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1.
Laccases- or laccase-like multicopper oxidases (LMCO) catalyze the oxidation of various substrates, such as phenols, diamines and metals, coupled with the reduction of molecular oxygen to water. Compared to studies on function and diversity of LMCO in plants and fungi, little is known about this enzyme type in bacteria and especially on their possible implication in degradation of organic matter in soils. This study presents a molecular investigation of the diversity and distribution of bacterial LMCO genes among three upper horizons of a forest Cambisol and in a grassland Cambisol. Some culture strains of soil bacteria were also analyzed at the molecular level and for their capability to oxidize naturally occurring 2,6-dimethoxyphenol, a LMCO substrate. A high LMCO gene diversity was found in the Cambisol soil samples with 16 distinct sequence type clades, of which approximately one half was not matching with any reference sequence of known bacteria. The highest richness of bacterial LMCO genes was observed in the organic horizon of the forest soil, which is concomitant with a previous analysis of the diversity of fungal laccase genes and corresponding soil laccase activity. Some clusters of sequence types showed a specific distribution in one of the soils or in horizons, while others appeared more ubiquist. Multiple bacterial LMCO genes were described in Agromyces salentinus and Sinorhizobium morelense, what so far was only known from fungi.  相似文献   

2.
Laccases of fungal origin have been intensively studied due to their importance in various biotechnological applications. There is a constant demand for new laccases with improved properties such as stability at higher temperatures or at an alkaline pH. Growing molecular evidence suggests that laccases may also be widespread in bacteria. While only a handful of bacterial laccases have been purified and characterized, several novel traits have already been discovered (e.g. pH-stability and 2-domain organization of the enzyme as opposed to the usual 3-domain structure of fungal laccases). The aim of this study was to examine the diversity of bacterial laccase-like genes in two types of high-organic peat soil using a cloning and sequencing approach. Gene libraries prepared of small fragments (150 base pairs) revealed an amazing diversity of bacterial laccases. The fragments clustered in 11 major lineages, and one third of the 241 sequences resembled laccase-like genes of Acidobacteria. Additionally, a new primer was used to retrieve several larger fragments of the putative bacterial laccase genes that spanned all four copper-binding sites. Both “conventional” 3-domain laccases and the recently described 2-domain small laccases have been obtained using this approach, demonstrating the potential of the primer. The present study thus contributes to the understanding of the diversity of bacterial laccases and provides a new tool for finding laccase-like sequences in bacterial strains and soil samples.  相似文献   

3.
土壤酸性磷酸酶影响土壤有机磷矿化。选择不同的缓冲液种类和不同p H值测定土壤酸性磷酸酶活性,可影响数据的可比性。为明确缓冲液及p H值对不同土壤磷酸酶活性的影响程度,选取p H值4～8的土壤样本,测定了在醋酸盐缓冲液、柠檬酸盐缓冲液和MUB缓冲液3种不同培养条件下酸性磷酸酶活性变化。结果表明,采用MUB缓冲液条件可获得较稳定的土样区分序列,且与缓冲液的p H值的变化无关。在醋酸盐缓冲液和柠檬酸盐缓冲液条件下,不同p H值范围的不同土样之间土壤酸性磷酸酶活性变异系数变化剧烈,而采用MUB缓冲液获得的变异系数较其他缓冲液大,且趋势平稳。综合考虑,进行土壤酸性磷酸酶活性测定,可优先选用MUB缓冲溶液。  相似文献   

4.
Anthropogenic metal contamination is a pervasive problem in many urban or industrial areas. The interaction of metals with native soil communities is an important area of research as scientists strive to understand effects of long-term metal contamination on soil properties. Measurements of free soil enzyme activities can serve as useful indicators of microbial metabolic potential. The goals of this study are to determine extracellular soil enzymatic activities with respect to corresponding metal concentrations within a site of long-term contamination. These data are examined to understand relationships between extracellular soil enzyme activities and persistent metal loads in situ. Here we present such results from a rare research opportunity at an un-remediated, urban brownfield in Jersey City, NJ, USA. The soils of the site developed over the last 150 years through the dumping of urban fill from New York City as well as industrial rail use. The site was abandoned and fenced in the late 1960s, and within it, there is a mapped gradient of metal concentration in the soils, including As, Pb, Cr, Cu, Zn, and V. We measured soil enzymatic potential (alkaline phosphatase, cellobiohydrolase, and l-leucine-amino-peptidase) across four plots within the site and at an uncontaminated reference site that is of the same successional age and geographic influence. We found the highest enzymatic activities for all three activities measured at the site with the greatest soil metal loads and a particularly strong relationship among enzyme activity and the metals V and Cr. Our results differ from many experimental studies that show decreased soil enzyme activity in soils experimentally treated with metals. The results may indicate the effects of long-term adaptation of soil communities within these metal contaminated soils.  相似文献   

5.
Lichens form the dominant plant cover in extreme environments and participate in mineral weathering, fine-earth stabilization and primary accumulation of soil organic matter. However, biochemical role of lichens in soil processes has never been investigated. Recently, laccases and tyrosinases have been discovered in representatives of the order Peltigerales (Laufer et al., 2006a, b; Zavarzina and Zavarzin, 2006). Laccases from most species had unusually large molecular weights (Laufer et al., 2009). Together with oligomeric laccases, we have found monomeric enzymes in Solorina crocea and Peltigera aphthosa (Lisov et al., 2007). In the present work we have purified homodimeric (large) and monomeric (small) laccases of the soil-stabilizing lichen S. crocea, determined their physico-chemical and catalytic properties and studied their reactions with soil humic acids. Our results suggest that oligomeric nature of lichen laccases can be artifactual, because homodimeric laccase was transformed into the monomeric form following hydrophobic interaction chromatography. We hypothesize that large laccase consists of two monomeric enzymes, each of which is bound with additional hydrophobic component(s). Small laccase is similar in its properties to the laccases of basidiomycetes. It is more resistant to elevated temperature and storage than the large form, showed a higher oxidation potential, had different pH-optima in oxidizing substrates and was less inhibited by humic acids. Despite these differences, both laccases depolymerized and decolorized humic acids from soils at comparable rates, with small laccase being slightly more effective. This finding suggests that lichens have a potential to participate in transformation of soil organic matter.  相似文献   

6.
Organo-mineral interactions have been hypothesized to play a major role in biogeochemical cycling and pedogenesis in some forest soils. These processes are likely to be controlled to some extent by their persistence in soil, however, the factors regulating their bioavailability remain poorly understood. Therefore, we investigated the microbial utilization of 14C-labeled citrate in glass bead filled bioreactors containing a biofilm developed from an inoculum from an acid forest soil. The removal of Al-citrate in the bioreactors was negligible compared to the rate of citrate removal in the absence of Al. There was no evidence that in the short-term the microbial community adapted to increase the utilization of Al-citrate. In bioreactors filled with a Picea abies forest soil (Haplic Arenosol) the rates of citrate utilization were always slightly higher than that of Al-citrate. We conclude that complexation of citrate by metals such as Al may have a significant effect on their role in soil biogeochemical cycles.  相似文献   

7.
The transformation of naturally occurring phenols to humic polymers through oxidative coupling reactions may involve oxidoreductive enzymes and soil minerals as catalysts. There is limited information on the possible inhibitory or synergistic interactions between oxidoreductases and mineral catalysts as they participate in oxidative coupling of phenolic substrates. In this study, a ternary system was investigated, in which a fungal enzyme (Trametes villosa laccase), birnessite (δ-MnO2), and a naturally occurring phenolic compound (catechol) were reacted together to model soil processes. Binary systems (catechol/laccase and catechol/birnessite) were included for comparison. In the absence of the mineral, T. villosa laccase (950 katal ml−1) transformed 31% of catechol, whereas birnessite (1 mg ml−1) in the absence of the enzyme showed a 24% catechol transformation. The percentages of catechol transformation in the binary systems did not accumulate in the ternary system; instead, birnessite and laccase tested together transformed only 36% of catechol. This suggested that birnessite had an inhibitory effect on substrate transformation by laccase catalysis. Enzyme assays indicated that inhibition was a result of enzyme deactivation by humic-like polymers produced by birnessite, and by Mn2+ ions released from the mineral. These observations underscore the importance of considering enzyme-soil mineral-organic matter interactions in studies of humus formation and contaminant removal.  相似文献   

8.
Recent studies have suggested that the organic matter contents of undisturbed soils (under natural vegetation) are in equilibrium with biological and biochemical properties. Accordingly, we hypothesised that such equilibria should be disrupted when soils are subjected to disturbance or stress, and that measurement of this disruption can be expressed mathematically and used as a soil quality index. In this study, we evaluated these hypotheses in soils from the H.J. Andrews Experimental Forest in Oregon. Both O and A horizons were sampled from nine sites in Spring 2005 and Fall 2006. Soil samples were analyzed for enzyme activities (phosphatase, β-glucosidase, laccase, N-acetyl-glucosaminidase, protease and urease), and other biological and chemical properties including N-mineralization, respiration, microbial biomass C (MBC), soil organic carbon (SOC) and total nitrogen content. In addition, soil samples from one old-growth site were manipulated in the laboratory to either simulate chemical stresses (Cu addition or pH alteration) or physical disturbances (wet-dry or freeze-thaw cycles). The results showed variation in biological and biochemical soil properties that were closely correlated with SOC. Multiple regression analysis of SOC levels against all soil properties showed that a model containing only MBC and phosphatase activity could account for 97% of the SOC variation among the sites. The model fit was independent of spatial and temporal variations because covariates such as site, stand age, sampling date, and soil horizon were found to be not statistically significant. Although the application of stress/disturbance treatments inconsistently affected most of the individual biochemical properties, in contrast, the ratio of soil C predicted by the model (Cp), and soil C measured (Cm) was consistently reduced in soils submitted to at least one level of stress and disturbance treatments. In addition, Cp/Cm was more affected in soils submitted to wet-dry cycles and Cu contamination than to freeze-thaw cycles or shifts in soil pH. Our results confirm previous evidence of a biochemical balance in high quality undisturbed soils, and that this balance is disrupted when the soil is submitted to disturbances or placed under stress conditions. The Cp/Cm ratio provides a simple reference value against which the degrading effects of pollutants or management practices on soil quality can be assessed.  相似文献   

9.
The impact of forest tree leaf litters on microbial activity and nutrient status of red laterite soil was tested for the ecological suitability of Cassia siamea, Shorea robusta, Acacia auriculiformes and Dalbergia sissoo, which are typically used for afforestation of wastelands in eastern India. The objectives were to compare seasonal variation in soil enzyme activity in 30-years old afforested sites, and to study nutrient status and microbial biomass and function during short-term in-situ incubation of litter in decomposition pits. In afforested soils, enzyme activities significantly varied between litters and seasons. All enzyme activity except invertase dominated in the soils containing Dalbergia and Cassia litters compared to the others. The seasonal effect was enzyme-dependent, with amylase and cellulase reaching peaks during the rainy season but invertase activity showed a reverse trend with lowest values in rainy season, except in Acacia soil, and protease activity was lowest in the soil containing Cassia and Dalbergia during the rainy season. Dehydrogenase activity was negligible in the soils containing Shorea and Acacia, but remained high with respect to Dalbergia and Cassia during all seasons. The decomposition pit study showed significant increase of soil nutrients with respect to litter types and intervals, except with respect to electrical conductivity. Cassia and Dalbergia litters enabled notable increase of soil nutrients than Shorea and Acacia. The soil enzyme activity, in general, increased with duration of litter decay, but microbial biomass C (MBC) decreased over time except in Shorea. Therefore, the enzyme rates normalized to the MBC indicated inverse relations for all enzymes, except in the soil containing Shorea. A positive relationship existed between MBC and soil respiration in Cassia, Acacia and Dalbergia. Analysis of variance revealed main effects of litter types for increasing protease, MBC and CO2 output, and a main effect of intervals for enhancing enzymes other than cellulase. Rates of soil respiration were greater in soils contain Cassia and Dalbergia, and showed significant differences between litters and between intervals. All enzymes were significantly correlated with electrical conductivity, organic carbon and available phosphorus contents, and all enzymes except invertase were correlated with nitrate concentrations. The acidic soil pH did not affect enzyme activities, and soil nutrients exerted only weak effect on MBC and respiration. Our study showed that leaf litters of Cassia and Dalbergia trees improved the nutrient status and microbial activity in soil more so than Shorea and Acacia litters, and therefore, afforestation using Cassia and Dalbergia trees may be particularly suitable for soil restoration in tropical laterite wastelands.  相似文献   

10.
There have been few investigations of the possible effects of genetically engineered plants on the microbiota and enzyme activities in flooded soil. We studied the influence of the transgenic rice KeMingDao (KMD) straw on the culturable microbiota and enzymatic activities in a flooded paddy soil under laboratory conditions. KMD contained a synthetic cry1Ab gene from Bacillus thuringiensis under the control of a maize ubiquitin promoter and linked in tandem with the gusA and hpt genes. The results showed that there were only some occasional significant differences (P<0.05) in the number of Colony forming units of aerobic bacteria, actinomycetes and fungi and in the number of anaerobic fermentative bacteria, denitrifying bacteria, hydrogen-producing acetogenic bacteria, and methanogenic bacteria between the paddy soil amended with Bt-transgenic rice straw and with the non-Bt parental rice straw during the early stages of incubation. From d14 to d84 there were significant increases (P<0.05) in soil dehydrogenase and soil neutral phosphatase activity in soils amended with rice straw compared to soil without added straw. The dehydrogenase activity was significantly greatly (almost 1.95-fold) in soil amended with Bt-transgenic straw from d7 to d14 but from d21 to d49 there was significantly greater activity (about 1.47-fold) in the soil amended with non-Bt-straw. There were no apparent differences between the activity of soil neutral phosphatase in the soils to which non-Bt-straw and Bt-straw had been added. However, both soils to which rice straws were added demonstrated significant differences in the number of microorganisms except for aerobic bacteria and enzymatic activities with respect to the control soil throughout the incubation. The above results indicated that the Bt-straw from KMD transgenic rice is not toxic to a variety of culturable microorganisms in the studied flooded paddy soil.  相似文献   

11.
The original focus of this study was an analysis of proteolytic enzyme activity of Alaskan arctic tundra soils, however initial results raised questions regarding the method (Watanabe and Hayano, 1995). Thus, the goals of the study changed to 1) an investigation of the method, and 2) a comparison of enzyme activities of two different soil layers from the arctic tundra. Methodological examination included the impact of toluene, used to prevent immobilization of the product, and blank correction on enzyme activity, and a search for a true 6-h linear rate of activity during a 48-h incubation. We measured native and potential, using casein as an artificial substrate, activities as net amino acid production in mineral and organic soil layer samples. Varying toluene concentration had no clear effect on activity; omitting toluene resulted in zero native activity and reduced potential for the organic samples, but not for the mineral. Comparison of activities with and without blank correction indicated, particularly for potential activity of samples with low native rates, that correction was required for accuracy. Native and potential activity of the organic samples, and native of the mineral were linear during the first 6 h of incubation; linearity was found during the 6-24 h incubation for potential activity of the mineral. Soil layer activity data indicated that native activity was higher in organic soils as compared with mineral. The organic layer potential activity was ten-fold greater than the native, suggesting substrate limitation; potential and native activities did not differ in the mineral layer, indicating substrate sufficiency. Casein addition changed the kinetic pattern for both layers from hyperbolic to sigmoidal for the mineral and linear for the organic, implying different enzyme pools or behavioral changes of existing pools. Native activity based on total soluble protein was higher for the mineral samples relative to the organic, reiterating substrate capacity differences and variations in enzyme/substrate interactions.  相似文献   

12.
Riparian buffers, located in the transition zone between terrestrial and aquatic ecosystems, are a hotspot for nitrogen (N) removal through denitrification. Earthworms are abundant in riparian buffers and may enhance denitrification. This study investigated earthworm demographics of three earthworm functional groups (anecic, epigeic, and endogeic) and denitrifier activity in temporarily flooded and non-flooded riparian soils from April to October 2012 in southern Quebec, Canada. Nine earthworm species, mostly endogeic, were found in the temporarily flooded soil, while only six earthworm species were found in the non-flooded soil. On average, there were 11.7 times more earthworms with 12.4 times greater biomass (P<0.05) found in the temporarily flooded soil than in the non-flooded soil. The denitrification enzyme activity (DEA) was of similar magnitude in temporarily flooded and non-flooded soils, with temporal variation associated with rainfall patterns. Endogeic earthworm biomass was positively correlated (P<0.05) with DEA, while epigeic earthworm biomass was positively correlated (P<0.05) with 16S rRNA gene copies and nosZ gene copies from bacteria, indicating an association between earthworm functional groups and denitrifier activity in riparian soils. Stepwise multiple regressions showed that DEA in riparian soils could be predicted using soil moisture, inorganic N concentration, and earthworm functional groups, suggesting that endogeic and epigeic earthworms contributed to denitrifier activity in riparian soils.  相似文献   

13.
Soil microbial communities and their activities are altered by land use change; however impacts and extent of these alterations are often unclear. We investigated the functional responses of soil microbes in agricultural soil under sugarcane and corresponding native soil under Eucalyptus forest to additions of contrasting plant litter derived from soybean, sugarcane and Eucalyptus in a microcosm system, using a suite of complimentary techniques including enzyme assays and community level physiological profiles (CLPP). Initially agricultural soil had 50% less microbial biomass and lower enzyme activities than forest soil, but significantly higher nitrification rates. In response to litter addition, microbial biomass increased up to 11-fold in agricultural soil, but only 1.8-fold in forest soil, suggesting a prevalence of rapidly proliferating ‘r’ and slower growing ‘K’ strategists in the respective soils. Litter-driven change in microbial biomass and activities were short lived, largely returning to pre-litter addition levels by day 150. Decomposition rates of sugarcane and soybean litter as estimated via CO2 production were lower in agricultural than in forest soil, but decomposition of more recalcitrant Eucalyptus litter was similar in both soils, contradicting the notion that microbial communities specialise in decomposing litter of the dominant local plant species. Enzyme activities and community level physiological profiles (CLPP) were closely correlated to microbial biomass and overall CO2 production in the agricultural soil but not the forest soil, suggesting contrasting relationships between microbial population dynamics and activity in the two soils. Activities of enzymes that break down complex biopolymers, such as protease, cellulase and phenol oxidase were similar or higher in the agricultural soil, which suggests that the production of extracellular biopolymer-degrading enzymes was not a factor limiting litter decomposition. Enzyme and CLPP analyses produced contrasting profiles of microbial activity in the two soils; however the combination of both analyses offers additional insights into the changes in microbial function and community dynamics that occur after conversion of forest to agricultural land.  相似文献   

14.
We studied the effects of a biochar made from fast pyrolysis of switchgrass on four soil enzymes (β-glucosidase, β-N-acetylglucosaminidase, lipase, and leucine aminopeptidase) to determine if biochar would consistently modify soil biological activities. Thus, we conducted a series of enzyme assays on biochar-amended soils. Inconsistent results from enzyme assays of char-amended soils suggested that biochar had variable effects on soil enzyme activities, thus we conducted a second experiment to determine if biochar reacts predictably with either enzyme or substrate in in vitro reactions. Both colorimetric and fluorescent assays were used for β-glucosidase and β-N-acetylglucosaminidase. Seven days after biochar was added to microcosms of 3 different soils, fluorescence-based assays revealed some increased enzyme activities (up to 7-fold for one measure of β-glucosidase in a shrub-steppe soil) and some decreased activities (one-fifth of the unamended control for lipase measured in the same shrub-steppe soil), compared to non-amended soil. In an effort understand the varied effects, purified enzymes or substrates were briefly exposed to biochar and then assayed. In contrast to the soil assays, except for β-N-acetylglucosaminidase, the exposure of substrate to biochar reduced the apparent activity of the enzymes, suggesting that sorption reactions between substrate and biochar impeded enzyme function. Our findings indicate that fluorometric assays are more robust to, or account for, this sorption better than the colorimetric assays used herein. The activity of purified β-N-acetylglucosaminidase increased 50-75% following biochar exposure, suggesting a chemical enhancement of enzyme function. In some cases, biochar stimulates soil enzyme activities, to a much greater degree than soil assays would indicate, given that substrate reactivity can be impeded by biochar exposure. We conclude that the effects of biochar on enzyme activities in soils are highly variable; these effects are likely associated with reactions between biochar and the target substrate.  相似文献   

15.
Rhodanese activity (RA) was studied in 4 soils, incubated under flooded and nonflooded (60% water-holding capacity) conditions. RA in 3 soils including an acid sulphate soil pokkali increased 2.5–6.0-fold (over respective nonflooded soils), while activity of the enzyme decreased markedly in flooded alluvial soil. Similarly, anaerobic incubation of nonflooded soils under N2 decreased RA in an alluvial soil, but increased it in pokkali soil. RA was negligible in soils, that had been reduced by flooding for 30 days and then sterilized by autoclaving. Rice rhizosphere soil exhibited significantly higher RA than the nonrhizosphere soil samples under flooded or nonflooded conditions. RA in aerobic soils was related to the microbial oxidation of S° to SO2?4. But, no relationship could be established between RA and S-oxidation in flooded soils and in rhizosphere soil suspensions of flooded rice plants.  相似文献   

16.
The aim of this study was to investigate the potentials and limitations in restoring soil suppressiveness in disturbed soils. Soils from three sites in UK and Switzerland (STC, REC, THE) differing in their level of suppressiveness to soil-borne and air-borne diseases were γ-irradiated and this soil matrix was re-inoculated with 1% (w/w) of either parent native soil or native soil from the other sites (‘soil inoculum’). Suppressiveness to air-borne and soil-borne diseases was quantified by means of the host-pathogen systems Lepidium sativum (cress)-Pythium ultimum, an oomycete causing root rot and seedling damping-off, and Arabidopsis thaliana-Hyaloperonospora parasitica, an oomycete causing downy mildew. Soil microbial biomass, activity and community structure, as determined by phospholipid fatty acid (PLFA) profiles, were measured in native, γ-irradiated, and re-inoculated soils. Both, L. sativum and A. thaliana were highly susceptible to the pathogens if grown on γ-irradiated soils. Re-inoculation completely restored suppressiveness of soils to the foliar pathogen H. parasitica, independently of soil matrix or soil inoculum, whereas suppressiveness to P. ultimum depended on the soil matrix and, to a lesser extent, on the soil inoculum. However, the soil with the highest inherent suppressiveness did not reach the initial level of suppressiveness after re-inoculation. In addition, native microbial populations as defined by microbial biomass, activity and community structure, could not be fully restored in re-inoculated soils. As for suppressiveness to P. ultimum, the soil matrix, rather than the source of soil inoculum was identified as the key factor for re-establishing the microbial community structure. Our data show that soils do not or only slowly fully recover from sterilisation by γ-irradiation, indicating that agricultural soil management practices such as soil fumigation or heat treatments frequently used in vegetable cropping should be avoided.  相似文献   

17.
Abstract

Total ionic strength buffers based on citrate at pH 7.5 and cyclohexylenediaminetetra‐acetic acid (CDTA) at pH 5.2 were evaluated for use when determining fluoride (F) concentration in soil solutions. Both buffers achieved acceptable recoveries of F added to ultra‐filtered (0.025 μm) solutions. Recovery of F added to unflltered and 0.22 μm filtered solution was low when the CDTA buffer was used. This effect was attributed to the adsorption of F by micro‐particulate materials present in these samples. The use of this buffer would need to be restricted to samples in which micro‐particulates were not present. The performance of the citrate buffer was not affected by the presence of microparticulates; however, electrode response times, and departure from Nernstian response at low F concentrations, were greater when this buffer was employed. Fluoride was found to be present at low concentrations in the soil solution of the 60 virgin highly weathered soils tested. The F concentration in 75% of the surface samples was <3 μM, while all subsoil samples had soil solution F concentrations <1 μM.  相似文献   

18.
The thermodynamic parameters of the enzymes catalase, dehydrogenase, casein-protease, α-N-benzoyl-l-argininamide (BAA)-protease, urease, Carboxymethyl (CM)-cellulase, invertase, β-glucosidase and arylsulphatase, were investigated in grassland soils from a European temperate-humid zone (Galicia, NW Spain). The effect of temperature on enzyme activity was determined at 5, 18, 27, 37, 57 and 70 °C. The temperature-dependence of the rate of substrate hydrolysis varied depending on the enzyme and soil. In general, the soil containing the least amount of organic matter (OM) showed the lowest enzyme activity for all temperatures and enzymes, whereas soils with similar OM contents showed similar levels of activity for the entire temperature range. Temperature had a noteworthy effect on the activity of oxidoreductases. Product formation in the reaction catalyzed by dehydrogenase increased with increasing temperature until 70 °C, which was attributed to chemical reduction of iodonitrotetrazolium violet (INT) at high temperatures. Catalase activity was not affected above 37 °C, which may be explained either by non-enzymatic decomposition of hydrogen peroxide or by the fact that catalase has reached kinetic perfection, and is therefore not saturated with substrate.The Arrhenius equation was used to determine the activation energy (Ea) and the temperature coefficient (Q10) for all enzymes. The values of Ea and Q10 for each enzyme differed among soils, although in general the differences were small, especially for those enzymes that act on substrates of low molecular weight. In terms of the values of Ea and Q10 and the differences established among soils, the results obtained for those enzymes that act on substrates of high molecular weight differed most from those corresponding to the other enzymes. Thus the lowest Ea and Q10 values corresponded to BAA-protease, and the highest values to CM-cellulase and casein-protease. Except for catalase in one of the soils, the values of Ea and Q10 for the oxidoreductases were similar to those of most of the hydrolases. In general, the effect of temperature appeared to be more dependent on the type of enzyme than on the characteristics of the soil.  相似文献   

19.
The purpose of this experiment was to evaluate whether soil storage and processing methods significantly influence measurements of potential in situ enzyme activity in acidic forest soils. More specifically, the objectives were to determine if: (1) duration and temperature of soil storage; (2) duration of soil slurry in buffer; and (3) age of model substrates significantly influence the activity of six commonly measured soil extracellular enzymes using methylumbelliferone (MUB)-linked substrates and l-dihydroxyphenylalanine (l-DOPA). Soil collected and analyzed for enzyme activity within 2 h was considered the best measure of potential in situ enzyme activity and the benchmark for all statistical comparisons. Sub-samples of the same soil were stored at either 4 °C or −20 °C. In addition to the temperature manipulation, soils experienced two more experimental treatments. First, enzyme activity was analyzed 2, 7, 14, and 21 days after collection. Second, MUB-linked substrate was added immediately (i.e. <20 min) or 2 h after mixing soil with buffer. Enzyme activity of soil stored at 4 °C was not significantly different from soil stored at −20 °C. The duration of soil storage was minimal for β-glucosidase, β-xylosidase, and peroxidase activity. N-acetyl-glucosaminidase (NAGase), phosphatase, and phenol oxidase activity appeared to change the most when compared to fresh soils, but the direction of change varied. Likewise, the activities of these enzymes were most sensitive to extended time in buffer. Fluorometric MUB and MUB-linked substrates generally had a 3-day shelf life before they start to significantly suppress reported activities when kept at 4 °C. These findings suggest that the manner in which acidic forest soils are stored and processed are site and enzyme specific and should not initially be trivialized when conducting enzyme assays focusing on NAGase, phosphatase, and phenol oxidase. The activities of β-glucosidase, β-xylosidase, and peroxidase are insensitive to storage and processing methods.  相似文献   

20.
Soil laccase was extracted with 0. 1 m sodium pyrophosphate and purified by Sephadex G-25 gel chromatography. Oxygen consumption during the enzymatic reaction was determined by a polarographic method with catechol or p-phenylenediamine as the substrates. The fraction showed laccase activity toward both the substrates. The activity toward p-phenylenediamine was six times higher than that toward catechol. The properties of soil laccase suggest that the enzyme is associated with humic compounds and forms a humus-laccase complex (Ruggiero and Radogna, 1984). The effect of three phenoxyacetic and three s-triazine herbicides on the activity of the humus-laccase complex was assessed. All the herbicides inhibited laccase activity to varying degrees. The inhibition of humus-laccase activity by 2,4-D was investigated in detail. The data, plotted in the form of a Lineweaver-Burk plot, showed that the 2,4-D acted as a non-competitive inhibitor. The apparent Km values were 28.7 and 6.0 nm for catechol and p-phenylenediamine, respectively. Some aspects of the relationship between herbicide, substrate and humus-laccase complex are discussed.  相似文献   

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