Isolation of a class IV chitin synthase gene from a zygomycete fungus, Rhizopus oligosporus

We found the presence of DNA sequence which shows sequence similarity to the class IV chitin synthase gene (CHS3) of Saccharomyces cerevisiae in the genome of 14 Rhizopus species which belong to zygomycetes. We cloned a gene (chs3), which might correspond to one of these homologous sequences, from Rhizopus oligosporus by low stringency plaque hybridization probed with CHS3. The deduced amino acid sequence of this gene showed highest similarity to the class IV chitin synthase of Neurospora crassa (46.7% identity over 1087 amino acids), showing that this gene encodes a class IV chitin synthase. Northern analysis revealed the differential expression pattern of this gene in the asexual life cycle with highest expression in the early stage of asexual spore formation. This is the first report of the isolation and analysis of a class IV chitin synthase gene from zygomycete fungi.     

Taxonomic investigation of five streptomycete cultures belonging to ISP standards

The taxonomic investigation of five streptomycete cultures belonging to the International Streptomyces Project (ISP) standards was carried out using the methods of population analysis, DNA-DNA hybridization, and multilocus DNA fingerprinting. Two species with names considered to be synonymous,S. alboviridis ISP 5326 andS. oligocarbophilus ISP 5589, were found to be actually identical. Three other species investigated,S. krainskii ISP 5321,S. craterifer ISP 5296, and 5.anulatus ISP 5361, whose names are usually referred to as synonymous, were shown to be different species.     

腐植酸生产菌株——链霉菌发酵条件研究

利用链霉菌ST菌株发酵生产腐植酸。在蔗渣发酵培养基的基础上,采用一次回归正交试验,对腐植酸发酵过程中接种量（I）、通气量（搅拌次数,A）、温度（T）及时间（D）4个因素对腐植酸产量的影响情况进行分析。结果表明发酵时间的影响最为显著,接种量影响显著;所得到的回归方程有效且可信度高,回归方程为Y=6．0356＋0．02501＋0．0033A＋0．0069T＋0．2425D,其中各因素均与腐植酸产量呈正相关性。该回归方程为腐植酸生产过程链霉菌ST菌株发酵条件的控制提供了很好的指导作用。     

Transposition and transduction of plasmid DNA inStreptomyces spp.

Summary To expand the application of molecular genetics to many different streptomycete species, we have been developing two potentially widely applicable methodologies: transposon mutagenesis and plasmid transduction. We constructed three transposons from theStreptomyces lividans insertion sequence IS493. Tn5096 and Tn5097 contain an apramycin resistance gene inserted in different orientations between the two open reading frames of IS493. These transposons transpose from different plasmids into many different sites in theStreptomyces griseofuscus chromosome and into its resident linear plasmids. Tn5099 contains a promoterlessxylE gene and a hygromycin-resistance gene inserted in IS493 close to one end. Tn5099 transposes inS. griseofuscus giving operon fusions in some cases that drive expression of thexylE gene product, catechol deoxygenase, giving yellow colonies in the presence of catechol. We have also developed plasmid vectors that can be transduced into many streptomycete species by bacteriophage FP43. We describe the characterization of FP43 and mapping of several bacteriophage functions. The region of cloned FP43 DNA essential for plasmid transduction includes the origin for headful packaging.     

Gene probes for the detection of 6-deoxyhexose metabolism in secondary metabolite-producing streptomycetes

DNA probes were designed from the streptomycin production genes strDELM of Streptomyces griseus involved in the biosynthesis of the 6-deoxyhexose (6DOH) dihydrostreptose which could detect the genomic fragments coding for 6DOH formation in other actinomycetes strains. In about 70% of the 43 strains tested at least one signal could be detected with strD-, strE- or strLM-specific probes. Evidence is presented that the hybridizing genes are mostly clustered and probably engaged in the formation of secondary metabolites. Because of the wide-spread use of 6DOH constituents in natural products these probes should allow to detect a vast array of different secondary metabolic gene clusters in actinomycetes.     

Kakadumycins,novel antibiotics from Streptomyces sp NRRL 30566, an endophyte of Grevillea pteridifolia

An endophytic streptomycete (NRRL 30566) is described and partially characterized from a fern-leaved grevillea (Grevillea pteridifolia) tree growing in the Northern Territory of Australia. This endophytic streptomycete produces, in culture, novel antibiotics - the kakadumycins. Methods are outlined for the production and chemical characterization of kakadumycin A and related compounds. This antibiotic is structurally related to a quinoxaline antibiotic, echinomycin. Each contains, by virtue of their amino acid compositions, alanine, serine and an unknown amino acid. Other biological, spectral and chromatographic differences between these two compounds occur and are given. Kakadumycin A has wide spectrum antibiotic activity, especially against Gram-positive bacteria, and it generally displays better bioactivity than echinomycin. For instance, against Bacillus anthracis strains, kakadumycin A has minimum inhibitory concentrations of 0.2-0.3 microg x ml(-1) in contrast to echinomycin at 1.0-1.2 microg x ml(-1). Both echinomycin and kakadumycin A have impressive activity against the malarial parasite Plasmodium falciparum with LD(50)s in the range of 7-10 ng x ml(-1). In macromolecular synthesis assays both kakadumycin A and echinomycin have similar effects on the inhibition of RNA synthesis. It appears that the endophytic Streptomyces sp. offer some promise for the discovery of novel antibiotics with pharmacological potential.     

The distribution of antibiotic resistance patterns within streptomycetes and their use in secondary metabolite screening

Summary A collection of 169 streptomycetes representing natural isolates and type strains were examined for resistance phenotypes to 11 antibiotics. A total of 84 profiles were obtained with 18 patterns being repeated in two or more strains. The most common pattern was resistance to penicillin in an otherwise sensitive phenotype and accounted for 51 strains. This data was used to cluster the strains and groups defined were examined for correlation with bioactivity. Antibiotic producers were found in clusters 1, 2, 4 and 5–10. Certain strains in these areas were highly bioactive and typically had multiple resistances. Almost half of the collection of strains examined grouped in cluster 3, and were characterized as having a sensitive phenotype and virtually no biological activity in agrochemical screens.     

海洋链霉菌S09的化学成分研究(英文)

从海洋链霉菌S09的发酵液提取物中分离得到6个化合物,通过波谱技术分别鉴定为:thymidine(1)、尿嘧啶(2)、3-甲基-2,5-哌嗪二酮(3)、2-吡咯甲酸(4)、卡拉霉素(5)和β-咔啉(6)。海虾致死实验结果显示:化合物5在10μg/mL的浓度下对丰年虾的致死率为87.6%,显示出较强的细胞毒活性。     

Isolation of Endophytic Actinomycetes from Different Cultivars of Tomato and their Activities Against Ralstonia solanacearum in Vitro

The populations of endophytic actinomycetes from healthy and wilting tomato plants (tomato cultivars resistant and susceptible to Ralstonia solanacearum) grown in three different sites from Guangzhou, Guangdong Province, South China were investigated by cultivation methods. Most of the isolates belonged to streptomycetes. The Aureus group of Streptomyces was the most frequently isolated group. The population composition of Streptomyces varied according to tomato cultivars, physiological status and soil types. The proportions of antagonistic Streptomyces strains from healthy plants were higher than that from wilting plants (P < 0.05), although the difference among the proportions of antagonistic Streptomyces strains from different cultivars of healthy tomato was not significant, the similar result was found from wilting plants. No significant difference was found in the proportions of siderophere-producing Streptomyces strains from the same site (P > 0.05), but the difference was found from the different sampling sites (P < 0.05). The percentage of bacterial cell wall-degrading streptomycetes from wilting tomato was higher than that from healthy plants (P < 0.05). These results indicated that the cultivar of the host plant, physiological status and sampling sites would influence the proportion of endophytic streptomycetes with different physiological traits. Diversity of endophytic Streptomyces and their physiological diversity should be involved in developing potential biocontrol agents.