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1.
长牡蛎碱性磷酸酶的分离纯化及部分性质研究   总被引:5,自引:1,他引:5  
本文以长牡蛎为酶源提取材料,用正丁醇抽提,硫酸铵分级分离,DEAE-52和Sephadex G-100柱层析,得到一定纯度的碱性磷酸酶,提纯倍数为61.96,比活力达1.326U/mg.该酶的最适pH值9.6,最适温度38℃,初速度6.8μmol/(dm~3·min),米氏常数Km值为1.30mmol/dm~3.实验结果表明,Mg~(2+)对该酶有明显的激活作用;Zn~(2+)、Cu~(2+)、Hg~(2+)对该酶有明显的抑制作用.  相似文献   

2.
以产物N-乙酰-β-D-氨基葡萄糖(NAG)及其类似物-葡萄糖(glu)、半乳糖(gal)、果糖(fru)和蔗糖(suc)为效应物,研究它们对锯缘青蟹N-乙酰-β-D-氨基葡萄糖苷酶(NAGase)活力的影响及抑制作用机理.结果表明,上述效应物对酶均表现为可逆的抑制作用.NAG有较强的抑制作用,其IC50为0.011 mol/dm3.但产物类似物对该酶的抑制作用均弱于NAG.NAG对该酶的抑制作用机理表现为反竞争性抑制,抑制常数KIS为4.37×10-3mol/dm3;glu、gal、suc则表现为非竞争性抑制,抑制常数KI分别为0.125、0.394、0.474 mol/dm3;而fru则表现为竞争性抑制,抑制常数KI为0.455mol/dm3.  相似文献   

3.
余群  王重刚  陈荣  郁昂  郑微云 《海洋科学》2008,32(10):56-61
探讨了微量硒(Se~(4+))对华贵栉孔扇贝(Chlamys nobilis(Reeve))碱性磷酸酶(AKP)的作用。以催化动力学原理研究了不同缓冲系统中微量硒时AKP的影响。结果表明,Se~(4+)对谈酶的作用与酶所处的环境有关。在0.05 mol/L Na_2CO_3-NaHCO_3(pH 9.0)缓冲溶液中Se~(4+)浓度小于12.5 mmol/L时呈现出激活作用;Se~(4+)浓度大于12.5 mmol/L时呈现出非竞争性的抑制作用,其抑制常数为6.81×10~(-2)mol/L。在0.05 mol/L Tris-HCl缓冲系统(pH 9.0)中,微量Se~(4+)对AKP具有强烈竞争性抑制作用,其抑制常数为3.79×10~(-1)mol/L。经紫外吸收光谱、荧光发射光谱和圆二色谱的实验表明,微量Se~(4+)与AKP作用后,该酶的构象已发生了变化。研究发现海水环境中若含有微量Se~(4+)(SeO_3~(2-)),可以促进扇贝AKP的水解。利于贝类的生长发育和外壳的形成,并有益于贝类的繁殖。  相似文献   

4.
采用新极谱技术(1.5次微分)的阳极溶出伏安滴定方法测定天然海水的铜络合容量和条件稳定常数。对测试条件、EDTA回收率和有关问题进行了探讨。青岛近岸水样的九次平行测定表明,铜的表现络合容量为3.01×10~(-7)mol/dm~3,条件稳定常数为7.34×10~7,相对标准偏差分别为9.2%和13.8%。  相似文献   

5.
通过一次培养、极谱滴定测定研究一次培养过程中叉边金藻细胞渗出物对铜、镉的络合容量及常数的变化;培养介质的营养水平;加入微量营养元素铁、锰和重金属铜对细胞生长和渗出物释放的影响。结果表明,在一次培养过程中,从接种到稳定生长期细胞渗出物的铜络合容量从1.49×10~(-7)mol/dm~3增加到8.34×10~(-7)mol/dm~3,条件稳定常数LgK′_(CuL)从7.57变化到7.11。培养介质的营养水平愈高,单个细胞渗出的有机物量增加,铁明显抑制细胞渗出物的释放,铜在细胞中可容忍的浓度范围内刺激细胞渗出物的释放。  相似文献   

6.
本文测定了一次培养实验和连续培养实验中三角褐指藻胞外产物的变化及其与汞的络合性能。28天的培养过程中测得胞外产物与汞的络合容量C_L为2.6×10~(-5)mol/dm~3;表观稳定常数K′为6.1×10~(10)。该结果解释了细胞培养过程中胞外有机物的释放对汞的解毒机理及对介质中汞形态的转化。  相似文献   

7.
报道了杂色鲍碱性磷酸酶(ALP)在甲醇、乙醇、丙醇、甲醛、丙酮等溶液中的失 活动力学.结果表明:酶的剩余活力随着有机溶剂浓度的增大而迅速下降.测得上述 5种有机溶剂对该酶的半抑制率(IC50)分别为5.41、3.07、1.80、31.0×10-3、2.30 mol/dm3.在这些有机溶剂溶液中酶的失活过程都是可逆反应.检测醇、醛、酮对该酶 的失活作用机理,结果表明甲醇、乙醇、丙醇对杂色鲍ALP的失活作用均为非竞争性 机制,其抑制常数分别为5.36、3.02、1.80mol/dm3.甲醛、丙酮对杂色鲍ALP的失活 作用都呈现为混合型抑制,其对游离酶的抑制常数(K1)分别为24.7×10-3、1.66 mol/dm3;对酶-底物络合物的抑制常数(KIS)分别为56.2×10-3、5.40mol/dm3.  相似文献   

8.
牟氏角毛藻的有机渗出物及其对铜的络合作用   总被引:1,自引:0,他引:1  
对牟氏角毛藻(chaetoceros muelleri)进行一次性的培养过程,不同时间取藻培养液过滤,测定滤液中的COD及铜络合容量CC_(cu)值。结果表明,牟氏角毛藻在生长过程中向体外渗出有机物,在指数生长期渗出的有机物量很少;细胞渗出有机物明显增加;出现在静止期之后。牟氏角毛藻的细胞渗出物对铜具有络合能力,CCcu值从初始到192小时,由1.65×10~(-7)mol/dm~3增加到2.20×10~(-7)mol/dm~3。铜络合物的条件稳定常数在整个培养过程中无明显改变,logKcuL变化范围从7.27到7.74。过滤藻液的COD值从初始到192小时由0.81mg/dm~3增加到1.58mg/dm~3。  相似文献   

9.
以中国鲎(Tachypleus tridentatus)内脏为材料,分别通过30%和80%饱和度(NH_4)_2SO_4沉淀分级分离、葡聚糖凝胶柱Sephadex G-200层析、以及离子交换柱DEAE-32层析,从而获得N-乙酰-β-D-氨基葡萄糖苷酶(EC3.2.1.52,NAGase),经PAGE检定达到电泳纯,酶的比活力为505.21 U/mg。SDS-PAGE显示一谱带,测得该酶蛋白亚基分子量为121.5kDa,等电聚焦电泳法测得酶的等电点pI为6.01。以pNP-β-D-GlcNAc为底物,研究NAGase催化反应的动力学参数。结果表明:中国鲎NAGase的最适pH、最适温度、K_m和V_(max)分别为5.4和55℃、0.421 mmol/L和13.158μmol/L·min~(-1);酶在pH=4.5~7.0范围内较稳定,在20~50℃之间具有较好的热稳定性。酶在276nm波长处有紫外吸收峰,在232.2nm波长的内源荧光激发下,酶内源荧光发射光谱峰在330.9nm波长处。Na~+,K~+和Li~+对NAGase活力没有影响,Ca~(2+)、Ba~(2+)和Co~(2+)对NAGase有不同程度的激活作用,Co~(2+)对NAGase的激活作用较大,5mmol/L Co~(2+)能使NAGase活力提高41.67%;Mg~(2+)、Cu~(2+)、Zn~(2+)、Mn~(2+)、Fe~(3+)、Al~(3+)、Pb~(2+)、Ag~+、Cd~(2+)和Hg~(2+)等10种金属离子对NAGase活力有不同程度的抑制作用,5mmol/L Cd~(2+)可使酶活力丧失85.60%,而Hg~(2+)对酶的抑制作用最强,0.1mmol/L Hg~(2+)可使酶活力丧失90.10%。EDTA对酶活力没有影响,推断中国鲎NAGase属于非金属酶类。  相似文献   

10.
从印度洋深海沉积物中分离出一株能够高效降解甲醛的假单胞菌(Pseudomonas sp.IOFA1),该菌株可在40 min内完全降解100 mg/cm~3的甲醛.通过阴离子交换柱层析对菌株破碎液中的粗酶液进行纯化,得到一条具有甲醛降解活性的单一蛋白条带.经MALDI-TOF/TOF质谱鉴定,该蛋白为甲醛歧化酶蛋白(FM11).序列分析表明FM11与其它物种已知醛脱氢酶的氨基酸序列最高相似性为65%;结构域分析表明FM11含有2个保守的锌结合位点和一个NAD结合位点.酶学性质分析结果显示,FM11的最适反应温度为40℃,在25~50℃范围内能够保留95%以上的酶活力,尤其是在高于70℃的高温环境下仍保有约50%的活力,表现出广泛的温度适应性.FM11具有非常广泛的最适反应pH值范围(pH值为5~9),且在pH值4和pH值10时仍分别保留50%和80%左右的酶活力,表现出广泛的pH适应性.在pH值为5~7的条件下孵育1 h后,FM11仍保有97%以上的酶活,且在pH值为7~10条件下孵育1 h后,仍保留90%以上的酶活,表现出极好的酸性和碱性稳定性.Ca~(2+)、Mg~(2+)在低浓度时(0.5 mmol/dm~3)对FM11有较明显的激活作用.这些研究为我们进一步开发Pseudomonas sp.IOFA1甲醛歧化酶在降解甲醛方面的用途奠定了基础.  相似文献   

11.
In order to assess the impact of deep-sea mining on the in situ benthic life, we measured the microbial standing stock and concentration of organic nutrients in the deep-sea sediments of the Central Indian Ocean Basin in the Indian pioneer area. Sediments were collected using box core and grab samples during September 1996. The total bacterial numbers ranged from 10 10 -10 11 cells per g -1 dry weight sediment. There was a marginal decrease in the number of bacteria from surface to 30 cm depth, though the subsurface section registered a higher number than did the surface. The highest numbers were encountered at depths of 4-8 cm. The retrievable number of bacteria were two orders less in comparison with the direct total counts of bacteria. An almost homogeneous distribution of bacteria, total organic carbon, living biomass, and lipids throughout the depth of cores indicates active microbial and benthic processes in the deep sea sediments. On the other hand, a uniform distribution of total counts of bacteria, carbohydrates, and total organic carbon in all the cores indicates their stable nature and suggests that they can serve as useful parameters for long-term monitoring of the area after the benthic disturbance. Further studies on temporal variability in this region would not only verify the observed norms of distribution of these variables but would also help to understand restabilization processes after the simulated benthic disturbance.  相似文献   

12.
Coastal inundation associated with extreme sea levels is the main factor which leads to the loss of life and property whenever a severe tropical cyclonic storm hits the Indian coasts. The Andhra and Orissa coasts are most vulnerable for coastal inundation due to extreme rise in sea levels associated with tropical cyclones. Loss of life may be minimized if extreme sea levels and associated coastal flooding is predicted well in advance. Keeping this in view, location specific coastal inundation models are developed and applied for the Andhra and Orissa coasts of India. Several numerical experiments are carried out using the data of past severe cyclones that struck these regions. The simulated inland inundation distances are found to be in general agreement with the reported flooding.  相似文献   

13.
This study tests the hypothesis that the in situ void ratio of surficial marine sediments may be predicted from shear wave velocity-depth data with a reliability equal to that of other methods currently available. Shear wave velocity is fundamentally controlled by the number of grain-to-grain contacts per unit volume of material and by the effective stress across those contacts. In this study, three previously established empirical formulae are used to predict void ratio from velocity-depth data. Field data were acquired along a transect off the northern Californian coast across which water depth increased from 35 to 70 m and seafloor sediment type varied from sand to silty-sand, respectively. A towed seafloor sled device was used to collect shear wave refraction data, and a marked, systematic decrease in velocity was observed along the line, ranging from 35-70 m/s for the coarse, near-shore material to 25-40 m/s for the finer, offshore deposits. Void ratios predicted from these velocities were compared with data measured directly from box-core samples. Of the formulae used for prediction, two agree remarkably well with the control data. Both predicted and control values increase from 0.6-0.8 for the sandy material to 1.1-1.5 for the silty-sand. Thus, this study does not disprove the hypothesis set and demonstrates the potential of field shear wave velocity-depth data as a means of delineating spatial variation in void ratio for surficial marine sediments in a remote, nondestructive manner.  相似文献   

14.
Grain size and water content in box-core sediments from the Clarion-Clipperton fracture zone (C-C zone) in the northeast equatorial Pacific were analyzed in detail to understand the downcore variations across a hiatus between Quaternary and Tertiary layers. Grain-size distributions in the topmost core sediments show two modes: a coarse mode (peaked at 50 μm) and a fine mode (at 2-25 μm). The coarse mode disappears gradually with depth accompanied by the dissolution of siliceous fossil tests, whereas the fine mode coarsens due to the formation of authigenic minerals. Water content increases abruptly across a color boundary between an upper pale brown layer and a lower dark brown layer that is the hiatus between Quaternary and Tertiary layers. Abundant smectites and microvoid molds, which are created by the prolonged fossil dissolution in the underlying sediment, are attributed for the abrupt downcore variation of water content. Overall variations in grain size and water content in the topmost core sediments in the western C-C zone are possibly constrained by the dissolution of biogenic siliceous fossils. Variations in geotechnical properties related to these changes must be considered in the design of nodule collectors.  相似文献   

15.
This article concerns an interrelation between the sea levels and the western boundary flow near a tectonic boundary in a local zone in the Northwestern Pacific. In this zone, sea level variations at stations located on the coast facing the Pacific are studied to find the interrelation between variations of the Kurosio flow as an index of the distance of the flow axis off a specific coast. The result is discussed after data processing of the monthly means of the sea levels, and a notice is taken of variations caused by active crustal upheavals during a seismic event, a local earthquake.  相似文献   

16.
Particulate samples were collected from the Changjiang river system during a flood period, in May 1997, and POC, stable isotope and lipids associated with particles were examined. Results showed the decrease (0.84% ~ 1.88%) of organic carbon content from the upper reaches to the estuary.δ13C values of particulate organic carbon was in the range of -24.9×10-3 to -26.6×10-3, which were close to the isotopic signature of continental C3 vegetation. Total particulate n-alkanes concentrations varied from 1.4 to 10.1μg/dm3,or from 23.7 to 107μg/g of total suspended matter. Fatty acids were present in all the samples, from 1.4 to 5.4μg/dm3, with saturated and unsaturated straight-chain and branched compounds in the carbon number range from C12 to C30. Both δ13C and the ratio of carbon content to nitrogen content indicate the predominance of terrestrial inputs (soil organic matter) among the particles. The biomarker approach has been used to identify the relative portion of terrigenous and autochthonous fraction in the particulate samples. The distribution of fatty acids suggests a striking phytoplanktonic and microbial signal in most particle samples. The terrestrial alkanes are used to estimate the contribution of terrestrial inputs along the mainstream.  相似文献   

17.
Current trends of development of satellite derived bathymetry (SDB) models rely on applying calibration techniques including analytical approaches, neuro-fuzzy systems, regression optimization and others. In most of the cases, the SDB models are calibrated and verified for test sites, that provide favorable conditions for the remote derivation of bathymetry such as high water clarity, homogenous bottom type, low amount of sediment in the water and other factors. In this paper, a novel 3-dimensional geographical weighted regression (3GWR) SDB technique is presented, it binds together methods already presented in other studies, the geographically weighted local regression (GWR) model, with depth dependent inverse optimization. The proposed SDB model was calibrated and verified on a relatively difficult test site of the South Baltic near-shore areas with the use of multispectral observations acquired by a recently launched Sentinel-2 satellite observation system. By conducted experiments, it was shown that the proposed SDB model is capable of obtaining satisfactory results of RMSE ranging from 0.88 to 1.23[m] depending on the observation and can derive bathymetry for depths up to 12m. It was also shown, that the proposed approach may be used operationally, for instance, in the continuous assessment of temporal bathymetry changes, for areas important in the context of ensuring local maritime safety.  相似文献   

18.
As a part of the environmental impact assessment studies, geotechnical properties of sediments were determined in the Central Indian Basin. The undrained shear strength and index properties of the siliceous sediments were determined on 20 box cores of uniform dimension collected from various locations in five preselected sites. The maximum core length encountered was 41 cm and most of the sediments were siliceous oozes consisting of radiolarian or diatomaceous tests. The shear strength measurements revealed that surface sediments deposited in recent times (0-10 cm) have a shear strength of 0-1 kPa; this value increases with depth, reaching 10 kPa at 40 cm deep. Older sediments have greater strength because of compaction. Water content varies in the wide range of 312-577% and decreases with depth. The clay minerals such as smectite and illite are dominant and show some control over water content. Wet density, specific gravity, and porosity do not indicate any notable variation with depth, thereby indicating a uniform, slow rate of sedimentation. The average porosity of sediments is 90.2%, specific gravity 2.18, and wet bulk density 1.12 g/cm 3 . Sediments exhibit medium to high plasticity characteristics, with the average plasticity index varying between 105% and 136%. Preliminary studies on postdisturbance samples showed an increase in natural water content and a decrease in undrained shear strength of sediments in the top 10- to 15-cm layer.  相似文献   

19.
An acoustic inversion method using a wide-band signal and two near field receivers is proposed and applied to multiple layered seabed models including a manganese sediment. The inversion problem can be formulated into a probabilistic model comprised of signals, a forward model, and additive noise. The forward model simulates wide-band signals, such as chirp signals, and is chosen to be the source-waveletconvolution plane wave modeling method. The wavelet matching technique, using weighted least-squares fitting, estimates the sediment sound-speed and thickness on which determination of the possible numerical ranges for a priori uniform distribution is based. The genetic algorithm is applied to a global optimization problem to find a maximum a posteriori solution for determined a priori search space. Here the object function is defined by an L 2 norm of the difference between measured and modeled signals. Not only the marginal pdf but also its statistics are calculated by numerical evaluation of integrals using the samples selected during importance sampling process of the genetic algorithm.  相似文献   

20.
Several major improvements to an existing method for calibrating satellite altimeters using tide gauge data are described. The calibration is in the sense of monitoring and correcting temporal drift in the altimetric time series, which is essential in efforts to use the altimetric data for especially demanding applications. Examples include the determination of the rate of change of global mean sea level and the study of the relatively subtle, but climatically important, decadal variations in basin scale sea levels. The improvements are to the method described by Mitchum (1998a), and the modifications are of two basic types. First, since the method depends on the cancellation of true ocean signals by differencing the altimetric data from the tide gauge sea level time series, improvements are made that produce a more complete removal of the ocean signals that comprise the noise for the altimetric drift estimation problem. Second, a major error source in the tide gauge data, namely land motion, is explicitly addressed and corrections are developed that incorporate space-based geodetic data (continuous GPS and DORIS measurements). The long-term solution, having such geodetic measurements available at all the tide gauges, is not yet a reality, so an interim solution is developed. The improved method is applied to the TOPEX altimetric data. The Side A data (August 1992?February 1999) are found to have a linear drift component of 0.55 + / 0.39 mm/yr, but there is also a significant quadratic component to the drift that is presently unexplained. The TOPEX Side B altimeter is estimated to be biased by 7.0 + / 0.7 mm relative to the Side A altimeter based on an analysis of the first 350 days of Side B data.  相似文献   

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