合浦珠母贝碱性磷酸酶的分离纯化与性质研究

以合浦珠母贝（Ｐｉｎｃｔａｄａ ｆｕｃａｔａ）为提取酶的原料,经Ｔｒｉｓ－ＨＣｌ缓冲液（ｐＨ７．５）抽提,正丁醇处理,硫酸铵分级沉淀分离,ＤＥＡＥ－纤维素离子交换树脂支析,ＤＥＡＥ－Ａ２５离子交换分子筛柱层析纯化,得到一定纯度的碱性磷酸酶,比活力达到１４４４Ｕ／ｍｇ。酶学性质和动力学性质研究表明,该酶催化对硝基苯磷酸（ｐＮＰＰ）的水解反应,最适ｐＨ值为９．７２,最适温度为４５℃,水解反应的活化能为     

Physico-chemical properties and enzyme activities of the arable soils in Lhasa, Tibet, China

An understanding of the physical,chemical,and biological properties of a soil provides a basis for soil use and management.This paper reports the major physico-chemical properties and enzyme activities of the soils of Lhasa’s main arable lands and the factors that influence these soil properties.Composite and core samples were taken from the three main arable soil types(alluvial soil,subalpine arable steppe soil,and subalpine arable meadow soil) and were analysed using standard methods.The bulk density and the ventilation porosity ratio of the soils were close to the recommended values for arable lands,and the dominant soil texture was sandy.The soil moisture release rates were arable steppe soil > alluvial soil > arable meadow soil.Soil organic matter content,Cation-Exchange Capacity(CEC),total and available nitrogen content,and catalase activity of the arable meadow soil were higher than those of the alluvial and the arable steppe soils,while soil pH in the arable meadow was lower.Most of the measured properties did not show a significant variance among these three soils.However,the measured indices(apart from the total potassium) indicate that there are notable differences among the three types of soil.The results implied that the utilisation patterns of the arable soil or human activities,such as tillage practices and fertiliser applications,have a substantialeffect on the soil properties in this region.Our results suggest that the cultivation practices in the region have apparently positive impact on the soil organic matter,nutrient contents and bulk density probably due to the sound fertiliser management such as the applications of farmyard manure and chemical fertilisers.However,intense cultivation practices lowered the activity of most soil enzymes.The results demonstrate that the choice of soil management strategy had a significant impact on the soil physicochemical and biological properties in the region studied.     

Einsatz eines Immunoassays als Screening-Verfahren für die Untersuchung von Grundwasserproben auf Mecoprop

Application of an Enzyme Immunoassay for the Detection of Mecoprop in a Ground water Screening Procedure A competitive solid-phase enzyme immunoassay based on rabbit polyclonal antibodies was tested for its reliability. This assay, used for the detection of the phenoxycarboxylic acid mecoprop [2-(4-chloro-2-methylphenoxy-)propionic acid], has already been characterized in terms of middle of the test (0.36 μg/L at 50% B/B₀) and detection limit (0.06 μg/L at 80% B/B₀) by Weber (Acta hydrochim. hydrobiol. 22, 53–59 (1994)). For the reliability test, a small number from a batch of one hundred groundwater samples were spiked. The analyst was neither aware of which nor of how many samples were spiked. The spiked samples were all detected as positive. There were no false negative results. From these findings, values of 1 were obtained for the diagnostic sensitivity and for the diagnostic specificity. Based on these results, the mecoprop contents of 228 groundwater samples from various waterlevel gauges of a test field were determined. The test site had previously been treated repeatedly with this herbicide. Confirmation of the enzyme immunoassay results was carried out by GC/MS. This showed that the enzyme immunoassay gave one false positive result only. All other results from immunoassay and GC/MS showed good correlation.     

盐度胁迫对大海马(Hippocampus kuda)幼体生长、组分及酶活力的影响

在盐度为25的条件下,以大海马幼体为实验材料,通过设置不同的盐度胁迫组(盐度从25胁迫至5、15和35)的方法,对其生长、生化组分以及酶活力的影响进行了研究。结果表明:15盐度组大海马幼体的体重、生化组分、能值与对照组(盐度25)相比差异不显著(P>0.05),体长、成活率指标则显著高于对照组(P<0.05),然而5、35盐度组的生长指标、成活率、生化组分等则显著低于对照组(P<0.05)。15盐度组的SOD、CAT酶活性低于对照组水平(P<0.05),MDA的含量变化不显著(P>0.05);而5、35盐度组SOD、CAT和MDA含量与对照组相比,随着时间的延长,呈现逐渐升高的趋势(P<0.05);随着盐度的升高,AKP酶活性具有逐渐升高的趋势,而ACP酶活性则呈现降低趋势。     

Relationship between differential retention of Escherichia coli and Enterococcus faecalis and variations in enzyme activity in the scallop Patinopecten yessoensis

Uptake of Escherichia coli and Enterococcus faecalis and variations of trypsin amylase activity acid phosphatase and superoxide dismutase in tissue of the scallop Patinopecten yessoensis were detected. The results showed that P. yessoensis accumulated E. faecalis in larger numbers and more rapidly than E. coli, both with the highest concentration in the digestive tract and lowest in hemolymph. Compared to E. coli, all scallops exposed to E. faecalis showed significantly higher trypsin and AMS activity. SOD activity in hemocytes and ACP activity in hemolymph was significantly higher in the treatments with 5 log₁₀CFU/ml E. coli than with E. faecalis. But no significant differences in ACP activity of P. yessoensis exposed to a 3 log₁₀CFU/ml inoculum of both bacteria were recorded. In conclusion, the mass retention of gut microflora in P. yessoensis is positively correlated with digestive enzymes activity and negatively correlated with ACP activity in the hemocyte.     

Sedimentary Enzyme Kinetics of Land/Water Ecotones with Reed Domination

Land/water ecotones, the transitional boundary zone between terrestrial and aquatic ecosystems, can effectively trap and assimilate nutrients from external sources. The changes of nutrient cycling will cause a response in enzyme activity. In this paper, the enzyme kinetics of land/water ecotones in a shallow eutrophic wetland in China, dominated by a reed (Phragmites australis) community, are studied. The results indicate that land/water ecotones exhibit a strong filtration function and nutrient factor is an important feature affecting the distribution of enzymes in the transect of the littoral zone. The obvious spatial differences of enzyme activities and nutrients are present in the transect of the reed beds. The land area is the hot zone, where significant changes in enzyme activity occur. T‐test results show that the landward spot, 1 m away from the water‐land interface, is quite different from other spots, and the 1 m spot is the hot spot where great biochemical change happens. The principal component analysis of the environmental variables shows that nutrient factors have an important affect on enzyme distribution in the transect of the littoral zone. The nutrient factor explains about 54% of the observed variance. In particular, TOC (total organic carbon) correlates with the other five factors (TN; total nitrogen, TP; total phosphorus, phosphatase, β‐glucosidase, and urease) with correlative coefficients of 0.932, 0.595, 0.488, 0.433, and –0.468, respectively. The results indicate that TOC is a very important factor in Baiyangdian wetlands.     

Improvement of a Monoclonal Antibody-based Immunoassay for the Determination of Terbutryn Verbesserung eines Immunassays mit monoklonalen Antikörpern zur Bestimmung von Terbutryn

The performance of an existing enzyme immunoassay (EIA) with monoclonal antibodies (mAb) for the determination of terbutryn was improved by the application of a new enzyme tracer. For this purpose the triazine derivative 6-chloro-2-(tert-butylamino)-1,3,5-triazine-4-(6-aminohexane carboxylic acid) was coupled to horseradish peroxidase (HRP). The competitive EIA on microwell plates made it possible to determine terbutryn in the range from 0.05 to 1 μg/L with a 50% B/B₀ value of the test at 0.2 μg/L. The application of the EIA to determine terbutryn in spiked surface waters provided good recoveries of terbutryn without matrix effects.     

牙鲆血清中免疫球蛋白M的分离及其测定

通过提取健康养殖牙鲆(Paraliehthys olivaceus)血清中的免疫球蛋白(Ig),并测定其相对分子质量,其中重链(H链)的相对分子质量为74．2ku,轻链(L链)的相对分子质量为23．8ku。用纯化的牙鲆血清中的Ig作抗原,制备多克隆抗体,确立牙鲆血清中免疫球蛋白的最佳测定方法。