紫杉醇产生菌HU1353的鉴定

通过对紫杉醇产生菌HU1 35 3群体形态的观察 ,以及个体形态包括菌丝、分生孢子梗、分生孢子、产孢细胞、是否具喙等特征的研究 ,确定了HU1 35 3的分类地位 ,为链格孢属一新种 ,命名为红豆杉链格孢。     

HU: promoting or counteracting DNA compaction?

The role of HU in Escherichia coli as both a protein involved in DNA compaction and as a protein with regulatory function seems to be firmly established. However, a critical look at the available data reveals that this is not true for each of the proposed roles of this protein. The role of HU as a regulatory or accessory protein in a number of systems has been thoroughly investigated and in many cases has been largely elucidated. However, almost 30 years after its discovery, convincing evidence for the proposed role of HU in DNA compaction is still lacking. Here we present an extensive literature survey of the available data which, in combination with novel microscopic insights, suggests that the role of HU could be the opposite as well. The protein is likely to play an architectural role, but instead of being responsible for DNA compaction it could be involved in antagonising compaction by other proteins such as H-NS.     

Nonspecific DNA binding and bending by HUαβ: interfaces of the three binding modes characterized by salt-dependent thermodynamics

Previous isothermal titration calorimetry (ITC) and Förster resonance energy transfer studies demonstrated that Escherichia coli HU_αβ binds nonspecifically to duplex DNA in three different binding modes: a tighter-binding 34-bp mode that interacts with DNA in large (> 34 bp) gaps between bound proteins, reversibly bending it by 140^o and thereby increasing its flexibility, and two weaker, modestly cooperative small site-size modes (10 bp and 6 bp) that are useful for filling gaps between bound proteins shorter than 34 bp. Here we use ITC to determine the thermodynamics of these binding modes as a function of salt concentration, and we deduce that DNA in the 34-bp mode is bent around—but not wrapped on—the body of HU, in contrast to specific binding of integration host factor. Analyses of binding isotherms (8-bp, 15-bp, and 34-bp DNA) and initial binding heats (34-bp, 38-bp, and 160-bp DNA) reveal that all three modes have similar log-log salt concentration derivatives of the binding constants (Sk_i) even though their binding site sizes differ greatly; the most probable values of Sk_i on 34-bp DNA or larger DNA are − 7.5 ± 0.5. From the similarity of Sk_i values, we conclude that the binding interfaces of all three modes involve the same region of the arms and saddle of HU. All modes are entropy-driven, as expected for nonspecific binding driven by the polyelectrolyte effect. The bent DNA 34-bp mode is most endothermic, presumably because of the cost of HU-induced DNA bending, while the 6-bp mode is modestly exothermic at all salt concentrations examined. Structural models consistent with the observed Sk_i values are proposed.     

朴树内生镰刀菌HU0174抗真菌次生代谢产物研究

通过硅胶柱层析、大孔吸附树脂柱层析和高效液相制备等色谱技术从一株朴树内生镰刀菌Fusarium sp.HU0174的大米发酵物中分离得到一个新的环酯肽:acuminatum D(1)和6个已知化合物:acuminatums A～C(2～4)、白僵菌素(5)、白僵菌酮(6)和吲哚-3-羧酸(7),运用NMR和MS等谱学方法鉴定了它们的化学结构。采用滤纸片琼脂扩散法测试表明环酯肽类化合物1～4对柑橘绿霉和新月弯孢霉两株植物致病真菌具有明显的体外抑制活性,初步阐明了镰刀菌HU0174的抗真菌活性物质基础。     

老鼠艻内生曲霉HU0206的抗细菌活性代谢成分

为阐明老鼠艻内生曲霉Aspergillus sp.HU0206发酵物的抗细菌物质基础，采用色谱分离法从其玉米发酵物中分离得到7个化合物，分别鉴定为6个甲基苯酚类化合物：sydowiol F (1)、5-methoxy-3,5''-dimethyl-2,3''-oxybiphenyl-1,1'',2''-triol (2)、corynesidone A (3)、鹿蹄草素(4)、3-羟基鹿蹄草素(5)、4-乙氧基-3-羟甲基苯酚(6)和1个甲基环己酮二聚体epoxydine A (7)，其中化合物1为新化合物。载样量为40 μg时，化合物1、2和4对金黄色葡萄球菌、耐甲氧西林金黄色葡萄球菌、黑胫病菌和青枯病菌抑菌效果较好，抑菌圈半径≥2 mm，最小抑菌浓度(MIC)为50或100μg/mL；化合物3对这4种菌虽有1~2 mm的抑菌圈，但MIC大于400 μg/mL。所有化合物对大肠杆菌均无抑制活性。     

酸模内生镰刀菌HU0298抗MRSA活性代谢成分

从酸模Rumex acetosa内生真菌Fusarium sp.HU0298玉米发酵物的活性流分中分离鉴定了一个主要成分:亚油酸;结合活性流分的气相色谱(GC)组分分析和活性评价发现亚油酸是该发酵物的主要抗耐甲氧西林金黄色葡萄球菌(MRSA)成分;通过对亚油酸和多个其他脂肪酸标准品的活性测试探究了脂肪酸化学结构和与抗M...     

CTXφ Replication Depends on the Histone-Like HU Protein and the UvrD Helicase

The Vibrio cholerae bacterium is the agent of cholera. The capacity to produce the cholera toxin, which is responsible for the deadly diarrhea associated with cholera epidemics, is encoded in the genome of a filamentous phage, CTXφ. Rolling-circle replication (RCR) is central to the life cycle of CTXφ because amplification of the phage genome permits its efficient integration into the genome and its packaging into new viral particles. A single phage-encoded HUH endonuclease initiates RCR of the proto-typical filamentous phages of enterobacteriaceae by introducing a nick at a specific position of the double stranded DNA form of the phage genome. The rest of the process is driven by host factors that are either essential or crucial for the replication of the host genome, such as the Rep SF1 helicase. In contrast, we show here that the histone-like HU protein of V. cholerae is necessary for the introduction of a nick by the HUH endonuclease of CTXφ. We further show that CTXφ RCR depends on a SF1 helicase normally implicated in DNA repair, UvrD, rather than Rep. In addition to CTXφ, we show that VGJφ, a representative member of a second family of vibrio integrative filamentous phages, requires UvrD and HU for RCR while TLCφ, a satellite phage, depends on Rep and is independent from HU.     

HU7251谷氨酸产生菌的筛选及发酵条件的初步试验

糖质发酵生产谷氨酸在本世纪六十年代全国普遍推广应用以后,给我国的味精生产工业带来了重大技术革新。这不但在生物化学研究上有重要意义,而且对于发展我国国民经济也有很大价值。近几年来,我国谷氨酸发酵工业,在毛主席革命路线指引下,有了很大的发展,取得了可喜的成绩。为进一步落实毛主席提出的“深挖洞、广积粮、不称霸”的伟     

HU binding to a DNA four-way junction probed by Förster resonance energy transfer

The Escherichia coli protein HU is a non-sequence-specific DNA-binding protein that interacts with DNA primarily through electrostatic interactions. In addition to nonspecific binding to linear DNA, HU has been shown to bind with nanomolar affinity to discontinuous DNA substrates, such as repair and recombination intermediates. This work specifically examines the HU-four-way junction (4WJ) interaction using fluorescence spectroscopic methods. The conformation of the junction in the presence of different counterions was investigated by Fo?rster resonance energy transfer (FRET) measurements, which revealed an ion-type conformational dependence, where Na(+) yields the most stacked conformation followed by K(+) and Mg(2+). HU binding induces a greater degree of stacking in the Na(+)-stabilized and Mg(2+)-stabilized junctions but not the K(+)-stabilized junction, which is attributed to differences in the size of the ionic radii and potential differences in ion binding sites. Interestingly, junction conformation modulates binding affinity, where HU exhibits the lowest affinity for the Mg(2+)-stabilized form (24 μM(-1)), which is the least stacked conformation. Protein binding to a mixed population of open and stacked forms of the junction leads to nearly complete formation of a protein-stabilized stacked-X junction. These results strongly support a model in which HU binds to and stabilizes the stacked-X conformation.     

Does the parallel evolution pattern between the replication-segregation proteins and HU have a biological significance?

The bacterial chromosome is a highly compacted nucleoproteic structure. Its apparent disordered morphology is difficult to conciliate with newly discovered mechanisms governing the propagation of genetic information between mother and daughter cells. Recent experiments in bacterial genetics, biochemistry and cytology from a number of laboratories are beginning to unravel how at each cell division, DNA replication and segregation proteins interact spatially with specific DNA motifs to orchestrate replication and movement of replication forks and chromosomes. We propose here a method to confirm and perhaps extend these experiments by in silico protein sequence comparisons and phylogeny. This analysis showed a parallel evolution between the histone-like protein HU and key protein factors involved in DNA replication and chromosome segregation.     

Ⅰ型大麻素受体介导大麻素HU210对星型胶质细胞CXCR4水平下调

目的探寻大麻素抑制中枢神经系统免疫反应的机制,为大麻素临床药物的合理应用提供实验依据。方法用不同浓度的HU210刺激培养的星形胶质细胞,利用Western blot检测并比较刺激组和未刺激组细胞CXCR4蛋白水平的差异,进而用Ⅰ型大麻素受体(type-1 cannabinoid receptor,CBIR)拮抗剂AM281刺激细胞后,观察HU210对CXCR4表达的影响。结果 Western blot检测结果显示,高浓度HU210能下调星形胶质细胞CXCR4表达,AM28能阻断HU210所致的CXCR4下调。结论 HU210经由CB1R下调CXCR4,这可能是大麻发挥免疫抑制作用的机理之一。     

A comparative analysis of karyotypes of three species of Macleaya—producers of a complex of isoquinoline alkaloids

Using a set of methods (C-banding, DAPI-staining, fluorescence hybridization in situ (FISH) with probes of 26S and 5S rDNA, and analysis of meiosis), the first comparative cytogenetic study of three species of Macleaya, producers of complex isoquinoline alkaloids, cordate Macleaya cordata (Willd.) R. Br. (2n = 20), small-fruited Macleaya microcarpa (Maxim.) Fedde (2n = 20) and Macleaya kewensis Turrill (2n = 20), was first carried out. On the basis of morphometric analysis, formulas of karyotypes were made for each species. Species ideograms for M. cordata, M. microcarpa, and M. kewensis were constructed taking into account the polymorphic variants of the C-banding patterns and indicating the location of 26S and 5S rDNA sites. A comparative study revealed that the karyotypes of M. microcarpa and M. kewensis have more in common with each other than with M. cordata. Analysis of meiotic chromosomes suggests of genetic stability of Macleaya genomes. The results of chromosome analysis were used to confirm the close relationship of Macleaya and to clarify their phylogenetic relationships.     

An alternative flexible conformation of the <Emphasis Type="Italic">E. coli</Emphasis> HUβ<Subscript>2</Subscript> protein: structural,dynamics, and functional aspects

The histone-like HU protein is the major nucleoid-associated protein involved in the dynamics and structure of the bacterial chromosome. Under physiological conditions, the three possible dimeric forms of the E. coli HU protein (EcHUα₂, EcHUβ₂, and EcHUαβ) are in thermal equilibrium between two dimeric conformations (N₂ ↔ I₂) varying in their secondary structure content. High-temperature molecular dynamics simulations combined with NMR experiments provide information about structural and dynamics features at the atomic level for the N₂ to I₂ thermal transition of the EcHUβ₂ homodimer. On the basis of these data, a realistic 3D model is proposed for the major I₂ conformation of EcHUβ₂. This model is in agreement with previous experimental data.     

Effect of γ-Irradiation on Development of Lachrymator of Onion

A thermosensitive uracil requiring mutant of Bacillus subtilis Marburg 168 thy trp₂ ts42 was examined as to the colony forming ability at the permissive and nonpermissive temperatures. The viability of the mutant cells decreased rapidly at the restrictive temperature in the modified Woese’s (MW) medium. However, the cells retained viability when sodium succinate or potassium chloride was added to the medium at that temperature although uracil deficiency was unchanged. A little but significant incorporation of adenine-8-¹⁴C into RNA still continued even after the incorporation of N-acetyl-³H-d-glucosamine into acid insoluble fraction of the cells terminated in the MW medium at 48°C. Both incorporations as well as increase of absorbance were slowed down in the presence of sodium succinate at 48°C. This mutant, ts42, was more sensitive to deoxycholate (DOC) than the parent strain. The restoration of colony forming ability after the temperature shift back from 48 to 37°C was suppressed by the addition of DOC to the medium. However, the cell became resistant to DOC when uracil was added to the medium prior to the temperature shift.     

Mechanisms of action of the α-amylase of Micromonospora melanosporea

The -amylase of Micromonospora melanosporea was produced extracellularly during batch fermentation in a 5.0-1 fermentor. The absence of an organic nitrogen source in its growth medium facilitated subsequent purification of the enzyme by ammonium sulphate fractionation and two consecutive Superose-12 gel-filtration steps. The enzyme exhibited maxima for activity at pH 7.0 and 55° C and was 72% stable at pH 6.0–12.0 for 30 min at 40° C. It had a relative molecular mass of 45 000 and an isoelectric point at pH 7.6. The enzyme catalyses the conversion of starch to maltose (53%, w/w) as the predominant final end-product. Initial hydrolysis of this substrate, however, gave rise to the formation of maltooligosaccharides in the range maltotriose to maltohexaose. Maximum yields of these intermediate sugars accumulated to between 31 and 42% (w/w) as the reaction proceeded. The action of the M. melanosporea amylase on high concentrations of saccharides larger than maltotriose resulted in the formation of mainly maltose and maltotriose without concomitant glucose production. A combination of hydrolytic and transfer events is postulated to be responsible for this phenomenon and for the high maltose levels achieved. Correspondence to: C. T. Kelly     

Selection of optimal variants of Gō-like models of proteins through studies of stretching

The Gō-like models of proteins are constructed based on the knowledge of the native conformation. However, there are many possible choices of a Hamiltonian for which the ground state coincides with the native state. Here, we propose to use experimental data on protein stretching to determine what choices are most adequate physically. This criterion is motivated by the fact that stretching processes usually start with the native structure, in the vicinity of which the Gō-like models should work the best. Our selection procedure is applied to 62 different versions of the Gō model and is based on 28 proteins. We consider different potentials, contact maps, local stiffness energies, and energy scales—uniform and nonuniform. In the latter case, the strength of the nonuniformity was governed either by specificity or by properties related to positioning of the side groups. Among them is the simplest variant: uniform couplings with no i, i + 2 contacts. This choice also leads to good folding properties in most cases. We elucidate relationship between the local stiffness described by a potential which involves local chirality and the one which involves dihedral and bond angles. The latter stiffness improves folding but there is little difference between them when it comes to stretching.     

Mechanism of action of α-galactosidase

The effect ofpH on K_m and V_max values of coconut α-galactosidase indicates the involvement of two ionizing groups with pK_a values of 3.5 and 6.5 in catalysis. Chemical modification has indicated the presence of two carboxyl groups, a tryptophan and a tyrosine, at or near the active site of α-galactosidase. Based on these facts a new mechanism of action for α-galactosidase is proposed in which the ionizing group with a pK_a of 3.5 is a carboxyl group involved in stabilizing a carbonium ion intermediate and the ionizing group with a pK_a of 6.5 is a carboxyl group perturbed due to the presence of a hydrophobic residues in its vicinity which donates a H⁺ ion in catalysis.