应用硫酸苯酚法建立一种甘露糖醛酸C-5差向异构酶活性检测方法的研究

建立了一种简便快捷的甘露糖醛酸C-5差向异构酶活性的测定方法。研究发现相同分子质量的聚甘露糖醛酸和聚古罗糖醛酸与苯酚硫酸试剂反应后显色程度不同,以此可以用来测定β-D-甘露糖醛酸/α-L-古罗糖醛酸(M/G)值的变化,从而计算甘露糖醛酸C-5差向异构酶的活性。利用本方法测定了已知结构的褐藻胶片断的M/G比,其结果与传统的1H-NMR测定值基本一致。     

Vibrio sp.510产褐藻胶裂合酶的底物专一性分析

Vibrio sp.51 0具有很强的产生褐藻胶裂合酶的能力。本实验对该菌发酵产生的褐藻胶裂合酶经疏水色谱除去杂蛋白 ,再经灌注色谱分离得到 3个酶组分峰。经底物专一性检测 ,峰 1和峰 3对聚甘露糖醛酸和聚古罗糖醛酸均具降解作用 ,峰 2只对聚甘露糖醛酸有降解作用。园二色谱测定酶的二级结构 :峰 1结构最为复杂 ,以β-转角为最高 ,占 31 .5% ,无规线团占 2 7% ,α-螺旋占 2 5.8% ;峰2为β-折叠 ,占 95.5% ;峰 3以 60 %的α-螺旋和 40 %的无规线团形式存在。 3个分离峰的底物专一性和二级结构的差异证明了褐藻胶裂合酶蛋白的结构与功能之间的相关性。     

运用酶法降解褐藻胶研究糖链序列结构的展望

多糖是一种生命的基础物质 ,其功能和作用日益受到重视 ,糖生物学与糖工程的研究必将成为基因工程、蛋白质工程及糖链结构分析技术之后新的研究热点。探讨多糖的研究方法、解决糖链的测序问题成为该研究领域的关键。褐藻胶是一种重要的海藻工业产品 ,是由古罗糖醛酸 (G)和甘露糖醛酸 (M)组成的一种典型的多糖类 ,广泛应用于工业、农业、医药、食品等各个领域。研究发现 ,不同的褐藻酸酶可以作用于褐藻酸的不同位点 ,形成不同的糖片段或寡糖 ,经过分离、纯化 ,应用电泳、色谱、质谱、核磁共振等方法可以确定糖链的结构、组成及取代基的位置…     

褐藻酸裂合酶的研究——Ⅱ.对底物的作用和性质

用两种酸局部降解法,制备了三种底物:短链多聚甘露糖醛酸(SM)、短链多聚古罗糖醛酸(SG)和短链多聚甘露糖醛酸-古罗糖醛酸(SMG)。用粘度下降法,证明酶Ⅰ为外切酶,酶Ⅱ和酶Ⅲ为内切酶。用Somogyi-Nelson法,测定酶解底物产生的还原性,表明三种酶对褐藻酸、SM和SMG的作用很强,而对SG的作用很弱。用紫外吸收法,测定酶解各种底物后,生成产物的吸收光谱,证明三种酶裂解SM和SMG后,产物在230nm处有强吸收峰;而三种酶裂解SG后,产物在230nm附近无明显峰值。以上证明,三种酶都是多聚甘露糖醛酸裂合酶(EC4·2·99·4)。三种酶裂解褐藻酸后,都能使M/G比值下降,其中酶Ⅰ降得最大,为3.65倍。因此,有可能用酶法制备阻吸90Sr的药物。     

富含古罗糖醛酸的褐藻多糖生产菌株的构建

为获得古罗糖醛酸(Guluronate)含量高的细菌胞外褐藻多糖,利用PCR从海洋细菌Pseudomonassp.QDA中克隆了其甘露糖醛酸C-5差向异构酶基因(algG),连接入质粒pMF 54Km,构建了重组表达载体pMF54 Km-algG。利用三亲接合法将pMF54 Km-algG转入菌株QDA中,获得algG过量表达重组菌株QDA-G1。H-NMR测定结果表明,QDA-G所产的褐藻多糖中β-D-甘露糖醛酸(M)与它的C-5差向异构体α-L-古罗糖醛酸(G)的比值为0.38,G的质量分数达到74.2%,比野生菌株QDA提高了26.4%。且重组菌株遗传稳定性良好,连续传代20代后,M/G的比值无明显变化。     

几种海藻多糖硫酸酯碱式铝盐的制备

本文以褐藻胶和κ-卡拉胶为原料通过酸水解分别获得了低聚的甘露糖醛酸、古罗糖醛酸和低聚 κ-卡拉胶 ,并采用 HPGPC法测定了这 3种低聚糖的重均分子量。在此基础上再经硫酸酯化和成盐修饰制备了各自的硫酸酯碱式铝盐。对制得的几种碱式铝盐进行了红外光谱 (IR)分析和有机硫含量 (S% )、铝含量 (Al% )及样品制酸力等部分理化性质的测定。测定结果初步显示出这几种海藻多糖硫酸酯碱式铝盐较好的抗消化性溃疡前景。     

某些金属离子对褐藻胶膜构体穿透行为研究

褐藻胶是一种多糖,为线性长链聚合物,呈大分子网状结构体,其基本单元是甘露糖醛酸共聚块(M-块)和古罗糖醛酸共聚块(G-块)及M者交替共聚块(MG-块),褐藻多糖是细胞壁的重要组成部分,并具有良好的胶体化学性能,如悬浮、乳化、增稠、絮凝等特性,被广泛应用于食品、医药等行业.本文应用汞膜电极反向极谱法以Ph+2等离子为例,研究和探讨了植物生长所必需的微量元素对褐藻胶结构的穿透性,为试制以褐藻多糖为一定组分的新型植物生长化控营养素提供理论参考.     

低硫酸化杂多糖水解单糖高效液相色谱指纹图谱研究

低硫酸化杂多糖(UF)是从海洋褐藻中分离的一种单糖组成复杂的硫酸化杂多糖,在细胞和动物实验中均表现出显著的神经保护活性,对帕金森病具有显著作用。本研究采用水提-醇沉-DEAE-Sepharose Fast Flow交换层析柱制备UF,经三氟乙酸(TFA)水解和1-苯基-3-甲基-5吡唑啉酮(1-phenyl-3-methyl-5-pyrazalone,PMP)衍生化,通过对水解条件、衍生化试剂使用量、衍生时间的考察,确定最佳衍生条件,通过高效液相色谱(HPLC)法测定UF的单糖组成,并建立UF的指纹图谱。在UF水解单糖的指纹图谱中,共标示出8个特征共有峰,经鉴别主要色谱峰8个,分别是D-甘露糖、D-甘露糖醛酸、L-鼠李糖、D-葡萄糖醛酸、D-葡萄糖、D-半乳糖、D-木糖及L-岩藻糖,其中岩藻糖含量最高,其次是半乳糖。该方法操作简便、快速、灵敏度高、重复性好,可作为UF检查和鉴别的方法,建立实验室UF质量控制标准,为临床用UF质量标准的建立奠定基础。     

GC-MS联合二维核磁对文蛤多糖Fr.2A组分结构的解析

采用GC-MS联合二维核磁的方法,对文蛤均多糖结构进行解析。运用双酶辅助水提的方法,提取文蛤粗多糖(MMPX),分离纯化得到文蛤均多糖,并通过气质联用技术(GC-MS)和核磁共振波谱(1D,2D NMR),对文蛤均多糖的结构进行了分析。结果表明,从文蛤中提取得到文蛤粗多糖(MMPX),经分离纯化得到高纯度文蛤均多糖Fr.2A,其单糖组成为葡萄糖(Glc)。GC-MS分析发现,Fr.2A存在三种残基单元1,4-Glcp、T-Glcp和1,4,6-Glcp,摩尔比为5.58︰1.26︰1.0;核磁共振波谱分析表明,Fr.2A是以α-(1→4)与少量α-(1→6)连接为主链,并存在少量的α-(1→6)和β-(1→4)支链的新型水溶性D型吡喃葡聚糖。     

红树林海洋细菌PLM4抑制肿瘤细胞生长活性多糖的纯化及结构分析

红树林海洋短小芽孢杆菌Bacillus pumilus（PLM4）的发酵液对人喉癌细胞Hep-2、肝癌细胞BEL-7404和舌癌细胞TCA8123的生长抑制活性分别为63．6％、59．2％和61．5％。PLM4发酵液经乙醇沉淀和凝胶柱层析纯化可得到多糖纯品。采用高效凝胶渗透色谱法测定其分子量和纯度,硫酸-咔唑法、2,4-二硝基苯肼比色法、氯化钡明胶比色法、茚三酮反应和气相色谱法等测定其组分。采用红外光谱分析、高碘酸氧化和Smith降解等方法研究其结构。结果表明,该多糖分子量为116kd,由己糖醛酸、甘露糖、葡萄糖和丙酮酸组成,其摩尔比为31：19：6：1,不含硫酸、乙酸和蛋白质。平均每100个己糖基中含有约42个1→3糖苷键、42个1→6糖苷键和／或非还原末端基、13．5个1→2糖苷键和2．5个1→4糖苷键。     

Microbial Biomass and Organic Nutrients in the Deep-Sea Sediments of the Central Indian Ocean Basin

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.     

Assessing the Validity of Seismo-Acoustic Predictor Equations for Obtaining Seabed and Sub-Surface Sediment Physical Properties

The interdependence between the seismo-acoustic properties of a marine sediment and its geotechnical/physical parameters has been known for many years, and it has been postulated that this should allow the extraction of geotechnical information from seismic data. Though in the literature many correlations have been published for the surficial layer, there is a lack of information for greater sediment depths. In this article, a desktop study on a synthetic seafloor model illustrates how the application of published near-surface prediction equations to subsurface sediments (up to several tens of meters burial depth) can lead to spurious predictions. To test this further, acoustic and geotechnical properties were measured on a number of sediment core samples, some of which were subjected to loading in acoustically-equipped consolidation cells (oedometers) to simulate greater burial depth conditions. For low effective pressures (representing small burial depths extending to around 10 meters subsurface), the general applicability of established relationships was confirmed: the prediction of porosity, bulk density, and mean grain size from acoustic velocity and impedance appears generally possible for the investigated sedimentary environments. As effective pressure increases through, the observed relationships deviate more and more from the established ones for the near-surface area. For the samples tested in this study, in some instances increasing pressure even resulted in decreasing velocities. There are several possible explanations for this abnormal behavior, including the presence of gas, overconsolidation, or bimodal grain size distribution. The results indicate that an appropriate depth correction must be introduced into the published prediction equations in order to obtain reliable estimates of physical sediment properties for greater subsurface depths.     

Geoacoustic Inversion via Genetic Algorithm and Its Application to Manganese Sediment Identification

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.     

海上大直径钢管桩打桩过程中桩周土强度弱化研究

海上大直径钢管桩打桩过程中,桩周土体受到强烈扰动而发生强度弱化,掌握桩周土体强度弱化规律对于准确预测打桩过程、保证工程安全具有重要意义。为研究土体强度弱化规律,开展了环剪试验模拟打桩对桩周土体的扰动,测试土体强度随剪切速率的变化规律,建立了描述土体强度弱化规律的拟合公式,引入到打桩分析软件中。研究结果表明：土体的强度折减程度不仅与土体本身的性质有关还受到土体的埋深和剪切速率的影响,埋深越深土体强度折减程度越低,剪切速率越高土体强度折减越高,在打桩分析中可采用这里推荐的线性折减方法来模拟不同深度处土体强度的折减规律。     

Interannual Sea Level Variations and Annual Tides in the Northwestern Pacific

Specific properties of the interannual sea level variations and annual tides in the Northwestern Pacific were studied. Several tide stations were monitored. The monthly mean sea level for the year of 1995 was analyzed at each tide station. A seismic event in 1995, some tectonic activity around the subject area, and the Kuroshio (the oceanic western boundary current) may possibly contaminate results which would have occurred from the astronomical annual tide alone.     

Flow Cytometric Analysis of Coastal Lagoon Bacterioplankton and Picophytoplankton: Fixation and Storage Effects

The effect of different fixation and storage protocols on the flow cytometric (FCM) simultaneous analysis of bacterioplankton and phytoplankton in coastal seawater samples (Mediterranean coastal lagoons) was investigated. FCM measurements (cell number, fluorescence and scatter characteristics) were obtained through DAPI staining. Three fixatives [glutaraldehyde (GA), formaldehyde (FA) and paraformaldehyde (PFA)] and two storage (3 months duration) methods (5 °C and −196 °C) were tested. Two dominant populations were detected in studied samples: bacteria and eukaryotic picophytoplankton. Adding fixatives (2% final concentration) appears necessary to obtain FCM exhaustive counts of all the bacteria and phytoplanktonic cells. This was related to the permeation effect of fixatives which allowed a better DAPI staining of the cells. Maximum fluorescence, i.e. optimal staining of the cells was obtained with FA or PFA, and significant lower fluorescences with GA. Fixed samples stored at 5 °C induced rapid cell loss. Only storage in liquid nitrogen of samples fixed with FA or PFA, allows mid-term (≥4 months) preservation of bacteria or picophytoplankton cell numbers, and limited evolution of DAPI-induced fluorescence and scatter characteristics.     

INFORMATION FOR AUTHORS

<正>Acta Oceanologica Sinica(AOS)is a comprehensive academic journal edited by the Editorial Committee of Acta Oceanologica Sinica and is designed to provide a forum for important research papers of the marine scientific community which reflect the information on a worldwide basis.The journal publishes scholarly papers on marine science and technology,including physics,chemistry,biology,     

Deep-Sea Sediment Compression Curves: Some Controlling Factors, Spurious Overconsolidation, Predictions, and Geophysical Reproduction

Oedometer tests have been carried out on 70 undisturbed surficial clays (at approximately 250 mm below the mudline), mostly collected by free-fall corers from sites widely scattered throughout the deep-sea North Atlantic. Acoustic measurements were also made, initially on contiguous samples and ultimately on the same sample using a geophysically instrumented oedometer which also collected electrical resistivity data. Apart from those quiescent areas below the carbonate compensation depth, such as north of the West Indies where very fine clays exist, most of the samples are silty clays whose geotechnical-geophysical properties are dependent on the type of clay minerals present (and their ability to take in moisture), the sand-size fraction, and the quantity of carbonate present. Thus the pure clays have high compressibilities which decrease on the addition of coarse particles, while the converse is true for the acoustic parameters, these increasing with the sand fraction. Using the notion of the intrinsic compression line for all samples, and comparison to it of the measured compression curves, it is clear that, contrary to some previously held ideas, most deep-sea clays are normally consolidated; the addition of carbonate has the effect of creating an open, stronger sediment skeleton. Interestingly, where information is available, the variation with depth of a sample's acoustic velocity follows the void ratio pressure relationship of the compression curve. This allows the construction of an in-situ sediment compression curve using the in-situ geophysical observations.