极地寡营养细菌(Alteromonas stellipolaris sp. nov.)色素的提取及其理化性质的研究

以分离自南极海洋的寡营养细菌所产的褐色素为试验材料,进行了该色素的提取和稳定性分析。结果表明,色素粗品至少含有3种以上不同的褐色素,该褐色素易溶于水和甲醇;最大吸收波长为214nm;在pH≥4.0、热、光照(自然光与紫外线)下都能表现出良好的稳定性;抗H2O2、HNO3氧化能力强,NaClO对其破坏较大;Na2SO3、抗坏血酸对其有护色作用;Fe3+、Fe2+和Pb2+则有一定的破坏作用,Mg2+对该色素有护色作用,其它金属离子影响不大。试验表明,寡营养细菌褐色素在稳定性上呈现出一定优势,具有作为一种天然色素资源的开发和应用价值。     

海洋细菌Pseudomonas sp.色素的提取及稳定性的研究

对分离自广东大亚湾表层海水的一株海洋细菌（Pseudomonassp.）所产的灵菌红素,作了提取和稳定性分析,探讨了酸度、温度、紫外线、金属离子对色素稳定性的影响.结果证明,灵菌红素在pH 2～pH 5之间稳定性很好,在碱性环境中稳定性差;在pH=3时该色素的热稳定性好,pH＞5时其热稳定性较差;紫外线随溶液的pH增大对色素的稳定性影响越大,pH=9时的影响最大;Zn^2＋对该色素有增色作用,Mg^2＋和Mn^2＋则有一定的破坏作用,Pb^2＋可以使色素产生沉淀,其它金属离子影响不大.总之,该灵菌红素各种性质比较稳定,具有作为一种天然色素资源的开发和应用价值.     

地衣芽孢杆菌胞外产物消化活性的研究

将地衣芽孢杆菌Bacillus licheniformis的胞外产物置于不同的pH、温度和金属离子条件下,分别测定其蛋白酶、淀粉酶、脂肪酶活性的变化。结果表明,当pH为7.5—8.5时胞外产物表现出较高的蛋白酶和淀粉酶活性,胞外产物消化脂肪的最适pH为9.0—10.0。胞外产物水解淀粉、脂肪和蛋白质最适温度分别为60—80、40—60和50—70℃,60℃时蛋白酶活力最大。试验测定了8种金属离子对胞外产物活性的影响,发现Mg2+、Zn2+Cu2+可抑制其水解蛋白质,而Ca2+、Fe3+、Mn2+和Ba2+却无此影响;在胞外产物消化脂肪底物时Co2+、Fe3+、Mn2+和Ba2+均能抑制其活力,而Mg2+、Zn2+能使其消化活力升高,与前者相比,金属离子对淀粉水解的影响相对较小,仅Cu2+有一定的抑制作用,其它离子对其均无明显的影响。还研究了不同胞外产物浓度与生物学活性的相关性,发现当反应体系中胞外产物浓度升高时水解蛋白质的活力会相应降低,但对脂肪的消化活力则与胞外产物的浓度呈正相关。反应体系中胞外产物的浓度在0—132.4μg.ml-1时其水解淀粉的活力随浓度的升高而升高,浓度为191.0μg.ml-1时活力最大,浓度大于191.0μg.ml-1时活力不再变化,稳定在一个较高的水平。     

中国明对虾体壁N-乙酰-β-D-氨基葡萄糖苷酶的分离纯化及酶学性质

为了探讨对虾N-乙酰-β-D-氨基葡萄糖苷酶(EC3.2.1.52,NAGase)的分离纯化及其酶学性质,作者以中国明对虾(Fenneropenaeus chinensis)体壁为材料,通过硫酸铵沉淀分级分离、SephadexG-100柱层析和DEAE-32离子交换柱层析纯化,获得聚丙烯酰胺凝胶电泳纯的N-乙酰-β-D-氨基葡萄糖苷酶酶制剂,纯化酶比活力为3938.56U/mg。通过SDS-PAGE凝胶电泳,测得该酶亚基分子量为48.88 kD。酶的最适pH为6.0,最适温度为45℃;该酶在pH 6.0~9.0区域较稳定,温度稳定性范围是20~35℃,45℃下处理1 h酶活力丧失65.04%。酶水解对硝基苯-N-乙酰-β-D-氨基葡萄糖苷的Km为0.229 mmol/L,Vmax为5.00μmol/(L·min)。进一步研究金属离子对酶活力的影响,结果表明:Li+、Na+和Ba2+对酶没有明显影响,Mg2+、Ca2+、Mn2+、Co2+、Fe3+和Al3+对酶均有不同程度的激活作用,Cu2+、Zn2+和Pb2+对酶呈抑制作用,1.0 mmol/L Hg2+对酶呈激活作用,而10 mmol/L Hg2+使酶活力降低42.37%。     

10种金属离子对三疣梭子蟹中肠腺消化酶活性的影响

以酶学分析方法研究了10种金属离子(Ag 、Cd2 、Ca2 、Cu2 、Fe3 、Hg2 、Mg2 、Mn2 、Pb2 和Zn2 )对三疣梭子蟹Portunus trituberculatus中肠腺蛋白酶、胰蛋白酶、胰凝乳蛋白酶、羧肽酶A、羧肽酶B、淀粉酶和纤维素酶活性的影响,10种金属离子浓度分别设置为5×10-4、1×10-3和5×10-3mol.L?13个梯度。结果表明,在实验浓度范围内低浓度(0.5mmol.L-1)Fe3 、Pb2 、Mn2 及0.5和1mmol.L-1的Ca2 对蛋白酶活性影响不显著,Mg2 对蛋白酶的激活作用随浓度的升高而升高,1mmol.L-1的Mn2 对蛋白酶表现为显著激活作用;3种浓度的Ca2 、Cd2 、Mg2 、Mn2 对胰蛋白酶均表现显著激活作用;Ca2 及低浓度的Mg2 和Cu2 对胰凝乳蛋白酶有激活作用;Hg2 及低浓度的Mg2 、Mn2 、Zn2 对羧肽酶A有激活作用;Ca2 及低浓度的Mg2 对羧肽酶B有激活作用;Ca2 、Cd2 、Mn2 和低浓度的Zn2 、Fe3 、Pb2 对淀粉酶表现为激活作用;Zn2 、Fe3 、Mn2 、Cu2 对纤维素酶表现为激活作用。除此之外,金属离子对这几种消化酶均表现为抑制作用,抑制作用随浓度的升高而升高。     

雨生红球藻虾红素的制备及其稳定性研究

以雨生红球藻提取物为原料,利用皂化法和溶剂分配法制备虾红素,并研究其结构及稳定性。通过优化皂化过程中的碱液浓度、反应时间和反应温度,得到最佳的工艺条件:碱液浓度7%,皂化时间120min,温度15℃。此条件下,虾红素的转化率达96.12%,制备的虾红素得率为62.16%,纯度达80.36%。用HPLC-DAD-(APCI)MS/MS分析反应产物,解析虾红素的质谱裂解规律,研究光照条件、金属离子和温度对虾红素稳定性的影响。结果表明:紫外光照处理对虾红素稳定性影响显著;Fe~(3+)、Ca~(2+)、Cu~(2+)对虾红素有不同程度的破坏作用,Zn~(2+)、Mg~(2+)、Na~+、Fe~(2+)对虾红素影响较小;pH在3~8范围内,随着pH升高,虾红素稳定性提高;随着温度的升高,虾红素的保留率下降。本研究为虾红素的活性、毒理及其在食品中的应用等研究提供了重要参考。     

红树林生境产嗜热纤维素酶芽孢杆菌的筛选及其活性分析

本研究从红树林沉积物样品中筛选获得一株具有较高纤维素酶活性的菌株ms-2,通过革兰氏染色、生理生化试验,以及16S r DNA基因片段比对鉴定该菌株为枯草芽孢杆菌(Bacillus subtilis)ms-2。对其产酶性质进行研究表明该菌株所产纤维素酶为胞外酶而非膜锚定酶,该酶具有良好的嗜热性,且在弱酸条件下酶活性较高,最适宜温度和pH分别为65℃和6. 0,在此条件下最高酶活性为0. 655 7 U/cm3;另外,在浓度为1 mmol/dm3Cu2+、Mg2+、Zn2+、Fe3+和Mn2+条件下,Cu2+对酶活性无显著影响,Mg2+、Zn2+、Fe3+和Mn2+对酶活性均有不同程度的抑制作用,其中Mn2+的抑制最为明显,酶活性下降了29. 96%。由于产业应用中酶解纤维素时通常需要在55～60℃的温度下进行,研究获得的嗜热纤维素酶不仅适合这样的温度环境,还能维持较高的酶活性水平,并且对Cu2+、Mg2+、Zn2+、Fe3+较不敏感,有利于产业开发应用。     

深海贝莱斯芽孢杆菌DH82的筛选、鉴定及其抗菌粗蛋白性质分析

本研究从西太平洋雅浦海沟 6 000 m 深的水样中分离得到 106 株深海细菌,其中筛选到一株能较强拮抗迟缓爱德华氏菌等水产致病菌的 DH82 菌株。采用 16S rDNA 和 gyrB 基因 （NCBI 登录号：MK203035 与 MK203036） 分子生物学分析, 结合形态学和生理生化特性,鉴定 DH82 菌株为贝莱斯芽胞杆菌 Bacillus velezensis。对其抗菌粗蛋白性质进行研究发现：该粗蛋白具有较好热稳定性,在 100 益,处理 30 min 后,仍有 85.5 %抑菌活性;木瓜蛋白酶、蛋白酶 K、胃蛋白酶及胰蛋白酶对粗蛋白影响不显著;粗蛋白在 pH7.0 条件下抑菌活性最佳;金属离子 K+和 Na+对粗蛋白抑菌活性有促进作用,Mg2+、Ca2+及 Fe3+等多价金属离子对粗蛋白抑菌活性有抑制作用。     

嗜热栖热菌重组锰超氧化物歧化酶的性质

嗜热栖热菌(Thermus thermophilus)wl的超氧化物歧化酶基因在Escherichia coli BL21(DE3)中进行表达,采用Ni-NTA agarose亲和层析获得重组超氧化物歧化酶,比活力达565 U/mg.重组酶的相对分子质量为110.9 kDa,亚基相对分子质量为28.3 kDa,该酶为同聚四聚体蛋白.重组酶对氯仿-乙醇和SDS敏感,对H2O2不敏感,并且它的紫外最大吸收波长位于279 nm,判断其属于锰超氧化物歧化酶类型.1.0 mmol/dm3Mn2+对重组酶有激活作用,同浓度的其他金属离子Mg2+、Ca2+、Ba2+、Co2+、Cu2+、Zn2+和Fe2+对重组酶活力都有抑制作用,其中以Co2+的作用最强,抑制了约64%的酶活性.重组酶在70℃下处理1 h,保持原有活力的60%,即使在90℃下处理1 h,仍保持原有活力的35%.在pH值为4.0～11.0的缓冲液中25℃下处理1 h,保持65%以上的原始活力.由此可见,重组锰超氧化物歧化酶具有高的热稳定性和宽pH稳定性,在工业上具有巨大的应用潜力.     

产碱性纤维素酶海洋细菌的筛选、鉴定及酶学性质研究

用CMC平板筛选方法,从青岛近海海域海水中分离出一株产碱性纤维素酶的海洋菌株QM11,经16S rDNA鉴定,该菌株为Cytophaga fucicola.对该菌的生物学特性研究表明,其最适生长温度为27℃,生长温度范围为4～48℃,为耐冷菌;在pH7.0～8.0、含3.0%NaC1的培养基条件下,最适宜菌株生长和产酶;QM11所产碱性纤维素酶最适反应温度为40℃,最适反应pH为9.0,在碱性条件下具有较高的酶活性和较好的稳定性.Mn2+、Fe3+对酶反应具有促进作用,Cu2+、Pb2+对酶反应具有抑制作用.     

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.     

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.     

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.     

Phylogenetic diversity and phenotypic characterization of cultivable bacterioplankton isolated from polar oceans

A set of 27 marine planktonic bacteria isolated from the polar regions was characterized by 16S rDNA sequencing and physiological and biochemical testing. More than half of these bacteria were positive for caseinase, gelatinase and 13-glucosidase, and could utilize glucose, maltose or malic acid as carbon source for cell growth. Twelve isolates expressed nitrate reduction activities. Except for one antarctic isolate BSwlO175 belonging to Actinobacteria phylum, these isolates were classified as γ-Proteobacteria, suggesting that γ-Proteobacteria dominated in cultivable marine bacterioplankton at both poles. Genus Pseudoalteromonas was the predominant group in the Chukchi Sea and the Bering Sea, and genus ShewaneUa dominated in cultivable bacterioplankton in the Prydz Bay. With sequence similarities above 97%, genus Psychrobacter was found at both poles. These 27 isolates were psychrotolerant, and significant 16S rDNA sequence similarities were found not only between arctic and antarctic marine bacteria （ 〉 99% ）, but also between polar marine bacteria and bacteria from other aquatic environments （ ≥ 98.8% ）, including temperate ocean, deep sea, pond and lake, suggesting that in the polar oceans less temperature-sensitive bacteria may be cosmopolitan and have a bipolar, even global, distribution at the species level.     

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.     

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.