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。     

南极树粉孢属真菌胞外多糖的分离纯化及结构表征

研究南极树粉孢属(Oidiodendron truncatum)真菌所产胞外多糖的理化性质和结构特征。采用阴离子交换柱层析和凝胶柱层析对南极树粉孢属(Oidiodendron truncatum)真菌胞外多糖进行了系统的分离纯化,从中得到4个均一的组分(AFW1,AFW2,AFS1和AFS2),并结合红外、气质及核磁等技术对它们的结构进行了解析。结果表明:AFW1和AFW2为中性多糖,AFS1和AFS2中含有少量的蛋白;AFW1和AFS1是由甘露糖、葡萄糖和半乳糖3种单糖组成的结构复杂的杂多糖;AFW2和AFS2则仅由葡萄糖组成,其中AFW2属线性(-1,6-葡聚糖,而AFS2属于C-3位有分支的α-1,6-葡聚糖。     

孔石莼热水提取多糖的研究

分析了孔石莼（Ulva pertusa Kjellm)热水提取得到的多糖的理化性质，并利用核磁共振碳谱解析其结构单元，推定其多糖主要由β-D-葡萄糖醛酸－（1→4）-3-硫酸基－α-L－鼠李糖和α-L－艾杜糖醛酸－1（1→4）－3－硫酸基α-L-鼠李糖结构单元组成。     

甲壳质/壳聚糖及其衍生物的应用化学

甲壳质(Chitin),也称几丁质、壳多糖,俗称甲壳素,是一种天然高分子聚合物,属于氨基多糖,学名为N-乙酰-2-氨基-2-脱氧-D-葡萄糖,分子式为(C8H13NO5)n,单体之间是以β(1→4)苷键连接,分子量一般在106左右,理论含氮量6.9%。甲壳质广泛存在于无脊椎动物的外壳,昆虫的外骨骼,内角质层,真菌的胞壁中。在自然界中,甲壳质的年生物合成量高达千亿吨之多,是仅次于纤维素的第2大自然资源。由于甲壳质天然、无毒(LD5016g/kg)和所具有的良好的生物相溶性、可降解性,以及独特的分子结构和物理、化学性质,使它在医学、药物制剂学、化工、食品、化妆品、…     

拉恩氏菌Rahnella sp. PJT09发酵产胞外多糖的研究

研究拉恩氏菌(Rahnella sp. PJT09)胞外多糖的发酵条件,并进行多糖结构分析。通过单因素试验及正交试验确定拉恩氏菌Rahnella sp. PJT09发酵产糖的最适培养基组成为(g/L):蔗糖40,酵母膏3,NaCl 0.5,ZnCl20.3,K2HPO41;最佳发酵条件为每250 mL三角瓶装液量为100 mL,接种量为3%(体积分数),初始pH值为6.0,28℃,160 r/min培养54 h。在此条件下,Rahnella sp. PJT09多糖产量可达18.51 g/L。采用高效凝胶渗透色谱技术(HPGPC)测定了多糖的分子量,13C-NMR技术确定多糖的结构。研究表明,Rahnella sp. PJT09多糖为重均分子量320 kDa的-β2→6-D-果聚糖。     

坛紫菜R-藻蓝蛋白和变藻蓝蛋白的亚基组成和发色团含量

坛紫菜(Porphyra haitanensis T.J.Chang et B.F.Zheng)中的R-藻蓝蛋白在Bio-Rex 70柱上用脲溶液(pH3.0)解离和层析,得到了α和β两个亚基,用SDS-PAGE测定α和β亚基的分子量分别为18 400和20 500。变藻蓝蛋白的α和β两亚基均分子量经测定,分别为18 800和19 700。R-藻蓝蛋白和变藻蓝蛋白的分子量分别确定为117 000和112 000,两藻胆蛋白中α和β两个亚基的摩尔比都为1:1,确定两藻胆蛋白的亚基组成都为(αβ)_3。各亚基的发色团含量为:R-藻蓝蛋白中,α亚基含1个藻蓝胆素,β亚基含1个藻蓝胆素和1个藻红胆素。变藻蓝蛋白中,α和β亚基各含1个藻蓝胆素。     

珊瑚藻硫酸多糖的提取分离及其结构表征

珊瑚藻(Corallina officinalis)经乙醇脱脂后,再依次经冷水、热水和热碱水溶液提取,从中获得了3种多糖(COW,COH和COA)。运用气相色谱法(GC)、高效凝胶渗透色谱法(HPGPC)、傅里叶变换红外光谱(FTIR)及核磁共振碳谱(13C-NMR)等方法对其单糖组成、重均分子量以及结构进行了表征。结果表明:COW与COH均由半乳糖(Gal)和3,6-内醚-半乳糖(AnG)组成,COA则主要由葡萄糖(Glu)组成;3种多糖的相对分子量分别为24.6,28.4和15.8 kD;硫酸基含量分别为19.0%,15.5%和12.6%。经FTIR和13C-NMR分析表明,COW和COH均属于含有κ-卡拉胶和硫琼胶的杂聚糖,COA为6-硫酸-α-1,4-葡聚糖。     

一株产褐藻酸多糖的海洋假单胞细菌Pseudomonas sp.QDA的筛选和鉴定

根据已获褐藻酸多糖生物合成基因簇中褐藻胶裂解酶基因(αlgL)的序列设计特异性引物，利用PCR技术从海洋微生物中筛选到1株能够分泌胞外多糖的细菌。采用形态学观察和16S rDNA序列分析鉴定该菌株，结果为假单胞属细菌，命名为Pseudomonas sp.QDA；系统发育树显示该菌株与P.putida亲缘关系最近。菌株产生的胞外多糖可被褐藻胶裂解酶(AlgL)降解，并在紫外234nm处检测到特征性吸收，初步证明含有褐藻酸多糖。     

4种海蕴多糖的提取分离及理化性质

以海蕴(Nemacystus decipiens)为研究对象,经85%乙醇脱脂后,通过热水和2%碳酸钠提取并结合乙醇分级法获得了4个多糖组分(NW1~2和NA1~2)。运用HPIC、HPGPC及红外光谱(IR)等方法对其单糖组成、相对分子量以及基本结构特征进行了比较。结果表明,NA1是褐藻胶,经1H-NMR分析,其M/G为0.34。NA2及NW1~2均属于岩藻聚糖硫酸酯,且岩藻糖含量均达80%以上。NW1和NW2中岩藻糖(Fuc)、葡萄糖(Glc)、半乳糖(Gal)、木糖(Xyl)和鼠李糖(Rha)摩尔比分别为100∶5∶3∶16∶1和100∶5∶4∶4∶1。NA2中不含葡萄糖,其Fuc、Gal、Xyl和Rha摩尔比为100∶13∶1∶2。多糖NW1~2和NA1~2的相对分子量分别为939.8,1001,223.6和402.3 kD。     

龙须菜(Gracilaria lemaneiformis)多糖的分离纯化及其组成研究

以龙须菜粗多糖为原料,利用离子交换层析分离纯化,紫外光谱、醋酸纤维薄膜电泳和凝胶过滤层析法鉴定多糖的纯度及其分子量,同时进一步采用气相色谱分析龙须菜多糖的单糖组成.结果表明,利用蒸馏水、0.3mol/L NaCl和1.2mol/L NaCl溶液分步洗脱,DEAE-Sephadex A-25离子交换层析纯化得到龙须菜多糖的三个组分G1、G2和G3,得率分别为3.61%、69.26%、18.70%:G1、G2和G3均显示为单一电泳谱带和单一洗脱峰,无蛋白、核酸杂质,分子量依次为21993、48460、61031U;G2和G3的单糖组成主要为半乳糖、3.6-内醚半乳糖及少量葡萄糖;G2糖链通过1→2键与1→6键方式连接,存在非还原末端基及连三羟基,G3糖链则通过1→3键与1→2键方式连接.     

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.     

A Review of Some Environmental Issues Affecting Marine Mining

This article reviews information recently available from existing marine and coastal mining for responses to environmental issues affecting marine mining at different depths. It is particularly but not exclusively concerned with those issues affecting seabed biodiversity impact and recovery. Much information has been gathered in the past 10 years from shallow mining operations for construction aggregate, diamonds, and gold, from coastal mines discharging tailings to shallow and deep water, and from experimental deep mining tests. The responses to issues identified are summarized in a series of eight tables intended to facilitate site-specific consideration. Since impacts can spread widely in the surface mixing layer SML, and can affect the biologically productive euphotic zone, the main issues considered arise from the depth of mining relative to the SML of the sea. Where mining is below the SML, the issue is whether it is environmentally better to bring the extraction products to the surface vessel for processing (and waste discharge), or to process the extraction products as much as possible on the seabed. Responses to the issues need to be sitespecific, and dependent on adequate preoperational environmental impact and recovery prediction. For deep tailings disposal from a surface vessel, there are four important environmental unknowns: (1) the possible growth of "marine snow" (bacterial flocs) utilizing the enormous quantities of fine tailings particles (hundreds or thousands of metric tons per day) as nuclei for growth, (2) the possibility that local keystone plankton and nekton species may migrate diurnally down to and beyond the depth of deep discharge and hence be subjected to tailings impact at depth, (3) the burrow-up capability of deep benthos and their ability to survive high rates of tailings deposition, and (4) the pattern and rate of dispersion of a tailings density current through the deep water column from discharge point to seabed. Actions to obtain relevant information in general and site-specifically are suggested.     

Increased Particle Fluxes at the INDEX Site Attributable to Simulated Benthic Disturbance

Particle fluxes were measured 7 m above the sea bottom during the predisturbance, disturbance, and postdisturbance periods by using time series sediment traps attached to seven deep-sea moorings deployed in the INDEX experiment site in the Central Indian Basin. The predisturbance particle fluxes varied between 22.3 to 55.1 mg m -2 day -1 . Increased and variable particle fluxes were recorded by the sediment traps during the disturbance period. The increase observed was 0.5 to 4 times more than the background predisturbance fluxes. The increases in particle fluxes (~4 times) recorded by the sediment trap located in the southwestern direction (DMS-1) were the greatest, which could be the result of preferential movement of resuspended particles generated during the deep-sea benthic disturbance along the general current direction prevailing in this area during the experimental period. Also, the traps located closer to the disturbance area recorded greater fluxes than did the traps far away, across the Deep Sea Sediment Resuspension System path. This variability in recorded particle fluxes by the traps around the disturbance area clearly indicates that physical characteristics such as grain size and density of the resuspended particles produced during the disturbance had an important effect on particle movement. The postdisturbance measurements during ~5 days showed a reduction in particle fluxes of ~50%, indicating rapid particle settlement.     

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。     

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.     

Determination of In Situ Sediment Shear Strength from Advanced Piston Corer Pullout Forces

The advanced piston corer (APC) has been used by the Ocean Drilling Program since 1985 for recovering soft sediments from the ocean floor. The pullout force measured on extracting the core barrel from the sediment is shown to correlate with the average shear strength of the sediment core measured in the ship's laboratory. A simple rule of thumb is derived relating the shear strength of the sediment to the pullout force. Multiple APC holes at individual sites allow the consistency of the pullout measurements to be assessed. The effects of different operational procedures during APC coring are also explored. Although generally applicable, the correlation between pullout force and laboratory measurements of shear strength breaks down for some APC holes, possibly because of the disturbance of some sediment types during the APC coring process. A better understanding of the physical process of APC coring, and its effect on the properties of the sediment both inside and immediately outside the core barrel, would indicate what confidence can be put on the measurement of pullout force as a way of evaluating the in situ shear strength of deep sea sediments.     

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.