阿佛曼链霉菌的~(12)C离子辐照效应及高产菌株选育；Research on Mutation Breeding of High-producing Strains from Streptomyces Avermitilis Irradiated by Ion Beam of ~(12)C~(6+)

选用12C6+离子辐照诱变阿维菌素B1a产生菌ZJAV-A1,研究其诱变效应。实验结果表明,12C6+离子辐照剂量50Gy时致死率97%,正突变率最高可达到34.2%。通过12C6+离子诱变处理,结合平板培养基及斜面培养基的正突变菌株筛选,最终获得一株稳定性良好,阿维菌素B1a组分产量稳定在4460—4588μg/ml之间,较出发菌株提高11.1%—14.7%的突变株ZJAV-Y1-203。

Complex organization of the Streptomyces avermitilis genes encoding the avermectin polyketide synthase

The avermectin (Av) polyketide synthase (PKS) and erythromycin (Er) PKS are encoded by modular repeats of DNA, but the genetic organization of the modules encoding Av PKS is more complex than Er PKS. Sequencing of several related DNA fragments from Streptomyces avermitilis that are part of the Av biosynthetic gene cluster, revealed that they encode parts of large multifunctional PKS proteins. The Av PKS proteins show strong similarity to each other, as well as similarity to Er PKS proteins [Donadio et al., Science 252 (1991) 675–679] and fatty acid synthases. Partial DNA sequencing of the 65-kb region containing all the related sequence elements in the avr genes provides evidence for twelve modular repeats encoding FAS-like domains. The genes encoding the Av PKS are organized as two sets of six modular repeats which are convergently transcribed.

T-Muurolol Sesquiterpenes from the Marine Streptomyces sp M491 and Revision of the Configuration of Previously Reported Amorphanes

Two new sesquiterpenes, 15-hydroxy-T-muurolol (3d) and 11,15-dihydroxy-T-muurolol (3e), along with the plant cadinenes T-muurolol (3f) and 3 alpha-hydroxy-T-muurolol (3g), were isolated from the marine-derived Streptomyces sp. M491. Their absolute configuration was established via NMR spectroscopy and X-ray crystallography of 3-oxo-T-muurolol (3a), which was reisolated from this strain. In addition, the absolute configuration of further sesquiterpenes previously reported from this strain was revised. These products were tested for their cytotoxicity against 37 human tumor cell lines using the MTT method. Only 3d was cytotoxic against a range of human tumor cell lines with a mean IC50 of 6.7 mu g/mL.

Chinikomycins A and B: Isolation, structure elucidation, and biological activity of novel antibiotics from a marine Streptomyces sp isolate M045

In our screening of marine Streptomycetes for bioactive principles, two novel antitumor antibiotics designated as chinikomycins A (2a) and B (2b) were isolated together with manumycin A (1), and their structures were elucidated by a detailed interpretation of their spectra. Chinikomycins A (2a) and B (2b) are chlorine-containing aromatized manumycin derivatives of the type 64-pABA-2 with an unusual para orientation of the side chains. They exhibited antitumor activity against different human cancer cell lines, but were inactive in antiviral, antimicrobial, and phytotoxicity tests.

Amorphane sesquiterpenes from a marine Streptomyces sp.

The chemical investigation of the crude extract of the marine-derived Streptomyces sp. M491 yielded three new sesquiterpenes, namely, 10 alpha,11-dihydroxyamorph-4-ene (4), 10 alpha,15-dihydroxyamorph-4-en-3-one (6), and 5 alpha,10 alpha,11-trihydroxyamorphan-3-one (7). In addition, the known compounds 10 alpha-hydroxyamorph-4-en-3-one (2), o-hydroxyacetanilide, genistein, prunetin, and indole-3-carbaldehyde and the macrolide antibiotic chalcomycin A were identified. The structures were determined on the basis of spectroscopic analysis, especially 1D and 2D NMR data. This is the first report of these sesquiterpenes from bacteria.

Intergeneric conjugation in holomycin-producing marine Streptomyces sp strain M095

Marine Streptomyces are potential candidates for novel natural products and industrial catalysts. In order to set up biosynthesis approach for a holomycin-producing strain M095 isotated from Jiaozhou Bay, China, a genetic transformation system was established using intergeneric conjugation. The plasmid pIJ8600 consists of an origin of replication for Escherichia coli, a phage integrase directing efficient site-specific integration in bacterial chromosome, thiostrepton-induced promoter and an attP sequence. Using E. coli ET12567 (pUZ8002) carrying pIJ8600 as a conjugal donor, while it was mated with strain M095, pIJ8600 was mobilized to the recipient and the transferred DNA was also integrated into the recipient chromosome. The frequency of exconjugants was 1.9 +/- 0.13 x 10(-4) per recipient cell. Analysis of eight exconjugants showed pIJ8600 was stable integrated at a single chromosomal site (attB) of the Streptomyces genome. The DNA sequence of the attB was cloned and shown to be conserved. The results of growth and antimicrobial activity analysis indicated that the integration of pIJ8600 did not seem to affect the biosynthesis of antibiotics or other essential amino acids. To demonstrate the feasibility of above gene transfer system, the allophycocyanin gene (apc) from cyanobacterium Anacystis nidulans UTEX625 was expressed in strain M095, and the results indicated heterologous allophycocyanin could be expressed and folded effectively. (c) 2006 Elsevier GmbH. All rights reserved.

5,7-Dihydroxy-5,6,7,8-tetrahydro-1H-azocin-2-one from a marine-derived Streptomyces sp.

In the course of a screening program, we have isolated the new natural product, 5,7-dihydroxy-5,6,7,8-tetrahydroazocin-2(IH)-one (1), from the staurosporine producing marine-derived Streptomyces sp. strain QD518. Here we report the isolation and structure elucidation of 1 and the artifacts 3 and 4 resulting from I by acid catalyzed intra- and inter-molecular reactions.

Heterologous expression and purification of recombinant allophycocyanin in marine Streptomyces sp isolate M097

Allophycocyanin is one of the most important marine active peptides. Previous studies suggested that recombinant allophycocyanin (rAPC) could remarkably inhibit the S-180 carcinoma in mice, indicating its potential pharmaceutical uses. Based on intergeneric conjugal transfer, heterologous expression of rAPC was first achieved in marine Streptomyces sp. isolate M097 through inserting the apc gene into the thiostrepton-induced vector pIJ8600. The transformation frequency for this system was approximately 10(-4) exconjugants/recipient. In the transformed Streptomyces sp. isolate M097, the yield of purified rAPC could amount to about 38 mg/l using a simple purification protocol, and HPLC analysis showed that the purity of the protein reached about 91.5%. In vitro activity tests also revealed that the purified rAPC had effective scavenging abilities on superoxide and hydroxyl radicals. This would widen the usefulness of the marine Streptomyces as a host to express the rAPC and to offer industrial strain for the production of rAPC.

软煤拉伸应力-应变关系的实验研究

提供一种用于测量软煤或土的抗拉强度、拉伸应力-应变关系等拉伸特性的实验装置。利用模具压制出均匀的煤环试件，对煤环内壁施加均布径向水压力使其拉伸变形和破坏。根据实验结果，提出了弹脆性拉断破坏的理论模型，并据此对煤的抗拉强度、弹性常数以及等效孔隙压力系数等进行了分析和讨论。实验数据稳定可靠，散布小，重复性好，说明所提供的实验装置和实验方法简单而有效。

疲劳裂纹扩展机制和塑性区的研究

本文用三点弯曲试样测定15MnVN钢在淬火和200℃低温回火状态下疲劳裂纹扩展速率。试验在循环载荷比R=0.5的恒幅条件下进行,并利用金属塑性变形在光滑表面上产生浮凸的现象,用相衬显微镜测定裂纹顶端单调塑性区尺寸。在平面应力状态下垂直于裂纹面方向的单调塑性区尺寸与相应的(K_(max)/σ_s)~2成正比,其比例系数为0.21。根据Rice循环塑性区尺寸与单调塑性区尺寸关系,得到垂直裂纹面方向的循环塑性区尺寸与相应裂纹扩展速率之间为一指数关系。在裂纹扩展过程中,相对裂纹尺寸a/W约等于0.687时裂纹顶端从小范围屈服进入大范围屈服;a/W<0.687裂纹顶端的塑性变形约在θ=30—60°方向角内向两翼伸展;当a/W>0.687,不仅两翼向更大的θ角展开,而且在θ=0°附近塑性变形也有相当发展。金相和扫描电镜观察结果,具有板条状马氏体组织的15MnVN钢疲劳裂纹扩展是以穿晶型的解理开裂方式进行的,是一种再生核扩展机制。

基于孔径耦合带状线的激光脉冲整形系统

实现了一种新型的激光脉冲整形系统，该系统使用了一个由孔径耦合带状线（ACSL）电脉冲整形器驱动的电光调制器。一个电脉冲整形器由两条通过其公共接地板上的耦合孔径发生耦合作用的带状传输线所组成的四端口装置。更换具有不同耦合孔径的公共接地板，该电脉冲整形发生器可以具有150ps时间结构的任意整形电脉冲。将任意整形的电脉冲输入到电光调制器上，就可以得到任意整形的激光脉冲。利用该系统，激光脉冲整形系统能够产生具有150ps前后沿，1～3ns脉冲宽度可调、高对比度、光滑过渡以及任意整形的激光脉冲。

Spectroscopic properties of Er3+ doped TeO2-BaO (Li2O, Na2O)-La2O3 glasses for 1.5-mu m optical amplifiers

This paper reports on the optical spectroscopic properties and thermal stability of Er3+-doped TeO2-BaO (Li2O,NaO)-La2O3 glasses for developing 1.5-mu m fiber amplifiers. Upon excitation at 977 nm laser diode, an intense 1.53-mu m infrared fluorescence has been observed with a broad full width at half maximum (FWHM) of about 60 nm for the Er3+-doped TeO2-BaO (Li2O, Na2O)-La2O3 glass with 10 mol% of BaO. The calculated fluorescence lifetime and the emission cross-sections of the 1.53-mu m transition are 2.91 ms and similar to 9.97 x 10(-21) cm(2), respectively. It is noted that the gain bandwidth, a, x FWHM, of the TeO2-BaO-La2O3Er2O3 glass is about 600, which is significantly higher than that in silicate and phosphate glasses. Meanwhile, it is interesting to note that the TeO2-BaO-La2O3-Er2O3 glass has shown a high glass thermal stability and good infrared transmittance. As a result, TeO2-BaO (Li2O, Na2O)-La2O3 glass with 10 mol% of BaO has been considered to be more useful as a host for broadband optical fiber amplifier. (c) 2005 Elsevier B.V. All rights reserved.

Identification of triosephosphate isomerase (TIM) genes from Microcystis (Cyanobacteria : Chroococcales) and theoretical analyses of the potential of cyanobacterial TIM as a target for designing specific inhibitors

The genes encoding triosephosphate isomerase (TIM) in three species of Microcystis (M. aeruginosa, M. viridis and M. wesenbergii) were investigated. Reverse transcriptase-polymerase chain reaction indicated that they were transcribed in the cells. Analyses showed that their DNA and deduced amino acid sequences were highly conserved between all the three species, only a single nonsynonymous substitution was seen at position 31, from an Asp in M. aeruginosa and M. viridis to Glu in M. wesenbergii. Sequence alignment of these with 12 other known cyanobacterial TIM sequences showed that all the cyanobacterial TIMs had a very high level of amino acid identity (over 50% between each two). Comparison of the cyanobacterial TIMs with other reported TIMs (from diverse lineages of the three Domains) showed that they possessed common active-site residues and sequence motifs. All cyanobacterial TIMs have two common cysteine residues (Cys127 and Cys176), and the Cys176 is almost cyanobacteria-specific with only one exception in Streptomyces coelicolor. Both secondary structure alignment and comparative modelling of Synechocystis sp. TIM showed that Cys176 was located at the hinge region of the flexible loop-6 and might therefore be critical to the movement of TIM's loop-6, which is important to the function of the enzyme. Thus, the cyanobacterial TIM-specific Cys176 may be a potential site for the discovery of suitable drugs against cyanobacteria, and such drugs may have utility in controlling water blooms due to cyanobacteria.

鱼腥藻Anabaena PCC7120原生质球和液泡的同步诱导

国家自然科学基金资助项目( 39870 0 83)

Actinomycetes and earthy-musty odorous compounds in brackish fishponds in Tianjin, China

The presence of the odorous compounds, 2-methylisoborneol (MIB) and geosmin, as well as causative microorganisms in brackish intensive cultivation fishponds in Tianjin, China that had a severe earthy-musty odor were evaluated. The results revealed that MIB was the primary odorous compound present in the Tianjin fishponds, with a concentration ranging from 0.53-5302.7 ng.L-1. Furthermore, the concentration of MIB was found to be closely correlated with the gross biomass of actinomycetes in the water, which ranged from 10.67-1528.24 x 10(6) cfu.ml(-1). Therefore, the sequences of the 16 SrRNA and morphological characteristics of the actinomycetes in the brackish fishponds were investigated. The results revealed that the actinomycetes in the brackish fishponds included 9 species of common and dominant actinomycetes belonging to 4 genera. Of these genera, Streptomyces were the dominant species, and Streptomyces, Nocardioides and Micromonospora were the most common species in the fishponds evaluated. Next, the ability of each of the isolated Streptomyces to produce MIB was measured under laboratory culture conditions. Streptomyces Sp2 was found to have a strong ability to produce MIB, which indicates that this strain may be the primary source of the earthy-musty odor reported in brackish intensive cultivation fishponds in Tianjin, China.