The isolation of prophyra-334 from marine algae and its UV-absorption behavior

1 INTRODUCTIONSolar ultraviolet radiation on the earth's surfaceincreases with the depletion of stratospheric ozone(Crutzen, 1992; Kerr and McElroy, 1993; Mad-ronich et al., 1998). Ultraviolet radiation is a highlyactive component of solar spectrum. The excessiveUV radiation has deleterious effects on all plant,animal and microbial groups exposed to solar radia-tion, including the destruction of DNA, proteins andother molecules (Harm, 1980), inhibition of photo-synthesis and growth (W…     

Alkaline Protease Production by a Strain of Marine Yeasts

Yeast strain 10 with high yield of protease was isolated from sediments of saltern near Qingdao, China. The protease had the highest activity at pH 9.0 and 45℃. The optimal medium for the maximum alkaline protease production of strain 10 was 2.5 g soluble starch and 2.0 g NaNO3 in 100 mL seawater with initial pH6.0. The optimal cultivation conditions for the maximum protease production were temperature 24.5 ℃, aeration rate 8.0 L min^- 1 and agitation speed 150 r min^-1 . Under the optimal conditions, 623.1 Umg^-1 protein of alkaline protease was reached in the culture within 30 h of fermentation.     

Preparation of cationic chitosan-polyacrylamide flocculant and its properties in wastewater treatment

Chitosan derived from crab shells,was used to prepare the graft polymer in aqueous solution with acrylamide (AM) and methacrylatoethyl trimethyl ammonium chloride (DMC) as raw materials and ceric ammonium nitrate (CAN) as initiator.The floc-culation ability of the resulting polymer (PCAD) was studied in waste water treatment experiments.Its properties were determined on the basis of the transmittance of waste water after flocculation.The effects of chitosan and DMC content on PCAD’s flocculation ability wer...     

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。     

Effects of temperature, pH and NaC1 on protease activity in digestive tract of young turbot, Scophthalmus maximus

The protease activity in digestive tract of young turbot Scophthalmus maximum was studied, and the optimal pH, temperature and NaCl concentration were determined for different portions of the fish＇s internal organs. The optimal activity in the fish＇s stomach was at pH of 2.2, while that in the intestinal extracts was within the alkaline range from 9.5 to 10.0. In hepatopancreas, the optimal pH was in low alkalinity at 8.5. The optimal reaction temperature was above 40℃ in stomach, intestine and hepatopancreas. With increasing temperature, the pH value increased in stomach, while in the intestine, an opposite tendency was observed due to combined effect of pH and temperature. NaCl concentration showed inhibitory impact on protein digestion in hepatopancreas. The main protease for protein digestion in turbot seemed to be pepsin. Moreover, the maximum protease activity in different segments of intestine existed in the hindgut.     

Purification and characterization of cold-active endo-1,4-β-glucanase produced by Pseudoalteromonas sp. AN545 from Antarctica

A bacterium hydrolyzing carboxymethylcellulose, isolated from Antarctic sea ice, was identified as Pseudoalteromonas sp. based on 16S rDNA gene sequences and named as Pseudoalteromonas sp. AN545. The extracellular endo-1,4-β-glucanase AN-1 was purified successively by ammonium sulfate precipitation, DEAE-Sepharose ion exchange chromatography and Sephadex G-75 gel filtration chromatography. The molecular mass of AN-1 was estimated to be 47.5 kDa utilizing SDS-PAGE and gel chromatography analysis. AN-1 could hydrolyze caboxymethylcellulose, avicel and β-glucan, but not cellobiose, xylan and p-Nitrophenyl-β-D-glucopyranoside. The optimal temperature and pH for the β-glucanase activity of AN-1 were determined to be at 30°C and pH 6.0, respectively. AN-1 was stable at acidic solutions of pH 5.0-6.5 and temperatures below 30°C for 1 h. Moreover, the specific activity was enhanced by Ca2+ and Mg2+, and inhibited by Cu2+. The kinetic parameters Michaelis constant (Km) and maximum velocity (Vmax) of AN-1 were 3.96 mg/mL and 6.06×10-2 mg/(min·mL), respectively.     

农用地土壤中7种重金属可提取态的测定

该文以氯化钙溶液进行提取,采用电感耦合等离子体质谱法和原子荧光光谱法,测定农用地土壤中Cd,As,Pb,Cr,Cu,Zn,Ni等7种重金属可提取态,检出限在0.003~0.06 mg/kg之间。该方法用于4个可提取态质控样品的测定,测定值与推荐值基本相符,测定的相对标准偏差(n=8)在3.3%~9.1%之间。基于800余件农用地土壤样品的测定数据,对7种重金属可提取态与土壤pH值及重金属总量之间的关系进行了初步考察。结果表明,7个重金属可提取态和总量关系无明显相关性,仅在土壤酸性条件下可提取态Cd与总量Cd呈正相关性。7种重金属提取率大小顺序为Cd>Zn>Ni>Cu,As>Pb>Cr,其中Cd,Zn,Ni,Cu,Pb的提取率随土壤pH值的增大而迅速下降,而As,Cr在土壤碱性条件下的提取率略高于中酸性条件下的提取率。     

Temperature effects on the mechanical properties of slates in triaxial compression test

High geothermal temperatures appear to be unfavorable for the construction of tunnels in slate rocks with high overburden. To investigate the mechanical characteristics of slates at various levels of geothermal temperature, conventional triaxial compression tests at different levels of confining stress were carried out at 4 different temperatures from 20℃ to 120℃. The obtained results show high confining pressures weaken the thermal effects on rock mechanical characteristics while higher temperatures enhance the effect of confining pressure.At higher levels of confining stress the thermal effects on the rock strength characteristics decrease. The higher the temperature, the larger is the effect of confining pressure on the mechanical characteristics of the slate. Increase of temperature leads to a decrease of the peak strength but increases the deformability and ductility of the slate, the thermo effect on the peak strength and Poisson's ratio is larger than on the elastic modulus. Higher temperatures reduce the shear strength of slate, the decrease is mainly caused by a decrease of the cohesion. In general, the slate samples fail in shear failure.     

Optimization of fermentation conditions for 1,3-propanediol production by marine Klebsiella pneumonia HSL4 using response surface methodology

The industrially important organic compound 1,3-propanediol(1,3-PDO) is mainly used as a building block for the production of various polymers. In the present study, response surface methodology protocol was followed to determine and optimize fermentation conditions for the maximum production of 1,3-PDO using marine-derived Klebsiella pneumoniae HSL4. Four nutritional supplements together with three independent culture conditions were optimized as follows: 29.3 g/L glycerol, 8.0 g/L K 2 HPO 4, 7.6 g/L(NH 4) 2 SO 4, 3.0 g/L KH 2 PO 4, pH 7.1, cultivation at 35°C for 12 h. Under the optimal conditions, a maximum 1,3-PDO concentration of 14.5 g/L, a productivity of 1.21 g/(L·h) and a conversion of glycerol of 0.49 g/g were obtained. In comparison with the control conditions, fermentation under the optimized conditions achieved an increase of 38.8% in 1,3-PDO concentration, 39.0% in productivity and 25.7% in glycerol conversion in flask. This enhancement trend was further confirmed when the fermentation was conducted in a 5-L fermentor. The optimized fermentation conditions could be an important basis for developing lowcost, large-scale methods for industrial production of 1,3-PDO in the future.     

Study on the Determination of Trace Ni（Ⅱ） by the Catalytic Kinetic Spectrophotometric Method

A new kinetic spectrophotometric method has been developed for the determination of trace Ni (II) in natural water.The method is based on the catalytic effect of Ni (II) on the oxidation of weak acid brilliant blue dye (RAWL) by KIO4 in acid medium.The concentration of nickel (II) can be determined spectrophotometrically by measuring the decrease of absorbance of RAWL at λ= 626 nm using the fix-time method.The influencing factors are investigated by the orthogonal experimental design.The obtained optimum analytical conditions are:pH=2.00,c RAWL=5.00×10-5 mol L-1,cKIO 4= 2.00×10-5 mol L-1,the reaction time t=10 min and the temperature T=25℃..Under the optimum conditions,the developed method allows the measurement of Ni (II) in a range of 0-40.0 ng mL-1.The standard deviation of eleven independent measurements of blank reaction is S=3.08×10-3 and the limit of detection is 2.20 ng mL-1.The relative standard deviations (RSDs) in six replicate determinations of 5 ng mL-1 and 8 ng mL-1 Ni (II) are 2.87% and 1.11%,respectively.Moreover,the experiments show few cations and anions can interfere with the measurement of Ni (II).The recovery efficiencies of this method are in a range of 97.0%-102.5% in freshwater samples.But there is a decreasing effect,which is about 0.2 times the added Ni (II) in seawater medium.After reasonable calibration this processing method is used for the determination of Ni (II) in seawater samples successfully.The results show this developed method has high accuracy and precision,high sensitivity,large range of linearity and high speed.The method can,therefore,be employed at room temperature.     

钻井液用高分子聚合物高温流变特性影响因素探究

聚合物对钻井液的高温流变特性具有重要影响。对比了聚合物种类、加量、剪切时间、盐、造浆黏土等对聚合物溶液高温流变性的影响，并对不同温度下的剪切速率-剪切应力关系进行了流变模型拟合。结果表明，温度升高、剪切时间及盐量增加均导致黏度降低，超过190℃后黏度下降速率加剧；含5%甲酸盐与5%卤盐的样品黏度在降温阶段的黏度恢复率分别为86.8%和2.7%；以宾汉模式对造浆黏土与聚合物混合液进行拟合，220℃时的动切力最高达到5.47 Pa。温度升高使得聚合物溶液由假塑性向牛顿性演变的趋势增强。高于130℃时，长时间剪切导致黏度下降的趋势明显，此时含甲酸盐的聚合物溶液黏度较含卤盐的高，且降温阶段的黏度恢复率也较高。黏土的存在增强了混合液的网间结构，有利于高温下携带岩屑。      

Purification and characterization of an alkaline protease from <Emphasis Type="Italic">Micrococcus</Emphasis> sp. isolated from the South China Sea

Protease is wildly used in various fields, such as food, medicine, washing, leather, cosmetics and other industrial fields. In this study, an alkaline protease secreted by Micrococcus NH54PC02 isolated from the South China Sea was purified and characterized. The growth curve and enzyme activity curve indicated that the cell reached a maximum concentration at the 30th hour and the enzyme activity reached the maximum value at the 36th hour. The protease was purified with 3 steps involving ammonium sulfate precipitation, ion-exchange chromatography and hydrophobic chromatography with 8.22-fold increase in specific activity and 23.68% increase in the recovery. The molecular mass of the protease was estimated to be 25 kDa by SDS-PAGE analysis. The optimum temperature and pH for the protease activity were 50°C and pH 10.0, respectively. The protease showed a strong stability in a wide range of pH values ranging from 6.0–11.0, and maintained 90% enzyme activity in strong alkaline environment with pH 11.0. Inhibitor trials indicated that the protease might be serine protease. But it also possessed the characteristic of metalloprotease as it could be strongly inhibited by EDTA and strongly stimulated by Mn²⁺. Evaluation of matrix-assisted laser desorption ionization/time-of-flight MS (MALDI-TOF-TOF/MS) showed that the protease might belong to the peptidase S8 family.     

Impacts of soil properties on phosphorus adsorption and fractions in purple soils

Information on phosphorus (P) adsorption and its impacts on the redistribution of the P fraction in soil profiles are important for environmental management under intensive agricultural practices. To clarify the dominant factors influencing soil phosphorus adsorption in an Entisol (locally known as purple soil), P adsorption experiments were conducted in Sichuan Basin of southwestern China for cropland and woodland soils with acidic, neutral and calcareous origins throughout their profile. After various doses of P were added during incubation experiments, soil P fractions were also analyzed. The results showed that there were no significant differences in Fe-oxides and P adsorption along the vertical gradients. Agricultural practices and lower pH conditions reduced the P adsorption capacity of purple soils throughout the soil profiles. For acidic and neutral purple soil profiles, the P adsorption capability was mainly influenced by Fe-oxides and soil texture. Ca-bound P and Fe-Al-bound P represented the majority of the total inorganic P of calcareous soils. There was a saturation of adsorption capacity by sesquioxide and a high risk of dissoluble reactive P (NH₄Cl-P) being released out of the soil profile in acidic and neutral purple soils after the greatest P addition, indicated by the higher proportions of NH₄Cl-P (over 40%) and decreasing Fe-Al-P fraction. P fractions migrated with greater difficulty in calcareous purple soil profiles as Ca-P fraction peaked over 65% when adding a P dose at or greater than 80 g P kg^-1, indicating the high potential of P adsorption. The X-Ray Diffraction analysis also verified the formation of brushite. Adaptive management practices should be designed to alleviate P losses for acidic and neutral purple soils.     

Determination of Trace Copper （Ⅱ） in Water Samples by Kinetic-spectrophotometry

A new kinetic-spectrophotometric method is proposed for the determination of copper （ Ⅱ ）. The method is based on the catalytic effect of copper （ Ⅱ ） on the oxidation of weak acid brilliant blue dye （RAWL） by hydrogen peroxide. The copper （ Ⅱ ） can be determined spectrophotometrically by measuring the decrease of absorbance of RAWL at λ = 626 nm using the fix-time method. The optimum reaction conditions are as follows： pH 7.20, buffer solution NaOH-KH2PO4, RAWL （200 mgL-1） 5.00 mL, H2O2 （30%） 0.50 mL, reaction temperature 80 ？？ and reaction time 20 min. The linear range of this method is between 0 μg L^-1 and 12 μg L^-1 and the limit of detection is 0.011 μg L-1, the relative standard deviation （RSD） in five replicate determinations for 2 and 8 μg L-1 copper （ Ⅱ ） are 3.2% and 2.3%, respectively. Twenty ions do not interfere in the determination of copper （ Ⅱ ）. The method has been applied satisfactorily to the determination of copper （ Ⅱ ） in freshwater samples （tap water and Yellow River water from Lijin, Shandong, China） and seawater samples （from the South China Sea）, the recovery rates are 98.0%, 102.5% and 96.0%, respectively.     

Effects of environment factors on initiation of sperm motility in sea cucumber Apostichopus japonicus (Selenka)

Sperm of sea cucumber Apostichopus japonicus (Selenka) were quiescent in electrolyte NaCl solution and artificial seawater (ASW) and nonelectrolyte glucose and mannitol solutions when the osmolality was less than 200 mOsm kg-1. The sperm started to be motile as a result of increased osmolality, indicating an osmolality-dependent initiation of sperm motility in sea cucumber. After a brief incubation in hypotonic NaCl and glucose solutions with osmolalities of 200 and 400 mOsm kg-1, sperm lost partial motile ability. Sperm became immobilized when pH was 6.0 in NaCl, glucose and mannitol solutions, suggesting that an H+ release is involved in sperm activation. The decreased pH had no effect on the percentage of motile sperm in ASW, whereas it delayed the time period to reach the maximum motility (motilitymax). Extracellular Ca2+ in electrolyte solutions was not essential for motility stimulation but shortened the time of reaching motilitymax. When Ca2+ was mixed in nonelectrolyte solutions the sperm motility was completely suppressed. The K+ channel blocker, quinine, suppressed the sperm motility in electrolyte solution, showing a possible involvement of K+ transport in the process. High K+ concentration did not affect the sperm motility in NaCl solution, but decreased it in ASW and almost entirely suppressed it in nonelectrolyte solutions. The different effects of pH and K+ in ASW and NaCl solution indicate that external ions may also regulate sperm motility.     

Dual filtration effect of geochemical and biogeochemical processes in the Changjiang Estuary

Dissolved nutrients (NO ₃ ⁻ , PO₄ ³⁻, SiO₃ ²⁻) and oxygen, chlorophyll- a, pH, and Eh were measured on board during a cruise in August 1988 in the Changjiang Estuary region. Heavy metals, organic matter and carbonate contents were analyzed in laboratory. The results show that geochemical processes in the Changjiang Estuary have dual filtration effect: on the one hand geochemical filtration effect, reflected by ferromanganese oxide flocculation and sedimentation, occurs near the turbidity maximum, and leads to enrichment of heavy metals in suspended matter and sediments; on the other hand biogeochemical filtration, reflected by nutrients consumption, organic matter and carbonate sedimentation and enrichment of trace elements in suspended matter, occurs outside the plume water front. The biogeochemical filtration affects the environmental conditions; the dissolved oxygen and pH increase, in surface water and decrease in bottom water. The biogeochemical filtration effect outside the plume front is more important than the geochemical filtration effect near the turbidity maximum.     

尼罗罗非鱼(Oreochromis niloticus Linn)胃酸的PH值,周日变化及其在消化兰藻中的作用

通过测定喂食水华微囊藻的尼罗罗非鱼胃PH值、不同颜色粪便的叶綠素/脱镁叶綠素比例,发现尼罗罗非鱼具有一个与摄食相关的胃酸分泌周期。摄食旺期胃平均PH值为1.47,最低值为0.8。停食后PH值升高,最大值为6.0。用不同浓度的盐酸处理水华微囊藻和对不同颜色粪便的营养分析结果表明,高浓度的胃酸(PH<2)溶解了水华微囊藻细胞壁,为肠消化酶对细胞内含物进行消化创造了先决条件。     

Diurnal and seasonal dynamics of soil respiration at temperate Leymus chinensis meadow steppes in western Songnen Plain,China

To evaluate the diurnal and seasonal variations in soil respiration(Rs) and understand the controlling factors, we measured carbon dioxide(CO2) fluxes and their environmental variables using a LI-6400 soil CO2 flux system at a temperate Leymus chinensis meadow steppe in the western Songnen Plain of China in the growing season(May–October) in 2011 and 2012. The diurnal patterns of soil respiration could be expressed as single peak curves, reaching to the maximum at 11:00–15:00 and falling to the minimum at 21:00–23:00(or before dawn). The time-window between 7:00 and 9:00 could be used as the optimal measuring time to represent the daily mean soil CO2 efflux. In the growing season, the daily value of soil CO2 efflux was moderate in late spring(1.06–2.51 μmol/(m2·s) in May), increased sharply and presented a peak in summer(2.95–3.94 μmol/(m2·s) in July), and then decreased in autumn(0.74–0.97 μmol/(m2·s) in October). Soil temperature(Ts) exerted dominant control on the diurnal and seasonal variations of soil respiration. The temperature sensitivity of soil respiration(Q10) exhibited a large seasonal variation, ranging from 1.35 to 3.32, and decreased with an increasing soil temperature. Rs gradually increased with increasing soil water content(Ws) and tended to decrease when Ws exceeded the optimum water content(27%) of Rs. The Ts and Ws had a confounding effect on Rs, and the two-variable equations could account for 72% of the variation in soil respiration(p 0.01).     

Effects of elevated air temperatures on soil thermal and hydrologic processes in the active layer in an alpine meadow ecosystem of the Qinghai-Tibet Plateau

In this study,effects of elevated air temperatures on thermal and hydrologic process of the shallow soil in the active layer were investigated. Open-top chambers(OTCs)were utilized to increase air temperatures 1-2℃ in OTC-1 and 3-5℃ in OTC-2 in the alpine meadow ecosystem on the Qinghai- Tibetan Plateau.Results show that the annual air temperatures under OTC-1 and OTC-2 were 1.21℃ and 3.62℃ higher than the Control,respectively.The entirely-frozen period of shallow soil in the active layer was shortened and the fully thawed period was prolonged with temperature increase.The maximum penetration depth and duration of the negative isotherm during the entirely-frozen period decreased, and soil freezing was retarded in the local scope of the soil profile when temperature increased.Meanwhile, the positive isotherm during the fully-thawed period increased,and the soil thawing was accelerated.Soil moisture under different manipulations decreased with the temperature increase at the same depth. During the early freezing period and the early fully- thawed period,the maximum soil moisture under the Control manipulation was at 0.2 m deep,whereas under OTC-1 and OTC-2 manipulations,the maximum soil moisture were at 0.4-0.5 m deep. These results indicate that elevated temperatures led to a decrease of the moisture in the surface soil.The coupled relationship between soil temperature and moisture was significantly affected by the temperature increase.During the freezing and thawing processes, the soil temperature and moisture under different manipulations fit the regression model given by the equationθV=a/{1+exp[b(TS+c)]}+d.     

新显色剂四-(3-磺基-4-氯苯基)卟啉分光光度法测定微量铜

研究了新显色剂四-(3-磺基-4-氯苯基)卟啉在微酸性介质中,表面活性剂OP与CPC存在下测定微量铜的显色反应。络合物在波长420 nm处有最大吸收峰,摩尔吸光度系数为3.44×105L·mol-1cm-1,用双峰双波长光度法测定摩尔吸光系数为6.72×105L·mol-1·cm-1,铜含量在0.5~4.5μg/25 m L范围内符合比耳定律。方法应用于矿样中微量铜的测定,结果令人满意。