多烯大环内酯类抗生素生物合成基因簇中调控因子的研究进展

多烯大环内酯类抗生素具有良好的抗真菌活性,广泛应用于医疗卫生、食品加工和农业生产领域。随着高通量测序技术和生物信息学技术的发展,越来越多的链霉菌抗生素生物合成基因簇被发现和鉴定,调控因子作为生物合成基因簇中的重要组成部分,在庞大复杂的调控网络中起着至关重要的作用。本文总结了链霉菌中重要的调控因子类型,综述了多烯大环内酯类抗生素生物合成基因簇中调控因子的生物学功能、结合位点、作用机制等研究进展,并展望了后续研究工作。     

硫链丝菌素诱导非小细胞肺癌细胞自噬性死亡

自噬在细胞的生命过程中起了非常重要的作用,它能通过清除受损的细胞器和过多的蛋白质以维持细胞内环境稳定基本能量代谢的稳定。然而,自噬的持续激活可导致细胞器以及必需的蛋白质的过度消耗,导致不依赖caspase的自噬性细胞死亡。因此,通过这种自噬途径诱导细胞死亡可能是癌症治疗的一种新方法。在本研究我们发现,硫链丝菌素能够减少非小细胞肺癌PC-9细胞的细胞活力和诱导细胞凋亡。另外,我们发现硫链丝菌素诱导了PC-9细胞自噬。此外,我们也发现硫链丝菌素诱导的细胞凋亡可以被自噬抑制剂3-甲基腺嘌呤(3-MA)阻止。这些结果表明,硫链丝菌素促进人体非小细胞肺癌细胞的自噬性死亡。在本研究我们的发现也提示硫链丝菌素联合自噬诱导剂可能在临床上对治疗人非小细胞肺癌有效。     

抗生素耐药性的研究进展与控制策略

抗生素是治疗细菌感染的有效药物,然而抗生素在人类医学及农业生产中的大规模使用催生了细菌耐药性在环境中的快速扩散和传播,特别是多种抗生素的联合使用更是促进了多重耐药性的产生,严重威胁着人类和动物健康及食品与环境安全,相关问题已经引起人们的警觉.因此新研究主要集中在以下几方面:利用组学及合成生物学等方法挖掘并合成新型抗生素...     

大环内酯类抗生素微生物降解的研究进展

大环内酯类抗生素是一类以大环内酯为母核的广谱抗生素。近些年,由于人们对其不规范的生产和使用,抗生素污染成为了重要的环境问题。大量研究表明,微生物降解是现阶段处理抗生素污染的最理想方法。为进一步推动大环内酯类抗生素生物降解的研究,文中概述了大环内酯类抗生素的环境污染现状、微生物降解菌株、降解酶、降解途径和降解大环内酯类抗生素的微生物处理方法,并对大环内酯类抗生素生物降解亟待解决的瓶颈问题进行了讨论,以期为微生物降解后续研究提供参考。     

Plant Terpenoids： Biosynthesis and Ecological Functions

Among plant secondary metabolites terpenolds are a structurally most diverse group; they function as phytoalexins In plant direct defense, or as signals In Indirect defense responses which involves herbivores and their natural enemies. In recent years, more and more attention has been paid to the Investigation of the ecological role of plant terpenolds. The biosynthesis pathways of monoterpenes, sesquiterpenes, and diterpenes Include the synthesis of C5 precursor isopentenyl diphosphate （IPP） and Its allylic isomer dlmethylallyl dlphosphate （DMAPP）, the synthesis of the immediate diphosphate precursors, and the formation of the diverse terpenoids. Terpene synthases （TPSs） play a key role In volatile terpene synthesis. By expression of the TPS genes, significant achievements have been made on metabolic engineering to Increase terpenoid production. This review mainly summarizes the recent research progress In elucidating the ecological role of terpenoids and characterization of the enzymes Involved in the terpenold biosynthesis. Spatial and temporal regulations of terpenoids metabolism are also discussed.     

一个硫链丝菌素抗性基因标记、用于DNA导入链霉菌的pSET152衍生载体

[目的]在链霉菌PCR-targeting系统中,安普霉素是使用最普遍的选择标记.然而在基因敲除阶段利用安普霉素选择标记后,在遗传补偿时就不能使用相同选择标记的许多重要载体,如pSET152.这常给研究带来不便,特别是当研究对象基因如一些调控基因,其生理功能对剂量敏感时更是如此.基于此,拟以pSET152为基础构建一个不以安普霉素为抗性标记的通用整合型载体.[方法]利用融合PCR和λRed重组等方法构建载体.[结果]来自pHZ1358上的硫链丝菌素抗性基因tsr和来自pCR2.1的氨苄抗性基因bla以“tsr在前bla在后”的次序融合.融合后的抗性片段替换pSET 152上的安普霉素抗性基因aac(3)-Ⅳ,从而获得新载体pGIM6626.利用该载体将删除的榴菌素最小聚酮合酶基因重新导入到Streptomyces vietnamensis突变株中,该突变株恢复了产榴菌素的能力,证实了该载体的有效性.[结论]构建了一个新的pSET152衍生载体pGIM6626.该载体包含氨苄和硫链丝菌素抗性基因,分别在大肠杆菌和链霉菌中作为选择标记.pGIM6626与pSET152用途相似,但前者由于与PCR-targeting系统不存在选择标记的冲突而与该系统更兼容.     

糖肽抗生素天然产物研究进展

糖肽抗生素(Glycopeptide antibiotics, GPA)是临床上用于治疗耐药革兰氏阳性病原菌感染的重要药物.不幸的是,随着GPA的应用,临床上也陆续发现了万古霉素耐药肠球菌与金黄色葡萄球菌感染的病例,使得相关病原菌感染的医治愈发困难.为了应对糖肽抗生素耐药的临床威胁,我们需要持续不断地去开发新一代高效GPA.微生物天然产物是临床药物的主要来源,其中GPA均来自放线菌天然产物.(宏)基因组测序显示,放线菌基因组中携带大量编码GPA生物合成的基因簇,是新一代GPA的资源宝库.本文系统性地回顾了GPA的发现史,总结了GPA的生物合成机制、作用机制、耐药机制以及最新的发掘方法与策略,以期为GPA的开发提供参考.     

A族乙型溶血性链球菌对大环内酯类抗生素耐药的研究进展

A族乙型溶血性链球菌（group A streptococcus pyogens,GAS）是链球菌中致病性最强的一种,广泛存在于自然界、人及动物粪便和健康人的鼻咽部,是急性呼吸道感染尤其是上呼吸道感染的重要细菌病原,可以引起儿童和成人急性咽炎和扁桃体炎,也可引起严重的侵袭性感染如坏死性筋膜炎和中毒休克综合征。感染后及时有效的治疗可避免给患者带来的严重伤害。     

宿主菌抗噬菌体机制的研究进展

噬菌体又称细菌病毒,是公认最丰富的微生物,也是最多样性的,这种多样性是适应所面对选择性压力例如普遍存在宿主菌的噬菌体抗性机制。噬菌体通过6步(吸附、注入、复制、转录翻译、组装和释放)侵入细菌并使之裂解,但是当噬菌体感染细菌,就会面临细菌抗噬菌体的机制,宿主菌能够进化出多种抗噬菌体的机制来避免噬菌体的侵染和裂解。本文就对宿主菌抗噬菌体各种机制作一综述。     

小麦赤霉病抗病机制研究进展

小麦赤霉病是一种小麦穗部病害,严重影响小麦的产量和品质。挖掘小麦赤霉病抗性基因,揭示其抗病机制,对于提高小麦赤霉病抗性,推动小麦赤霉病抗性育种进程具有重要的意义。系统阐述了抗赤霉病相关QTL、多组学研究、细胞壁防卫、信号转导、次生代谢物合成、识别应答等小麦赤霉病抗性机制的研究进展,并对未来小麦赤霉病抗性机制的研究方向进行了探讨。希望以此加深研究者对小麦赤霉病抗性机制的了解,为未来小麦抗赤霉病分子机制研究提供理论基础,为小麦抗赤霉病遗传改良提供丰富的基因资源。     

Distribution of streptothricin acetyltransferase encodi determinants among environmental bacteria

In this paper we report about a screening for streptothricin- (St)-resistant phenotypes and genotypes among environmental bacteria from a St virgin area. St-resistant bacteria were isolated from river water, sewage, manure and soil by selective plating. The resistance quotient was typical of an area without selective pressure. The occurrence of streptothricin acetyltransferase-encoding determinants and their localization on a Tn7-like transposon was tested by the application of a set of gene probes. Sat genes could be detected in 22.5% of the tested St-resistant bacteria but in 100% of the checked Enterobacteriaceae. However, we could not detect sat genes in St-resistant bacteria from soil samples. Surprisingly the sat genes were found to be located on conjugative or mobilizable plasmids for a rather high number of strains. The determined plasmid species and their restriction patterns showed a high degree of similarities to those observed from an area of strong selective pressure.     

Nucleotide Sequence and Genetic Characterization of the Novel IncQ-like Plasmid pIE1107

The analysis of the complete nucleotide sequence of the small resistance plasmid pIE1107 revealed a close similarity to the well-known IncQ plasmids. Highly conserved replication proteins and nearly identical origins of replication (oriV) suggest equivalent functions in the related replication systems. However, pIE1107 contains two copies of IncQ-oriV-like DNA which are slightly different regarding the iterons. Upon deletion of a silent copy of IncQ-oriV-like DNA the resulting plasmid is fully compatible with IncQ plasmids, indicating that there is no mutual communication between the replication control of the respective replicons. Experiments with clonedoriV DNA strongly suggest that the replication initiation protein of pIE1107 has specialized into the distinct target-iterons of its ownoriV which differs only by a few nucleotides from theoriV of IncQ plasmids. Implications from the apparent highly specific protein–DNA recognition and from the incompatibility properties of pIE1107 for the evolution of a family of compatible, IncQ-like plasmids are discussed.     

The Biological Function of the Bacterial Isochorismatase-Like Hydrolase SttH

The streptothricin hydrolase (SttH), which is a member of the isochorismatase-like hydrolase (ILH) super-family, catalyzes the hydrolysis of the streptolidine lactam group in streptothricin (ST) antibiotics, thereby inactivating them. In this study we identified a novel homologous gene (sttH-sn) and sequenced the flanking regions of the sttH and sttH-sn genes. The organization of genes around the sttH, sttH-sn, and ILH genes revealed that a number of the genes were clustered with genes encoding oxidoreductases with molybdopterin binding subunits, suggesting that the true role of these gene products (SttHs and a number of ILHs) might have to do with the chemical modification of molybdopterin, rather than ST-resistance. In addition, mutant enzymes were constructed in which Ser was substituted for highly conserved Cys-176 and Cys-158 of SttH and SttH-sn respectively, and no enzyme activities were detected. Thus, biochemically, these ILHs were found to be “cysteine hydrolases.”     

Sesquiterpenoid metabolites from Stereum complicatum

A new sesquiterpene antibiotic, complicatic acid, isolated from cultures of Stereum complicatum (Fr.)Fr. has been shown to be dehydrohirsutic acid C. Hirsutic acid C was also isolated from the same fungus. [2-¹⁴C]-MVA was incorporated into both metabolites and complicatic acid has been shown to be formed from hirsutic acid C both in vivo and in vitro.     

Noureseothricin (streptothricin) inactivated by a plasmid pIE636 encoded acetyl transferase of Escherichia coli: Location of the acetyl group

Abstract Escherichia coli strains harbouring the plasmid pIE636 are able to synthesize acetylcoenzyme A: streptothricin acetyltransferase (ACSAT). The (enzymatic) N -acetylation of streptothricin F is known to contribute significantly towards the loss of antibacterial activity. ¹³C-NMR analysis of [¹⁴C]N-acetyl-labelled streptothricin F, produced by ACSAT-catalysed acetylation of streptothricin F and subsequent purification by various chromatographical steps, unequivocally revealed streptothricin F to be acetylated at the β-amino group (C16) (and not at the ε-amino group (C19)).