Contparison of the influence of Cu,Zn, and Cd on the activity and kinetics of free and intntobilized acid phosphatase

The influence of Cu, Zn, and Cd on the activity and kinetics of acid phosphatase immobilized by two soil clays, kaolin or goethite was investigated. The ability of Cu to inhibit the enzyme activity was higher than that of Zn in all the enzyme complexes examined. The ability of Cd was negligible. The inhibitory effects of Cu and Zn on the two soil clay- and kaolin-enzyme complexes were much stronger than those on the goethite-enzyme complex. The V _max and K _m values of the enzyme complexes indicated that both Cu and Zn decreased the maximum reaction velocity of the enzymes, but increased the affinity of the enzymes for the substrate. The degree of the decrease and increase was higher in the Cu systems than in the Zn systems.     

柠檬酸盐和磷酸盐存在下酸性磷素酶在矿物和土壤胶体上的吸附

The aim of this work was to study the influence of phosphate and citrate, which are common inorganic andorganic anions in soils, on the adsorption of acid phosphatase by kaolin, goethite and the colloids separatedfrom yellow-brown soil (YBS) and latosol (LS) in central-south China. The YBS colloid has the major claymineral composition of 1.4 nm mineral, illite and kaolinite while the LS colloid mainly contains kaolinite andoxides. The adsorption isotherm of acid phosphatase on the examined soil colloids and minerals fitted tothe Langmuir model. The amount of enzyme adsorbed in the absence of ligands was in the order of YBScolloid ＞LS colloid＞kaolin≈goethite. In the presence of phosphate or citrate, the amounts of the enzymeadsorbed followed the sequence YBS colloid＞kaolin＞LS colloid＞goethite. The presence of ligands alsodecreased the binding energy between the enzyme and soil colloids or minerals. With the increase of ligandconcentration from 10 mmol L-1 to 400 m mol L-1, different behaviors for the adsorption of enzyme werefound in the colloid and mineral systems studied. A sharp decrease in enzyme adsorption was observed ongoethite while gradual decreases of enzyme adsorption were recorded in the two soil colloid systems. However,no any decrease was found for the amount of enzyme adsorbed on kaolin at higher ligand concentrations.When phosphate or citrate was introduced to the system before the addition of enzyme, the ligands usuallyenhanced the adsorption of enzyme. The results obtained in this study suggested the important role ofkaolinite mineral in the adsorption of enzyme molecules in acidic soils in the presence of various ligands.     

应用反射光谱法对土壤粘土矿物的鉴定的初步研究

The characteristics of the reflectance spectra of clay minerals and their influences on the reflectance spectra of soils are dealt with in the paper.The results showed that dominant clay minerals in soils could be distinguished in light of the spectral -form parameters of the reflectance spectra of soils,thus making it possible to develop a quick method to determine clay minerals by means of reflectance spectra of soils in the lab.and providing a theoretic basis for remote sensing of clay minerals in soils with a high resolution imaging spectrometer.     

磁性壳聚糖微球吸附苹果汁有机酸的动力学及热力学特性

为充分利用中国丰富的苹果资源,开发多品类的苹果深加工产品,以磁性壳聚糖微球为吸附剂,通过磁分离技术,吸附获得苹果汁中的天然有机酸,并对其吸附过程进行研究。利用Lagergren准一级动力学方程、准二级动力学方程、Elovich方程及内扩散方程对吸附反应动力学过程进行拟合;利用Langmuir等温吸附模型、Freundlich等温吸附模型及Temkin等温吸附模型对吸附等温数据进行拟合,并对其吸附反应热力学特性进行分析。通过比较线性拟合方程的决定系数,发现磁性壳聚糖微球吸附苹果汁中有机酸的动力学过程更加符合Lagergren准二级动力学模型,吸附温度越高,吸附速率常数和初始吸附速率越大,但平衡吸附量越低。等温吸附过程更加符合Langmuir等温吸附模型,表明该吸附过程更趋向于单分子层的化学吸附。298 K时,有机酸的饱和吸附量可达到188.679 2 mg/g,表明磁性壳聚糖微球是苹果汁中有机酸的1种高效吸附剂。热力学参数ΔG°0,ΔH°0,ΔS°0,表明磁性壳聚糖微球对苹果汁有机酸的吸附过程为熵增加的可自发进行的放热过程。动力学及热力学结果为磁性壳聚糖微球吸附苹果汁有机酸的研究提供了理论基础与技术支持。     

根瘤菌存在下土壤胶体和矿物对镉的吸附

Experiments were conducted to study the adsorption of Cd on two soil collids(red soil and yellowbrown soil) and three variable-charge minerals (goethite,noncrystalline Fe oxide and kaolin) in the absence and presence of rhizobia.The tested strain Rhizobium fredii C6,tolerant to 0.8 mmol L^-1 Cd,was selected from 30 rhizobial strains.Results showed that the isotherms for the adsorption of Cd by examined soil colloids and minerals in the presence of rhizobia could be described by Langmuir equation.Within the range of the numbers of rhizobial cells studied,the amount of Cd adsorbed by each system increased with increasing rhizobial cells,Greater increases for the adsorption of Cd were found in red soil and kaolin systems.Rhizobia influence on the adsorption of Cd by examined soil colloids and minerals was different from that on the adsorption of Cu.The presence of rhizobia increased the adsorption affinity of soil colloids and minerals for Cd,particularly for the goethite and kaolin systems.The discrepancies in the influence of rhizobia on the adsorbability and affinity of selected soil colloids and minerals for Cd suggesed the different interactions of rhizobia with various soil components.It is assumed that bacterial biomass plays an important role in controlling the mobility and bioavailability of Cd in soils with kaolinite and goethite as the major colloidal compnents,such as in variable-charge soil.     

缓冲液种类对土壤酸性磷酸酶活性的影响

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。     

Invertase and amylase activities as influenced by clay minerals,soil-clay fractions and topsoils under grassland

The effects of five clay minerals, three monominerallic clay fractions from soil, and two topsoils from tussock grasslands on the activity of added invertase and α- and β-amylase were determined. The activity levels of the added invertase were similar to those found in many grassland soils: those of the added amylases were somewhat higher. Significant decreases in activity were found in unbuffered aqueous suspensions of the clay minerals, partly because of the instability of the enzymes in water, and in buffered clay suspensions, in which adsorption of the enzymes was generally incomplete. The influence of clay minerals and clays on inhibition of invertase and α-amylase activity was in the order muscovite < allophane < illite < montmorillonite; in the presence of clay fractions from soil, the order was muscovite ≈ mica-vermiculite < mica-beidellite. Kaolinite had only a weak effect on invertase activity but a strong one on α-amylase activity. All of the clay samples inhibited β-amylase activity. In contrast, the topsoils, containing mica-vermiculite or mica-beidellite as the main clay component, did not adsorb the added enzymes or inhibit their activity. Under field conditions, it seems unlikely that the clay fraction in these well-developed topsoils would have any direct influence of the activity of freshly acquired invertase or amylase.     

Adsorption of DNA on clay minerals and various colloidal particles from an Alfisol

Adsorption and desorption of salmon sperm DNA on four different colloidal fractions from Brown Soil and clay minerals were studied. The adsorption isotherms of DNA on the examined soil colloids and minerals conformed to the Langmuir equation. The amount of DNA adsorbed followed the order: montmorillonite?fine inorganic clay>fine organic clay>kaolinite>coarse inorganic clay>coarse organic clay. A marked decrease in the adsorption of DNA on organic clays and montmorillonite was observed with the increase of pH from 2.0 to 5.0. Negligible DNA was adsorbed by organic clays above pH 5.0. As for inorganic clays and kaolinite, a slow decrease in DNA adsorption was found with increasing pH from 2.0 to 9.0. The results implied that electrostatic interactions played a more important role in DNA adsorption on organic clays and montmorillonite. Magnesium ion was more efficient than sodium ion in promoting DNA adsorption on soil colloids and minerals. DNA molecules on soil colloids and minerals were desorbed by sequential washing with 10 mM Tris, 100 mM NaCl and 100 mM phosphate at pH 7.0. A percentage of 53.7-64.4% of adsorbed DNA on organic clays and montmorillonite was released, while only 10.7-15.2% of DNA on inorganic clays and kaolinite was desorbed by Tris and NaCl. The percent desorption of DNA from inorganic clays, organic clays, montmorillonite and kaolinite by phosphate was 39.7-42.2, 23.6-28.8, 29.7 and 11.4%, respectively. Data from this work indicated that fine clays dominate the amount of DNA adsorption and coarse clays play a more important role in the binding affinity of DNA in soil. Organic matter may not favor DNA adsorption in permanent-charge soil. The information obtained is of fundamental significance for the understanding of the ultimate fate of extracellular DNA in soil.     

Adsorption of pure and dirty bacterial DNA on clay minerals and their transformation frequency

Several studies have investigated the adsorption of pure DNA on soil particles and its transformation ability. However, the presence of not purified (dirty) rather than pure DNA is more common in the extracellular soil environment. For this reason, we have investigated the adsorption and binding of dirty DNA on montmorillonite (M) and kaolinite (K) and their transforming ability in comparison to pure DNA. After lysis of Bacillus subtilis cells, induced by KCl, dirty DNA was characterized by the presence of cellular wall debris (cwd) and other constitutional organic components (coc). The dirty DNA was then divided into two fractions, one with cellular wall debris (DNA +cwd) and the other without cellular wall debris (DNA −cwd). B. subtilis BD 1512 (Cm^r) and BD 170 (Cm^s) were selected as donor and recipient bacteria, respectively, for adsorption and transformation studies. Both cwd and coc seem to facilitate the adsorption of DNA to clay minerals, whereas only cwd promote the DNA binding on clays, protecting also the DNA fragments below 400 bp against nucleases. Both dirty DNA fractions adsorbed and bound on clay minerals were able to transform competent cells.     

Kinetic properties and stability of potato acid phosphatase immobilized on Ca-polygalacturonate

 Acid phosphatase from potato was adsorbed and immobilized on a pre-formed network of Ca-polygalacturonate, a substrate which has a composition and morphology similar to the mucigel present at the root-soil interface. The influence of different types of organic buffers on enzyme adsorption and activity was investigated. The highest enzyme activity, for free and adsorbed enzyme, was obtained with Na-maleate buffer at pH 6.0, which was used for all subsequent experiments. The Michaelis-Menten kinetic parameters, V_max and K_m, were determined for free and adsorbed phosphatase. V_max showed a 60% decrease upon adsorption (2.09±0.30 U/mg, for the soluble form and 0.84±0.15 U/mg, for the adsorbed enzyme), whereas K_m increased from 0.49±0.15 mM for the free enzyme to 0.99±0.20 mM for adsorbed phosphatase. Phosphatase adsorption decreased as the concentration of NaCl increased, indicating that the enzyme is bound to the carrier gel through coulombic interactions. Adsorption increased stability of the enzyme as compared with the free enzyme (t _1/2 of the activity was 9.4 days and 5.8 days, respectively), but increased thermal and proteolytic inactivation. The pH/activity profile revealed no change in terms of shape or optimum pH (4.5) upon adsorption of the enzyme. These results indicate that adsorption of acid phosphatase on Ca-polygalacturonate induces changes in the kinetic properties and stability of the enzyme, and the same type of response can be extrapolated from these results for acid phosphatases of the rhizosphere. Received: 1 July 1997     

中国福建省土壤的主要粘土矿物

The clay minerals of more than 200 soil samples collected from various sites of Fujian Province were studied by the X-ray diffraction method and transmission electron microscopy to study their distribution and evolution.Montmorillonite was found in coastal solonchak,paddy soils derived from marine deposit,lacustrine deposit and river deposit,and some lateritic red soil,red soil and yellow soil with a low weathering degree.Chlorite existed mainly in coastal solonchak and paddy soil developed from marine deposit.1.4nm intergradient mineral appeared frequently in yellow soil,red soil and lateritic red soil.The content of 1.4nm intergradient mineral increased with the decrease of weathering degree from lateritic red soil to red soil to yellow soil.Hydrous micas were more in coastal solonchak,paddy soils derived from marine deposit,lacustrine deposit and river deposit.and puple soil from purple shale than in other soils.Kaolinte was the most important clay mineral in the soils iun this province.The higher the soil weathering degree,the more the kaolinite existed.From yellow soil to red soil to lateritic red soil,kaolinite increased gradually,Kaolinite was the predominant clay mineral accompanied by few other minerals in typical lateritic red soil. Tubular halloysite was a widespread clay mineral in soils of Fujian Province with varying quantities.The soil derived from the paent rocks rich in feldspar contained more tubular halloysite.Spheroidal halloysite was found in a red soil and a paddy soil developed from olivine basalt gibbsite in the soils in this district was largely“primary gibbsite” which formed in the early weathering stage.Gibbsite decreased with the increase of weathering degree from yellow soil to red soil to lateritic red soil.Goethite also decreased in the same sequence while hematite increased.     

磷酸盐在水铁矿及水铁矿-胡敏酸复合体表面的吸附

The adsorption of phosphate onto ferrihydrite (FH) and two FH-humic acid (HA) complexes, obtained by co-precipitating FH with low (FH-HA1) and relatively high amounts of humic acid (FH-HA2), was studied through kinetics and isotherm experiments to determine the differences in phosphate adsorption between FH-HA complexes and FH and to reveal the mechanism of phosphate adsorption onto two soil compositions. The isoelectric point (IEP) and the specific surface area (SSA) of the mineral decreased as the particle porosity of the mineral increased, which corresponded to an increase in the amount of organic carbon. The adsorption capacity of phosphate was higher on FH than on FH-HA1 and FH-HA2 at the scale of micromoles per kilogram. The initial adsorption rate and adsorption affinity of phosphate decreased with an increase in the amount of HA in the mineral. The sensitivity of phosphate adsorption to the change in the pH was greater for FH than for FH-HA complexes. Ionic strength did not affect the adsorption of phosphate onto FH and FH-HA1 at a lower pH, and the increase in the ionic strength promoted phosphate adsorption at a higher pH. However, for the FH-HA2 complex, the increase in the ionic strength inhibited the adsorption of phosphate onto FH-HA2 at a lower pH and increased the adsorption at a higher pH.     

Effects of Addition Sequence and pH of Oxalate on the Adsorption and Activity of Acid Phosphatase on Soil Colloids and Minerals

The adsorption and specific activities of acid phosphatase on soil colloids and minerals in oxalate systems with different pH values were studied using the batch method. Results indicated that the amount of phosphatase adsorption decreased in the following order: goethite ? yellow-brown soil (YBS) > kaolinite > latosol, and the specific activities of immobilized phosphatase were goethite > latosol > kaolinite > YBS. The profiles of enzymatic adsorption and specific activity on soil minerals were bell shaped, and their change tendencies were out of sync. The optimal pH for phosphatase adsorption was located between the isoelectric point of phosphatase and the zero point of charge of the studied soil minerals, and the optimal pH for the specific activity of the immobilized enzyme either did not change or shifted toward alkalinity. Enzymatic adsorption amounts and specific activities were greater when the enzyme was added before oxalate than when enzyme was added after oxalate.     

Aspects of the phosphorus nutrition of white clover populations II. Root exocellular acid phosphatase activity

The effect of phosphorus (P) level, solution pH, aluminium (Al) level on root exocellular acid phosphatase activity of white clover (Trifolium repens L.) populations was investigated in three experiments. Lotus pedunculatus Cav. cv. Maku was also included in one experiment.

Phosphatase activity increased with decreasing P and increasing Al supply, but was unaffected by the solution pH at which populations had been grown.

There was usually a significant negative correlation between acid phosphatase activity and parameters measuring plant size, for plants grown at adequate P levels, but in only one of the three experiments were there significant differences between white clover populations for phosphatase activity. There was no clear correlation between the acid phosphatase activity of populations and the P status of soil from which they were collected.

We conclude that root exocellular acid phosphatase activity is unlikely to be a useful character in screening for tolerance of low‐P conditions.     

胡敏酸对铵钾在粘土矿物上交互作用的影响

Interaction of ammonium (NH+4) and potassium (K+) is typical in field soils. However, the effects of organic matter on interaction of NH+4 and K+have not been thoroughly investigated. In this study, we examined the changes in major physicochemical properties of three clay minerals (kaolinite, illite, and montmorillonite) after humic acid (HA) coating and evaluated the influences of these changes on the interaction of NH+4 and K+on clay minerals using batch experiments. After HA coating, the cation exchange capacity (CEC) and specific surface area (SSA) of montmorillonite decreased significantly, while little decrease in CEC and SSA occurred in illite and only a slight increase in CEC was found in kaolinite. Humic acid coating significantly increased cation adsorption and preference for NH+4, and this effect was more obvious on clay minerals with a lower CEC. Results of Fourier transform infrared spectrometry analysis showed that HA coating promoted the formation of H-bonds between the adsorbed NH+4 and the organo-mineral complexes. HA coating increased cation fixation capacity on montmorillonite and kaolinite, but the opposite occurred on illite. In addition, HA coating increased the competitiveness of NH+4 on fixation sites. These results showed that HA coating affected both the nature of clay mineral surfaces and the reactions of NH+4 and K+with clay minerals, which might influence the availability of nutrient cations to plants in field soils amended with organic matter.     

粘土矿吸附过程中四环素基团变化研究

湖南红壤主要由富含硅铝的高岭土和蒙脱石组成,选取这两种粘土矿作为吸附介质在实验室环境下模拟了四环素的吸附过程,通过高效液相色谱和红外光谱检测分析了四环素的主要基团在不同吸附阶段的变化,结果表明-OH是影响吸附速率和吸附量的主要基团,发生的吸附在解吸过程中也难于解吸;C=O主要在吸附后期起作用,吸附能力不强,易解吸,可能与其吸电子能力较弱有关。     

基于不同方法测定土壤酸性磷酸酶活性的比较

土壤酸性磷酸酶与有机磷的矿化及植物的磷素营养关系最为密切。目前国内学者在测定酸性磷酸酶活性时主要参照关松荫《土壤酶及其研究法》中以磷酸苯二钠为基质的测定方法,而国外学者主要参照Dick《Methods of Soil Enzymology》中以对硝基苯磷酸二钠为基质的测定方法(PNPP)。但是,在以磷酸苯二钠为基质测定生成物的过程中,常出现显色程度不明显的问题;另外,采用不同基质测定酸性磷酸酶活性也造成了测定方法选择的困难。为合理选择土壤酸性磷酸酶活性的测定方法,本研究选用酸性、中性和碱性土壤各10个土样,分别采用以磷酸苯二钠为基质,且在显色阶段分别加入pH5.0醋酸盐缓冲液(DPP 1)和pH9.4硼酸盐缓冲液(DPP 2)的方法,以及PNPP方法测定土壤酸性磷酸酶活性。同时也研究了不同pH缓冲液和苯酚浓度对生成物显色反应的影响。结果表明:以磷酸苯二钠为基质、在显色反应阶段加入pH≤6的缓冲液时,苯酚和2,6-二溴苯醌氯亚胺不显色;当加入pH≥8的缓冲液时,两者之间显色且苯酚浓度和吸光值的Pearson相关系数极显著。这说明pH低是导致高苯酚浓度和2,6-二溴苯醌氯亚胺显色效果差的一个主要原因。此外,采用PNPP方法测定时,在酸性、中性和碱性土壤中,10个样本酸性磷酸酶活性的变异系数分别较DPP 2增加了70.04%、42.44%和21.17%;极差分别是DPP 2的27.18倍、26.85倍和39.43倍。总之,如果选用磷酸苯二钠为基质测定土壤酸性磷酸酶活性,应在显色阶段加入碱性硼酸盐缓冲液;选用对硝基苯磷酸二钠为基质,是更为简单和灵敏的方法。     

受钙、锌影响硼在酸性紫色土中的吸附特征

酸性紫色土对硼的等温吸附-解吸和吸附动力学试验表明，硼的等温吸附符合Frendlich方程，根据方程的K值（0．441），供试土壤对硼的吸附较弱。锌与硼复合，对土壤硼的吸附量和解吸量影响均不大，但吸附强度下降。钙则大大地提高了土壤对硼的吸附量和吸附常数，使土壤硼平均吸附量提高191．9％，并使平均解吸率下降6．4％。土壤吸附硼的动力学以Elovich方程（Ct=a＋blnt）描述最佳，反映土壤对硼吸附速率大小的方程参数b值以B＋Ca〉B＋Ca＋Zn〉B＋Zn〉B，钙对硼的吸附速率影响比锌的作用更强。     

Soil‐aggregate formation as influenced by clay content and organic‐matter amendment

Naturally occurring wetting‐and‐drying cycles often enhance aggregation and give rise to a stable soil structure. In comparatively dry regions, such as large areas of Australia, organic‐matter (OM) contents in topsoils of arable land are usually small. Therefore, the effects of wetting and drying are almost solely reliant on the clay content. To investigate the relations between wetting‐and‐drying cycles, aggregation, clay content, and OM in the Australian environment, an experiment was set up to determine the relative influence of both clay content (23%, 31%, 34%, and 38%) and OM amendments of barley straw (equivalent to 3.1 t ha^–1, 6.2 t ha^–1, and 12.4 t ha^–1) on the development of water‐stable aggregates in agricultural soil. The aggregate stability of each of the sixteen composite soils was determined after one, three, and six wet/dry cycles and subsequent fast and slow prewetting and was then compared to the aggregate stabilities of all other composite soils. While a single wet/dry cycle initiated soil structural evolution in all composite soils, enhancing macroaggregation, the incorporation of barley straw was most effective for the development of water‐stable aggregates in those soils with 34% and 38% clay. Repeated wetting‐and‐drying events revealed that soil aggregation is primarily based on the clay content of the soil, but that large straw additions also tend to enhance soil aggregation. Relative to untreated soil, straw additions equivalent to 3.1 t ha^–1 and 12.4 t ha^–1 increased soil aggregation by about 100% and 250%, respectively, after three wet/dry cycles and fast prewetting, but were of less influence with subsequent wet/dry cycles. Straw additions were even more effective in aggregating soil when combined with slow prewetting; after three wet/dry cycles, the mean weight diameters of aggregates were increased by 70% and 140% with the same OM additions and by 160% and 290% after six wet/dry cycles, compared to samples without organic amendments. We suggest that in arable soils poor in OM and with a field texture grade of clay loam or finer, the addition of straw, which is often available from preceding crops, may be useful for improving aggregation. For a satisfactory degree of aggregate stability and an improved soil structural form, we found that straw additions of at least 6.2 t ha^–1 were required. However, rapid wetting of straw‐amended soil will disrupt newly formed aggregates, and straw has only a limited ability to sustain structural improvement.