放线菌次级代谢途径的设计

近来有关放线菌次线产物生物合成的分子遗传学和生物化学方面的进展为我们改造其代谢途径提供了一个明确的方向。近年来,对微生物的初级代谢途径进行基因改造取得了成功,但放线菌的次级代谢工程产物却都没有达到中试或生产规模。     

极地放线菌次级代谢产物及药理活性研究进展CSCD

极地微生物处在南北极严峻、极端的自然环境下,体内的一些“沉默基因”可能会被激活,形成了独特的生存机制和生物合成途径,能够产生化学结构多样、活性显著的次级代谢产物。近年来,从极地放线菌中分离鉴定出一系列结构新颖、活性突出的次级代谢产物。本文综述了1999至2021年间的极地放线菌次级代谢产物及药理活性的研究进展,将极地放线菌来源的104个次级代谢产物按肽类、生物碱类、萜类、大环内酯类、聚酮类、大环内酰胺类等化学类别,着重介绍结构新颖和活性突出的次级代谢产物,为进一步开发利用极地放线菌、研发创新药物提供参考。     

培菌昆虫相关放线菌的次级代谢产物研究进展

昆虫共生微生物是一种特殊的微生物资源,其中放线菌在昆虫肠道、体表和巢穴中广泛分布。近年来,人们从培菌昆虫来源的放线菌中分离得到多种新型化合物,可以选择性抑制菌圃的致病真菌,部分还对植物致病真菌、昆虫致病真菌、人类病原菌和癌细胞有抑制活性。因此,研究培菌昆虫相关微生物不仅有助于了解宿主与微生物的共生机制,还能发掘新的活性物质,用于生物农药、生物医药的开发。本文对培菌昆虫来源放线菌次级代谢产物的研究进展进行了综述。     

非定向激活沉默基因簇挖掘链霉菌活性次级代谢产物研究进展

链霉菌(Streptomyces spp.)是活性次级代谢产物的主要生产菌,在发现结构新颖化合物上具有重大潜力。随着对链霉菌基因组数据的深入分析,发现大量产次级代谢产物合成基因簇处于沉默状态,因此利用非定向和定向策略激活链霉菌中沉默基因簇、挖掘结构新颖化合物成为当前的主要手段。本文主要综述了培养基组成改变或培养条件优化、共培养、添加化学激发子及全局性调控等非定向策略在挖掘链霉菌次级代谢产物中的应用以及取得的进展,以期为链霉菌次级代谢产物高效开发提供参考。     

利用合成生物学技术深入挖掘放线菌中活性次级代谢产物

放线菌是一类能够产生丰富生物小分子药物的微生物, 对人类的健康事业做出了杰出的贡献。但近几十年来, 来源于微生物并最终上市的药物越来越少, 而病原菌的抗药性问题却越发严重, 人们对新药的期待越来越迫切。本文介绍了近十年里发展迅速的合成生物学对微生物次级代谢产物研发的促进作用。合成生物学以工程化的思想对生命系统进行设计与改造, 使传统方法难以获取的放线菌次级代谢产物通过外源宿主得以产生, 充分利用了自然界的资源; 此外, 对次级代谢基因簇的合理设计和对生物元件的应用不仅使人们获得了自然界中原本不存在的新化合物, 还能使某些具有广泛应用价值药物的产量得以显著提高; 最后, 本文还介绍了合成生物学领域近些年DNA组装的新技术和新方法, 为从事次级代谢产物研发的工作者提供便利。     

提高放线菌次级代谢产物产量方法的研究进展

放线菌的次级代谢产物一直是抗生素等药物开发的重要来源。通过基因技术来提高放线菌次级代谢产物产量的方法已被广泛接受。结合近年的研究成果,概述了利用基因技术过表达或者敲除代谢途经关键基因及加倍基因簇,诱变核糖体相关蛋白的基因,以及异源表达外源基因簇等提高次级代谢产物产量的方法。旨在为寻找新药及改造高产菌株提供相应的技术参考。     

海洋放线菌Streptomyces variabilisstrain 6-1次级代谢产物的研究

目的：对来自海洋软珊瑚的链霉菌6-1（Streptomyces variabilisstrain6-1）进行次级代谢产物的分离和鉴定,寻找具有生物活性的化合物,为人类健康服务。方法：采用液体培养基对分自海洋软珊瑚Scleronephthya sp中的链霉菌6．1（Streptomyces vafiabilisstrain6-1）进行发酵培养,用乙酸乙酯对发酵液进行萃取;采用半制备高效液相色谱（semi-preparative HPLC）分离方法对乙酸乙酯萃取物进行分离纯化,得到单体化合物;运用电喷雾质谱（ESI—MS）、核磁共氢振（1HNMR）、核磁共振碳谱（13C NMR）和物理性质对所得单体化合物进行结构鉴定。结果：从海洋链霉菌6-1（strain6-1）发酵液的乙酸乙酯萃取物中分离得到3个单体化合物,分别鉴定为：7,4’-二羟基异黄酮（1）、5,7,4’-三羟基异黄酮（2）和丁烯酸内酯-I（3）。结论：丁烯酸内酯．I是从链霉菌属首次分离得到,化合物1和2均是从Streptomyces variabilis中首次分离得到;变异链霉菌6-1（Stmptomyces variabilis strain6-1）可以作为活性化合物3（丁烯酸内酯-I）的重要来源。     

放线菌JXJ-45固体发酵产物化学成分研究

通过对放线菌JXJ-45菌株固体发酵物的初步分离纯化,得到3个化合物,由波谱解析鉴定为:2,6-二甲基-2,6-二羟基-庚-4-酮(1),N-[2-(4-羟苯基)-乙基]乙酰胺(2),熊果苷(3).化合物2首次在放线菌中发现,化合物3首次在微生物中发现,是国际公认的高效祛斑美白剂.     

海洋放线菌Streptomyces variabilis strain 6-1次级代谢产物的研究

目的:对来自海洋软珊瑚的链霉菌6-1(Streptomyces variabilis strain 6-1)进行次级代谢产物的分离和鉴定,寻找具有生物活性的化合物,为人类健康服务。方法:采用液体培养基对分自海洋软珊瑚Scleronephthya sp中的链霉菌6-1(Streptomyces variabi-lis strain 6-1)进行发酵培养,用乙酸乙酯对发酵液进行萃取;采用半制备高效液相色谱(semi-preparative HPLC)分离方法对乙酸乙酯萃取物进行分离纯化,得到单体化合物;运用电喷雾质谱(ESI-MS)、核磁共氢振(1H NMR)、核磁共振碳谱(13C NMR)和物理性质对所得单体化合物进行结构鉴定。结果:从海洋链霉菌6-1(strain 6-1)发酵液的乙酸乙酯萃取物中分离得到3个单体化合物,分别鉴定为:7,4'-二羟基异黄酮(1)、5,7,4'-三羟基异黄酮(2)和丁烯酸内酯-Ⅰ(3)。结论:丁烯酸内酯-Ⅰ是从链霉菌属首次分离得到,化合物1和2均是从Streptomyces variabilis中首次分离得到;变异链霉菌6-1(Streptomyces variabilis strain 6-1)可以作为活性化合物3(丁烯酸内酯-Ⅰ)的重要来源。     

放线菌次级代谢途径的设计

近来有关放线菌次级产物生物合成的分子遗传学和生物化学方面的进展为我们改造其代谢途径提供了一个明确的方向。近年来,对微生物的初级代谢途径进行基因改造取得了成功,但放线菌的次级代谢工程产物却都没有达到中试或生产规模。进展如此缓慢的主要原因是放线菌自身复杂的代谢途径以及细胞循环中复杂的调节方式及其特异性。目前人们着力于通过基因操作改造酶,从而重新设计以其催化产物为基本骨架的代谢途径,最终产生修饰的或新的天然终产物。本文将讨论达到此目的的几种设计策略。     

植物内生放线菌多样性及其活性代谢产物的研究进展

植物内生放线菌是一类能与宿主植物长期协同生长且不引起植物感染的特殊微生物.其作为植物内生微生物资源的重要组成部分,不仅分布广泛,资源丰富,绝大部分可产生与宿主植物相同或相似的次生代谢产物,而且这些次级代谢产物往往丰富多样并具有多种生物活性,是寻找和开发新型先导化合物的重要资源,具有重要的研究意义和潜在的应用价值.植物内...     

极端环境放线菌TRM45037鉴定及次生代谢产物分析

为了明确新疆罗布泊极端环境放线菌TRM45037的分类,挖掘其次生代谢产物及其生物活性,本研究通过形态学观察、生物学特征和16S rDNA序列分析等方法,确定TRM45037为中度嗜盐性拟诺卡氏菌,与菌株Nocardiopsis dassonvilei subsp. dassonvillei DSM 4311(T) 的相似度为99.2%。采用硅胶柱色谱、凝胶柱色谱和高效液相色谱等方法对次生代谢产物进行了分离纯化,并通过核磁共振谱（1H NMR,13C NMR）结合相关文献确定化合物结构。从TRM45037发酵液中分离出4个化合物,分别鉴定为3, 4-二氢-6, 8-二羟基-3-甲基异香豆素、2-甲基-1, 4-苯二醇、邻苯二甲酸（2-乙基己基）二酯和1, 6-二羟基吩嗪。抑菌活性测定表明, 3, 4-二氢-6, 8-二羟基-3-甲基异香豆素和2-甲基-1, 4-苯二醇对金黄色葡萄球菌具有抑制作用,其最小抑菌浓度(minimal inhibitory concentration, MIC)值分别为0.78 mg/mL和0.39 mg/mL,最小杀菌浓度(minimum fugicide concentration, MFC)值分别为1.56 mg/mL和0.78 mg/mL。本研究结果可为嗜盐放线菌活性次生代谢产物挖掘提供理论依据,对新疆极端环境放线菌资源保护与利用具有切实意义。     

海草生物活性成分的研究进展

本文综述了海草中生物活性成分的研究进展,其化学成分类型主要是苯酚类、黄酮类和二萜类化合物,生物活性主要包括抗菌、抗氧化、抗病毒、细胞毒性、抑制海藻生长和抗污损等.     

Red Soils Harbor Diverse Culturable Actinomycetes That Are Promising Sources of Novel Secondary Metabolites

Red soils, which are widely distributed in tropical and subtropical regions of southern China, are characterized by low organic carbon, high content of iron oxides, and acidity and, hence, are likely to be ideal habitats for acidophilic actinomycetes. However, the diversity and biosynthetic potential of actinomycetes in such habitats are underexplored. Here, a total of 600 actinomycete strains were isolated from red soils collected in Jiangxi Province in southeast China. 16S rRNA gene sequence analysis revealed a high diversity of the isolates, which were distributed into 26 genera, 10 families, and 7 orders within the class Actinobacteria; these taxa contained at least 49 phylotypes that are likely to represent new species within 15 genera. The isolates showed good physiological potentials for biosynthesis and biocontrol. Chemical screening of 107 semirandomly selected isolates spanning 20 genera revealed the presence of at least 193 secondary metabolites from 52 isolates, of which 125 compounds from 39 isolates of 12 genera were putatively novel. Macrolides, polyethers, diketopiperazines, and siderophores accounted for most of the known compounds. The structures of six novel compounds were elucidated, two of which had a unique skeleton and represented characteristic secondary metabolites of a putative novel Streptomyces phylotype. These results demonstrate that red soils are rich reservoirs for diverse culturable actinomycetes, notably members of the families Streptomycetaceae, Pseudonocardiaceae, and Streptosporangiaceae, with the capacity to synthesize novel bioactive compounds.     

红树林放线菌研究进展

红树林是生长在热带和亚热带海岸潮间带的一种独特植物群落,构成陆地与海洋间的过渡,具有水分高、盐分高、耐缺氧等特征的特殊生态系统及很高的生物多样性,是筛选和发现放线菌新物种和药用生物活性次级代谢产物的宝贵资源。在物种鉴定方面,放线菌分类学是目前红树林放线菌物种鉴定的重要手段。本文综述了红树林放线菌物种鉴定主要方法、红树林放线菌生态环境和物种分布特征、生物学活性及其次级代谢产物等,并展望了该领域研究与发展的重要问题与趋势。     

Pathway Engineering in Secondary Metabolite-Producing Actinomycetes

Abstract

Actinomycetes represent the microbial group richest in production of variable secondary metabolites. These mostly bioactive molecules are the end products of complex multistep biosynthetic pathways. Recent progress in the molecular genetics and biochemistry of the biosynthetic capacities of actinomycetes enables first attempts to redesign these pathways in a directed fashion. However, in contrast to several examples of designed biochemical improvement of primary metabolic processes in microorganisms, none of the products or strains derived from pathway engineering in actinomycetes discussed herein have reached pilot or production scale. The main reasons for this slow progress are the complicated pathways themselves, their complex regulation during the actinomycete cell cycle, and their uniqueness, as most pathways and products are specific for a strain rather than for a given species or larger taxonomic group. However, the modular use of a minimum of very similar enzymes and their conversion of similar intermediates to form the building blocks for the production of a maximum of divergent end products gives hope for the future application of these genetic models for the redesign of complex pathways for modified or new natural products. Several strategies that can be followed to reach this aim are discussed, mainly for the variable 6-deoxyhexose metabolism as an ubiquitously applicable example.     

海洋沉积物来源链霉菌属次生代谢产物及其生物活性研究进展

海洋沉积物是营养较为丰富的微生物栖息地,近年来从海洋沉积物中分离培养出了大量海洋链霉菌,从中还发现了一些新的属种。人们已从海洋沉积物来源链霉菌属中发现了许多具有药用价值的活性化合物,有力推动了海洋天然产物化学的发展,并为新药研发提供基础。本文就海洋沉积物来源链霉菌属次生代谢产物的结构类型及其生物活性进行简要综述。     

Use of Direct-Infusion Electrospray Mass Spectrometry To Guide Empirical Development of Improved Conditions for Expression of Secondary Metabolites from Actinomycetes

A major barrier in the discovery of new secondary metabolites from microorganisms is the difficulty of distinguishing the minor fraction of productive cultures from the majority of unproductive cultures and growth conditions. In this study, a rapid, direct-infusion electrospray mass spectrometry (ES-MS) technique was used to identify chemical differences that occurred in the expression of secondary metabolites by 44 actinomycetes cultivated under six different fermentation conditions. Samples from actinomycete fermentations were prepared by solid-phase extraction, analyzed by ES-MS, and ranked according to a chemical productivity index based on the total number and relative intensity of ions present in each sample. The actinomycete cultures were tested for chemical productivity following treatments that included nutritional manipulations, autoregulator additions, and different agitation speeds and incubation temperatures. Evaluation of the ES-MS data from submerged and solid-state fermentations by paired t test analyses showed that solid-state growth significantly altered the chemical profiles of extracts from 75% of the actinomycetes evaluated. Parallel analysis of the same extracts by high-performance liquid chromatography–ES-MS–evaporative light scattering showed that the chemical differences detected by the ES-MS method were associated with growth condition-dependent changes in the yield of secondary metabolites. Our results indicate that the high-throughput ES-MS method is useful for identification of fermentation conditions that enhance expression of secondary metabolites from actinomycetes.