186 resultados para STREPTOMYCES CLAVULIGERUS


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A Gram-staining positive, non-motile, rod-shaped, catalase positive and oxidase negative bacterium, designated NCCP-1331(T), was isolated from a hot water spring soil collected from Tatta Pani, Kotli, Azad Jammu and Kashmir, Pakistan. The isolate grew at a temperature range of 18-40 °C (optimum 30 °C), pH 6.0-9.0 (optimum 7.0) and with 0-6 % NaCl (optimum 2 % NaCl (w/v)). The phylogenetic analysis based on 16S rRNA gene sequence revealed that strain NCCP-1331(T) belonged to the genus Streptomyces and is closely related to Streptomyces brevispora BK160(T) with 97.9 % nucleotide similarity, followed by Streptomyces drosdowiczii NRRL B-24297(T) with 97.8 % nucleotide similarity. The DNA-DNA relatedness values of strain NCCP-1331(T) with S. brevispora KACC 21093(T) and S. drosdowiczii CBMAI 0498(T) were 42.7 and 34.7 %, respectively. LL-DAP was detected as diagnostic amino acid along with alanine, glycine, leucine and glutamic acid. The isolate contained MK-9(H8) as the predominant menaquinone. Major polar lipids detected in NCCP-1331(T) were phosphatidylethanolamine, phosphatidylinositol and unidentified phospholipids. Major fatty acids were iso-C16: 0, summed feature 8 (18:1 ω7c/18:1 ω6c), anteiso-C15:0 and C16:0. The genomic DNA G + C content was 69.8 mol %. On the basis of phylogenetic, phenotypic and chemotaxonomic analysis, it is concluded that strain NCCP-1331(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces caldifontis sp. nov. is proposed. The type strain is NCCP-1331(T) (=KCTC 39537(T) = CPCC 204147(T)).

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Rifampicin and its derivatives are at the forefront of the current standard chemotherapeutic regimen for active tuberculosis; they act by inhibiting the transcription activity of prokaryotic RNA polymerase. Rifampicin is believed to interact with the beta subunit of RNA polymerase. However, it has been observed that protein-protein interactions with RNA polymerase core enzyme lead to its reduced susceptibility to rifampicin. This mechanism became more diversified with the discovery of RbpA, a novel RNA polymerase-binding protein, in Streptomyces coelicolor that could mitigate the effect of rifampicin on RNA polymerase activity. MsRbpA is a homologue of RbpA in Mycobacterium smegmatis. On deciphering the role of MsRbpA in M. smegmatis we found that it interacts with RNA polymerase and increases the rifampicin tolerance levels, both in vitro and in vivo. It interacts with the beta subunit of RNA polymerase. However, it was found to be incapable of rescuing rifampicin-resistant RNA polymerases in the presence of rifampicin at the respective IC50.

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FERROVERDIN, a green iron-containing pigment, was isolated in 1955 by Chain, Tonolo and Carilli1 from an unidentified species of Streptomyces. It was at first assigned the formula C30H24O8N2Fe and the iron was shown by measurements of magnetic susceptibility to be in the ferrous state2. Later the ligand present was proved to be the p-vinyl phenyl ester of 3-nitroso-4-hydroxy-benzoic acid3,4. X-ray crystallographic measurements were undertaken to find the atomic arrangement in this unusual complex; they show, in two different crystal structures, that each iron atom is attached to three nitrosophenyl ligands and that the charge is balanced by sodium ions.

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Much of the chemical structure of thiostrepton, a sulphur containing metabolic product of the microorganism Streptomyces azureus, has been determined by X-ray crystallographic techniques.

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The discovery of GH (Glycoside Hydrolase) 19 chitinases in Streptomyces sp. raises the possibility of the presence of these proteins in other bacterial species, since they were initially thought to be confined to higher plants. The present study mainly concentrates on the phylogenetic distribution and homology conservation in GH19 family chitinases. Extensive database searches are performed to identify the presence of GH19 family chitinases in the three major super kingdoms of life. Multiple sequence alignment of all the identified GH19 chitinase family members resulted in the identification of globally conserved residues. We further identified conserved sequence motifs across the major sub groups within the family. Estimation of evolutionary distance between the various bacterial and plant chitinases are carried out to better understand the pattern of evolution. Our study also supports the horizontal gene transfer theory, which states that GH19 chitinase genes are transferred from higher plants to bacteria. Further, the present study sheds light on the phylogenetic distribution and identifies unique sequence signatures that define GH19 chitinase family of proteins. The identified motifs could be used as markers to delineate uncharacterized GH19 family chitinases. The estimation of evolutionary distance between chitinase identified in plants and bacteria shows that the flowering plants are more related to chitinase in actinobacteria than that of identified in purple bacteria. We propose a model to elucidate the natural history of GH19 family chitinases.

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Background: mIHF belongs to a subfamily of proteins, distinct from E. coli IHF. Results: Functionally important amino acids of mIHF and the mechanism(s) underlying DNA binding, DNA bending, and site-specific recombination are distinct from that of E. coli IHF. Conclusion: mIHF functions could contribute beyond nucleoid compaction. Significance: Because mIHF is essential for growth, the molecular mechanisms identified here can be exploited in drug screening efforts. The annotated whole-genome sequence of Mycobacterium tuberculosis revealed that Rv1388 (Mtihf) is likely to encode for a putative 20-kDa integration host factor (mIHF). However, very little is known about the functional properties of mIHF or the organization of the mycobacterial nucleoid. Molecular modeling of the mIHF three-dimensional structure, based on the cocrystal structure of Streptomyces coelicolor IHF duplex DNA, a bona fide relative of mIHF, revealed the presence of Arg-170, Arg-171, and Arg-173, which might be involved in DNA binding, and a conserved proline (Pro-150) in the tight turn. The phenotypic sensitivity of Escherichia coli ihfA and ihfB strains to UV and methyl methanesulfonate could be complemented with the wild-type Mtihf but not its alleles bearing mutations in the DNA-binding residues. Protein-DNA interaction assays revealed that wild-type mIHF, but not its DNA-binding variants, binds with high affinity to fragments containing attB and attP sites and curved DNA. Strikingly, the functionally important amino acid residues of mIHF and the mechanism(s) underlying its binding to DNA, DNA bending, and site-specific recombination are fundamentally different from that of E. coli IHF. Furthermore, we reveal novel insights into IHF-mediated DNA compaction depending on the placement of its preferred binding sites; mIHF promotes DNA compaction into nucleoid-like or higher order filamentous structures. We therefore propose that mIHF is a distinct member of a subfamily of proteins that serve as essential cofactors in site-specific recombination and nucleoid organization and that these findings represent a significant advance in our understanding of the role(s) of nucleoid-associated proteins.

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The annotated whole-genome sequence of Mycobacterium tuberculosis indicated that Rv1388 (Mtihf) likely encodes a putative 20 kDa integration host factor (mIHF). However, very little is known about the functional properties of mIHF or organization of mycobacterial nucleoid. Molecular modeling of the mIHF three-dimensional structure, based on the cocrystal structure of Streptomyces coelicolor IHF-duplex DNA, a bona fide relative of mIHF, revealed the presence of Arg170, Arg171, and Arg173, which might be involved in DNA binding, and a conserved proline (P150) in the tight turn. The phenotypic sensitivity of Escherichia coli Delta ihfA and Delta ihfB strains to UV and methylmethanesulfonate could be complemented with the wild-type Mtihf, but not its alleles bearing mutations in the DNA-binding residues. Protein DNA interaction assays revealed that wild-type mIHF, but not its DNA-binding variants, bind with high affinity to fragments containing attB and attP sites and curved DNA. Strikingly, the functionally important amino acid residues of mIHF and the mechanism(s) underlying its binding to DNA, DNA bending, and site-specific recombination are fundamentally different from that of E. coli IHF alpha beta. Furthermore, we reveal novel insights into IHF-mediated DNA compaction depending on the placement of its preferred binding sites; mIHF promotes compaction of DNA into nucleoid-like or higher-order filamentous structures. We hence propose that mIHF is a distinct member of a subfamily of proteins that serve as essential cofactors in site-specific recombination and nucleoid organization and that these findings represent a significant advance in our understanding of the role(s) of nucleoid-associated proteins.

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Con el propósito de responder a preguntas de orden nutricional y biológico del suelo con un enfoque agroecológico se desarrolló el presente trabajo, en un terreno calcáreo, no salino, que permaneció en barbecho cinco años antes de iniciado el estudio. Se evaluó la implementación de una diversificación de cultivos (guayaba, nopal, piña y papaya) y tres dosis de vermicompost (a 1 :10, a 2 :15 y a 3 :20 t (ha año) - 1 . La investigación se inició en mayo del 2009 hasta diciembre del 2011 y el diseño usado fue cuasiexperimental en franjas pareadas. Las características físicas - químicas de suelo analizadas fueron: pH en H 2 O, materia orgánica , N, arena, limo y arcilla en %; P disponible, Fe, Cu, Mn y Zn en ppm; K disponible, Ca y Mg en meq, conductividad eléctrica en μS/cm y la relación C/N. Desde el punto de vista biológico del suelo se determinaron presencia de hongos, bacterias y actinomicetos. Antes de iniciar el estudio en 2009, y durante el ensayo, se determinaron las variables físicas, químicas y biológicas del suelo. Para la interpretación agronómica de la nutrición para cada cultivo se hizo un análisis combinado entre los resultados de las propiedades físico - químicas y los rangos definidos para cada cultivo por la metodología propuesta por Quintana (1983) . La diversificación de cultivos, influyó sobre las propiedades físicas, químicas y biológicas del suelo. En el cultivo de guayaba si se aplica 20 t (ha año) - 1 hay que realizar aplicaciones suplementarias de Cu. Aplicaciones de a 1 y a 2 en nopal y papaya respectivamente, además de aplicaciones complementarias de Cu, también necesitan suplemento de P. Sin efecto antropogénico los microorganismos que predominaron fueron bacterias del género Bacillus y esporádicamente Pseudomona y Sarcina . Con la diversificación de cultivos y aplicaciones de vermicompost se incrementó la diversidad de géneros de microorganismos. El total de géneros de microorganismos, al concluir el estudio fueron 13; de éstos 10 fueron diferentes (Cinco hongos: Curvularia, A spergillus, Pestalotia, Fusarium, Penicillium ; cuatro bacterias: Serratia, Erwinia, Sporolactobacillus y Caryophanon y un actinomiceto: Streptomyces). Variables del crecimiento en el cultivo de guayaba demostraron que con un mayor número de ramas terciarias existe un 70 % de probabilidad de tener más frutos comerciales. En nopal, el tratamiento con los mejores resultados para variables del crecimiento es 15 t (ha año) - 1 cosechado cada 90 días. Para piña, se encontró resultados superiores con 10 t (ha año) - 1 . Papaya obtuvo los mejores resultados con 20 t (ha año) - 1 . Con el cultivo de guayaba el mejor rendimiento se alcanzó utilizando 20 t (ha año) - 1 . En nopal un mayor número de cladodios se alcanza aplicando 15 t (ha año) - 1 cosechado cada 30 días. Un mayor peso en el cladodio se obtiene al aplicar 20 t (ha año) - 1 , cosechado cada 90 días. La piña obtiene mayor número de frutos aplicando 15 t (ha año) - 1 . En el cultivo de papaya un mayor número de frutos cosechados fue alcanzado con 15 - 20 t (ha año) - 1.

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Os bastonetes Gram positivos irregulares (BGPIs) compõem um grupo de espécies bacterianas com ampla diversidade fenotípica e que podem estar presente no meio ambiente, na microbiota humana e de animais. A identificação acurada de BGPIs em nível de gênero e espécie empregando métodos bioquímicos convencionais é bastante limitada, sendo recomendado, portanto, o uso de técnicas moleculares. No presente estudo, foram identificadas amostras de BGPIs oriundas de espécimes clínicos de humanos, de produtos farmacêuticos e de áreas limpas através da análise de sequencias do gene 16S rRNA e de outros genes conservados (housekeeping genes). Os resultados obtidos pelo sequenciamento dos genes 16S rRNA e rpoB demonstraram C. striatum multi-resistente (MDR) como responsável por surto epidêmico em ambiente hospitalar da cidade do Rio de Janeiro. Quinze cepas de C. striatum foram isoladas em cultura pura a partir de secreção traqueal de pacientes adultos submetidos a procedimentos de entubação endotraqueal. A análise por eletroforese em gel de campo pulsado (PFGE) indicou a presença de quatro perfis moleculares, incluindo dois clones relacionados com cepas MDR (PFGE I e II). Os dados demonstram a predominância de PFGE I entre cepas MDR isoladas de unidades de terapia intensiva e enfermarias cirúrgicas. Uma potencial ligação causal entre a morte e a infecção por C. striatum MDR (PFGE tipos I e II) foi observada em cinco casos. Adicionalmente, acreditamos que este seja o primeiro estudo de identificação de espécies de Nocardia relacionadas com infecções humanas pela análise da sequencia multilocus (MLSA) no Brasil. Diferente dos dados observados na literatura (1970 a 2013) e obtidos pelos testes fenotípicos convencionais, a caracterização molecular de quatro lócus (gyrB-16S-secA1-hsp65) permitiu a identificação das espécies N. nova, N. cyriacigeorgica, N. asiatica e N. exalbida/gamkensis relacionadas com quadros de nocardiose em humanos. Cepas de N. nova isoladas de diferentes materiais clínicos de um único paciente apresentaram padrões de susceptibilidade antimicrobianos idênticos e dois perfis PFGE, indicando a possibilidade de quadros de co-infecção por N. nova em humanos. Em outra etapa da investigação, amostras de BGPIs obtidos de ambientes de salas limpas que não puderam ser identificadas por critérios convencionais foram submetidas a análise da sequência do gene 16S rRNA e caracterizadas 95,83% em nível de gênero e 35,42% em espécies. Para gêneros mais encontrados no estudo, foram analisados os genes rpoB e recA de dezessete cepas de Microbacterium e utilizado o MLSA para a identificação de sete cepas identificadas como Streptomyces. Os ensaios permitiram a identificação de três cepas de Microbacterium e de uma única amostra de Streptomyces ao nível de espécie. A análise da sequencia do gene rpoB também se mostrou eficaz na identificação de espécies de cepas de Corynebacterium. Finalmente, para as cepas ambientais pertencentes à classe Actinobacteria os dados morfológicos, bioquímicos e genotípicos permitiram documentar a cepa 3117BRRJ como representante de uma nova espécie do gênero Nocardioides, para o qual o nome Nocardioides brasiliensis sp. nov. e as cepas 3712BRRJ e 3371BRRJ como representante de um novo gênero e espécie para o qual o nome Guaraldella brasiliensis nov. foi proposto.

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In the present investigation the marine bacteria isolated from corals, sponges sea water and sediments of coral regions in the larak Island located in the Persian Gulf and were examined for ability to produce cytotoxic metabolits in order to use as an anticancer compounds. Cytotoxic effect were isolated bacteria from different samples and were examined by Artemia Cytotoxic Bioassay test, in which 4.5 percent of sea waters, 12 percent of sediments and 28 percent of marine invertebrat showed cytotoxic activity, using Brine Shrimp Bioassay test. Streptomyces S-2004 isolated from soft coral specified as Sinularia erecta had LC50=0.5mg/m1 in Brine Shrimp Bioaassay test. The streptomyces S-2004 produced cytotoxic metabolits in low nutrient condition and sea water medium after 7 days on 250 rpm shaken in vitro condition. The extract partially were semipurified. Then ethyl acetate extraction from aceton extracted of bacterial plate had cytotoxic effect (LC50=4.19ktg/m1) in Human epidermoid carcinoma of mouth cells (KB) by using neutral red assay. Morphological effects of this extract on KB cells showed turgescence, cellular blebs and apoptosis which was a proof for anticancer compounds of the extract. It is seems that streptomyces S-2004 is a new strain and could be introduced as a talented bacteria, which produced cytotoxic metabolits.

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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.

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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.

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放线菌是一种重要的微生物资源,广泛分布于自然界中。已经发现的微生物来源的生物活性物质中,由放线菌产生的约占三分之二。从我国云南省不同地方的土壤样品中分离到多株有潜在生物活性的放线菌,本文对从西双版纳原绐森林土壤样品中分得的JX-47菌株着重进行了研究,用形态观察、化学分类及分子分类等多项分类的方法确定了该菌株的分类地位,为链霉菌一新种,命名为Streptomyces autolyticus sp nov.。从该菌株的发酵液中分离得到三个化合物,用各种光谱实验鉴定了它们的结构式。这三个化合物均属于Ansamycin类抗生素;其中一个为新化合物,命名为autolytimycin,并申请了国家发明专利(申请号为:00102982. 7)。初步的实验结果证明这些化合物具有细胞毒性和一定的抗病毒活性。同时,对分离到的JX-42、JX-45、JX-46菌性的分类地位和次生代谢产物进行了初步研究。根据形态特征和细胞壁的化学组成,JX-42和JX-46菌株被确定属于链霉菌属;JX-45菌株被确定属于小单孢菌属。从它们的发酵液中,共分离出10个化合物,并鉴定了结构。

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本文综述了放线菌分类学研究的目的和作用,分析了放线菌分类学的历史和现状,介绍了当前放线菌多相分类研究中所采用的技术方法及适用范围。同时还重点介绍了极端高温、低温、高盐放线菌分离及分类研究的进展。从云南采集高温温泉水样、火山口土样,从云南、新疆等地采集雪山土样,从新疆、青海等地采集盐碱土样进行放线菌分离,对不同极端环境下的放线菌分离方法进行探讨,并对分离到的部分典型放线菌菌株采用形态特征、培养特征,生理生化测定,细胞化学组份分析,DNA G+C mol%和DNA同源性测定,以及16SrDNA全序列分析等相结合的多相分类技术进行系统的分类研究。从表型、基因型及系统发育三个不同层次对其分类地位进行了最终确定。其中,分离自云南洱源温泉的菌株YIM60013和腾冲火山口的菌株YIM60032分别确定为高温放线菌属的两个新种:白色高温放线菌(Thermoactznomyces albus sp. nov.)和云南高温放线菌(Termoactomyces yunnanensis sp. nov.);分离自新疆北疆地区的一株低温放线菌菌株,结合其形态特征、细胞化学组份及16S rDNA序列分析将其鉴定为链霉菌的一个新种,北疆链霉菌(Streptomyces beijiangensis sp. nov.);来自新疆盐碱土样的6株嗜盐放线菌菌株YIM90001-90006中,菌株YIM90001被命名为嗜盐普氏菌新种(Prauserella halophila sp. nov.),菌株YIM90005被 命名为脱卤普氏菌新种(Prauserella dehalogenans sp. nov.),菌株YIM90002和YIM90003鉴定为拟诺卡氏菌科中的链单抱菌新属Streptomonospora gen. nov.)和它的两个新种:菌株YIM90002定为盐生链单抱菌新种(Streptomonospora saline sp. nov.),菌株YIM90003定为白色链单抱菌新种(Streptomonospora alba sp. nov.);菌株YIM90004和YIM90006分别被确定为拟诺卡氏菌属的一个新种和一个亚种:新疆拟诺卡氏菌新种(Nocardopsi sxiniangensis sp. nov.)和嗜阿拉伯糖新疆拟诺卡氏菌亚种( Noocardiopsi sxiniangensis subsp.arabicus subsp.nov,)。

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为了筛选对靶基因LDLR和VCAM-1的表达具有调节作用的生物活性物质,建立了两个基于重组人细胞系的高通量的筛选模型,使用荧光素酶在96-孔版上来筛选对上述靶基因的表达具有调节作用的微生物代谢产物。模型之一是来自于人肝HepG2细胞系的重组L39细胞,用于筛选增加LDLR报告基因表达的生物活性物质,以期发现新的具有降胆固醇作用的药物。筛选之二为来源于细胞系ECV304的重组细胞株Nl-14,用于筛选抑制VCAM-1基因表达的活性物质,以期发现治疗风湿性关节炎等免疫性疾病治疗的药物。上述筛选系统均是稳定转染的细胞系,分别含有与荧光素酶报告基因相融合的LDLR或VCAM-1基因的转录调节元件。通过对6300株微生物的总计12600个样品的筛选,共发现和分离了17个活性化合物并进行了结构解析。其中两个被命名为Cladospolede D和Zelkovamycin的化合物被确定为新的化合物。由真菌 FO-6605的发酵液提取得到的一个化合物对LDLR报告基因的表达具有很强的上调作用,其SC200为1 Onmol/L a使用荧光标记的LDL检测到该化合物对于HepG2细胞膜上LDLR具有剂量依赖的增强作用。由真菌FO-5897的发酵液中分离到了一个已知的化合物Ascofuranone,该化合物曾经被报道具有降血脂抗肿瘤的活性。值得注意的是我们首次发现了该化合物同时具有抑制 VCAM-1报告基因表达和增强LDLR报告基因表达的作用,该发现有可能会对其降血脂作用的深入研究提供帮助。由海洋真菌FT-0012产生的化合物Cladospolede D为一个12-员环的大环内酷类的化合物,该化合物对两个测活系统均显示出无选择性的抑制作用形态学研究显示该真菌属于Cladosporiun属。另外一个由土壤放线菌K96-670产生的新化合物为一个环八肤类的化合物,经~1H~1-H COSY,~(13)C-H COSY,~(13)C-~1H HMQC, ~(15)N-~1H HMQC,~(15)-~1N HHMBC等波谱学研究得知该化合物的分子结构中含有六个非普通的氨基酸和两个普通氨基酸。该化合物对VCAM-I报告基因的表达显示出非常好的选择性的抑制活性,其IC50值为9.5ug/ml.形态学的研究表明该菌株属于链霉菌属。 在筛选过程中从来源于云南省西双版纳的土壤中分离到了一株编号为YIM1272的放线菌,经包括形态学、生理一生化和16S rDNA在内的分类学研究,确定该菌株为链霉菌属的一个新种,被命名为佩版纳链霉菌,(Streptomyces.bannaensis.sp.nov)。