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。     

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。     

微生物来源天然产物中的β-甲基色氨酸单元及其生物合成研究进展

微生物在次级代谢过程中通常会产生结构复杂、活性多样的天然产物。这些天然产物是新药发展的基础,亦可作为先导化合物或重要的药效基团用于药物研发。结构多样的氨基酸单元是参与合成复杂多样天然产物的重要前体。天然产物中的β-甲基氨基酸单元不仅可以赋予其生物活性,还能增强其生物稳定性而不被肽酶水解。本文综述了含有β-甲基氨基酸单元的天然产物,尤其对含有β-甲基色氨酸单元的天然产物生物合成途径进行了阐释。对β-甲基色氨酸单元生物合成途径的理解结合基因组数据有助于进行新结构天然产物的挖掘,并为运用代谢科学理念和合成生物学技术开发含有该单元的新化合物提供理论基础和可操作遗传元件。     

微生物来源天然产物中的β-甲基色氨酸单元及其生物合成研究进展

微生物在次级代谢过程中通常会产生结构复杂、活性多样的天然产物。这些天然产物是新药发展的基础,亦可作为先导化合物或重要的药效基团用于药物研发。结构多样的氨基酸单元是参与合成复杂多样天然产物的重要前体。天然产物中的β-甲基氨基酸单元不仅可以赋予其生物活性,还能增强其生物稳定性而不被肽酶水解。本文综述了含有β-甲基氨基酸单元的天然产物,尤其对含有β-甲基色氨酸单元的天然产物生物合成途径进行了阐释。对β-甲基色氨酸单元生物合成途径的理解结合基因组数据有助于进行新结构天然产物的挖掘,并为运用代谢科学理念和合成生物学技术开发含有该单元的新化合物提供理论基础和可操作遗传元件。     

核糖体工程与微生物次级代谢产物合成

核糖体工程通过对微生物次级代谢产物合成相关基因表达的两个重要元件——核糖体或RNA聚合酶进行修饰和改造,带来其结构和功能上的改变,进而影响次级代谢产物的合成。因此,向核糖体组成元件中引入特定的突变就能够有效地调节次级代谢产物的合成,通过该技术改造有重要商业价值的工业微生物,提高其次级代谢产物(如抗生素)的合成能力,对于微生物次生代谢产物研发及产业化具有重要的科研与经济价值。     

微生物次级代谢产物抗辣椒疫霉的研究进展CSCD

辣椒疫霉(Phytophthora capsici)是一种破坏性极强的蔬菜作物病原菌,会使植物患疫病,已对农业生产造成巨大的经济损失。微生物次级代谢产物可通过破坏细胞膜通透性、干扰蛋白质合成以及诱导植物产生抗性等机制来抑制辣椒疫霉,在防治辣椒疫霉和其他植物病原菌中发挥着重要作用。微生物源次级代谢产物如吩嗪-1-羧酸是我国自主创制的绿色杀菌剂申嗪霉素(shenqinmycin)的主要成分,对包括辣椒疫病在内的多种植物病害有良好的防治效果。因此,微生物次级代谢产物的应用是生物防治中控制植物病害的有效手段,也是实现农业绿色发展的有效策略。本文以微生物类型(细菌、放线菌和真菌)为主线,简要综述了近二十年来94种具有抗辣椒疫霉活性的微生物次级代谢产物的来源、抗菌效果和部分次级代谢产物的抗菌机理,以期为微生物源次级代谢产物抗辣椒疫霉的研究与开发提供参考。     

抗生素生物合成基因簇克隆策略的研究进展

抗生素是一类具有多种生物活性的微生物次级代谢产物,自然界中60%以上的抗生素由放线菌所产生。克隆抗生素生物合成基因簇可为阐明其生物合成机制,结构改造,产量提高提供重要信息。在后基因组时代,新的抗生素基因簇克隆策略不断涌现,大大提高了微生物重要次级代谢产物生物合成基因簇的克隆效率,并缩短了抗生素的发现周期。本文综述了抗生素生物合成基因簇克隆策略的研究进展,包括近年来发展的后基因组的克隆策略。     

贝莱斯芽孢杆菌生防次级代谢产物研究进展

贝莱斯芽孢杆菌（Bacillus velezensis）是生防芽孢杆菌中的重要代表,作为微生物农药的核心菌种,已被广泛应用于作物病害生物防治。贝莱斯芽孢杆菌具有植物内生性,其生防作用机制主要包括产生次级代谢产物对抗植物病原物;改善宿主植物根际微生物群落,促进宿主营养和生长;激发宿主植物产生防御反应,诱导植物获得系统抗性。其中,产生次级代谢产物是其最重要的生防作用机制。贝莱斯芽孢杆菌含有多个编码生物合成次级代谢产物的基因簇,其中包括编码聚酮化合物合酶（PKS）和非核糖体肽合成酶（NRPS）的基因簇,同时存在核糖体途径合成次级代谢产物基因簇。通过非核糖体途径可产生脂肽类化合物、聚酮类化合物、二肽和铁载体;通过核糖体途径产生小菌素、细菌素、羊毛硫抗生素。这些具有生物活性的次级代谢产物成为了天然新药和候选抗生素的储存库,对于解析生防菌作用机制具有重要意义。本文综述了贝莱斯芽孢杆菌的命名与更迭,产生次级代谢产物的类型、合成与调控基因以及靶标病原菌,以期为生防菌株的改良和生物农药的研发提供参考。     

海绵共附生真菌多样性及其次级代谢产物的研究进展

海绵由于其独特的生理结构、进食方式使其体内部聚集了大量的微生物,这些微生物产生了多种结构新颖的生物活性物质,因此海绵及其共附生微生物的研究成为了海洋药物研发的热点。就海绵中共附生真菌的分布情况,新技术的应用及其生物次级代谢产物的生物活性展开综述。     

真菌诱导子在发酵工业中的应用现状及展望

真菌诱导子是一类能诱导植物和微生物产生次级代谢产物的活性物质,它一经识别,将通过信号转导途径,引起相关基因表达发生变化,从而调节次级代谢产物合成途径中相关酶的活性,诱导特定次级代谢产物的积累。近年来国内外在真菌诱导子诱导途径及机制方面进行了深入研究,同时在生物工业领域,尤其在发酵工业中的应用也引起了广泛关注。以下结合本实验室的研究工作,重点介绍了真菌诱导子在植物和微生物细胞次级代谢产物合成方面的应用现状、诱导机制和存在的问题及展望。     

海洋动物来源微生物的共培养研究进展

近年来,由于一些新疾病的发生和细菌耐药性的出现,微生物来源次级代谢产物的筛选重复率越来越高,微生物一些代谢基因在现有实验室条件下无法表达,所以需要发现新的微生物资源,同时找到激活微生物代谢产物基因的方法。海洋动物体内蕴含着大量的共附生微生物资源,可以产生很多具有生物活性的化合物,是潜在的药用资源。本文综述了近年来海洋动物(海鞘、海绵、珊瑚和海葵等)来源的微生物进行共培养的研究策略,包括共培养菌株的选择、共培养条件、群体感应和信号分子对共培养菌株的影响,以及不同种类微生物间的共培养实例。共培养与单培养相比,增加了次级代谢产物的种类,提高了次级代谢产物的生物活性或产量。共培养的研究有助于发现新的海洋动物来源微生物的活性天然产物,为海洋药物的开发提供新思路。     

Bioactive secondary metabolites produced by microorganisms associated with plants

In the past few decades groups of scientists have focused their study on relatively new microorganisms called endophytes. By definition these microorganisms, mostly fungi and bacteria, colonise the intercellular spaces of the plant tissues. The mutual relationship between endophytic microorganisms and their host plants, taxanomy and ecology of endophytes are being studied. Some of these microorganisms produce bioactive secondary metabolites that may be involved in a host-endophyte relationship. Recently, many endophytic bioactive metabolites, known as well as new substances, possesing a wide variety of biological activities as antibiotic, antitumor, antiinflammatory, antioxidant, etc. have been identified. The microorganisms such as endophytes may be very interesting for biotechnological production of bioactive substances as medicinally important agents. Therefore the aim of this review is to briefly characterize endophytes and summarize the structuraly different bioactive secondary metabolites produced by endophytic microorganisms as well as microbial sources of these metabolites and their host plants.     

Current strategies to induce secondary metabolites from microbial biosynthetic cryptic gene clusters

The genome of actinomycetes and several other microorganisms are endowed with many cryptic gene clusters that can code for previously undetected, a plethora of complex secondary metabolites. Under standard laboratory controlled conditions, the genes regulating these biosynthetic clusters are expressed at very low levels or remain phenotypically cryptic (silent). Over the past several decades, multi-drug-resistant bacteria have been observed with increased frequency, posing a significant threat to human health worldwide. The present alarming situation urgently calls for concerted global efforts for the discovery of new antimicrobials. The present situation, if not controlled, will take us again to the pre-antibiotic era. Today, in the post-genomic era, various new strategies such as the activation of cryptic gene clusters in microorganisms rejuvenate a new conviction in the field of natural product research that may lead to the identification of yet-unidentified novel secondary metabolites of therapeutic and other use. Decryptification of this versatile endogenous genetic reservoir may provide in the near future the more concrete rationale for antibiotic discovery. The present review is an attempt to provide a comprehensive detail, outlining current strategies that have been shown successful to activate cryptic biosynthetic gene clusters in microorganisms.     

Application of molecular biology for the discovery of biosynthetic genes of polyketide and peptide antibiotics produced by actinomycetes

Actinomycetes are currently the main source of antibiotics. Genome sequencing reveals the presence in these organisms of multiple gene clusters for the synthesis of yet unidentified secondary metabolites. Technological advances in DNA isolation, cloning and sequencing, as well as development of bioinformatics, facilitate large scale search for new gene clusters in organisms with unknown genome sequence and in environmental DNA. Methods used for detection of polyketide synthase (PKS) and non-ribosomal peptide synthetase (NRPS) genes are described in this article. New PKS and NRPS genes give access to new biologically active natural products which can become drugs or substrates for chemical modifications. Even more inspiring is their use in combinatorial biosynthesis to produce a variety of compounds with rationally designed structures.     

Bioactive compounds from marine actinomycetes

Actinomycetes are one of the most efficient groups of secondary metabolite producers and are very important from an industrial point of view. Among its various genera, Streptomyces, Saccharopolyspora, Amycolatopsis, Micromonospora and Actinoplanes are the major producers of commercially important biomolecules. Several species have been isolated and screened from the soil in the past decades. Consequently the chance of isolating a novel actinomycete strain from a terrestrial habitat, which would produce new biologically active metabolites, has reduced. The most relevant reason for discovering novel secondary metabolites is to circumvent the problem of resistant pathogens, which are no longer susceptible to the currently used drugs. Existence of actinomycetes has been reported in the hitherto untapped marine ecosystem. Marine actinomycetes are efficient producers of new secondary metabolites that show a range of biological activities including antibacterial, antifungal, anticancer, insecticidal and enzyme inhibition. Bioactive compounds from marine actinomycetes possess distinct chemical structures that may form the basis for synthesis of new drugs that could be used to combat resistant pathogens.     

Importance of microbial natural products and the need to revitalize their discovery

Microbes are the leading producers of useful natural products. Natural products from microbes and plants make excellent drugs. Significant portions of the microbial genomes are devoted to production of these useful secondary metabolites. A single microbe can make a number of secondary metabolites, as high as 50 compounds. The most useful products include antibiotics, anticancer agents, immunosuppressants, but products for many other applications, e.g., antivirals, anthelmintics, enzyme inhibitors, nutraceuticals, polymers, surfactants, bioherbicides, and vaccines have been commercialized. Unfortunately, due to the decrease in natural product discovery efforts, drug discovery has decreased in the past 20 years. The reasons include excessive costs for clinical trials, too short a window before the products become generics, difficulty in discovery of antibiotics against resistant organisms, and short treatment times by patients for products such as antibiotics. Despite these difficulties, technology to discover new drugs has advanced, e.g., combinatorial chemistry of natural product scaffolds, discoveries in biodiversity, genome mining, and systems biology. Of great help would be government extension of the time before products become generic.     

Harnessing the potential of chemical defenses from antimicrobial activities

Resistance to the drugs used in the treatment of many infectious diseases is increasing, while microbial infections are being found to be responsible for more life-threatening diseases than previously thought. Despite a large investment in the invention and application of high-throughput screening techniques involving miniaturization and automation, and a diverse array of strategies for designing and constructing various chemical libraries, relatively few new drugs have resulted. Natural products, however, have been a major source of drugs for centuries. Since some of them are produced by organisms as a result of selection in favour of improved defense against competing deleterious microorganisms, in principle they would be less likely to incur resistance. Furthermore, the production of those defensive secondary metabolites is inducible because their original function is a response to environmental challenges. Moreover, symbioses, co-habitation associations between two or more different species of organisms, are universal in nature, and the production of secondary metabolites by symbiotic microbes may be an important adaptation allowing microbes to affect their hosts. Therefore, co-culture strategies, using combinations of plant cell-pathogenic microbes, plant cell-endophytes (or symbionts), and symbiont-pathogenic microbes, based on the principles of chemical defense and the known mechanisms of organism interactions, may be an efficient general approach in the search for new anti-microbial drugs.     

嗜盐微生物在生物医药领域的应用研究进展

近年来抗生素耐药性问题日趋严重,患癌人数也在逐年增加,亟需开发新型药物。嗜盐微生物作为一类特殊的极端环境微生物,具有代谢多样性丰富、营养需求较低和能适应恶劣条件等特点,是发现新型药物的希望。目前,国内外学者已从嗜盐微生物中分离出了多种代谢产物和酶,具有明显的抗菌和/或抗肿瘤等活性。文中综述了嗜盐微生物及其相关产物在抗菌、抗炎、抗肿瘤、抗氧化、生物医学材料以及药物载体等生物医学方面的作用,尤其对近年来在嗜盐微生物中发现的新型抗菌和抗肿瘤物质以及嗜盐微生物特有的代谢产物四氢嘧啶等进行了总结,并对其后续在生物医药领域的开发和产业化应用进行了展望。     

Marine Sponges as Pharmacy

Marine sponges have been considered as a gold mine during the past 50 years, with respect to the diversity of their secondary metabolites. The biological effects of new metabolites from sponges have been reported in hundreds of scientific papers, and they are reviewed here. Sponges have the potential to provide future drugs against important diseases, such as cancer, a range of viral diseases, malaria, and inflammations. Although the molecular mode of action of most metabolites is still unclear, for a substantial number of compounds the mechanisms by which they interfere with the pathogenesis of a wide range of diseases have been reported. This knowledge is one of the key factors necessary to transform bioactive compounds into medicines. Sponges produce a plethora of chemical compounds with widely varying carbon skeletons, which have been found to interfere with pathogenesis at many different points. The fact that a particular disease can be fought at different points increases the chance of developing selective drugs for specific targets.     

Endophytic Penicillium species secretes mycophenolic acid that inhibits the growth of phytopathogenic fungi

The worldwide demand for reduced and restricted use of pesticides in agriculture due to serious environmental effects, health risks and the development of pathogen resistance calls for the discovery of new bioactive compounds. In the medical field, antibiotic-resistant microorganisms have become a major threat to man, increasing mortality. Endophytes are endosymbiotic microorganisms that inhabit plant tissues without causing any visible damage to their host. Many endophytes secrete secondary metabolites with biological activity against a broad range of pathogens, making them potential candidates for novel drugs and alternative pesticides of natural origin. We isolated endophytes from wild plants in Israel, focusing on endophytes that secrete secondary metabolites with biological activity. We isolated 302 different endophytes from 30 different wild plants; 70 of them exhibited biological activity against phytopathogens. One biologically active fungal endophyte from the genus Penicillium, isolated from a squill (Urginea maritima) leaf, was further examined. Chloroform-based extraction of its growth medium was similarly active against phytopathogens. High-performance liquid chromatography separation followed by gas chromatography/mass spectrometry analysis revealed a single compound—mycophenolic acid—as the main contributor to the biological activity of the organic extract.