A fungal enzyme with the ability of aflatoxin B₁ conversion: purification and ESI-MS/MS identification

Armillariella tabescens, a Chinese edible and medicinal fungus, whose multienzyme exist ability of AFB₁-converting, and ADTZ (aflatoxin-detoxizyme) had previously purified from the A. tabescens multienzyme monitored by AFB₁ conversion_. However, the enzyme now confirmed an oxidase and renamed aflatoxin-oxidase (AFO). In this paper, AFO was purified by an economical and practical three-step procedure monitored by AFB₁ conversion. And ESI-MS/MS analysis was done for identification of AFO. The following database searching (Protein Blast on NCBI) results did not show any homologous oxidase protein, which implied that AFO was mostly a new oxidase differing from other reported aflatoxin-converting enzymes such as fungal laccase and horse radish peroxidase. HPTLC analysis of the purified AFO activity suggested that the enzyme reacted at the bisfuran ring of AFB₁ which was the key toxic structure. Therefore, all these investigations implied a new choice for biodegradation of aflatoxin in foods and feeds with the practical application of AFO.     

A water-soluble 1,8-naphthalimide-based 'turn on' fluorescent chemosensor for selective and sensitive recognition of mercury ion in water

A water-soluble 1,8-naphthalimide-based fluorescent chemosensor 1, bearing two acetic carboxylic moieties, exhibited high selectivity and sensitivity for recognition of Hg(2+) ion in water over other heavy and transition metal (HTM) ions with fluorescent enhancement. An increase in the fluorescent intensity at 562 nm was due to the formation of a 1:1 1-Hg(2+) inclusion complex.     

Effective charge measurements reveal selective and preferential accumulation of anions, but not cations, at the protein surface in dilute salt solutions

Specific-ion effects are ubiquitous in nature; however, their underlying mechanisms remain elusive. Although Hofmeister-ion effects on proteins are observed at higher (>0.3M) salt concentrations, in dilute (<0.1M) salt solutions nonspecific electrostatic screening is considered to be dominant. Here, using effective charge (Q*) measurements of hen-egg white lysozyme (HEWL) as a direct and differential measure of ion-association, we experimentally show that anions selectively and preferentially accumulate at the protein surface even at low (<100 mM) salt concentrations. At a given ion normality (50 mN), the HEWL Q* was dependent on anion, but not cation (Li⁺, Na⁺, K⁺, Rb⁺, Cs⁺, GdnH⁺, and Ca²⁺), identity. The Q* decreased in the order F⁻ > Cl⁻ > Br⁻ > NO₃⁻ ∼ I⁻ > SCN⁻ > ClO₄⁻ ≫ SO₄²⁻, demonstrating progressively greater binding of the monovalent anions to HEWL and also show that the SO₄²⁻ anion, despite being strongly hydrated, interacts directly with the HEWL surface. Under our experimental conditions, we observe a remarkable asymmetry between anions and cations in their interactions with the HEWL surface.     

The Cus efflux system removes toxic ions via a methionine shuttle

Gram-negative bacteria, such as Escherichia coli, frequently utilize tripartite efflux complexes in the resistance-nodulation-cell division (RND) family to expel diverse toxic compounds from the cell. These efflux systems span the entire cell envelope to mediate the phenomenon of bacterial multidrug resistance. The three parts of the efflux complexes are: (1) a membrane fusion protein (MFP) connecting (2) a substrate-binding inner membrane transporter to (3) an outer membrane-anchored channel in the periplasmic space. One such efflux system CusCBA is responsible for extruding biocidal Cu(I) and Ag(I) ions. We recently determined the crystal structures of both the inner membrane transporter CusA and MFP CusB of the CusCBA tripartite efflux system from E. coli. These are the first structures of the heavy-metal efflux (HME) subfamily of the RND efflux pumps. Here, we summarize the structural information of these two efflux proteins and present the accumulated evidence that this efflux system utilizes methionine residues to bind and export Cu(I)/Ag(I). Genetic and structural analyses suggest that the CusA pump is capable of picking up the metal ions from both the periplasm and cytoplasm. We propose a stepwise shuttle mechanism for this pump to extrude metal ions from the cell.     

Acid mediated formation of an N-acyliminium ion from tubulysins: a new methodology for the synthesis of natural tubulysins and their analogs

Tubuylsins are extremely potent cytotoxic agents which inhibit tubulin polymerization and lead to cell cycle arrest and apoptosis. Tubulysins have been isolated from fermentation mixtures and have been chemically synthesized; however, these efforts have been hampered by poor yields and arduous purifications. In contrast, treatment of a mixture of natural tubulysins A, B, C, G, and I, obtained from a fermentation batch with trifluoroacetic acid results in the formation of a single N-acyliminium ion. Subsequent addition of butyric, isopentyl, or acetic acid results in the formation of tubulysin B, A, or I, respectively, as a single species. New tubulysin analogs can be formed upon treatment of the acyliminium ion with other nucleophiles such as alcohols, thiols, and nitriles, resulting in corresponding N-acyl-N,O-acetals, N-acyl-N,S-thioacetals, and N,N'-diacyl-aminals. Carbon-carbon bond formation is also possible with a modification of this protocol. The cytotoxicies of the natural tubulysins and tubulysin analogs synthesized by this method were evaluated on KB cells.     

Development of a potassium ion-selective fluorescent sensor based on 3-styrylated BODIPY

We have developed a red-emitting fluorescent K(+) probe, B3TAC, which also shows a wavelength shift upon binding to K(+). The probe was synthesized by conjugating a cryptand-based chelator, 2-triazacryptand [2,2,3]-1-(2-methoxyethoxy)benzene (TAC), to position 3 of the BODIPY fluorophore through a styryl linker. In water-acetonitrile mixed solvent, it responded to K(+) in the physiological concentration range with high selectivity over Na(+) and other metal ions. B3TAC is potentially useful for measuring cellular K(+) ion concentration, as well as for simple, naked-eye detection of K(+) in solution.     

空气重金属元素在悬铃木叶中的亚细胞分布及其区隔化效应

以南京市常见行道树二球悬铃木为试材,研究了交通繁忙区和相对清洁区道路两边悬铃木叶内6种重金属元素的亚细胞分布及其区隔化效应.结果显示:交通污染区悬铃木叶内各亚细胞组分中Cr、Cu、Ni、Pb和Zn 5种重金属元素的含量均明显高于对照区,交通空气污染是影响其含量增加的主要原因之一.相对清洁区和交通污染区5种重金属元素在悬铃木叶片、叶柄的细胞壁组分中含量最高,胞外隔离系数和污染指数均大于0.900,细胞壁是大气重金属元素重要的吸滞器官,并对重金属有明显的阻隔效应;胞内细胞器对Pb和Cu的隔离系数和污染指数最大,细胞器双层膜能在一定程度上抵御重金属元素进入细胞内.悬铃木叶片和叶柄亚细胞组分的污染指数表现为胞质组分＞细胞壁组分＞细胞器组分,即包括液泡液在内的胞质组分是囤积重金属元素的场所.研究表明,悬铃木叶片、叶柄各亚细胞组分对重金属均有不同程度的累积能力,叶内胞质组分的囤积作用以及细胞壁、质膜与细胞器双层膜的区隔化作用可能是悬铃木叶解除重金属元素毒害的重要原因.     

基于城市土地利用类型的地表温度与植被指数的关系

以沈阳市三环作为研究区域,基于QuickBird、Landsat/TM遥感影像和GIS空间分析技术,获取城市土地利用信息,陆地表面温度(LST)以及归一化植被指数(NDVI),定量分析了LST与NDVI在不同城市土地利用类型之间的差异及空间关系.结果表明:LST和NDVI具有明显相反的变化趋势,城市不同土地利用类型的LST与NDVI平均值也具有显著的差异性;多重比较发现,LST和NDVI在两两土地利用类型之间的差异不同;LST与NDVI相关性的显著程度受空间尺度的影响,LST与NDVI的相关性随空间尺度变化呈现出先增加后降低再逐渐增加的趋势.研究结果可为城市规划以及城市绿地系统规划中缓解城市热岛效应提供科学依据.     

重金属污染土壤稳定/固化修复技术研究进展

修复重金属污染土壤一直是国际上的难点和热点研究课题.目前常用的污染场地修复技术主要包括挖掘、稳定/固化(solidification/stabilization,S/S)、化学淋洗、气提、热处理、生物修复等.本文在参考美国环境保护署(EPA)、英国环境署的S/S技术规范、国内外发明专利基础上,对S/S的概念、国内外发展现状及今后的发展方向进行了系统论述.固定化技术通过把污染物囊封入惰性基材中,或在污染物外面加上低渗透性的材料,来减少污染物暴露的淋滤面积以达到限制污染物迁移的目的.稳定化技术是从改变污染物的有效性出发,将污染物转化为不易溶解、迁移能力或毒性更小的形式.S/S技术包括:水泥固化、石灰火山灰固化、塑性材料包容固化、玻璃化技术、药剂稳定化.在稳定化技术中,加入药剂的目的是改变土壤的物理、化学性质,通过pH控制技术、氧化还原电势技术、沉淀技术、吸附技术、离子交换技术等改变重金属在土壤中的存在状态,从而降低其生物有效性和迁移性.本文还论述了S/S修复效果评价方法,并指出需加强S/S技术中的分子键合技术、土聚合物以及我国的S/S技术导则制定等工作.     

低能氮离子注入对决明子的生物效应研究

用低能氮离子注入决明子种胚,种子浸泡液电导率显著增加,发芽势和发芽指数显著降低,幼苗主根长度减少,平均鲜重和活力指数显著降低.直接接受离子注入的种胚,其SOD酶活性随注入剂量的增加呈现出先下降再轻微上升的趋势.而没有直接接受离子注入的胚乳,其SOD酶活随注入剂量的增加呈马鞍形曲线.