赤子爱胜蚓核酸酶样蛋白的纯化及其活性

目的纯化赤子爱胜蚓核酸酶样蛋白,并鉴定其活性。方法将制备的赤子爱胜蚓核酸酶样蛋白粗提液透析后,经Bio-CAD CM柱层析系统非连续梯度洗脱,收集洗脱峰,对有活性的梯度洗脱峰样品进行单向聚丙烯酰胺凝胶电泳(1D-PAGE)及切胶纯化,将切胶回收纯化的蛋白样品进行双向聚丙烯酰胺凝胶电泳(2D-PAGE),回收蛋白,进行活性和浓度检测。结果透析后的蛋白粗提液经Bio-CAD CM柱层析分离,得到3个不同洗脱梯度的可降解DNA的活性峰,3个梯度蛋白的PAGE带型基本一致,经PAGE分离获得3个核酸酶样蛋白纯品EWD1、EWD2和EWD3,得率和纯化倍数分别为:27.9%,11.6;16.7%,11.2;16.7%,18.6。结论成功纯化了赤子爱胜蚓核酸酶样蛋白,并保持了其良好的生物活性。该纯化方法简单、快速、经济、实用,为该蛋白结构及功能的进一步研究奠定了基础。     

Earthworm coelomocytes as nanoscavenger of ZnO NPs

Earthworms can ‘biotransform’ or ‘biodegrade’ chemical contaminants, rendering them harmless in their bodies, and can bioaccumulate them in their tissues. They ‘absorb’ the dissolved chemicals through their moist ‘body wall’ due to the interstitial water and also ingest by ‘mouth’ while soil passes through the gut. Since the advent of the nanotechnology era, the environmental sink has been continuously receiving engineered nanomaterials as well as their derivatives. Our current understanding of the potential impact of nanomaterials and their natural scavenger is limited. In the present investigation, we studied the cellular uptake of ZnO nanoparticles (NPs) by coelomocytes especially by chloragocytes of Eisenia fetida and their role as nanoscavenger. Results from exposure to 100- and 50-nm ZnO NPs indicate that coelomocytes of the earthworm E. fetida show no significant DNA damage at a dose lower than 3 mg/l and have the potential ability to uptake ZnO NPs from the soil ecosystem and transform them into microparticles.     

Metal availability in heavy metal-contaminated open burning and open detonation soil: Assessment using soil enzymes, earthworms, and chemical extractions

The effects of heavy metal contamination on soil enzyme activity and earthworm health (bioaccumulation and condition) were studied in contaminated soils collected from an formerly open burning and open detonation (OBOD) site. Soil extraction methods were also evaluated using CaCl₂ and DTPA solutions as surrogate measures of metal bioavailability and ecotoxicity. Total heavy metal content of the soils ranged from 0.45 to 9.68 mg Cd kg⁻¹, 8.96 to 5103 mg Cu kg⁻¹, 40.21 to 328 mg Pb kg⁻¹, and 56.61 to 10,890 mg Zn kg⁻¹. Elevated metal concentrations are assumed to be primarily responsible for the reduction in enzyme activities and earthworm health indices. We found significant negative relationships between CaCl₂- and DTPA-extractable metal content (Cd, Cu, and Zn) and soil enzyme activity (P < 0.01). Therefore, it could be concluded that soil enzyme activity and metal bioaccumulation by earthworms can be used as an ecological indicator of metal availability. Furthermore, CaCl₂ and DTPA extraction methods are proved as promising, precise, and inexpensive surrogate measures of Cd, Cu, Pb, and Zn bioavailability from heavy metal-contaminated soils.     

Amino acids,fatty acids,and dietary fibre in edible seaweed products

The nutritional compositions of 34 edible seaweed products of the Laminaria sp., Undaria pinnatifida, Hizikia fusiforme and Porphyra sp. varieties were analyzed.     

应用滤纸接触法甄别[BMIM]PF6对赤子爱胜蚓的急性毒性效应

文章选用咪唑类离子液体1-丁基-3-甲基咪唑六氟磷酸盐（[BMIM]PFe）作为目标污染物,采用赤于爱胜蚓作为受试动物,应用滤纸接触法,通过测试不同暴露浓度下蚯蚓在60h、72h和84h时的死亡率,研究其对受试动物的急性毒性效应。实验发现,[BMIM]PF6虽然是新型绿色溶剂．但其仍存在一定程度的环境璺眸,带要笋基事性和生态效应做进一步研究。     

Vermistabilization of municipal sewage sludge amended with sugarcane trash using epigeic Eisenia fetida (Oligochaeta)

Efforts have been made in this study to stabilize the sewage sludge mixed with sugarcane trash in four different proportions: 20% (T(1)); 40% (T(2)); 60% (T(3)) and 80% (T(4)), under laboratory conditions using epigeic earthworm (Oligochaeta) Eisenia fetida. The composting potential of worm was also evaluated in 100% sewage sludge treatment (T(5)). The changes in chemical properties of substrate was measured at the end. The vermicomposted material showed decrease in organic C (4.8-12.7%) and exchangeable K (3.2-15.3%) content, whereas increase in total N (5.9-25.1%) and available P (1.2-10.9%), exchangeable Ca (2.3-10.9%) and exchangeable Mg (4.5-14.0%) contents. Vermicomposting process caused considerable reduction in concentration of diethylene-triaminepentaacetic acid (DTPA) extractable metals: Cu (4.98-30.5%), Fe (5.08-12.64%), Mn (3.31-18.0%), Zn (2.52-15.90%) and Pb (2.38-20.0%). E. fetida showed the better growth performances in first three treatments (T(1)-T(3)) possibly due to higher content of organic matter (supplied by bulking agent, i.e. sugarcane trash). The earthworm mortality was higher in vermibeds those contained more sludge proportions. Study revealed that vermicomposting might be an efficient technology to convert negligible municipal sewage sludge into value-added products. The feasibility of earthworms to mitigate the metal toxicity and to enhance the nutrient profile might be useful to convert noxious sludge into useful products, at low-input basis.     

Determination of major phlorotannins in Eisenia bicyclis using hydrophilic interaction chromatography: Seasonal variation and extraction characteristics

In this study, a hydrophilic interaction chromatography (HILIC) condition was developed for the simultaneous determination of five major phlorotannins from an extract of Eisenia bicyclis (Kjellman) Setchell with good linearity (r² > 0.999). Based on this method, the seasonal variations and extraction characteristics, in terms of total extraction yield and the content of the phlorotannins, were investigated under various extraction conditions. In results, the yields and phlorotannins were increased two-to-four times in summer (June–October) and then, were decreased to normal levels in winter (November–March). In the extraction of E. bicyclis, ethanol percentage in water, extraction time and washing time significantly affected the yield of the extract and the phlorotannins, whereas the temperature and the sample/solvent ratio impacted the extraction to a lesser degree. These results will be useful information in the application of this macroalga in the commercial areas related to nutraceuticals, pharmaceuticals, and cosmeceuticals.     

赤子爱胜蚓Calreticulin的cDNA和氨基酸序列分析

目的分析赤子爱胜蚓Calreticulin的cDNA和氨基酸序列。方法从本院构建的赤子爱胜蚓cDNA文库中经测序获得其Calreticulin cDNA序列。采用生物信息学分析工具DNAMAN、NCBI ORF finder、ExPASy Protparam、ExPASY Protscale、NCBI Conserved Domains、SignalP 4.1 Server、TMHMM Server v.2.0、SOSUI、NetPhos 3.1 Server、PSORT、SOPMA、ExPASy COILS、SWISS-MODEL分别对Calreticulin的cDNA序列、氨基酸序列、一级结构基本理化性质、亲/疏水性、保守结构域、信号肽、跨膜区、可溶性、磷酸化修饰、亚细胞定位、二级结构、卷曲螺旋域以及三级结构进行预测及分析。结果赤子爱胜蚓Calreticulin cDNA序列长476 bp,其中包含354 bp的开放阅读框,编码118个氨基酸残基;Calreticulin由19种氨基酸组成,呈酸性,为稳定的可溶性亲水蛋白;具有1段Calreticulin超家族保守结构域;不存在信号肽和跨膜结构;具有12个潜在的磷酸化位点;主要存在于细胞核中;二级结构元件主要由α-螺旋(29.66%)和无规则卷曲(50.85%)组成,具有卷曲螺旋;三级结构预测结果与二级结构一致。结论掌握了赤子爱胜蚓Calreticulin的理化性质,预测了其潜在的特征,并分析了其结构,为后续深入开展其功能及分子作用机制的研究提供了理论依据。