合浦珠母贝基质蛋白KRMP-3对二价金属离子选择性的研究

用大肠杆菌表达含有GST标签的基质蛋白KRMP-3。利用圆二色谱(Circular Dichroism, CD)研究不同浓度钙离子和镁离子对基质蛋白 KRMP-3二级结构的影响。结果表明,钙离子对其二级结构的变化远大于镁离子;同时,采用荧光淬灭法研究 KRMP-3对钙,镁,锶,钡等二价金属离子的选择性,结果表明, KRMP-3对钙离子有特异性选择性,钙离子与KRMP-3的结合常数K约为103 L/mol,结合位点数 n 近似为1,表明 KRMP-3与钙离子的结合能力适中,推测基质蛋白 KRMP-3对合浦珠母贝( Pinctada fucata)棱柱层形成起到促进作用。     

宁波沿海陆源排污口细菌结构多样性和铁代谢相关的毒力基因的研究

应用多重PCR技术对肠杆菌铁代谢相关毒力基因fhuA、irp2、sltA和sodB进行同步检测, 研究宁波沿海陆源排污口人畜共患致病菌群落结构在人类活动影响下的分布差异及对环境因子变化的响应趋势。通过16S rRNA 基因的PCR扩增和限制性片段长度多态性(restriction fragment length polymorphism, RFLP)分析技术, 研究了东海重要陆源排污口的细菌群落多样性; 建立肠杆菌铁载体外膜受体蛋白fhuA、铁调节蛋白irp2、类志贺样毒素sltA、含铁超氧物歧化酶sodB基因多重PCR体系, 并用该体系检测铁代谢相关毒力基因在陆源排污口的分布情况。2011年3、5、8、10月份, 在宁波沿海陆源排污口中共分离纯化1000余株、98种细菌, 16S rRNA 基因序列分析表明, 它们分属于4个门5个纲13个科21个属, 其中?-变形杆菌纲(67株, 68.4%)和芽孢杆菌纲(16株, 16.3%)分别为变形菌门、厚壁菌门中主要纲。98株细菌中, fhuA+irp2+sodB+基因型检出率为35.71%; fhuA+irp2+ sltA+的基因型检出率为25.27%; irp2的基因型检出率为13.19%; fhuA基因型检出率为12.09%; fhuA+ irp2+ 基因型检出率为9.89%; sltA+sodB+基因型检出率为8.79%; fhuA+irp2+sodB+sltA+基因型检出率为8.79%。     

Distribution and Variation of Ribonucleic Acid （RNA） and Protein and Its Hydrolysis Products in Lake Sediments

Protein and RNA in lake sediments tend to be decomposed progressively with time and sedimentation depth. Their concentrations tend to decrease starting from the sedimentation depth of 17 cm and that of 19 cm, respectively. However, the products of their decomposition —amino acids and nucleotides show different rules of variation. At the depth from 27 cm to 30 cm the amino acids are most abundant in the pore waters of lake sediments. Such variation tendency seems to be related to the extent to which microbes utilize amino acids and nucleotides. Due to polymerization in the geological processes and the adsorption of protein on minerals and organic polymers, below the sedimentation depth of 17 cm there is still a certain amount of protein in the sediments. With the time passing by, protein has been well preserved in various sediment layers, indicating that its decomposition is relatively limited. The peak values of protein content in the sediments of the two lakes are produced in the surface layers at the depth of 10 cm, implicating that the surface sediments are favorable to the release of protein. The contents of amino acids in the pore waters of lake sediments are closely related to the activities of microbes. Below the depth of 27 cm, the amino acids are significantly accumulated in Lake Aha sediments, probably indicating the weakening of microbial activities. The research project was financially supported jointly by the Ntional Natural Science Foundation of China (No. 40173038) and the Guizhou Provincial Foundation (No. 3090) and the Chinese Academy of Sciences Knowledge-Innovation Foundation (KZCX2-105).     

Microbial Diversity in Hydrothermal Systems and Their Influence on Geological Environments

Abstract: Microorganisms found at terrestrial and seafloor hydrothermal systems are classified based on their optimum growth temperatures. Prokaryotes exhibiting optimum growth temperatures above 45C are defined as thermophiles, while those with optimum growth temperatures above 80C are called hyperthermophiles. Hyperthermophiles have chemically and thermodynamically stable cytoplasmic‐membrane consisting of ether–lipids. Hyperthermophiles observed at temperature up to 113C in seafloor hydrothermal areas should have thermostable proteins. Presumably, the thermostable proteins are replacing certain amino acid residue by proline instead of alanine in order to increase hydrogen bonds and salt bridges. Microorganisms inhabiting terrestrial and seafloor hydrothermal areas obtain energy mainly by use of chemical disequilibrium, so that their habitat may be restricted to the redox boundary between ambient water and reduced hydrothermal fluids. Among more than 200 species of cultivable thermophiles and hyperthermophiles, hyperthermophiles mainly obtain energy by sulfur reduction, because this reaction can produce energy efficiently at > 80C. Some hyperthermophilic sulfur reducers can reduce Cr(VI), Mn(IV), Fe(III), Co(III), As(V), Se(VI), Tc(VII), Au(III) and U(VI) through hydrogen or organic compounds that act as reducing agents for yielding energy. Some hyperther‐mophiles use W instead of Mo for activating their enzymes. Bacteria can pump out toxic metal ions from their cells, however toxic metal resistance systems of archaea, including hyperthermophiles, have not yet been studied. Methane producing hyperthermophiles can yield energy by the autotrophic reaction, when sufficient carbon dioxide and molecular hydrogen can be supplied and methane can be removed from hydrothermal systems. If nitrogen can be supplied into a hydrothermal system, hydrogen oxidization by nitrogen species (e.g. NO₃^‐, NO₂^‐, NO and N₂O) may be expected through the metabolism of some hyperthermophiles.     

Seasonal nutritive value of native grasses of Argentine Calden Forest Range

This study assessed the nutritive value of the most important forage species of the Calden forest (central semi-arid La Pampa, Argentina), for samples collected in fall, winter and spring, under grazing conditions and during two successive years, for ranges of good and fair conditions. The crude protein concentration (CP) of short-winter grasses (Piptochaetium napostaense, Poa ligularis, Stipa clarazii and Hordeum stenostachys) was about 10%. Mid-winter grasses (S. tenuissima and S. gynerioides) never reached 6% CP. Summer grasses (Digitaria californica and Trichloris crinita) ranged from 7% to 9% CP. In vitro dry matter digestibility (IVDMD) was similar among short-winter and summer grasses (40–50%). Mid-winter grasses had the lowest IVDMD for all seasons (<40%). Effects of sampling year and range condition on CP were consistently significant only for short-winter grasses. Good condition ranges provide a more acceptable forage supply than fair condition ranges.     

Proline with or without Hydroxyproline Influences Collagen Concentration and Regulates Prolyl 4-Hydroxylase α(I) Gene Expression in Juvenile Turbot(Scophthalmus maximus L.)

This study was conducted to investigate the effect of dietary proline(Pro), and Pro and hydroxyproline(Hyp) in combination on the growth performance, total Hyp and collagen concentrations of tissues, and prolyl 4-hydroxylase α(I)(P4H α(I)) gene expression in juvenile turbot feeding high plant protein diets. A diet containing 50% crude protein and 12% crude lipid was formulated as the basal and control, on which other two protein and lipid contents identical experimental diets were formulated by supplementing the basal with either 0.75% Pro(Pro-0.75) or 0.75% Pro and 0.75% Hyp(Pro+Hyp). Four groups of fish in indoor seawater recirculating systems, 35 individuals each, were fed twice a day to apparent satiation for 10 weeks. The results showed that dietary Pro and Hyp supplementation had no significant effect on growth performance and feed utilization of juvenile turbot(P 0.05). Total Hyp and collagen concentrations in muscle were significantly increased when dietary Pro and Hyp increased(P 0.05), and fish fed diet Pro+Hyp showed significantly higher free Hyp content in plasma than those fed other diets(P 0.05). The expression of P4 H α(I) gene in liver and muscle was significantly up regulated in fish fed diet Pro-0.75 in comparison with control(P 0.05); however the gene was significantly down regulated in fish fed diet Pro+Hyp in muscle in comparison with fish fed diet Pro-0.75(P 0.05). It can be concluded that supplement of crystal L-Pro and L-Hyp to high plant protein diets did not show positive effects on growth performance of juvenile turbot, but enhanced total collagen concentrations in muscle.     

环渤海湿地沉积物球囊霉素相关土壤蛋白分布特征及其对湿地土壤风化的影响

【研究目的】球囊霉素作为丛枝菌根真菌（AMF）分泌的一种难降解土壤蛋白,广泛分布于陆地生态系统中,是长期碳贮的重要组成。当前鲜有研究涉及其在滨海湿地中的分布。基于此,本文对中国环渤海主要滨海湿地表层沉积物中球囊霉素相关土壤蛋白（GRSP）的空间分布进行表征,探讨不同湿地生境下GRSP分布及其对沉积物风化的指示意义。【研究方法】本文选取辽河三角洲、北大港和黄河三角洲湿地作为研究区,对166个表层样（0~5 cm）和4个柱状样（0~35 cm）的GRSP、粒度以及常量元素进行测定,并计算化学蚀变系数（CIA）。【研究结果】研究区GRSP在空间上的动态变化受植被类型影响显著,范围在0.06~11.31 mg/g,均值为（2.35±0.16） mg/g;沉积物以粉砂质砂和砂为主,CIA值分布范围为44.79~69.59,部分区域达到中等化学风化;CIA与GRSP呈显著相关（R=~0.49,p<0.01）,总体上CIA随GRSP的增加呈现先增加后减少的趋势。【结论】GRSP在滨海湿地沉积物中的分布受到生境差异性的影响,其与CIA的相关性表征AMF及其代谢产物在地质体风化过程中发挥了潜在的生态功能。创新点：揭示了球囊霉素相关土壤蛋白与地质过程的互馈作用;阐明了丛枝菌根真菌潜在的生态功能。     

Testing the effect of oxidizing pre-treatments on amino acids in benthic and planktic foraminifera tests

Amino acid racemization (AAR) is a geochronological method that uses the ratio of D- to L-configurations in optically active amino acids from carbonate fossils to determine the time elapsed since the death of an organism. Although AAR techniques have been widely applied to foraminiferal tests, there have been limited dedicated assessments of the potential of isolating a bleach-resistant, intra-crystalline fraction of proteins to improve the reliability of AAR in this biomineral system. In this study, we evaluate the effect of two oxidative pre-treatments (hydrogen peroxide and bleach) on amino acid concentrations and D/L values in sub-modern benthic foraminifers (Ammonia spp. and Haynesina germanica) and well-preserved mid Holocene and mid Pleistocene planktic foraminifers (Pulleniatina obliquiloculata, Globorotalia truncatulinoides, and Globorotalia tumida). The oxidative pre-treatments successfully reduced the amino acid content of the foraminiferal tests to a residual fraction, and with the exception of Ammonia spp., neither pre-treatment substantially affected the relative proportion of individual amino acids. The bleaching pre-treatment does not consistently alter D/L values when compared to peroxide pre-treatment, but it does tend to reduce the subsample variability in D/L values, albeit only to a small degree in an inconsistent fashion. Therefore, we recommend that a relatively weak oxidative pre-treatment with 3% hydrogen peroxide is sufficient for foraminifera-based AAR applications.     

微酸性电解水结合复合保鲜剂对凡纳滨对虾冷藏期间蛋白特性影响

【目的】研究微酸性电解水(SAEW)结合迷迭香提取物(RE)-柠檬酸(CA)复合保鲜剂处理对凡纳滨对虾(Litopenaeus vannamei)冷藏期间蛋白特性变化影响,探讨此法用于虾类贮藏保鲜的效果。【方法】将样品分别经SAEW、质量分数4%RE+质量分数1%CA(RC)、SAEW结合RC(SAEW+RC)和无菌水(CK)处理10 min后,自然沥干15 min,装入PE保鲜袋于(4±1)℃冰箱中保藏,每2 d测定样品的pH值、总挥发性盐基氮(TVB-N)、总巯基含量、Ca²⁺-ATPase活性、化学键、肌原纤维小片化指数(MFI)与内源荧光强度(IFI)等指标,并结合聚丙烯酰胺凝胶电泳(SDS-PAGE)进行综合分析。【结果】复合处理能延缓凡纳滨对虾贮藏期间pH与TVB-N值的升高,抑制总巯基含量下降,CK、RC、SAEW与SAEW+RC组样品的Ca²⁺-ATPase活力分别由初始0.16、0.17、0.17与0.17μmol/(mg·min)降至0.12、0.13、0.12与0.14μmol/(mg·min),MFI值较初始值上升1.88、1.73、1.76和1.54倍,可见SAEW+RC对延缓Ca²⁺-ATPase活性下降与MFI值的上升有较好作用。同时,SAEW+RC处理还能减缓蛋白质分解、氧化和变性,保持蛋白质化学键与IFI值的相对稳定。【结论】微酸性电解水结合复合保鲜剂处理能较好抑制凡纳滨对虾冷藏期间的蛋白质氧化分解,延缓其品质劣变。