金属离子对黄柄曲霉生长和抗真菌抗生素合成的影响

不同的金属离子对兼性海洋霉菌黄柄曲霉179的生长有不同的影响。在0．002mol/L的浓度下,Hg^2 、Ag^ 和Cr^3 能强烈抑制该菌的生长,Pb^2 、Sr^2 、Co^2 、Al^3 对其生长有一定的抑制,生长量低于对照;Mn^2 、Ba^2 、Zn^2 对其生长没有明显影响;Cu^2 对该菌的生长有微弱的促进作用。菌体生物量略高于对照;不同的金属离子对曲霉179真菌抗生素179M合成有不同的影响。与对照相比,Ba^2 对共产量没有影响;Al^3 、Zn^2 、Sr^2 有一定的抑制作用,其发酵相对效价分别为对照的81．4％、55．5％和65％;0．002mol／L斩Mn^2 和Pb^2 能强烈抑制此抗生素的合成,在添加0．002mol/LMn^2 和Pb^2 的培养基中,虽然菌体生长良好,但无179M产生;Co^2 和Cu^2 则有明显的促进作用,当培养基中添加0．002ml／L的Co^2 时其发酵相对效价提高到261．4％;当培养基中添加0．003mol／L的Cu^2 时其发酵相对效价提高到350．2％。Co62 和Cu62 对发酵的促进效应是相互拮抗的。     

透明颤菌血红蛋白在肉桂地链霉菌中的表达对基因细胞生长及抗生素合成的影响

肉桂地链霉菌（S.cinnamonensis)是莫能菌素（Monensin)的产生菌,大肠杆菌－链霉菌穿梭表达载体pHZ1252中的透明颤菌血红蛋白基因（vhb)位于硫链丝菌素诱导启动子ＰtipA之下,它在肉桂地链霉菌中的结构不稳定,,发生了重组缺失,缺失的片段包括大肠杆菌质粒部分vhb基因。但来自阿维链霉菌(S.avermitilis)中缺失了大肠杆菌质粒部分却保留了完整的vhb基因及tipA启动子的pHZ1252,可在肉桂地链霉菌中稳定复制,不再发生缺失,经硫链丝菌素诱导表达出了有生物活性的ＶＨb蛋白,摇瓶发酵实验证明,ＶＨb蛋白在氧限条件下可明显促进肉桂地链霉菌的菌体生长和抗生素合成。     

绿色产色链霉菌突变株发酵菌丝体对青脚土杂肉鸡生长性能和肌肉品质的影响

在青脚土杂肉鸡口粮中按照600mg／kg的比例添加绿色产色链霉菌诱变菌株W26菌株的液体深层发酵菌丝体,考察茼丝体别‘肉鸡的生产性能和肌肉品质的影响。实验结果表明：日粮中添加600mg／kg的绿色产色链霉菌菌丝体,叮以显著提高肉鸡的活体重,动物日增重与对照组相比提高了26．84％,显著改善了饲料转化率（P〈0．05）。菌丝体的添加可以提高肉鸡肌肉中必需氨基酸、蛋白质、灰分的含量,降低肌肉肌间脂肪含量,但效果不显著。综合实验结果说明,绿色产色链霉菌诱变菌株的液体深层发酵菌丝体可以显著提高青脚土杂肉鸡的生长性能。     

枸杞子对云芝菌丝体生长和多糖含量的影响

本文主要研究了枸杞子水煎液的不同浓度对云芝菌丝体生长和云芝多糖积累的影响,试验结果表明,低浓度的枸杞子水煎液有明显的促进作用,高浓度的枸杞子水煎液不表现过强的抑制作用.     

克隆链霉菌的抗生素生物合成基因;产生杂种抗生素

人们最初克隆了一些识别相对简便的,在受体中以“显性”可选择的表型出现的链霉菌的一些基因,例如下列一些基因;决定次甲基霉素抗性的(8),硫链菌素和新霉素的抗性(19)以及紫霉素和红霉素的抗性(18)。     

抗生素对耐药菌生长繁殖影响的观察

目的：观察耐药菌株在含抗生素的培养基中生长情况与规律,进一步探讨抗生素应用的某些方法以及细菌耐药性形成的机制。方法：将多重耐药性的金黄色葡萄球菌、肺炎克雷伯菌、铜绿假单胞菌接种于含临床剂量的头孢唑啉、庆大霉素、红霉素、呋喃妥因培养基内;置温箱内37℃培养并在不同时间检测各菌的数量及培养基抗生素活性。结果：临床治疗浓度的头孢唑啉或庆大霉素对金黄色葡萄球菌,铜绿假单胞菌及肺炎克雷伯菌的耐药性菌株的生长繁殖具有短暂的抑制作用,随后培养基内虽然仍保留较高的抗生素活性,但细菌的数量却逐渐增多。结论：临床治疗浓度的抗生素对耐药性细菌的生长繁殖仍然具有短暂的抑制作用,以致经验性用药能够使感染症患者的症状缓解,经验性用药不能有效杀灭病原菌,易造成病原菌在宿主体内长期携带和疾病的慢性过程。     

抗生素对大豆愈伤组织的诱导和生长的影响

用红霉素、头孢唑唑钠、头孢拉定、头孢霉素（国产和进口）等5种抗生素对农杆菌LBA4404进行抑菌试验,以头孢霉素的抑菌效果最好。头孢霉素作为抑菌剂用大于豆遗传转化试验时,在下胚轴浓度以300mg/L,在子叶节以500mg/L。大豆品种对卡那霉素的反应在出愈率上表现相似,在褐化率上表现有些不同。大豆不同外植体对卡那霉素的反应存在较大差异,以真叶反应最敏感,下胚轴反应最迟钝。在以卡那霉素作为抗性选择标记时,选择压力真叶和子叶节以50－100mg/L为好,下胚轴以100－200mg/L为宜。     

天蓝色链霉菌分化调控基因scrX的功能研究

克隆天蓝色链霉菌中一个新基因scrX并进行了序列分析, 利用基因破坏策略进行了该基因的功能研究. 结果表明, scrX基因由660个碱基组成, 编码产物是一个220个氨基酸残基的蛋白质;该基因含有3个在链霉菌中的稀有密码子--AAA, AAA和ATA, 是典型的在翻译水平上受到严紧调控的分化调控基因. 氨基酸序列同源性比较结果表明, scrX编码蛋白属于原核生物转录调控蛋白IclR家族.基因功能研究结果揭示, scrX基因在天蓝色链霉菌孢子形成中可能起正调控作用.     

天蓝色链霉菌调控基因tcrA功能的初步研究

天蓝色链霉菌的开放阅读框SCO5433编码一个含有TPR(Tetratricopeptide repeat)结构域的调控蛋白。该基因的阻断突变株表现出孢子颜色加深和色素产量增加的表型变化。孢子颜色的加深在以葡萄糖或甘露醇为碳源的MM培养基上表现明显;色素产量的增加在以甘露醇为碳源的MM培养基和MS培养基上表现最为明显;插片培养结合光学显微镜观察并没有发现突变株在形态分化上有显著变化;这些发现预示着可能存在一个SCO5433参与的调控途径,在一定条件下,这一途径对天蓝色链霉菌次级代谢可能起着负调控作用,而与形态分化无关。     

Combinatorial biosynthesis of lipopeptide antibiotics in Streptomyces roseosporus

Daptomycin is a cyclic lipopeptide antibiotic produced by Streptomyces roseosporus. Cubicin (daptomycin-for-injection) was approved in 2003 by the FDA to treat skin and skin structure infections caused by Gram-positive pathogens. Daptomycin is particularly significant in that it represents the first new natural product antibacterial structural class approved for clinical use in three decades. The daptomycin gene cluster contains three very large genes (dptA, dptBC, and dptD) that encode the nonribosomal peptide synthetase (NRPS). The related cyclic lipopeptide A54145 has four NRPS genes (lptA, lptB, lptC, and lptD), and calcium dependent antibiotic (CDA) has three (cdaPS1, cdaPS2, and cdaPS3). Mutants of S. roseosporus containing deletions of one or more of the NRPS genes have been trans-complemented with dptA, dptBC, and dptD by inserting these genes under the control of the ermEp* promoter into separate conjugal cloning vectors containing phiC31 or IS117 attachment (attP int) sites; delivering the plasmids into S. roseosporus by conjugation from Escherichia coli; and inserting the plasmids site-specifically into the chromosome at the corresponding attB sites. This trans-complementation system was used to generate subunit exchanges with lptD and cdaPS3 and the recombinants produced novel hybrid molecules. Module exchanges at positions D: -Ala(8) and D: -Ser(11) in the peptide have produced additional novel derivatives of daptomycin. The approaches of subunit exchanges and module exchanges were combined with amino acid modifications of Glu at position 12 and natural variations in lipid side chain starter units to generate a combinatorial library of antibiotics related to daptomycin. Many of the engineered strains produced levels of novel molecules amenable to isolation and antimicrobial testing, and most of the compounds displayed antibacterial activities.     

Proteomic analysis of the GlnR-mediated response to nitrogen limitation in Streptomyces coelicolor M145

GlnR is the global regulator of nitrogen assimilation in Streptomyces coelicolor M145 and other actinobacteria. Two-dimensional polyacrylamide gel electrophoresis analyses were performed to identify new GlnR target genes by proteomic comparison of wild-type S. coelicolor M145 and a ΔglnR mutant. Fifty proteins were found to be differentially regulated between S. coelicolor M145 and the ΔglnR mutant. These spots were identified by nanoHPLC–ESI-MS/MS and classified according to their cellular role. Most of the identified proteins are involved in amino acid biosynthesis and in carbon metabolism, demonstrating that the role of GlnR is not restricted to nitrogen metabolism. Thus, GlnR is supposed to play an important role in the global metabolic control of S. coelicolor M145.     

Identification of DNA amplifications near the center of the Streptomyces coelicolor M145 chromosome

Linear streptomycete chromosomes frequently undergo spontaneous gross DNA rearrangements at the terminal regions. Large DNA deletions of the chromosome ends are in many cases associated with tandemly reiterated DNA amplifications, found at the border of the deletable areas. In contrast to previous reports, we have discovered amplifications near the center of the Streptomyces coelicolor M145 chromosome. The detected amplified units of DNA are 19.9 kb and 16 kb in length and exist in copy numbers of 30 and 40, respectively. Both amplifications were located in the same region and share at least 3.6 kb.     

Pleiotropic effects of cAMP on germination, antibiotic biosynthesis and morphological development in Streptomyces coelicolor

In wild-type Streptomyces coelicolor MT1110 cultures, cyclic adenosine 3′,5′ monophosphate (cAMP) was synthesized throughout the developmental programme with peaks of accumulation both during germination and later when aerial mycelium and actinorhodin were being produced. Construction and characterization of an adenylate cyclase disruption mutant (BZ1) demonstrated that cAMP facilitated these developmental processes. Although pulse-labelling experiments showed that a similar germination process was initiated in BZ1 and MT1110, germ-tube emergence was severely delayed in BZ1 and never occurred in more than 85% of the spores. Studies of growth and development on solid glucose minimal medium (SMMS, buffered or unbuffered) showed that MT1110 and BZ1 produced acid during the first rapid growth phase, which generated substrate mycelium. Thereafter, on unbuffered SMMS, only MT1110 resumed growth and produced aerial mycelium by switching to an alternative metabolism that neutralized its medium, probably by reincorporating and metabolizing extracellular acids. BZ1 was not able to neutralize its medium or produce aerial mycelium on unbuffered SMMS; these defects were suppressed by high concentrations (>1 mM) of cAMP during early growth or on buffered medium. Other developmental mutants (bldA, bldB, bldC, bldD, bldG) also irreversibly acidified this medium. However, these bald mutants were not suppressed by exogenous cAMP or neutralizing buffer. BZ1 also differentiated when it was cultured in close proximity to MT1110, a property observed in cross-feeding experiments between bald mutants and commonly thought to reflect diffusion of a discrete positively acting signalling molecule. In this case, MT1110 generated a more neutral pH environment that allowed BZ1 to reinitiate growth and form aerial mycelium. The fact that actinorhodin synthesis could be induced by concentrations of cAMP (< 20 μM) found in the medium of MT1110 cultures, suggested that it may serve as a diffusible signalling molecule to co-ordinate antibiotic biosynthesis.     

Accumulation of S-adenosylmethionine induced oligopeptide transporters including BldK to regulate differentiation events in Streptomyces coelicolor M145

S-Adenosylmethionine (SAM), the major methyl donor in diverse biological processes, was recently found to be involved in the regulation of differentiation in streptomycetes. Exogenous SAM, in a quantity as low as 2muM, enhanced antibiotic production and inhibited morphological development of Streptomyces coelicolor M145. Total protein profiling of S. coelicolor M145 revealed that SAM enhanced the expression of oligopeptide-binding components related to ABC transporters that included BldK, a well-known regulatory factor in S. coelicolor differentiation. A radiolabeled SAM feeding experiment verified that exogenous SAM can be imported into the cell, which is under the control of the bld cascade. This study substantiated that BldK serves as a transducer of the SAM signal and uncovered the possible role of oligopeptide import in the regulation of Streptomyces differentiation, particularly in relation to SAM.     

Protoplast formation from submerged mycelium and from spore germinants of Streptomyces coelicolor

Stages in the formation of protoplasts from S. coelicolor strain A3(2) have been studied by transmission electron microscopy. Protoplasts liberated from submerged mycelial growth were variable in size and were released when digestion of the cell wall by lysozyme had completely or almost completely taken place. Protoplasts did not fully adopt the typical rounded shape until after release. A single region of cytoplasm gave rise to more than one protoplast unit. Protoplasts released from spore germinants escaped from the tip of the germ tube, which was the region of the cell wall most susceptible to digestion. Protoplasts derived from spore germinants were more consistent in size and rounded up more rapidly. If a cross-wall had formed in a germinant then it gave rise to separate protoplasts from each cellular compartment. Protoplasts of either type contained a single DNA region. These studies give an indication of the cellular organization of a streptomycete colony, which can be visualized as a multinucleated assemblage of cellular units in a common cytoplasm. The assembly of units separates into a number of protoplasts on digestion of the cell wall.