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PubMed2004

Arsenic complexes in the arsenic hyperaccumulator Pteris vittata (Chinese brake fern).

Pteris vittata (Chinese brake fern), the first reported arsenic (As) hyperaccumulating plant, can be potentially applied in the phytoremediation As-contaminated sites. Understanding the mechanisms of As tolerance and detoxification in this plant is critical to further enhance its capability of As hyperaccumulation. In this study, an unknown As species, other than arsenite (AsIII) or arsenate (AsV) was found in leaflets by using anion-exchange chromatography-hydride generation-atomic fluorescence spectroscopy and size-exclusion chromatography-atomic fluorescence spectrometry. The chromatographic behavior of this unknown As species and its stability suggest that it is likely an As complex. Although phytochelatin with two subunits (PC2) was the only major thiol in P. vittata under As exposure, this unknown As complex was unlikely to be an AsIII-PC2 complex by comparison of their chromatographic behaviors, stability at different pHs and charge states. The complex is sensitive to temperature and metal ions, but relatively insensitive to pH. In buffer solution of pH 5.9, it is present in a neutral form.

Arsenicals↗
PubMed2004

Antioxidative responses to arsenic in the arsenic-hyperaccumulator Chinese brake fern (Pteris vittata L.).

This study measured antioxidative responses of Chinese brake fern (Pteris vittata L.) upon exposure to arsenic (As) of different concentrations. Chinese brake fern was grown in an artificially-contaminated soil containing 0 to 200 mg As kg(-1) (Na2HAsO4) for 12 weeks in a greenhouse. Soil As concentrations at < or =20 mg kg(-1) enhanced plant growth, with 12-71% biomass increase compared to the control. Such beneficial effects were not observed at >20 mg As kg(-1). Plant As concentrations increased with soil As concentrations, with more As being accumulated in the fronds (aboveground biomass) than in the roots and with maximum frond As concentration being 4675 mg kg(-1). Arsenic uptake by Chinese brake enhanced uptake of nutrient elements K, P, Fe, Mn, and Zn except Ca and Mg, whose concentrations mostly decreased. The contents of non-enzymatic antioxidants (glutathione, acid-soluble thiol) followed similar trends as plant As concentrations, increasing with soil As concentrations, with greater contents in the fronds than in the roots especially when exposed to high As concentrations (>50 mg kg(-1)). The activities of enzymatic antioxidants (superoxide dismutase, catalase, ascorbate peroxidase, guaiacol peroxidase) in Chinese brake followed the same trends as plant biomass, increasing with soil As up to 20 mg kg(-1) and then decreased. The results indicated though both enzymatic and non-enzymatic antioxidants played significant roles in As detoxification and hyperaccumulation in Chinese brake, the former is more important at low As exposure (< or =20 mg kg(-1)), whereas the latter is more critical at high As exposure (50-200 mg kg(-1)).

Antioxidants↗
PubMed2004

Low molecular weight thiols in arsenic hyperaccumulator Pteris vittata upon exposure to arsenic and other trace elements.

Low molecular weight thiol-containing compounds have been reported to play an important role in metal detoxification and accumulation in some higher plants. The formation of these low molecular weight thiols in the recently discovered arsenic hyperaccumulator, Chinese Brake fern (Pteris vittata) upon exposure to arsenic and other trace metals was investigated. In addition to cysteine and glutathione, an unidentified thiol was observed in the plants exposed to arsenic, which was not found in the control. The concentration of the unidentified thiol showed a very strong and positive correlation with arsenic concentration in the leaflets. The unidentified thiol was low in rachises and undetectable in the roots for As-treated plants. Total and acid-soluble thiols were also measured and the results indicated that arsenic mainly stimulated the synthesis of acid-soluble thiol in Chinese Brake. The investigations of other trace elements (Cd, Cu, Cr, Zn, Pb, Hg, and Se) showed that these elements were not accumulated in Chinese Brake to high levels and the synthesis of the unidentified thiol in the plant was not observed. Our study suggests that the unidentified thiol was induced specifically by arsenic and the distribution patterns of the unidentified thiol and arsenic in the plant were consistent, indicating that the synthesis of this compound was related to As exposure.

Arsenic↗
PubMed2004

Thiol synthesis and arsenic hyperaccumulation in Pteris vittata (Chinese brake fern).

Pteris vittata (Chinese brake fern) has potential for phytoremediation of As-contaminated sites. In this study, the synthesis of total thiols and acid-soluble thiols in P. vittata was investigated under arsenic exposure. The strong and positive correlation between As concentration and acid-soluble thiols in plant leaflets suggests that acid-soluble thiols may play a role in As detoxification. A major As-induced thiol was purified and characterized. A molecular ion (M + 1) of 540 m/z suggests that the thiol was a phytochelatin (PC) with two base units (PC(2)). However, the ratios of acid-soluble thiols to As in leaflets exposed to As ranged from 0.012 to 0.026, suggesting that only a very small part of As is complexed by PC(2). PCs could play a minor detoxification role in this hyperaccumulator. A PC-independent mechanism appears to be mainly involved in As tolerance, while PC-dependent detoxification seems to be a supplement.

Arsenic↗
PubMed2004

Arsenic hyperaccumulation by Pteris vittata from arsenic contaminated soils and the effect of liming and phosphate fertilisation.

Pot experiments were carried out to investigate the potential of phytoremediation with the arsenic hyperaccumulator Pteris vittata in a range of soils contaminated with As and other heavy metals, and the influence of phosphate and lime additions on As hyperaccumulation by P. vittata. The fern was grown in 5 soils collected from Cornwall (England) containing 67-4550 mg As kg(-1) and different levels of metals. All soils showed a similar distribution pattern of As in different fractions in a sequential extraction, with more than 60% of the total As being associated with the fraction thought to represent amorphous and poorly-crystalline hydrous oxides of Fe and Al. The concentration of As in the fronds ranged from 84 to 3600 mg kg(-1), with 0.9-3.1% of the total soil As being taken up by P. vittata. In one soil which contained 5500 mg Cu kg(-1) and 1242 mg Zn kg(-1), P. vittata suffered from phytotoxicity and accumulated little As (0.002% of total). In a separate experiment, neither phosphate addition (50mg P kg(-1) soil) nor liming (4.6 g CaCO3 kg(-1) soil) was found to affect the As concentration in the fronds of P. vittata, even though phosphate addition increased the As concentration in the soil pore water. Between 4 and 7% of the total soil As was taken up by P. vittata in 4 cuttings in this experiment. The results indicate that P. vittata can hyperaccumulate As from naturally contaminated soils, but may be suitable for phytoremediation only in the moderately contaminated soils.

Arsenic↗
PubMed2005

Effects of arsenic on concentration and distribution of nutrients in the fronds of the arsenic hyperaccumulator Pteris vittata L.

Pteris vittata was the first terrestrial plant known to hyperaccumulate arsenic (As). However, it is unclear how As hyperaccumulation influences nutrient uptake by this plant. P. vittata fern was grown in soil spiked with 0-500 mg As kg(-1) in the greenhouse for 24 weeks. The concentrations of essential macro- (P, K, Ca, and Mg) and micro- (Fe, Mn, Cu, Zn, B and Mo) elements in the fronds of different age were examined. Both macro- and micronutrients in the fronds were found to be within the normal concentration ranges for non-hyperaccumulators. However, As hyperaccumulation did influence the elemental distribution among fronds of different age of P. vittata. Arsenic-induced P and K enhancements in the fronds contributed to the As-induced growth stimulation at low As levels. The frond P/As molar ratios of 1.0 can be used as the threshold value for normal growth of P. vittata. Potassium may function as a counter-cation for As in the fronds as shown by the As-induced K increases in the fronds. The present findings not only demonstrate that P. vittata has the ability to maintain adequate concentrations of essential nutrients while hyperaccumulating As from the soil, but also have implications for soil management (fertilization in particular) of P. vittata in As phytoextraction practice.

Arsenic↗
PubMed2004

Effects of heavy metals on growth and arsenic accumulation in the arsenic hyperaccumulator Pteris vittata L.

The effects of Cd, Ni, Pb, and Zn on arsenic accumulation by the arsenic hyperaccumulator Pteris vittata were investigated in a greenhouse study. P. vittata was grown for 8 weeks in an arsenic-contaminated soil (131 mg As kg(-1)), which was spiked with 50 or 200 mg kg(-1) Cd, Ni, Pb, or Zn (as nitrates). P. vittata was effective in taking up arsenic (up to 4100 mg kg(-1)) and transporting it to the fronds, but little of the metals. Arsenic bioconcentration factors ranged from 14 to 36 and transfer factors ranged from 16 to 56 in the presence of the metals, both of which were reduced with increasing metal concentration. Fern biomass increased as much as 12 times compared to the original dry weight after 8 weeks of growth (up to 19 g per plant). Greater concentrations of Cd, Ni, and Pb resulted in greater catalase activity in the plant. Most of the arsenic in the plant was present as arsenite, the reduced form, indicating little impact of the metals on plant arsenic reduction. This research demonstrates the capability of P. vittata in hyperaccumulating arsenic from soils in the presence of heavy metals.

Arsenic↗
PubMed2005

Interactions of mycorrhizal fungi with Pteris vittata (As hyperaccumulator) in As-contaminated soils.

A greenhouse trial was conducted to investigate the role of arbuscular mycorrhizas (AM) in aiding arsenic (As) uptake and tolerance by Pteris vittata (As hyperaccumulator) and Cynodon dactylon (a multi-metal root accumulator). Plants inoculated with lived and killed native mycorrhizas isolated from an As mine site were grown in a sterile and slightly acidic soil. The infectious percentage of mycorrhizas (0 mg/kg As: 26.4%, 50 mg/kg As: 30.3%, 100 mg/kg As: 40.6%) and the average biomass of shoots in infected P. vittata increased (0 mg/kg As: 2.45 g/pot, 50 mg/kg As: 2.48 g/pot, 100 mg/kg As: 10.9 g/pot) according to the increase of As levels when compared to control. The indigenous mycorrhizas enhanced As accumulation (0 mg/kg As: 3.70 mg/kg, 50 mg/kg As: 58.3 mg/kg; 100 mg/kg As: 88.1 mg/kg) in the As mine populations of P. vittata and also sustained its growth by aiding P absorption. For C. dactylon, As was mainly accumulated in mycorrhizal roots and translocation to shoots was inhibited.

Arsenic↗
PubMed2005

Factors influencing arsenic accumulation by Pteris vittata: a comparative field study at two sites.

This study compared the factors influencing arsenic (As) accumulation by Pteris vittata at two sites, one containing As along with Au mineralization and the other containing Hg/Tl mineralization. The soils above these two sites contained high As concentrations (26.8-2955 mg kg(-1)). Although the As concentration, pH, soil cation exchange capacity and plant biomass differed significantly between the two sites, no differences were observed in the As concentrations in the fronds and roots, or the translocation factors, of P. vittata, suggesting that this species has consistent As hyperaccumulation properties in the field. The As concentration in the fronds was positively related to phosphorus (P) and potassium (K), but negatively related to calcium (Ca), at one site. This suggested that P, K and Ca influenced As accumulation by P. vittata in the field.

Arsenic↗
PubMed2006

Arsenic chemistry in the rhizosphere of Pteris vittata L. and Nephrolepis exaltata L.

This greenhouse experiment evaluated the influence of arsenic uptake by arsenic hyperaccumulator Pteris vittata L. and non-arsenic hyperaccumulator Nephrolepis exaltata L. on arsenic chemistry in bulk and rhizosphere soil. The plants were grown for 8 weeks in a rhizopot with a soil containing 105 mg kg(-1) arsenic. The soil arsenic was fractionated into five fractions with decreasing availability: non-specifically bound (N), specifically bound (S), amorphous hydrous-oxide bound (A), crystalline hydrous-oxide bound (C), and residual (R). P. vittata produced larger plant biomass (7.38 vs. 2.32 mg plant(-1)) and removed more arsenic (2.61 vs. 0.09 mg pot(-1) arsenic) than N. exaltata. Plant growth reduced water-soluble arsenic, and increased soil pH (P. vittata only) in the rhizosphere soil. P. vittata was more efficient than N. exaltata to access arsenic from all fractions (39-64% vs. 5-39% reduction). However, most of the arsenic taken up by both plants was from the A fraction (67-77%) in the rhizosphere soil, the most abundant (61.5%) instead of the most available (N fraction).

Arsenic↗
PubMed2005

Immunomodulatory mechanism of the aqueous extract of sword brake fern (Pteris ensiformis Burm.).

Sword brake fern (Pteris ensiformis Burm.) is an ingredient in most of the traditional herbal beverage formulas in Taiwan; however, no information is available to explain its bioactivity. The aim of this study is to elucidate the molecular pharmacological activity in the aqueous extract of sword brake fern (SBF). We found that SBF (0.05-0.25 mg/ml) slightly induced TNF-alpha, IL-6, NO (nitric oxide) and PGE2 (prostaglandin E2) production in unstimualted murine macrophages, RAW264.7 cells. Furthermore, SBF (0.05-0.25 mg/ml) dose-dependently suppressed LPS-induced TNF-alpha, IL-1beta, IL-6, NO and PGE2 in activated RAW264.7 cells without exerting cytotoxicity. Further analysis of molecular mechanisms revealed that SBF prominently repressed LPS-induced iNOS (inducible nitric oxide synthase) and COX-2 (cyclooxygenase-2) promoter activities. Activation of the transcription factor NF-kappaB, which is one of the important pathways for transduction of LPS-stimulated inflammatory mediator producing signals, was suppressed by SBF in a dose-dependent manner, as demonstrated by both electrophoretic mobility shift assay (EMSA) and transfection with pNF-kappaB-Luc plasmid. These results suggest that SBF attenuates inflammatory mediator synthesis of activated macrophages partially through a NF-kappaB-dependent pathway. The immunomodulatory activity of SBF supports its traditional health promotion effect.

Animals↗
PubMed2004

Absorption of foliar-applied arsenic by the arsenic hyperaccumulating fern (Pteris vittata L.).

The fact that heavy metals can enter various domains of the plant system through foliar pathways spurred us to explore if the fronds of the Chinese brake fern (Pteris vittata L.), a hyperaccumulator of arsenic, a carcinogenic metalloid, was proficient in absorbing arsenic in the form of sprays. The specific objective of this study was to investigate the impact of frond age, form of arsenic, and time of application on the absorption of foliar-applied arsenic by the brake fern; also examined were the effects of foliar sprays on surface ultrastructure and arsenic speciation in the frond following absorption. Foliar sprays of different arsenic concentrations (0, 50, 100, 200, and 400 ppm) were applied to young and fertile fronds. A positive linear relationship existed between arsenic concentration and absorption; the arsenic concentration of fronds increased from 50 to 200 ppm. Time-course analysis with excised pinnae indicated an initial linear increase followed by a plateau at 48 h. The young fronds with immature sori absorbed more arsenic (3100 ppm) than the fertile mature fronds (890 ppm). In the frond, the arsenic absorption was greatest in the lamina of the pinnae followed by the sori and the rachis. Applying arsenic during night (20:00-22:00 h) or afternoon (12:00-14:00 h) resulted in greater absorption of arsenic than the application in the morning (08:00-10:00 h). The arsenic absorption was greater through abaxial surfaces than through adaxial surfaces. The brake fern absorbed more arsenic when it was applied in the form of arsenite. Regardless of the form of arsenic and the surface it was applied to, arsenic occurred as arsenite, the reduced and the most toxic form of arsenic, after having been absorbed by the fronds. Scanning electron microscopy revealed no surface morphological alterations following all arsenic sprays. The study unequivocally illustrated that the Chinese brake fern absorbed foliar-applied arsenic with great efficiency. Consequently, the arsenic concentrations in the fronds transcended the levels of hyperaccumulation; such a characteristic could be exploited in the phytoremediation of groundwater contaminated with arsenic.

Absorption↗
PubMed2005

Using phosphate rock to immobilize metals in soil and increase arsenic uptake by hyperaccumulator Pteris vittata.

This greenhouse experiment evaluated the effects of phosphate rock (PR) on arsenic and metal uptake by the arsenic hyperaccumulator Pteris vittata in a soil spiked with arsenic and heavy metals Cd, Pb and Zn. Five soil treatments were used, 1) control with no arsenic, 2) spiked with 50 mg kg(-1) As (As) as Na2H AsO4, 3) spiked with 50 mg kg(-1) As and P as PR (AsP), 4) spiked with 50 mg kg(-1) As, Pb, Cd, and Zn (AsM), and 5) spiked with 50 mg kg(-1) As, Pb, Cd, Zn and P (AsMP). The plants were harvested after growing in the soil for five weeks. Compared to the As treatment, the presence of heavy metals (AsM) reduced arsenic concentrations in the fronds from 1631 to 608 mg kg(-1). However, this effect was mitigated by PR (AsMP), with arsenic concentrations in the fronds increased from 608 to 1046 mg kg(-1). Phosphate rock also significantly reduced Pb (13.5 to 4.10 mg kg(-1)) and Cd (13.0 to 3.45 mg kg(-1)) concentrations in the fronds. Most of the arsenic in P. vittata was accumulated in the fronds (89-93%). Compared to the control, P was more concentrated in the roots along with less P being translocated to the fronds in the treatments with arsenic. While in those same treatments higher Ca concentrations in both the fronds and roots were observed. This research shows that PR was effective in increasing arsenic uptake and decreasing metal uptake by P. vittata and thus can be used as a cost-effective amendment for phytoremediation of arsenic and metal polluted soils.

Arsenic↗
PubMed2003

Arsenic species and leachability in the fronds of the hyperaccumulator Chinese brake (Pteris vittata L.).

Arsenic speciation is important not only for understanding the mechanisms of arsenic accumulation and detoxification by hyperaccumulators, but also for designing disposal options of arsenic-rich biomass. The primary objective of this research was to understand the speciation and leachability of arsenic in the fronds of Chinese brake (Pteris vittata L.), an arsenic hyperaccumulator, with an emphasis on the implications for arsenic-rich biomass disposal. Chinese brake was grown for 18 weeks in a soil spiked with 50 mg As kg(-1) as arsenate (AsO4(3-)), arsenite (AsO3(3-)), dimethylarsinic acid (DMA), or methylarsonic acid (MMA). Plant samples were extracted with methanol/water (1:1) and arsenic speciation was performed using high performance liquid chromatography coupled with atomic fluorescence spectrometry. The impacts of air-drying on arsenic species and leachability in the fronds were examined in the laboratory. After 18 weeks, water-soluble arsenic in soil was mainly present as arsenate with little detectable organic species or arsenite regardless of arsenic species added to the soil. However, arsenic in the fronds was primarily present as inorganic arsenite with an average of 94%. Arsenite re-oxidation occurred in the old fronds and the excised dried tissues. Arsenic species in the fronds were slightly influenced by arsenic forms added to the soil. Air-drying of the fronds resulted in leaching of substantial amounts of arsenic. These findings can be of significance when looking at disposal options of arsenic-rich biomass from the point of view of secondary contamination.

Arsenates↗
PubMed2003

Effects of compost and phosphate amendments on arsenic mobility in soils and arsenic uptake by the hyperaccumulator, Pteris vittata L.

Chinese brake fern (Pteris vittata L.), an arsenic (As) hyperaccumulator, has shown the potential to remediate As-contaminated soils. This study investigated the effects of soil amendments on the leachability of As from soils and As uptake by Chinese brake fern. The ferns were grown for 12 weeks in a chromated-copper-arsenate (CCA) contaminated soil or in As spiked contaminated (ASC) soil. Soils were treated with phosphate rock, municipal solid waste, or biosolid compost. Phosphate amendments significantly enhanced plant As uptake from the two tested soils with frond As concentrations increasing up to 265% relative to the control. After 12 weeks, plants grown in phosphate-amended soil removed >8% of soil As. Replacement of As by P from the soil binding sites was responsible for the enhanced mobility of As and subsequent increased plant uptake. Compost additions facilitated As uptake from the CCA soil, but decreased As uptake from the ASC soil. Elevated As uptake in the compost-treated CCA soil was related to the increase of soil water-soluble As and As(V) transformation into As(III). Reduced As uptake in the ASC soil may be attributed to As adsorption to the compost. Chinese brake fern took up As mainly from the iron-bound fraction in the CCA soil and from the water-soluble/exchangeable As in the ASC soil. Without ferns for As adsorption, compost and phosphate amendments increased As leaching from the CCA soil, but had decreased leaching with ferns when compared to the control. For the ASC soil, treatments reduced As leaching regardless of fern presence. This study suggest that growing Chinese brake fern in conjunction with phosphate amendments increases the effectiveness of remediating As-contaminated soils, by increasing As uptake and decreasing As leaching.

Adsorption↗
PubMed2003

Rhizosphere characteristics of the arsenic hyperaccumulator Pteris vittata L. and monitoring of phytoremoval efficiency.

Recently discovered As-hyperaccumulator ferns hold promise for phytoremediation of As-polluted soils. We investigated changes in the rhizosphere characteristics of Pteris vittata (Chinese Brake fern) relevant for its use in phytoextraction. Plants were grown in rhizoboxes filled with soil containing 2270 mg kg(-1) As. Dissolved organic carbon (DOC) concentrations in rhizosphere soil solution were increased by 86% and appeared to enhance total Fe solubility due to complexation reactions. Despite substantial removal of As by the fern, As was not significantly decreased in the rhizsophere soil solution after one cropping, apparently due to the large buffer capacity of the soil and possibly because of ion competition with DOC. However, the difference between 0.05 M (NH4)2SO4-extractable labile As in bulk and rhizosphere soil accounted for 8.9% of total As accumulated in the fern, indicating that As was mainly acquired from less available pools. Moreover, As depletion in the rhizosphere and limited resupply from less available pools were indicated by a 19.3% decreased As flux, measured using the technique of diffusive gradients in thin films (DGT). Modeling of the DGT-soil system was able to show that the rate of release from solid phase to solution in the rhizosphere was one-third of that in the bulk soil. Applying the remedial strategy of bioavailable contaminant stripping, which aims at diminishing the phytoavailable pollutant fraction, DGT can be used as a monitoring tool to evaluate the efficiency of phytoextraction and to study the potential resupply of bioavailable pools after phytoextraction has ceased.

Arsenic↗
PubMed2004

Over-expression of Bcl-2 against Pteris semipinnata L-induced apoptosis of human colon cancer cells via a NF-kappa B-related pathway.

Ent-11 alpha-hydroxy-15-oxo-kaur-16-en-19-oic-acid (5F), an antitumor component, is a chemical compound isolated from Pteris semipinnata L (PsL), a Chinese traditional herb. We examined whether 5F could affect apoptosis in human colon cancer HT-29 cells, and test whether and how the over-expression of Bcl-2 and Bcl-xL could offset the effect of 5F on cell growth. The result demonstrated that 5F significantly induced apoptosis of HT-29, as shown by MTT assay and DNA fragmentation measurement. Treatment of HT-29 with 5F increased both p38 and iNOS levels, suggesting these two molecules may contribute to the apoptotic effect of 5F. Over-expression of Bcl-2 or Bcl-xL attenuated the increase of p38 and iNOS induced by 5F. The cells with Bcl-2 or Bcl-xL over-expression showed an elevation of nuclear factor kappa B (NF-kappa B) activity, accompanying a significant reduction of 5F-induced apoptosis. Furthermore, inhibition of NF-kappa B by I k B alpha SR, which is a powerful inhibitor of NF-kappa B, restored the ability of 5F to induce apoptosis in the cells transfected with Bcl-2. These data strongly indicated that the apoptotic effect of 5F on HT-29 was closely associated with the activity of NF-kappa B, which was up-regulated by Bcl-2 and Bcl-xL. In conclusion, 5F induced apoptosis in HT-29 cells and this apoptotic effect was associated with the high level of p38 and iNOS expression. The apoptotic effect of 5F could be significantly offset by over-expression of either Bcl-2 or Bcl-xL. Bcl-2, and to the less extent, Bcl-xL, were able to increase the activity of NF-kappa B, which was a known anti-apoptotic molecule in human colon cancer cells.

Apoptosis↗
PubMed2001

An invertase inhibitory protein from Pteris deflexa link fronds.

Plant invertases play important roles in sucrose metabolism. Cell wall invertase was reported to participate in phloem loading and unloading. Soluble invertases would be involved in hexose level regulation in mature tissues and in stored sucrose utilization within vacuoles. Invertase inhibitory proteins were described as one of the possible mechanisms for invertase activity regulation in some plant species; nevertheless, these proteins were found only in sink tissues, suggesting that this mechanism would not be relevant in the sucrose turnover of leaves. This report describes the purification of invertase from Pteris deflexa fronds and the occurrence of an invertase inhibitory protein in this fern organ, as well as its purification and invertase-inhibitor interactions. The Mr of the invertase and of its inhibitory protein were 90,000 and 18,000, respectively. SDS-PAGE in the presence of 2-mercaptoetanol gave two subunits for the enzyme (Mr=66,000 and 30,000) and only one for the inhibitor. The inhibitor protein is a glycoprotein (12% w/w of neutral sugars) that did not show agglutinating activity like some others, and also showed a high heat stability at pH 5.0. The optimum pH of invertase activity is 5.0, while invertase inhibitory protein caused maximal inhibition at the same pH value. Invertase-inhibitor complex formation occurs in an immediate manner and a protease activity was discarded. The inhibition is non-competitive (Ki=1.5 x 10(-6) M) without interactions among the binding sites. The complex is slightly dissociable and sucrose was able to partially reduce the inhibitory effect. Up to the present, invertase inhibitory proteins have been found solely in heterotrophic tissues. In this work we demonstrate that this protein is also present in an autotrophic tissue of a lower vascular plant.

Enzyme Inhibitors↗