Antioxidant properties of recombinant allophycocyanin expressed in Escherichia coli

Allophycocyanin (A-PC) is the main core component of phycobilisome found in blue-green algae. The apo-allophycocyanin and its subunits were expressed in Escherichia coli and their antioxidant properties were evaluated using deoxyribose assay. The result showed that both recombinant allophycocyanin fused with maltose binding protein (MBP) tag and 6 x His-tag and their alpha or beta subunits can scavenge hydroxyl radicals successfully, and the separated g or beta subunits had a higher inhibition effect on hydroxyl radicals than that when they combined together. The scavenging effects increased with the increasing concentration. These results clearly suggested that apo-allophycocyanin is involved in the antioxidant and radical scavenging activity of phycocyanin, and the antioxidant activity may be partially responsible to the anti-tumor effect of the recombinant allophycocyanin. (c) 2006 Elsevier B.V. All rights reserved.

Heterologous expression and purification of recombinant allophycocyanin in marine Streptomyces sp isolate M097

Allophycocyanin is one of the most important marine active peptides. Previous studies suggested that recombinant allophycocyanin (rAPC) could remarkably inhibit the S-180 carcinoma in mice, indicating its potential pharmaceutical uses. Based on intergeneric conjugal transfer, heterologous expression of rAPC was first achieved in marine Streptomyces sp. isolate M097 through inserting the apc gene into the thiostrepton-induced vector pIJ8600. The transformation frequency for this system was approximately 10(-4) exconjugants/recipient. In the transformed Streptomyces sp. isolate M097, the yield of purified rAPC could amount to about 38 mg/l using a simple purification protocol, and HPLC analysis showed that the purity of the protein reached about 91.5%. In vitro activity tests also revealed that the purified rAPC had effective scavenging abilities on superoxide and hydroxyl radicals. This would widen the usefulness of the marine Streptomyces as a host to express the rAPC and to offer industrial strain for the production of rAPC.

The cDNA cloning and mRNA expression of heat shock protein 70 gene in the haemocytes of bay scallop (Argopecten irradians, Lamarck 1819) responding to bacteria challenge and naphthalin stress

Heat shock protein 70 (HSP70) is an important member of the heat shock protein superfamily, and it plays a key role in the process of protecting cells, facilitating the folding of nascent peptides and responding to stress. The cDNA of bay scallop Argopecten irradians HSP70 (designated AIHSP70) was cloned by the techniques of homological cloning and rapid amplification of cDNA end (RACE). The full length of AIHSP70 cDNA was 2651 bp in length, having a 5' untranslated region (UTR) of 96 bp, a 3' UTR of 575 bp, and an open reading frame (ORF) of 1980 bp encoding a polypeptide of 659 amino acids with an estimated molecular mass of 71.80 kDa and an estimated isoelectric point of 5.26. BLAST analysis revealed that the AIHSP70 gene shared high identity with other known HSP70 genes. Three classical HSP signature motifs were detected in AIHSP70 by InterPro, analysis. 3-D structural prediction of AIHSP70 showed that its N terminal ATPase activity domain and,C terminal substrate-binding domain shared high similarity with that in human heat shock protein 70. The results indicated that the AIHSP70 was a member of the heat shock protein 70 family. A semi-quantitive RT-PCR method was used to analyse the expression of AIHSP70 gene after the treatment of naphthalin which is one kind of polycyclic aromatic hydrocarbon (PAH) and the challenge of bacteria. mRNA expression of AIHSP70 in scallop was up-regulated significantly after the stimulation of naphthalin and increased with increasing naphthalin concentration. A clearly time-dependent expression pattern of AIHSP70 was observed after the scallops were infected by Vibrio anguillarum, and the mRNA expression reached a maximum level at 8 h and lasted to 16 h, and then dropped progressively. The results indicated that AIHSP70 could play an important role in mediating the environmental stress and immune response in scallop. (c) 2006 Elsevier Ltd. All rights reserved.

Characteristics and Formation Mechanism of Polygonal Faults in Qiongdongnan Basin, Northern South China Sea

Based on high-resolution 3D seismic data, we document the polygonal faults within the Miocene Meishan (sic) Formation and Huangliu (sic) Formation of the Qiongdongnan (sic) basin, northern South China Sea. Within the seismic section and time coherent slice, densely distributed extensional faults with small throw and polygonal shape were identified in map view. The orientation of the polygonal faults is almost isotropic, indicating a non-tectonic origin. The deformation is clearly layer-bounded, with horizontal extension of 11.2% to 16%, and 13.2% on average. The distribution of polygonal faults shows a negative correlation with that of gas chimneys. The development of polygonal faults may be triggered by over-pressure pore fluid which is restricted in the fine-grained sediments of bathyal facies when the sediments is compacted by the burden above. The polygonal faults developed to balance the volumetric contraction and restricted extension. The product of hydrocarbon in the Meishan Formation may have contributed to the development of the polygonal faults. In the study area, it was thought that the petroleum system of the Neogene post-rift sequence is disadvantageous because of poor migration pathway. However, the discovery of polygonal faults in the Miocene strata, which may play an important role on the fluid migration, may change this view. A new model of the petroleum system for the study area is proposed.

Petroleum System in Deepwater Basins of the Northern South China Sea

The northern South China Sea margin has experienced a rifting stage and a post-rifting stage during the Cenozoic. In the rifting stage, the margin received lacustrine and shallow marine facies sediments. In the post-rifting thermal subsidence, the margin accumulated shallow marine facies and hemipelagic deposits, and the deepwater basins formed. Petroleum systems of deepwater setting have been imaged from seismic data and drill wells. Two kinds of source rocks including Paleogene lacustrine black shale and Oligocene-Early Miocene mudstone were developed in the deepwater basin of the South China Sea. The deepwater reservoirs are characterized by the deep sea channel rill, mass flow complexes and drowned reef carbonate platform. Profitable capping rocks on the top are mudstones with huge thickness in the post-rifting stage. Meanwhile, the faults developed during the rifting stage provide a migration path favourable for the formation of reservoirs. The analysis of seismic and drilling data suggests that the joint structural and stratigraphic traps could form giant hydrocarbon fields and hydrocarbon reservoirs including syn-rifting graben subaqueous delta, deepwater submarine fan sandstone and reef carbonate reservoirs.

Hydrogeochemistry of formation water in relation to overpressures and fluid flow in the Qikou Depression of the Bohai bay basin, China

The Qikou Depression is the largest hydrocarbon bearing depression in the western part of the Bohai bay basin, dominated by fan delta and lacustrine strata with volcanic and volcaniclastic rocks. In this study, the formation pressures and hydrochemistry of the formation water in the Qikou depression are investigated. It is found that a significant overpressure occurs in the Dongying (Ed) Formation and the first member (Est), the second member (Es2), the third member (Es3) of the Shahejie Formation. The pressure coefficients commonly range from 1.2 to 1.6 with the highest pressure coefficient being 1.7. The analysis of hydrochemistry data shows that the whole depression is dominated by NaHCO3 water type. The concentration of total dissolved solid (TDS) ranges from 2.13 to 53.16 g/L and shows a distinct vertical variation of salinity and ion ratios. High salinity water (TDS> 10 g/L) occurs below a depth of 2500 m, which coincides with the presence of the overpressured system. However, the increasing trend of TDS is diminished below 3500 m because the generation of organic acids in Qikou Depression is inhibited in the presence of overpressure. The analysis of the relationship among different ions indicates that the present-day characteristics of the formation water result from the albitization of feldspar and the dissolution of sodium-rich silicate minerals and halite in the different hydrochemical and pressure systems. (C) 2009 Elsevier B.V. All rights reserved.

Geophysical evaluation methods for buried hill reservoirs in the Jiyang superdepression of the Bohai Bay basin, eastern China

The Jiyang superdepression is one of the richest hydrocarbon accumulations in the Bohai Bay basin, eastern China. Comprehensive seismic methods have been used in buried hill exploration in Jiyang to describe these fractured reservoirs better. Accurate seismic stratigraphic demarcation and variable-velocity mapping were applied to reveal the inner structure of the buried hills and determine the nature of the structural traps more precisely. Based on the analysis of rock properties and the characteristics of well-developed buried hill reservoirs, we have successfully linked the geology and seismic response by applying seismic forward technology. Log-constrained inversion, absorption coefficient analysis and tectonic forward-inversion with FMI loggings were applied to analyse and evaluate the buried hill reservoirs and gave satisfying results. The reservoir prediction was successful, which confirmed that the comprehensive utilization of these methods can be helpful in the exploration of buried hill reservoirs.

渤海湾盆地济阳坳陷东部走滑构造特征及其对油气成藏的影响研究

研究区位于郯庐断裂中段与济阳坳陷的构造结合部，区内走滑构造广泛发育，主要的走滑断裂有7条，分别是郯庐断裂带的东西两支、垦东断层、孤东断层、长堤断层、埕东断层和发育于垦东凸起中部的浅层走滑构造带。走滑构造带与油气富集带有着明显的对应关系。 通过对研究区内二维、三维地震测线和平面构造图的精细解释和分析，分别揭示了各走滑断裂在平面、剖面和三维空间上的构造形态。根据走滑断裂及其伴生构造的平面和剖面上的几何学特征，将研究区内的走滑断裂划分为三种类型：成熟型走滑断裂、隐伏型走滑断裂、不连续型的走滑断裂。 从理论模式研究入手，推导了拉分盆地中盆地的走滑速率与沉降速率之间的关系，证实了走滑速率同盆地的几何形状参数、最大沉降深度和盆地的沉降速率存在着稳定的数值关系。通过对莱州湾地区潍北凹陷基底沉降历史的分析，建立了潍北凹陷沉降速率与郯庐断裂中段走滑速率之间的经验关系式，进而求出郯庐断裂中段新生代右行走滑位移量的大小为40km。 运用2DMove软件，对研究区内四条典型剖面进行构造复原，计算出了各条剖面每个时期的伸展参数，对研究区构造活动强度进行了定量分析，揭示了研究区的构造演化规律。通过运用Ansys软件进行有限元模拟，恢复了晚白垩世晚期-古近纪早期研究区内的构造应力场和应变场，揭示了扭张作用是研究区内走滑断层开始走滑的主要原因。 通过上述分析，结合对究区内近几年勘探开发成功和失败的实例分析，全面探讨了走滑活动对于油气成藏“生”、“储”、“盖”、“圈”、“运”、“保”各因素的影响。

东海沿岸贝类体内石油烃和有机氯农药的分布特征及评价

本研究分别于2006、2007年5月在东海沿岸海域22个站位采集牡蛎、贻贝、四角蛤蜊、文蛤、花蛤5种贝类样品，采用荧光分光光度法和气相色谱法检测贝类体内的石油烃（TPHs）和有机氯农药(HCHs和DDTs) 含量，分析东海沿岸海域贝类体内的TPHs、HCHs和DDTs的时空分布特征和HCHs、DDTs的组分特征，比较不同贝类体内污染物含量的差异，探讨HCHs和DDTs的来源，并对贝类进行初步海洋生物质量评价和人体健康风险评价。研究结果表明： （1）东海沿岸海域贝类体内TPHs含量范围为0.40 74.40 mg kg-1，平均含量为9.42 mg kg-1；HCHs含量为ND～5.42×10-3mg kg-1，平均含量为1.47×10-3 mg kg-1；DDTs含量为2.09×10-3～197.66×10-3mg kg-1，平均含量为38.05×10-3 mg kg-1。其中，TPHs含量在长江口及邻近海域较高， HCHs和DDTs含量在江苏省的如东、海门和浙江省的各海域含量较高。与2006年相比，2007年贝类体内TPHs含量与2006年基本持平，HCHs含量有稍微降低，但是DDTs含量有明显升高趋势。 （2）HCHs和DDTs组分分析表明，α-HCH和β-HCH占HCHs总量的绝大多数，γ-HCH和δ-HCH仅有极少量检出，大多数区域的α-HCH/γ-HCH比值在4-7之间，说明研究区域内有新的HCHs污染源； DDTs 中O,P’-DDT/P,P'-DDT的比值远高于工业DDTs中O,P’-DDT/P,P'-DDT的比值，分析认为其与周围区域使用三氯杀螨醇密切有关。 （3）贝类体内的TPHs、HCHs和DDTs与水体、沉积物中的相应有机物存在正相关性。相对于TPHs和HCHs，DDTs更容易在贝类体内富集；牡蛎对TPHs和HCHs、DDTs的累积能力明显高于四角蛤蜊和贻贝。 （4）质量评价显示TPHs在局部海域超标，HCHs未超标，DDTs含量超标严重。人体健康风险评估结果显示，东海沿岸海域贝类体内HCHs和DDTs的致癌风险和暴露风险均在可接受的范围之内，消费者因食用监测海域的贝类而受到HCHs和DDTs危害的可能性不大。 （5）与世界其他海域比较表明，东海海域贝类TPHs和HCHs含量处于中等水平， DDTs含量处于比较高的水平。

南海北部陆缘深水区构造演化及其资源效应

南海北部陆缘深水区（水深>300m）蕴藏着丰富的资源，我国对深水区的地质研究刚刚起步，但相关领域已成为科研热点。深水油气盆地的构造演化是油气勘探中最重要的基础性研究之一，因此针对我国南海北部陆缘深水区开展构造演化及其资源效应的研究具有重要的理论意义和实际意义。 本文利用钻井和地震资料并结合区域地质资料，重点研究了珠江口盆地深水区的结构和构造演化，取得如下创新性成果：1）首次利用半地堑分析方法系统解剖了研究区的结构、各构造单元发育特征，在此基础上指出五个有利油气运聚带；2）采用回剥法并利用最新资料进行校正，得到了研究区更为可靠的构造沉降曲线，重新划分了裂陷期和裂后期的分界，认为32Ma南海海底扩张之后裂陷作用仍在持续，直到23Ma左右才开始大规模裂后热沉降，并进一步解释了裂陷期延迟的形成机制；3）应用非连续拉张模型计算拉张系数的方程计算了研究区的壳幔拉张系数，指出了深水区地幔相对于地壳的优势伸展作用；首次运用平衡剖面技术重建了研究区的构造发育史，计算了各构造期的拉张率和沉积速率，指出研究区新生代整体呈现持续拉张，拉张系数在1.1-1.24之间；4）精细刻画了水合物钻采区的地质构造特征，建立了该区天然气水合物成藏的概念模式；建立了一套根据地震叠加速度计算流体势的方法，为水合物成藏规律的研究提供了新的思路。

Accumulation of petroleum hydrocarbons and heavy metals in clams (Ruditapes philippinarum) in Jiaozhou Bay, China

Accumulation and distributions of aliphatic and polyaromatic hydrocarbons (PAHs) and heavy metals were measured in tissues of the clam Ruditapes philippinarum collected from 5 sites in Jiaozhou Bay, Qingdao, China. The concentrations of total aliphatic hydrocarbon and PAHs ranged from 570 to 2 574 ng/gdw (gram dry weight) and from 276 to 939 ng/gdw, in the most and least polluted sites, respectively. The bio-accumulation of hydrocarbons and PAHs in the clams appeared to be selective. Aliphatic hydrocarbons were predominantly represented by short chain (< nC(23)) n-alkanes, suggesting that petroleum hydrocarbons were likely the major contamination source. The selective uptake of 3 and 4 ring PAHs, such as naphthalene, fluorene, phenanthrene, fluoranthene and pyrene, by the clams was probably related to the physiological and bio-kinetic processes that were energetically favorable for uptake of compounds with fewer rings. Accumulation of the metals Cd, Cu, Zn, Pb, Cr, Hg, and As in the clam tissues also showed high variability, ranging from 0.043 to 87 A mu g/gdw. Among the 7 detected metals, Zn, Cd, Cu, and As had a particularly high potential of accumulation in R. philippinarum. In general, a positive correlation was found between the tissue concentrations and sediment concentrations of hydrocarbons and of some metals. Our study suggests that moderate contamination with polyaromatic hydrocarbons, and low to moderate contamination with metals, currently exists for clam R. philippinarum in Jiaozhou Bay, in comparison with other regional studies. A long-term monitoring program is certainly needed for assessment of the potential ecological influence and toxicity of these contaminants of R. philippinarum in Jiaozhou Bay.

Preparation and in vitro antioxidant activity of kappa-carrageenan oligosaccharides and their oversulfated, acetylated, and phosphorylated derivatives

In order to study the relationship between chemical structure and properties of modified carrageenans versus antioxidant activity in vitro, K-carrageenan oligosaccharides were prepared through mild hydrochloric acid hydrolysis of the polysaccharide, and these were used as starting materials for the partial synthesis of their oversulfated, acetylated, and phosphorylated derivatives. The structure and substitution pattern of the oligosaccharides and their derivatives were Studied using FTIR and C-13 NMR spectroscopy, and their in vitro antioxidant activities were investigated. Certain derivatives of the carrageenan oligosaccharides exhibited higher antioxidant activity than the polysaccharides and oligosaccharides in certain antioxidant systems. The oversulfated and acetylated derivatives, which scavenge superoxide radicals, the phosphorylated and low-DS acetylated derivatives, which scavenge hydroxyl radicals, and the phosphorylated derivatives, which scavenge DPPH radicals, all exhibited significant antioxidant activities it, the systems examined. The effect of the molecular weight of the carrageenan on antioxidant activities, however, is not obvious from these studies. (c) 2005 Elsevier Ltd. All rights reserved.