-NPs restores the aggregation state of Au-NPs because PPase catalyzes the hydrolysis of PPi into orthophosphate and thus consumes PPi in the reaction system. In this study, we utilize this reversibility of the change between the aggregation/dispersion states of Au-NPs for real-time colorimetric monitoring of PPase activity by continuously measuring the ratio of absorbance at the wavelength of 650 nm (A650) to that at 522 nm (A522) in the time-dependent UV-vis spectra of Au-NP dispersions containing different activities of PPase. To calculate the kinetics of the PPase-catalyzed hydrolysis of PPi, the A650/A522 values are converted into PPi concentrations to obtain the time-dependent changes of PPi concentrations in the dispersions containing different activities of PPase. The initial reaction rates (v0) are thus achieved from the time-dependent logarithm of PPi concentrations with the presence of different PPase activities. Under the experimental conditions employed here, the v0 values are linear with the PPase activity within a range from 0.025 to 0.4 U with a detection limit down to 0.010 U (S/N = 3). Moreover, the colorimetric method developed here is also employed for PPase inhibitor evaluation. This study offers a simple yet effective method for real-time PPase activity assay.