漆酶介体催化的研究进展

漆酶是一种含铜多酚氧化酶,可催化氧化酚型底物,不能氧化降解非酚型结构单元,但介体的加入可实现漆酶与非酚类底物间的电子传递,实现漆酶对非酚型底物的最终氧化。文章综述漆酶介体催化常用介体及其研究进展,并针对合成介体有毒价高、天然介体提取纯化复杂的特点,提出一种新型介体——木质素降解产物中的漆酶介体。     

真菌漆酶在绿色化学中的研究进展

漆酶主要是由真菌分泌的一类胞外多铜氧化酶,因其具备多功能特性而被广泛地应用于绿色环境化学。漆酶能够以分子氧作为电子接受体,催化氧化多种酚类和非酚类底物形成自由基中间体,随后这些自由基中间体涉及到氧化耦合或键断裂,最终导致底物发生氧化、分解或聚合反应。目前,主要采用包埋、吸附、共价绑定和交联结合等方法制备固定化漆酶,以增强漆酶在实际环境中的催化活性、稳定性和循环再利用功效。概述了真菌漆酶的分子组成、结构特性及其催化氧化不同底物的作用机理,系统地比较了漆酶的固定化技术及其利弊,重点阐述了漆酶在有机污染物去除、合成染料脱色、造纸废水、食品加工、生物传感器和环境质量指示器等领域的研究进展,旨在为拓展和开发真菌漆酶在绿色环境化学中的多功能应用提供新的理论依据和指导思想。     

漆酶的来源及固定化漆酶载体研究进展

漆酶是一种多酚氧化酶,可催化氧化多种难降解有机污染物,在环境污染防治领域具有良好的应用前景。总结了植物漆酶、动物漆酶以及微生物漆酶的研究现状,详细讨论了漆酶固定化载体的研究进展,进一步指出了目前漆酶研究存在的问题,并提出未来的研究方向,旨在为漆酶的开发与应用研究提供参考。     

天然介体在产漆酶真菌氧化蒽和芘中的重要作用（英文稿）

【目的】研究了氧化还原介体在产漆酶真菌氧化蒽和芘的作用。【方法】通过非变性电泳和酶活力分析。【结果】发现血红密孔菌Z-1和木蹄层孔菌Z-5只产漆酶,其最大酶产量分别为11.90 U/mL和4.83 U/mL,不产木质素过氧化酶和锰过氧化物酶。木蹄层孔菌Z-5的胞外液尽管具有较低的漆酶活性,但是氧化了74.3%的蒽和12.4%的芘,高于血红密孔菌Z-1对蒽和芘的氧化率,提示天然介体可能存在于真菌胞外液中并且影响了漆酶对多环芳烃的氧化。实验进一步表明,木蹄层孔菌Z-5灭活和不灭活的超滤液以及灭活的胞外液对纯漆酶氧化多环芳烃的促进作用均大于血红密孔菌Z-1,说明木蹄层孔菌Z-5的天然介体比血红密孔菌Z-1能够更为有效地促进多环芳烃氧化。【结论】氧化还原结体在产漆酶真菌降解底物过程中发挥了重要作用,这也解释了木蹄层孔菌Z-5胞外液尽管漆酶活性不高,但是具有较大多环芳烃氧化率的原因。     

漆酶的固定化及其在生物传感器方面的应用

漆酶是一类含铜的多酚氧化酶,它能够催化许多酚类和非酚类物质的氧化.固定化漆酶能改善漆酶的稳定性,实现酶制剂的重复连续使用,具有重要意义.该文综述了漆酶固定化的各种方法,阐述了漆酶相关活性、机械性能和功能等内容,并对漆酶固定化在生物传感器方面的应用作了介绍.     

天然介体在产漆酶真菌氧化蒽和芘中的重要作用(英文)

【目的】研究氧化还原介体在产漆酶真菌氧化蒽和芘的作用。【方法】通过非变性电泳和酶活力分析。【结果】发现血红密孔菌Z-1和木蹄层孔菌Z-5只产漆酶,其最大酶产量分别为11.90 U/mL和4.83 U/mL,不产木质素过氧化酶和锰过氧化物酶。木蹄层孔菌Z-5的胞外液尽管具有较低的漆酶活性,但是氧化了74.3%的蒽和12.4%的芘,高于血红密孔菌Z-1对蒽和芘的氧化率,提示天然介体可能存在于真菌胞外液中并且影响了漆酶对多环芳烃的氧化。实验进一步表明,木蹄层孔菌Z-5灭活和不灭活的超滤液以及灭活的胞外液对纯漆酶氧化多环芳烃的促进作用均大于血红密孔菌Z-1,说明木蹄层孔菌Z-5的天然介体比血红密孔菌Z-1能够更为有效地促进多环芳烃氧化。【结论】氧化还原结体在产漆酶真菌降解底物过程中发挥了重要作用,这也解释了木蹄层孔菌Z-5胞外液尽管漆酶活性不高,但是具有较大多环芳烃氧化率的原因。     

一色齿毛菌漆酶的酶学特性及染料脱色研究

染料由于具有复杂的化学结构通常难以降解。本文从白腐菌一色齿毛菌LS0547中纯化出胞外漆酶并用于染料脱色实验。SDS-PAGE结果显示纯化的漆酶分子量大小为63.7kDa。漆酶氧化底物ABTS的最适pH为2.2,最适温度为50℃。叠氮钠可强烈抑制漆酶活性,半胱氨酸和二硫苏糖醇可部分抑制漆酶活性。漆酶氧化ABTS,丁香醛连氮和2,6-二甲氧基苯酚的米氏常数分别为0.217,0.306和0.199mmol/L。粗酶和纯化的漆酶用于不同化学结构的染料的脱色研究,结果表明一色齿毛菌纯化漆酶可快速对RB亮蓝进行脱色,偶氮胭脂红和结晶紫的脱色效果低于RB亮蓝,测试的三种染料均可在没有介体存在的条件下被漆酶脱色,显示出一色齿毛菌漆酶在染料废水处理中的应用前景。     

愈创木酚法快速筛选漆酶产生菌

目的:寻找漆酶产生菌实用、简便的筛选方法,以提高筛选的效率。方法:用6个不同浓度的9种试剂,在3种不同的固体培养基上对23株真菌进行了试验研究,并将其所形成的氧化带与液体发酵培养产生漆酶的活力进行了比较。结果:愈创木酚作底物能形成鲜艳而均匀的红棕色氧化带,且其颜色深浅、带宽能较好地反应漆酶活性。结论:含有0.04%愈创木酚的PDA培养基,在48~72h内产生的氧化带与漆酶活力有明显的相关性,此法能快速筛选产生漆酶的菌株。     

细菌漆酶的研究及应用进展北大核心CSCD

漆酶作为一种绿色环保的多酚氧化酶类,目前被广泛地应用于染料降解、造纸等领域。细菌漆酶与真菌漆酶相比,有更好的热稳定性和更宽的最适pH范围。因此,在工业应用方面更具优势与潜力。综述细菌漆酶的来源、分布、分子结构等基本信息,以及目前细菌漆酶发酵生产水平,并对固定化细菌漆酶进行总结。此外,对细菌漆酶在染料废水、电化学应用及造纸等工业生产方面的应用作简要的介绍。     

漆酶可降解底物种类的研究进展

漆酶是一种含铜离子的多酚氧化酶,广泛存在于植物及真菌中。漆酶含特有的铜离子,其功能为传递结构中的电子,使漆酶具有了较强的氧化还原能力,能与木质素、胺类化合物、芳香化合物等底物发生作用,且大多数反应的唯一产物为水。目前,漆酶在降解多种有毒物质和有害污染物方面表现出高效、成本较低的特性,如白腐真菌所产的高水平漆酶已广泛成熟应用在工业废水处理等生物整治和修复领域。近年来最新研究利用载体固定化酶的技术使漆酶能够在使用后回收反复利用且更具有稳定性,这降低成本的同时还保持了漆酶催化氧化的特性,克服了不少漆酶在解决环境污染中出现的问题。利用介体的介导作用解决了漆酶氧化还原电势较低的问题,大量增多了可降解底物的种类,使其在废水处理、污染物降解、土壤修复、工业染料漂白等领域的应用前景更广阔。对现有漆酶应用于各领域进行研究总结,综述了降解各领域中的有害污染物等底物种类,提出了利用漆酶的降解过程中的现有不足和改进方向,以期为生物法降解环境污染物的研究提供参考。     

On the mechanism of non-endosomial peptide-mediated cellular delivery of nucleic acids

Recently, we described a new strategy for the delivery of nucleic acids into mammalian cells, based on an amphipathic peptide of 27 residues called MPG, which was designed on the basis of a hydrophobic domain derived from a fusion sequence associated with a nuclear localization sequence and separated by a linker. This peptide carrier constitutes a powerful tool for the delivery of nucleic acids in cultured cells, without requiring any covalent coupling. We have examined the conformational states of MPG in its free form and complexed with a cargo, as well as its ability to interact with phospholipids, and have investigated the structural consequences of these interactions. In spite of its similarity to the similarly designed cell-penetrating peptide Pep-1, MPG behaves significantly differently from the conformational point of view. Circular dichroism (CD) analysis reveals a transition from a nonstructured to a β-sheet conformation upon interaction with phospholipids. We propose that the membrane crossing process involves formation of a transient transmembrane pore-like structure. Partial conformational change of MPG is associated with formation of a complex with its cargo, and an increase in sheet content occurs upon association with the cell membrane.     

1590名农村中老年人生存质量调查分析(英文)

目的:采用SF-36量表中文版评价农村中老年人生存质量,并为提高农村中老年人的生存质量和健康水平提供科学依据和建议。方法:采用随机抽样的方法,采用面对面访谈的形式,记分方法参照国际生存质量评价计划(IQOLA)的制定标准,评价其生理和心理健康状况。结果:1590名农村中老年人男性生存质量得分高于女性。经济水平对农村中老年人的生理健康的影响大于对其心理健康的影响。配偶健在的农村中老年人的各维度得分均高于无配偶者。农村中老年人的各维度得分随教育水平的升高呈升高趋势。结论:农村中老年人的生存质量与年龄成负相关,同时也受文化程度和经济水平等多个因素的影响。因而,对农村中老年人这一弱势群体应该给予更多关注。     

The importance of microscopic examination in the management of desquamative diseases of the scalp

After determining the usual malassezic biota of the scalp in adult, normal persons, 259 patients with different desquamative diseases were studied by a simple adhesive tape technique. The main purpose of this study was to investigate the utility of this technique to improve the diagnosis and treatment of patients. Most patients with seborrhoeic dermatitis and sebopsoriasis demonstrated large numbers of {it Malassezia} spp. cells corresponding to the so called ``pityrosporosis'. Only 43.6% of patients with pityriasis capitis (dandruff) presented with such a diagnosis. Symptomatic pityrosporosis of the scalp should be treated with imidazolic derivatives or other antifungal substances. Patients with psoriasis of the scalp showed a typical microscopic picture represented by parakeratosic (nucleated) keratinocytes with absence of lipophilic yeasts which should be attributed to the usual dryness of the scales .Microbial epidermitis (eczema) of the scalp revealed another characteristic picture constituted by abundant leukocytes and bacteria without the presence of yeasts. The different microscopic pictures seen with this simple technique for diseases of the scalp, offer an adjunct to make a proper diagnosis and to establish a convenient treatment in cases which are not clinically well defined. This revised version was published online in June 2006 with corrections to the Cover Date.     

Kinetics of lipid-membrane binding and conformational change of L-BABP

We designed an experimental approach to differentiate the kinetics of protein binding to a lipid membrane from the kinetics of the associated conformational change in the protein. We measured the fluorescence intensity of the single Trp6 in chicken liver bile acid-binding protein (L-BABP) as a function of time after mixing the protein with lipid membranes. We mixed the protein with pure lipid membranes, with lipid membranes in the presence of a soluble quencher, and with lipid membranes containing a fluorescence quencher attached to the lipid polar head group. We fitted simultaneously the experimental curves to a three-state kinetic model. We conclude that in a first step, the binding of L-BABP to the interfacial region of the anionic lipid polar head groups occurred simultaneously with a conformational change to the partly unfolded state. In a second slower step, Trp6 buried within the polar head group region, releasing contacts with the aqueous phase.     

Semiquantitative assessment of the distribution of Salmonella in the environment of caged layer flocks

AIMS: To evaluate a semiquantitative technique for the enumeration of Salmonella in the environment of layer flocks and to compare findings with those of a standard qualitative technique. METHODS AND RESULTS: Samples were taken from faeces, floor dust, dust on cages, feeders and egg belts. After mixing with buffered peptone water, serial dilutions were prepared and culture was performed using pre-enrichment, then plating on semisolid selective and solid isolation media. Comparison with a qualitative pre-enrichment technique indicated a similar sensitivity for both methods despite smaller sample sizes. The numbers of Salmonella detected for a site or sample type did not correlate closely with the prevalence of positive samples. CONCLUSIONS: The sensitive detection and quantification of Salmonella in the flock environment is practicable with the technique described. Quantitative data in many cases do not correlate with qualitative findings. SIGNIFICANCE AND IMPACT OF THE STUDY: The significance of certain environmental factors and interventions in the maintenance and dissemination of Salmonella in poultry houses may be over- or under-represented by prevalence data alone. The technique described allows the issue of poultry house contamination to be examined from a new perspective.     

Self-assembly of mixtures of a dendrimer and lipids: effects of hydrophobicity and electrostatics

Self-assemblies of mixtures of a G7 dendrimer and dimyristoylphosphatidylcholine (DMPC) lipids were simulated using the coarse-grained force fields. A single G7 dendrimer, which consists of either a hydrophilic or a hydrophobic interior, was simulated with the randomly distributed zwitterionic DMPC or anionic lipids. For the dendrimer with hydrophilic interior, its mixture with anionic lipids self-assembles to the dendrimer-encasing liposome, but the one with zwitterionic lipids does not. The liposome diameter agrees with experiment, which is smaller than the typical liposome without dendrimer. This indicates that the strong electrostatic interactions between dendrimer terminals and lipid phosphates induce high curvature of the bilayer, leading to such a small liposome. For the dendrimer with hydrophobic interior, lipids penetrate the dendrimer interior because of the hydrophobic interactions between the dendrimer interior and lipid tails, leading to the dendrimer–lipid micelle with increased dendrimer size. These simulation findings agree qualitatively with the experimentally proposed liposome and micelle models, and indicate that these proposed conformations of the dendrimer–lipid complexes are significantly modulated by dendrimer hydrophobicity and lipid charge.     

Embryos of therizinosauroid theropods from the Upper Cretaceous of China: diagnosis and analysis of ossification patterns

Exceptionally complete, in ovo dinosaur embryos from the Upper Cretaceous of China are analysed. Ossification patterns of these embryos suggest that they died during the final third of their development. The therizinosauroid identity of the embryos follows from: (1) an edentulous premaxilla with a sharp downturned edge; (2) dentary with a lateral shelf; (3) teeth with fan-shaped crowns, with a few marginal cusps; (4) humerus with a massive deltopectoral crest extending proximally, with a pointed proximomedial tuberosity; (5) ilium with an expanded and hooked preacetabular process; (6) strongly curved hypertrophied manual unguals tapering to sharp points. These embryos are closest to two Chinese therizinosauroids, Neimongosaurus yangi Zhang et al . 2001 and Erliansaurus bellamanus Xu et al . 2002 . An elongated narial opening, reduced basipterygoid process, low cervical neural spines, a transversely narrow pubic apron, and a pubic foot expanded anteriorly are found in these embryos and are synapomorphies uniting the Therizinosauroidea and the Oviraptorosauria. Fusion of cervical and caudal neural arches and centra, complete ossification of thoracic ribs and ilium, possible co-ossification of tibia and fibula, fused pubes, complete meta- and acropodial elements, together with small portions of unossified epiphyses of long bones suggest an advanced precociality of these embryos.     

Synthesis of mono- and disaccharide analogs of moenomycin and lipid II for inhibition of transglycosylase activity of penicillin-binding protein 1b

Three types of mono- and disaccharides 3a,b, 4a–c, 5, and some chaetomellic acid A analogs 6 and 42–44 were synthesized as potential inhibitors of the transglycosylase activity of penicillin-binding protein 1b (PBP1b), a key bacterial enzyme responsible for the formation of the polysaccharide backbone of peptidoglycan as well as for cross-linking of its peptide portions. The target compounds combine structural features of both the active portion of moenomycin and the natural PBP1b substrate, lipid II. The desired skeletons were obtained in a convergent fashion involving attachment of the lipid-alkylated glyceric acid moieties 11a,b to the corresponding carbohydrate-containing phosphonic acids 23, 24a, and 24b. Compounds 3a,b were prepared to verify the distance requirements between the sugar and the noncleavable C-phosphonate moieties. Compounds 4a–c were synthesized to examine the importance of the first sugar unit of moenomycin, a known inhibitor of transglycosylase catalysis by PBP1b, with respect to antibiotic activity. These were prepared by condensation of 11a,b with 28a and 28c, which were made by glycosylation of 3-bromopropanol with oxazolines 25a,b, and Arbuzov reaction with triethyl or trimethyl phosphite, followed by dealkylation with bromotrimethylsilane. Compound 5 was generated to verify the possibility of using a dicarboxylate group to mimic the diphosphate of lipid II. It was synthesized by coupling of alcohol 31 with -trichloroacetimidate 34. Chaetomellic acid A analogs were prepared by a Michael addition to dimethyl acetylenedicarboxylate. With the exception of 3b, all of the target compounds were found to inhibit PBP1b, albeit with modest potency.     

Diurnal rhythm of the total antioxidant activity of saliva of children with chronic viral hepatitis

The diurinal rhythms of a total antioxidant activity of a saliva were investigated depending on activity of the inflammatory process in a liver of children with chronic virus hepatitis HBV and HCV. The circadian rhythms were found desynchronized, which was showed in lowering of the mesor value, oscillations amplitude, and a rhythm inversion and transformation, as an additional biorhythmological criterion of the presence of the chronic hepatitis of a viral etiology. A tendency of a circadian rhythm to remain with a daily acrophasis but with a law level of amplitude oscillations and a mesor was to be observed after the treatment of children with inactive HBV. By an active HBV a circadian rhythm with it's inversion has been renewed. It was determined the remaining of an ultradian rhythm with increasing level of amplitude oscillations after the treatment of inactive HCV. The law amplitude oscillations and a insignificant increasing of a meror level were characteristic for the circadian rhythm of total AOA of the children with active HBV. It was proved the possibility of usage of the biorhythmological parameters of diurinal rhythms of total AOA as a criteria of efficiency of the conducted therapy for the given group with disease children.     

蚀斑减少中和试验检测抗天花免疫球蛋白效价

为了建立测定抗天花免疫球蛋白效价的检测方法,采用抗天花免疫球蛋白50%中和100PFU痘苗病毒的稀释度来确定其效价。通过与NIBSC提供的抗天花人免疫球蛋白标准品进行的对比试验,结果显示基本一致,证明该方法是可行的。