Plant carbon substrate supply regulated soil nitrogen dynamics in a tallgrass prairie in the Great Plains,USA: results of a clipping and shading experiment

Aims Land use management affects plant carbon (C) supply and soil environments and hence alters soil nitrogen (N) dynamics, with consequent feedbacks to terrestrial ecosystem productivity. The objective of this study was to better identify mechanisms by which land-use management (clipping and shading) regulates soil N in a tallgrass prairie, OK, USA.Methods We conducted 1-year clipping and shading experiment to investigate the effects of changes in land-use management (soil microclimates, plant C substrate supply and microbial activity) on soil inorganic N (NH 4 + ? N and NO 3 ? ? N), net N mineralization and nitrification in a tallgrass prairie.Important findings Land-use management through clipping and/or shading significantly increased annual mean inorganic N, possibly due to lowered plant N uptake and decreased microbial N immobilization into biomass growth. Shading significantly increased annual mean mineralization rates (P < 0.05). Clipping slightly decreased annual mean N nitrification rates whereas shading significantly increased annual mean N nitrification rates. Soil microclimate significantly explained 36% of the variation in NO 3 ? ? N concentrations (P = 0.004). However, soil respiration, a predictor of plant C substrate supply and microbial activity, was negatively correlated with NH 4 + ? N concentrations (P = 0.0009), net N mineralization (P = 0.0037) and nitrification rates (P = 0.0028) across treatments. Our results suggest that change in C substrate supply and microbial activity under clipping and/or shading is a critical control on NH 4 + ? N, net N mineralization and nitrification rates, whereas clipping and shading-induced soil microclimate change can be important for NO 3 ? ? N variation in the tallgrass prairie.     

Landscape Distribution of Microbial Activity in the McMurdo Dry Valleys: Linked Biotic Processes, Hydrology, and Geochemistry in a Cold Desert Ecosystem

In desert ecosystems, microbial activity and associated nutrient cycles are driven primarily by water availability and secondarily by nutrient availability. This is especially apparent in the extremely low productivity cold deserts of the McMurdo Dry Valleys, Antarctica. In this region, sediments near streams and lakes provide the seasonally wet conditions necessary for microbial activity and nutrient cycling and thus transfer energy to higher organisms. However, aside from a few studies of soil respiration, rates of microbial activity throughout the region remain unexplored. We measured extracellular enzyme activity potentials (alkaline phosphatase, leucine-aminopeptidase, beta-glucosidase, phenol oxidase, and peroxidase) in soils adjacent to lakes and streams, expecting activity to be primarily related to soil water content, as well as time of season and organic matter supply. Phosphatase and beta-glucosidase activities were higher in shoreline than upland soils; however, potential rates were not correlated with soil water content. Instead, soil organic matter, salinity, and pH were the best predictors of microbial activity. Microbial nutrient limitation metrics estimated from extracellular enzyme activity were correlated with pH and salinity and exhibited similar patterns to previously published trends in soil P and N content. Compared to other terrestrial ecosystems, organic matter specific rates for leucine-aminopeptidase and oxidative enzyme activities were high, typical of alkaline desert soils. Phosphatase activity was close to the global mean whereas beta-glucosidase activity was extremely low, which may reflect the lack of vascular plant derived organic matter in the Dry Valleys. In this cold desert ecosystem, water availability promotes microbial activity, and microbial nutrient cycling potentials are related to soil geochemistry. Author contributions:   LHZ performed research, analyzed data, and wrote the paper; RLS contributed new methods and wrote the paper; JEB conceived/designed study, performed research and analyzed data; MNG conceived/designed study and performed research; CTV conceived/designed study and performed research.     

Methane consumption by montane soils: implications for positive and negative feedback with climatic change

We report here three years of field observations of methane uptake, averaging 1.2 mg CH₄ m^–2 d^–1 in montane meadow soils. Surface soil moisture influenced diffusion of substrate while in deeper soil, where methane oxidation was maximum, moisture influenced both diffusion and microbial activity. Microbial oxidation of methane was maximum at an intermediate level of soil moisture, at this site at about 25% moisture by weight (50% water holding capacity). Laboratory incubations also showed inhibition below 20% moisture. These results provide in situ characterization of moisture limitation of methanotroph activity and evidence that soil drying may diminish the methane sink strength. The microbial limitation to methane consumption at low soil moisture provides a mechanism for positive feedback between methane flux and climate warming, as suggested by ice core data (Blunier et al. 1993; Chappellaz et al. 1990; Stauffer et al. 1985).     

Response of microbial activity and community structure to decreasing soil osmotic and matric potential

Low soil water content (low matric potential) and salinity (low osmotic potential) occur frequently in soils, particularly in arid and semi-arid regions. Although the effect of low matric or low osmotic potential on soil microorganisms have been studied before, this is the first report which compares the effect of the two stresses on microbial activity and community structure. A sand and a sandy loam, differing in pore size distribution, nutrient content and microbial biomass and community structure, were used. For the osmotic stress experiment, salt (NaCl) was added to achieve osmotic potentials from ?0.99 to ?13.13 MPa (sand) and from ?0.21 to 3.41 MPa (sandy loam) after which the soils were pre-incubated at optimal water content for 10d. For the matric stress experiment, soils were also pre-incubated at optimal water content for 10d, after which the water content was adjusted to give matric potentials from ?0.03 and ?1.68 MPa (sand) and from ?0.10 to 1.46 MPa (sandy loam). After amendment with 2% (w/w) pea straw (C/N 26), soil respiration was measured over 14d. Osmotic potential decreased with decreasing soil water content, particularly in the sand. Soil respiration decreased with decreasing water potential (osmotic?+?matric). At a given water potential, respiration decreased to a greater extent in the matric stress experiment than in the osmotic stress experiment. Decreasing osmotic and matric potential reduced microbial biomass (sum of phospholipid fatty acids measured after 14 days) and changed microbial community structure: fungi were less tolerant to decreasing osmotic potential than bacteria, but more tolerant to decreasing water content. It is concluded that low matric potential may be more detrimental than a corresponding low osmotic potential at optimal soil water content. This is likely to be a consequence of the restricted diffusion of substrates and thus a reduced ability of the microbes to synthesise osmolytes to help maintain cell water content. The study also highlighted that it needs to be considered that decreasing soil water content concentrates the salts, hence microorganisms in dry soils are exposed to two stressors.     

水肥处理对黄瓜土壤养分、酶及微生物多样性的影响

以津优1号黄瓜为试材,设3个土壤相对含水量水平（50%～60%、70%～80%、90%～100%）和2个肥料追施量（600 kg N·hm^-2和420 kg P₂O₅·hm^-2,420 kg N·hm^-2和294 kg P₂O₅·hm^-2）处理,研究了不同水肥供应对日光温室黄瓜土壤养分、酶活性及微生物多样性的影响.结果表明：土壤中NH₄⁺-N含量随施肥量的增加而提高,随土壤相对含水量的增加而降低;水肥供给的增加有利于提高土壤中速效磷含量和蔗糖酶活性;肥料增加使土壤中蛋白酶活性降低,而水分降低使土壤中脲酶活性提高.土壤中微生物多样性与土壤中养分含量无显著相关性,与土壤脲酶活性呈显著正相关,与蔗糖酶活性呈显著负相关.土壤相对含水量70%～80%、氮肥追施量600 kg N·hm^-2和420 kg P₂O₅·hm^-2处理的土壤养分含量、蔗糖酶、磷酸酶和脲酶活性较高,且土壤中微生物多样性和均匀度显著高于其他处理,土壤生产潜力最优.     

Influence of soil water potential on respiration and nitrogen fixation ofAzotobacter vinelandii

Summary Respiration and N₂-fixation (acetylene reduction) ofAzotobacter vinelandii have been studied at a variety of soil water potentials. Both processes were strictly linked and strongly reduced at water potentials between –0.6 and –1.3 MPa. Complete inhibition occurred below –2.1MPa. Osmotic potentials in soil compared to matric potentials of the same value were less inhibitory to respiration and acetylene reduction by Azotobacter. The N₂-fixing efficiency (mg N/g glucose) was not influenced by water potentials ranging from –0.1 to –2.1 MPa.     

Capacity for Methane Oxidation in Landfill Cover Soils Measured in Laboratory-Scale Soil Microcosms

Laboratory-scale soil microcosms containing different soils were permeated with CH(inf4) for up to 6 months to investigate their capacity to develop a methanotrophic community. Methane emissions were monitored continuously until steady states were established. The porous, coarse sand soil developed the greatest methanotrophic capacity (10.4 mol of CH(inf4) (middot) m(sup-2) (middot) day(sup-1)), the greatest yet reported in the literature. Vertical profiles of O(inf2), CH(inf4), and methanotrophic potential in the soils were determined at steady state. Methane oxidation potentials were greatest where the vertical profiles of O(inf2) and CH(inf4) overlapped. A significant increase in the organic matter content of the soil, presumably derived from methanotroph biomass, occurred where CH(inf4) oxidation was greatest. Methane oxidation kinetics showed that a soil community with a low methanotrophic capacity (V(infmax) of 258 nmol (middot) g of soil(sup-1) (middot) h(sup-1)) but relatively high affinity (k(infapp) of 1.6 (mu)M) remained in N(inf2)-purged control microcosms, even after 6 months without CH(inf4). We attribute this to a facultative, possibly mixotrophic, methanotrophic microbial community. When purged with CH(inf4), a different methanotrophic community developed which had a lower affinity (k(infapp) of 31.7 (mu)M) for CH(inf4) but a greater capacity (V(infmax) of 998 nmol (middot) g of soil(sup-1) (middot) h(sup-1)) for CH(inf4) oxidation, reflecting the enrichment of an active high-capacity methanotrophic community. Compared with the unamended control soil, amendment of the coarse sand with sewage sludge enhanced CH(inf4) oxidation capacity by 26%; K(inf2)HPO(inf4) amendment had no significant effect, while amendment with NH(inf4)NO(inf3) reduced the CH(inf4) oxidation capacity by 64%. In vitro experiments suggested that NH(inf4)NO(inf3) additions (10 and 71 (mu)mol (middot) g of soil(sup-1)) inhibited CH(inf4) oxidation by a nonspecific ionic effect rather than by specific inhibition by NH(inf4)(sup+).     

Microbial communities in terrestrial surface soils are not widely limited by carbon

Microbial communities in soils are generally considered to be limited by carbon (C), which could be a crucial control for basic soil functions and responses of microbial heterotrophic metabolism to climate change. However, global soil microbial C limitation (MCL) has rarely been estimated and is poorly understood. Here, we predicted MCL, defined as limited availability of substrate C relative to nitrogen and/or phosphorus to meet microbial metabolic requirements, based on the thresholds of extracellular enzyme activity across 847 sites (2476 observations) representing global natural ecosystems. Results showed that only about 22% of global sites in terrestrial surface soils show relative C limitation in microbial community. This finding challenges the conventional hypothesis of ubiquitous C limitation for soil microbial metabolism. The limited geographic extent of C limitation in our study was mainly attributed to plant litter, rather than soil organic matter that has been processed by microbes, serving as the dominant C source for microbial acquisition. We also identified a significant latitudinal pattern of predicted MCL with larger C limitation at mid- to high latitudes, whereas this limitation was generally absent in the tropics. Moreover, MCL significantly constrained the rates of soil heterotrophic respiration, suggesting a potentially larger relative increase in respiration at mid- to high latitudes than low latitudes, if climate change increases primary productivity that alleviates MCL at higher latitudes. Our study provides the first global estimates of MCL, advancing our understanding of terrestrial C cycling and microbial metabolic feedback under global climate change.     

The effects of different soil water potentials, temperature and salinity on germination of seeds of the desert shrub Zygophyllum dumosum

The effects of different soil water potentials, temperature and NaCl concentration on seed germination of Zygophyllum dumosum Boiss., a common shrub in Israeli deserts, was investigated. Seeds had to be exposed to constant field capacity conditions (–0.0316 MPa) for a minimal period of two days before germination could start. Maximal germination under such conditions occurred after four days or more. Under simulated conditions of gradual dehydration of the soil, seeds were inhibited either at low soil water potentials (–0.10 to –10.00 MPa) or at high ones (–0.002 to –0.0398 MPa). Germination of Z. dumosum was independent of temperature in the range of 10–25°C, but strongly inhibited at 30 and 35°C. At 20°C germination was inhibited by salinity of the medium but still occurred (0.5%) even at a concentration of 0.5 M NaCl.     

Effect of Water Diffusion Limitations on the Thermostability of Enzymes in Non-Aqueous Environments

The thermostability of anhydrous α-chymotrypsin has been analysed both in air and in organic solvents, with regard to the effect of the protein water-content on the course of deactivation. A higher initial water content increases the rate of inactivation.

Deactivation tests carried out under a constant thermodynamic activity of water indicate that reductions in dehydration rate lead to lower stability.

The effect of water diffusion phenomena has also been studied. Protein aggregates of larger size are less thermostable, thus indicating that diffusional limitations to water transfer can play a significant role in thermoinactivation.

The effect of water content on enzyme thermostability was also measured in the presence of two organic solvents of different hydrophobicity. In both cases, the resulting increased thermolability can be explained in terms of a limitation in water transfer towards the non-aqueous environment.     

The biogeochemical controls of N2O production and emission in landfill cover soils: the role of methanotrophs in the nitrogen cycle

Emissions of N2O from cover soils of both abandoned (> 30 years) and active landfills greatly exceed the maximum fluxes previously reported for tropical soils, suggesting high microbial activities for N2O production. Low soil matrix potentials (<-0.7 MPa) indicate that nitrification was the most likely mechanism of N2O formation during most of the time of sampling. Soil moisture had a strong influence on N2O emissions. The production of N2O was stimulated by as much as 20 times during laboratory incubations, when moisture was increased from -2.0 MPa to -0.6 MPa. Additional evidence from incubation experiments and delta13C analyses of fatty acids (18:1) diagnostic of methanotrophs suggests that N2O is formed in these soils by nitrification via methanotrophic bacteria. In a NH3(g)-amended landfill soil, the rate of N2O production was significantly increased when incubated with 100 ppmv methane compared with 1.8 ppmv (atmospheric) methane. Preincubation of a landfill soil with 1% CH4 for 2 weeks resulted in higher rates of N2O production when subsequently amended with NH3(g) relative to a control soil preincubated without CH4. At one location, at the soil depth (9-16 cm) of maximum methane consumption and N2O production, we observe elevated concentrations of organic carbon and nitrogen and distinct minima in delta15N (+1.0%) and delta13C (-33.8%) values for organic nitrogen and organic carbon respectively. A delta13C value of -39.3% was measured for 18:1 carbon fatty acids in this soil, diagnostic of type II methanotrophs. The low delta15N value for organic nitrogen is consistent with N2 fixation by type II methanotrophs. These observations all point to a methanotrophic origin for the organic matter at this depth. The results of this study corroborate previous reports of methanotrophic nitrification and N2O formation in aqueous and soil environments and suggest a predominance of type II rather than type I or type X methanotrophs in this landfill soil.     

Controls over nitric oxide and ammonia emissions from Mojave Desert soils

Emissions of reactive N compounds produced during terrestrial N cycling can be an important N loss pathway from ecosystems. Most measurements of this process focus on NO and N(2)O efflux; however, in alkaline soils such as those in the Mojave Desert, NH(3) production can be an important component of N gas loss. We investigated patterns of NO and NH(3) emissions in the Mojave Desert and identified seasonal changes in temperature, precipitation and spatial heterogeneity in soil nutrients as primary controllers of soil efflux. Across all seasons, NH(3) dominated reactive N gas emissions with fluxes ranging from 0.9 to 10 ng N m(-2) s(-1) as compared to NO fluxes of 0.08-1.9 ng N m(-2) s(-1). Fluxes were higher in April and July than in October; however, a fall precipitation event yielded large increases in both NO and NH(3) efflux. To explore the mechanisms driving field observations, we combined NO and NH(3) soil flux measurements with laboratory manipulations of temperature, water and nutrient conditions. These experiments showed a large transient NH(3) pulse (~70-100 ng N m(-2) s(-1)) following water addition, presumably driven by an increase in soil NH(4) (+) concentrations. This was followed by an increase in NO production, with maximum NO flux rates of 34 ng N m(-2) s(-1). Our study suggests that immediately following water addition NH(3) volatilization proceeds at high rates due to the absence of microbial competition for NH(4) (+); during this period N gas loss is insensitive to changes in temperature and soil nutrients. Subsequently, NO emission increases and rates of both NO and NH(3) emission are sensitive to temperature and nutrient constraints on microbial activity. Addition of labile C reduces gaseous N losses, presumably by increasing microbial immobilization, whereas addition of NO(3) (-) stimulates NO and NH(3) efflux.     

Contribution of osmotic adjustment to the dehydration tolerance of water-stressed pigeon pea (Cajanus cajan (L.) millsp.) leaves

Abstract Water-stressed pigeonpea leaves have high levels of osmotic adjustment at low leaf water potentials. The possible contribution of this adjustment of dehydration tolerance of leaves was examined in plants grown in a controlled environment. Osmotic adjustment was varied by withholding water from plants growing in differing amounts of soil, which resulted in different rates of decline of leaf water potential. The level of osmotic adjustment was inversely related to leaf water potential in all treatments. In addition, at any particular water potential, plants that had experienced a rapid development of stress exhibited less osmotic adjustment than plants that experienced a slower development of stress. Leaves with different levels of osmotic adjustment died at water potentials between –3.4 and –6.3 MPa, but all leaves died at a similar relative water content (32%). Consequently, leaves died when relative water content reached a lethal value, rather than when a lethal leaf water potential was reached. Osmotic adjustment delayed the time and lowered the leaf water potential when the lethal relative water content occurred, because it helped maintain higher relative water contents at low leaf water potentials. The consequences of osmotic adjustment for leaf survival in water-stressed pigeonpea are discussed.     

The Dual Arrhenius and Michaelis–Menten kinetics model for decomposition of soil organic matter at hourly to seasonal time scales

Decomposition of soil carbon stocks is one of the largest potential biotic feedbacks to climate change. Models of decomposition of soil organic matter and of soil respiration rely on empirical functions that relate variation in temperature and soil water content to rates of microbial metabolism using soil‐C substrates. Here, we describe a unifying modeling framework to combine the effects of temperature, soil water content, and soluble substrate supply on decomposition of soluble soil‐C substrates using simple functions based on process concepts. The model's backbone is the Michaelis–Menten equation, which describes the relationship between reaction velocity and soluble organic‐C and O₂ substrate concentrations at an enzyme's reactive site, which are determined by diffusivity functions based on soil water content. Temperature sensitivity is simulated by allowing the maximum velocity of the reaction (V_max) to vary according to Arrhenius function. The Dual Arrhenius and Michaelis–Menten kinetics (DAMM) model core was able to predict effectively observations from of laboratory enzyme assays of β‐glucosidase and phenol‐oxidase across a range of substrate concentrations and incubation temperatures. The model also functioned as well or better than purely empirical models for simulating hourly and seasonal soil respiration data from a trenched plot in a deciduous forest at the Harvard Forest, in northeastern United States. The DAMM model demonstrates that enzymatic processes can be intrinsically temperature sensitive, but environmental constrains of substrate supply under soil moisture extremes can prevent that response to temperature from being observed. We discuss how DAMM could serve as a core module that is informed by other modules regarding microbial dynamics and supply of soluble‐C substrates from plant inputs and from desorption of physically stabilized soil‐C pools. Most importantly, it presents a way forward from purely empirical representation of temperature and moisture responses and integrates temperature‐sensitive enzymatic processes with constraints of substrate supply.     

Nitrogen Transformations in Wetland Soil Cores Measured by (sup15)N Isotope Pairing and Dilution at Four Infiltration Rates

The effect of water infiltration rate (IR) on nitrogen cycling in a saturated wetland soil was investigated by applying a (sup15)N isotope dilution and pairing method. Water containing [(sup15)N]nitrate was infiltrated through 10-cm-long cores of sieved and homogenized soil at rates of 72, 168, 267, and 638 mm day(sup-1). Then the frequencies of (sup30)N(inf2), (sup29)N(inf2), (sup15)NO(inf3)(sup-), and (sup15)NH(inf4)(sup+) in the outflow water were measured. This method allowed simultaneous determination of nitrification, coupled and uncoupled denitrification, and nitrate assimilation rates. From 3% (at the highest IR) to 95% (at the lowest IR) of nitrate was removed from the water, mainly by denitrification. The nitrate removal was compensated for by the net release of ammonium and dissolved organic nitrogen. Lower oxygen concentrations in the soil at lower IRs led to a sharper decrease in the nitrification rate than in the ammonification rate, and, consequently, more ammonium leaked from the soil. The decreasing organic-carbon-to-nitrogen ratio (from 12.8 to 5.1) and the increasing light A(inf250)/A(inf365) ratio (from 4.5 to 5.2) indicated an increasing bioavailability of the outflowing dissolved organic matter with increasing IR. The efflux of nitrous oxide was also very sensitive to IR and increased severalfold when a zone of low oxygen concentration was close to the outlet of the soil cores. N(inf2)O then constituted 8% of the total gaseous N lost from the soil.     

Responses of soil nitrogen dynamics in a Mojave Desert ecosystem to manipulations in soil carbon and nitrogen availability

We investigated the effects of changes in soil C and N availability on N mineralization, nitrification, denitrification, NH(3) volatilization, and soil respiration in the Mojave Desert. Results indicate a C limitation to microbial N cycling. Soils from underneath the canopies of Larrea tridentata (DC.) Cov., Pleuraphis rigida Thurber, and Lycium spp. exhibited higher rates of CO(2 ) flux, lower rates of NH(3) volatilization, and a decrease in inorganic N (NH(4)(+)-N and NO(3)(-)-N) with C addition. In addition to C limitation, soils from plant interspaces also exhibited a N limitation. Soils from all locations had net immobilization of N over the course of a 15-day laboratory incubation. However, soils from interspaces had lower rates of net nitrification and potential denitrification compared to soils from under plant canopies. The response to changes in C availability appears to be a short-term increase in microbial immobilization of inorganic N. Under controlled conditions, and over a longer time period, the effects of C and N availability appear to give way to larger differences due to spatial location. These findings have implications for ecosystems undergoing changes in soil C and N availability due to such processes as desertification, exotic species invasions, or elevated atmospheric CO(2) concentration.     

Solid-state culture of Geotrichum candidum and Penicillium camembertii on a glutamate and lactate based medium

Geotrichum candidum and Penicillium camembertii have been cultivated at the surface of a glutamate, lactate-based medium. Glutamate has been chosen since it is a convenient carbon source for both fungi, in addition to a nitrogen source.The surface growth of both fungi induced the diffusion of substrates from the core to the rind. However, significant substrate concentrations (glutamate and lactate) always remained at the top of the gel, showing the absence of diffusional limitation of growth, in addition to the absence of substrate limitation. An absence of diffusional limitation, when the medium contained peptones instead of glutamate, was also indirectly deduced from the comparison of both media. Indeed, peptones are too complex for a possible identification of diffusional limitation from gradients analysis.An inhibitory effect of pH limited growth: at the end of the linear growth (oxygen limitation), inhibitory pH values were observed at the surface of the medium, even if it was not the case for the mean pH values. Since growth was limited by the alkaline pH at the surface of the gel, to account for this inhibition, an additional term has been introduced in the expression for the partial linking between consumption or production and growth. The diffusion coefficients for glutamate, lactate and ammonium have been also considered. Then, the concentration gradients for glutamate, lactate and ammonium have been calculated (second Fick law) and found to match with the experimental gradients.     

砂仁光合作用的CO2扩散限制与气孔限制分析

目前常用从气体交换参数计算的胞间CO₂浓度(C_i)来计算气孔限制值(L_s),但由于胁迫情况下计算的C_i偏高常导致结果不准确。该文引入扩散限制分析概念,以砂仁为例介绍了一种不需 C_i的计算扩散限制值(L_d)的新方法。同时通过叶绿素荧光参数间接估算受干旱胁迫植株的C_i(用C_i'表示)计算气孔限制值(L_s')。采用这3种方法分析了生长在100%和40%土壤相对湿度(RSM)下的砂仁(Amomum villosum)净光合速率的限制因素。结果表明两种水分状况下砂仁午后净光合速率的限制因素不同。100%RSM下,午后砂仁L_s没有升高,说明光合作用气孔限制并未增强;午后其L_d升高表明光合作用的CO₂扩散限制增强,这主要是由叶肉阻力相对增大所致。40%RSM下,午后砂仁L_s'升高比L_d升高明显,说明气孔阻力在所有扩散阻力中占主导作用,是限制净光合速率的主要原因;而其 L_s午后并未升高,暗示传统的气孔限制分析会得出非气孔限制的错误结论。C_i'低于C_i,说明干旱胁迫时传统的气体交换方法高估了C_i。上述结果都证明水分胁迫情况下传统方法不可靠,该文介绍的两种新方法比较准确可靠,同时使用两种新方法还可定性推测叶肉阻力的变化方向。     

Neurotoxic Organophosphate Degradation with Polyvinyl Alcohol Gel-Immobilized Microbial Cells

A genetically engineered strain of Escherichia coli that expresses organophosphorus hydrolase (OPH) was immobilized in a polyvinyl alcohol (PVA) cryogel to form a porous biocatalyst that successfully degrades organophosphorus (OP) neurotoxins. The impacts of both diffusion and reaction on biocatalyst efficiency were determined to enable prediction and optimization of the biocatalyst performance. The kinetic rate parameters and activation energies of pure OPH, free cell suspensions, and the immobilized cell biocatalyst were compared. Diffusion was a determining factor for paraoxon hydrolysis because of the very rapid OPH kinetics for its model substrate. Both the paraoxon diffusion through the PVA matrix and the diffusion associated with microbial transport of paraoxon were shown to impact the biocatalyst reaction. However, the enhancement in storage stability resulting from diffusional limitations provides an advantage to diffusion-limited operation. This research may serve as a guide to define the influence of diffusion in biological reaction systems. The broad substrate specificity and hydrolytic efficiency of OPH coupled with the ability to genetically engineer the enzyme for specific target OP neurotoxins enhance the suitability of OPH-based technologies for detoxification of these compounds. Cryoimmobilization provides a suitable vehicle as a cost-effective, efficient technology for bioremediation of environmental media contaminated with OP compounds.     

Kinetic aspects of the bioconversion of L-tyrosine into L-DOPA by cells of Mucuna pruriensL. Entrapped in different matrices

Plant cells of Mucuna pruriens L. entrapped In calcium alginate, calcium pectinate, agarose, or gelatine were able to convert L-tyrosine to L-DOPA, which was released Into the medium. Michaelis-Menten kinetics could be applied on the entrapped cells, based on the measurement of initial rates of L-DOPA production. The calculated apparent affinity constants were comparable with the affinity constants obtained with enzyme preparations. Comparison of the apparent maximum rate of bioconversion of the entrapped cells and the maximum rate of bioconversion of a derived cell homogenate indicated that the systems were not operating optimally. Measurement of the effective diffusion coefficients of L-tyrosine pointed out that this substrate could diffuse freely into the matrices. From the initial rates of bioconversion and the effective diffusion coefficients, the observable modulus was calculated for each system. The obtained values confirmed that the diffusional supply rate of L-tyrosine was not the limiting factor. For oxygen, which was utilized for byconversion as well as for cell respiration, the calculated observable moduli was directed toward strong oxygen transfer limitations. The values found for the oxygen consumption indicated that the entrapped cells remained partly or totally viable in the four matrices tested. Based on the highest viability and the highest rates of bioconversion, it was concluded that alginate-entrapped cells of M. pruriens formed the most suitable biocatalytic system for the production of L-DOPA from L-tyrosinre.