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Nanomolar Responsiveness of Marine Phaeobacter inhibens DSM 17395 toward Carbohydrates and Amino Acids 海洋嗜酸杆菌抑制剂DSM 17395对碳水化合物和氨基酸的纳摩尔响应性
IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2022-04-25 DOI: 10.1159/000524702
Arne Weiten, K. Kalvelage, Meina Neumann-Schaal, Ramona Buschen, Sabine Scheve, M. Winklhofer, R. Rabus
Phaeobacter inhibens DSM 17395 is a heterotrophic member of the ubiquitous, marine Roseobacter group and specializes in the aerobic utilization of carbohydrates and amino acids via pathways widespread among roseobacters. The in vivo responsiveness of P. inhibens DSM 17395 was studied with nonadapted cells (succinate-grown), which were exposed to a single pulse (100–0.01 µM) each of N-acetylglucosamine, mannitol, xylose, leucine, phenylalanine, or tryptophan (effectors). Responsiveness was then determined by time-resolved transcript analyses (quantitative reverse transcription-PCR) of “degradation” and “uptake” genes selected based on previously reported substrate-specific proteome profiles. The transcriptional response thresholds were: 50–100 nM for nagK (N-acetylglucosamine kinase), paaA (ring 1,2-phenylacetyl-CoA epoxidase), and kynA (tryptophan 2,3-dioxygenase), 10–50 nM for xylA (xylose isomerase), and around 10 nM for mtlK (mannitol 2-dehydrogenase). A threshold for leucine could not be determined due to the elevated intrinsic presence of leucine in the exometabolome of succinate-grown cells (no effector addition). Notably, the response thresholds for presumptive carbohydrate-binding proteins of ABC-transporters were in the same range or even lower: 0.1–1 µM for c27930 (N-acetylglucosamine) and even below 10 nM for c13210 (mannitol) and xylF (xylose). These results shed new light on the sensory/regulatory sensitivity of a well-studied roseobacter for recognizing potential substrates at low ambient concentrations and on the concentration threshold below which these might escape biodegradation (“emergent recalcitrance” concept of dissolved organic matter persistence).
抑黄杆菌DSM 17395是普遍存在的海洋玫瑰菌群中的异养成员,专门通过玫瑰菌中广泛存在的途径对碳水化合物和氨基酸进行有氧利用。用非适应细胞(琥珀酸盐生长)研究了抑制素DSM 17395的体内反应性,这些细胞暴露于N-乙酰葡糖胺、甘露醇、木糖、亮氨酸、苯丙氨酸或色氨酸(效应物)的单个脉冲(100-0.01µM)。然后通过基于先前报道的底物特异性蛋白质组图谱选择的“降解”和“摄取”基因的时间分辨转录物分析(定量逆转录PCR)来确定反应性。转录反应阈值为:nagK(N-乙酰葡糖胺激酶)、paaA(环1,2-苯乙酰-CoA环氧化物酶)和kynA(色氨酸2,3-双加氧酶)为50–100 nM,xylA(木糖异构酶)为10–50 nM,mtlK(甘露醇2-脱氢酶)约为10 nM。由于亮氨酸在琥珀酸盐生长的细胞的外代谢组中的固有存在增加(没有添加效应物),因此无法确定亮氨酸的阈值。值得注意的是,ABC转运蛋白的假定碳水化合物结合蛋白的反应阈值在相同的范围内,甚至更低:c27930(N-乙酰葡糖胺)为0.1–1µM,c13210(甘露醇)和xylF(木糖)甚至低于10 nM。这些结果为研究充分的玫瑰菌在低环境浓度下识别潜在底物的感官/调节敏感性以及这些底物可能逃脱生物降解的浓度阈值(溶解有机物持久性的“紧急顽固性”概念)提供了新的线索。
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引用次数: 1
Interplay between the Conserved Pore Residues Thr-91 and His-209 Controls Formate Translocation through the FocA Channel 保守孔隙残基Thr-91和His-209之间的相互作用控制了甲酸酯通过FocA通道的转运
IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2022-04-07 DOI: 10.1159/000524454
Michelle Kammel, Oliver Trebbin, R. Sawers
The formate channel A (FocA) belongs to the formate-nitrite transporter (FNT) family, members of which permeate small monovalent anions. FocA from Escherichia coli translocates formate/formic acid bi-directionally across the cytoplasmic membrane during fermentative growth. Two residues are particularly well-conserved within the translocation pores of FNTs: threonine-91 and histidine-209, based on E. coli FocA numbering. These residues are located at the tips of two broken transmembrane helices and control anion passage. H209 is the only charged residue within the pore and interacts with T91. Here, we addressed the role of the T91-H209 interaction network in the permeation of formate in vivo through FocA by performing an extensive amino acid-exchange study. Monitoring changes in intracellular formate using a formate-responsive fdhFP::lacZ reporter system revealed that T91 is essential for the ability of FocA to translocate formate bi-directionally. Only exchange for serine was partially tolerated, indicating that the hydroxyl group of T91 is mechanistically important. Substitution of H209 with N or Q was previously shown to convert FocA into a formate efflux channel. We show here that residue exchanges A, I, and T at this position resulted in a similar phenotype. Moreover, efflux function was confirmed for these FocA variants by measuring excreted formate in the culture medium. Substitution of bulky or charged residues for H209 prevented bi-directional formate passage. Studies using hypophosphite, a toxic analogue of formate taken up by FocA, and which causes impaired growth, confirmed that T91 and H209 substitutions essentially abolished, or drastically reduced, FocA’s translocation activity, as shown by effects on growth rate. The exceptions were T91S- and T91Y-exchange variants that retained partial ability to take up inhibitory hypophosphite. Together, our findings indicate that T91 is essential for formate permeation in both directions; however, it is particularly important to allow anion efflux. Moreover, H209 is essential for formate uptake by FocA, strongly suggesting that protonation-deprotonation of this residue plays a role in formate uptake. Finally, our results substantiate the premise that efflux and influx of formate by FocA are mechanistically distinct processes that are controlled by the interplay between T91 and H209.
甲酸通道A (FocA)属于甲酸亚硝酸盐转运体(FNT)家族,其成员渗透小的单价阴离子。在发酵过程中,来自大肠杆菌的FocA在细胞质膜上双向转运甲酸/甲酸。根据大肠杆菌FocA编号,两个残基在fnt的易位孔中保存得特别好:苏氨酸-91和组氨酸-209。这些残基位于两个断裂的跨膜螺旋的尖端,并控制阴离子的通过。H209是孔内唯一带电的残基,并与T91相互作用。在这里,我们通过进行广泛的氨基酸交换研究,解决了T91-H209相互作用网络在甲酸盐通过FocA在体内渗透中的作用。利用fdhFP::lacZ报告系统监测细胞内甲酸的变化,发现T91对FocA双向转运甲酸的能力至关重要。只有丝氨酸的交换是部分耐受的,这表明T91的羟基在机械上很重要。先前的研究表明,用N或Q取代H209可以将FocA转化为甲酸外排通道。我们在这里展示了A, I和T在这个位置上的残基交换导致了相似的表型。此外,通过测量培养基中分泌的甲酸,证实了这些FocA变体的外排功能。大块或带电荷的残基取代H209阻止了双向甲酸通道。利用次磷酸酯(FocA吸收的甲酸的有毒类似物,会导致生长受损)进行的研究证实,T91和H209取代基本上会消除或大幅降低FocA的转运活性,这可以从对生长速度的影响中看出。例外的是T91S-和t91y -交换变体,它们保留了部分吸收抑制性次磷酸盐的能力。总之,我们的研究结果表明,T91对甲酸盐在两个方向的渗透都是必不可少的;然而,允许阴离子外排是特别重要的。此外,H209对于FocA摄取甲酸至关重要,这强烈表明该残基的质子化-去质子化在甲酸摄取中起作用。最后,我们的研究结果证实了一个前提,即FocA的甲酸外排和内流是由T91和H209之间的相互作用控制的机制不同的过程。
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引用次数: 4
Acknowledgement to Reviewers 对评审员的确认
IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2022-04-01 DOI: 10.1159/000520900
© 2022 The Author(s) Published by S. Karger AG, Basel Karger Publishers and the editors of Microbial Physiology would like to thank the reviewers for the ongoing support in reviewing manuscripts for our Journal in 2021. This year we have chosen not to disclose the names of our reviewers to preserve the principle of anonymity inherent to the single-blind peer-review we follow. Even so, this should not be in our way to sincerely thank all contributing reviewers who have volunteered their time, effort, and expertise in benefit of the quality of the manuscripts we received and published in 2021. Individual reviewers can still claim their personal “Certificate of Review” via the Journal’s manuscript submission system.”
©2022作者由s . Karger AG出版,Basel Karger Publishers和《微生物生理学》的编辑感谢审稿人对我们期刊2021年审稿的持续支持。今年,我们选择不披露审稿人的姓名,以维护我们所遵循的单盲同行评审固有的匿名原则。即便如此,这也不应该妨碍我们真诚地感谢所有贡献的审稿人,他们自愿付出时间、精力和专业知识,以提高我们在2021年收到和发表的稿件的质量。个人审稿人仍然可以通过期刊的稿件提交系统申请他们的个人“审稿证书”。
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引用次数: 0
Luciferase-Based Determination of ATP/NAD(H) Pools in a Marine (Environmental) Bacterium 基于萤光素酶的海洋(环境)细菌ATP/NAD(H)库测定
IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2022-03-25 DOI: 10.1159/000522414
Daniel Wünsch, Sabine Scheve, Arne Weiten, K. Kalvelage, R. Rabus
In all living organisms, adenosine triphosphate (ATP) and NAD(H) represent universal molecular currencies for energy and redox state, respectively, and are thus widely applicable molecular proxies for an organism’s viability and activity. To this end, corresponding luciferase-based assays in combination with a microplate reader were established with the marine model bacterium Phaeobacter inhibens DSM 17395 (Escherichia coli K12 served as reference). Grey multiwell plates best balanced sensitivity and crosstalk, and optimal incubation times were 5 min and 30 min for the ATP and NAD(H) assay, respectively, together allowing limits of detection of 0.042, 0.470 and 0.710 nM for ATP, NAD+, and NADH, respectively. Quenching of bacterial cell samples involved Tris-EDTA-DTAB and bicarbonate base-DTAB for ATP and NAD(H) assays, respectively. The ATP and NAD(H) yields determined for P. inhibens DSM 17395 at ¼ ODmax were found to reside well within the range previously reported for E. coli and other bacteria, e.g., 3.28 µmol ATP (g cellsdry)−1. Thus, the here described methods for luciferase-based determination of ATP/NAD(H) pools open a promising approach to investigate energy and redox states in marine (environmental) bacteria.
在所有生物体中,三磷酸腺苷(ATP)和NAD(H)分别代表能量和氧化还原状态的通用分子货币,因此是生物体生存能力和活性的广泛适用的分子替代物。为此,用海洋模式细菌Phaeobacter inhibbens DSM 17395(作为参考的大肠杆菌K12)建立了相应的基于荧光素酶的测定法,并结合酶标仪。灰色多孔板对ATP和NAD(H)测定的灵敏度和串扰的最佳平衡以及最佳孵育时间分别为5分钟和30分钟,使ATP、NAD+和NADH的检测限分别为0.042、0.470和0.710nM。细菌细胞样品的猝灭分别涉及用于ATP和NAD(H)测定的Tris-EDTA DTAB和碳酸氢盐碱DTAB。在¼ODmax下,为P.inhibbens DSM 17395测定的ATP和NAD(H)产量被发现在先前报道的大肠杆菌和其他细菌的范围内,例如3.28µmol ATP(g cellsdry)−1。因此,本文所述的基于荧光素酶的ATP/NAD(H)池测定方法为研究海洋(环境)细菌的能量和氧化还原状态开辟了一种很有前途的方法。
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引用次数: 1
Assembly of Bacillus subtilis Dynamin into Membrane-Protective Structures in Response to Environmental Stress Is Mediated by Moderate Changes in Dynamics at a Single Molecule Level 枯草芽孢杆菌Dynamin在环境胁迫下组装成膜保护结构是由单分子水平上的适度动力学变化介导的
IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2022-03-10 DOI: 10.1159/000521585
Laura Sattler, P. Graumann
Dynamin-like proteins are membrane-associated GTPases, conserved in bacteria and in eukaryotes, that can mediate nucleotide-driven membrane deformation or membrane fusion reactions. Bacillus subtilis’ DynA has been shown to play an important role in protecting cells against chemicals that induce membrane leakage, and to form an increased number of membrane-associated structures after induction of membrane stress. We have studied the dynamics of DynA at a single molecule level in real time, to investigate how assembly of stress-induced structures is accompanied by changes in molecule dynamics. We show that DynA molecule displacements are best described by the existence of three distinct populations, a static mode, a low-mobility, and a fast-mobile state. Thus, DynA is most likely freely diffusive within the cytosol, moves along the cell membrane with a low mobility, and arrests at division sites or at stress-induced lesions at the membrane. In response to stress-inducing membrane leakage, but not to general stress, DynA molecules become slightly more static, but largely retain their mobility, suggesting that only few molecules are involved in the repair of membrane lesions, while most molecules remain in a dynamic mode scanning for lesions. Our data suggest that even moderate changes in single molecule dynamics can lead to visible changes in protein localization patterns.
Dynamin样蛋白是膜相关的GTP酶,在细菌和真核生物中保守,可以介导核苷酸驱动的膜变形或膜融合反应。枯草芽孢杆菌的DynA已被证明在保护细胞免受诱导膜渗漏的化学物质的侵害方面发挥着重要作用,并在诱导膜应激后形成数量增加的膜相关结构。我们在单分子水平上实时研究了DynA的动力学,以研究应力诱导结构的组装如何伴随分子动力学的变化。我们表明,DynA分子位移最好用三种不同种群的存在来描述,即静态模式、低迁移率和快速迁移状态。因此,DynA很可能在胞质溶胶内自由扩散,以低迁移率沿着细胞膜移动,并在分裂位点或膜上应激诱导的损伤处停止。作为对应力诱导的膜渗漏的反应,而不是对一般应力的反应,DynA分子变得稍微更静态,但在很大程度上保持了它们的流动性,这表明只有少数分子参与了膜损伤的修复,而大多数分子仍处于动态模式扫描损伤。我们的数据表明,即使单分子动力学的适度变化也会导致蛋白质定位模式的明显变化。
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引用次数: 1
Heterogeneity of Subcellular Diffusion in Bacteria Based on Spatial Segregation of Ribosomes and Nucleoids. 基于核糖体和类核空间分离的细菌亚细胞扩散异质性。
IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2022-01-01 DOI: 10.1159/000526846
Simon Dersch, Daniel A O Rotter, Peter L Graumann

It has long become clear that in spite of generally lacking internal membrane systems, bacteria contain well-structured subcellular structures of usually filamentous proteins, and a preferred 3D arrangement of their chromosome(s). Some of these systems are set up by so-called cytoskeletal elements, or by polar landmark proteins, but the mechanism of specific localization is still unclear in most cases. Intriguingly, apart from such spatially organizing systems, the bacterial cytoplasm has unusual properties in terms of the diffusion of molecules, which varies between different sites within the cell. In many bacteria, chromosomes are compacted into centrally located nucleoids, being orderly folded as opposed to consisting of random coils of DNA. In these bacteria, there is a separation of transcription and translation, such that transcription by RNA polymerase occurs on the nucleoids, and translation takes place mostly at the cell poles and directly underneath the cell membrane, because 70S ribosomes accumulate at sites surrounding the nucleoids. Interestingly, accumulation of ribosomes appears to slow down diffusion of enzymes, noticeable for larger enzyme complexes, while nucleoids provide areas of confined motion for DNA-binding proteins, yet acceleration zones for non-DNA-binding proteins. Crowded regions at the cell poles set up zones of higher concentration of the translation machinery, shortening diffusion distances for rate-limiting translation factor/ribosome interactions, and of metabolic enzymes, possibly speeding up pathways containing low concentrations of metabolites. Thus, heterogeneous diffusion adds another layer of subcellular organization on top of cytoskeletal elements.

长期以来,人们已经清楚,尽管细菌通常缺乏内部膜系统,但细菌含有结构良好的亚细胞结构,通常是丝状蛋白质,以及它们的染色体的首选3D排列。其中一些系统是由所谓的细胞骨架元件或极性地标蛋白建立的,但在大多数情况下,特异性定位的机制仍不清楚。有趣的是,除了这种空间组织系统外,细菌细胞质在分子扩散方面具有不同寻常的特性,这在细胞内不同部位之间是不同的。在许多细菌中,染色体被压缩成位于中心的类核,有序折叠,而不是由随机的DNA线圈组成。在这些细菌中,转录和翻译是分离的,因此RNA聚合酶的转录发生在类核上,而翻译主要发生在细胞极点和直接在细胞膜下面,因为70S核糖体聚集在类核周围的位置。有趣的是,核糖体的积累似乎减缓了酶的扩散,这对于较大的酶复合物来说是显而易见的,而类核为dna结合蛋白提供了限制运动的区域,但为非dna结合蛋白提供了加速区。细胞两极的拥挤区域建立了翻译机制的高浓度区域,缩短了限速翻译因子/核糖体相互作用和代谢酶的扩散距离,可能加速了含有低浓度代谢物的途径。因此,异质扩散在细胞骨架元素之上增加了另一层亚细胞组织。
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引用次数: 3
Discovery and Characterization of the Phospholemman/SIMP/Viroporin Superfamily. 磷蛋白/SIMP/病毒蛋白超家族的发现与表征。
IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2022-01-01 DOI: 10.1159/000521947
Daniel Tyler, Kevin J Hendargo, Arturo Medrano-Soto, Milton H Saier
Using bioinformatic approaches, we present evidence of distant relatedness among the Ephemerovirus Viroporin family, the Rhabdoviridae Putative Viroporin U5 family, the Phospholemman family, and the Small Integral Membrane Protein family. Our approach is based on the transitivity property of homology complemented with five validation criteria: (1) significant sequence similarity and alignment coverage, (2) compatibility of topology of transmembrane segments, (3) overlap of hydropathy profiles, (4) conservation of protein domains, and (5) conservation of sequence motifs. Our results indicate that Pfam protein domains PF02038 and PF15831 can be found in or projected onto members of all four families. In addition, we identified a 26-residue motif conserved across the superfamily. This motif is characterized by hydrophobic residues that help anchor the protein to the membrane and charged residues that constitute phosphorylation sites. In addition, all members of the four families with annotated function are either responsible for or affect the transport of ions into and/or out of the cell. Taken together, these results justify the creation of the novel Phospholemman/SIMP/Viroporin superfamily. Given that transport proteins can be found not just in cells, but also in viruses, the ability to relate viroporin protein families with their eukaryotic and bacterial counterparts is an important development in this superfamily.
利用生物信息学方法,我们提出了蜉蝣病毒毒蛋白家族、横纹肌病毒科推定病毒毒蛋白U5家族、磷蛋白家族和小整合膜蛋白家族之间的远亲关系的证据。我们的方法是基于同源性的传递性,并补充了五个验证标准:(1)显著的序列相似性和比对覆盖率,(2)跨膜段拓扑的兼容性,(3)亲水性谱的重叠,(4)蛋白质结构域的保守性,(5)序列基序的保守性。我们的研究结果表明,Pfam蛋白结构域PF02038和PF15831可以在所有四个家族的成员中找到或投射到Pfam蛋白结构域。此外,我们还发现了一个在整个超家族中保守的26个残基基序。该基序的特点是疏水残基有助于将蛋白质固定在膜上,而带电残基则构成磷酸化位点。此外,具有注释功能的四个家族的所有成员都负责或影响离子进出细胞的运输。综上所述,这些结果证明了创建新的磷蛋白/SIMP/病毒蛋白超家族的合理性。考虑到运输蛋白不仅可以在细胞中发现,也可以在病毒中发现,将病毒孔蛋白家族与其真核和细菌对应物联系起来的能力是这个超家族的一个重要发展。
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引用次数: 0
Comparative Analyses of the Transport Proteins Encoded within the Genomes of nine Bifidobacterium Species. 9种双歧杆菌基因组中转运蛋白编码的比较分析。
IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2022-01-01 DOI: 10.1159/000518954
Hassan Zafar, Milton H Saier

The human microbiome influences human health in both negative and positive ways. Studies on the transportomes of these organisms yield information that may be utilized for various purposes, including the identification of novel drug targets and the manufacture of improved probiotic strains. Moreover, these genomic analyses help to improve our understanding of the physiology and metabolic capabilities of these organisms. The present study is a continuation of our studies on the transport proteins of the major gut microbes. Bifidobacterium species are essential members of the human gut microbiome, and they initiate colonization of the gut at birth, providing health benefits that last a lifetime. In this study we analyze the transportomes of nine bifidobacterial species: B. adolescentis, B. animalis, B. bifidum, B. breve, B. catenulatum, B. dentium, B. longum subsp. infantis, B. longum subsp. longum, and B. pseudocatenulatum. All of these species have proven probiotic characteristics and exert beneficial effects on human health. Surprisingly, we found that all nine of these species have similar pore-forming toxins and drug exporters that may play roles in pathogenesis. These species have transporters for amino acids, carbohydrates, and proteins, essential for their organismal lifestyles and adaption to their respective ecological niches. The strictly probiotic species, B. bifidum, however, contains fewer such transporters, thus indicative of limited interactions with host cells and other gut microbial counterparts. The results of this study were compared with those of our previous studies on the transportomes of multiple species of Bacteroides, Escherichia coli/Salmonella, and Lactobacillus. Overall, bifidobacteria have larger transportomes (based on percentages of total proteins) than the previously examined groups of bacterial species, with a preference for primary active transport systems over secondary carriers. Taken together, these results provide useful information about the physiologies and pathogenic potentials of these probiotic organisms as reflected by their transportomes.

人类微生物组对人类健康的影响既有消极的一面,也有积极的一面。对这些生物的转运体的研究产生的信息可用于各种目的,包括鉴定新的药物靶点和制造改进的益生菌菌株。此外,这些基因组分析有助于提高我们对这些生物的生理和代谢能力的理解。本研究是我们对主要肠道微生物转运蛋白研究的延续。双歧杆菌是人类肠道微生物群的重要成员,它们在出生时就开始在肠道定植,提供持续一生的健康益处。本研究分析了9种双歧杆菌的转运体:青春期双歧杆菌、动物双歧杆菌、两歧双歧杆菌、短双歧杆菌、链状双歧杆菌、牙状双歧杆菌、长双歧杆菌。婴儿,B.长亚种。长,和b伪atenulatum。所有这些物种都具有已证实的益生菌特性,并对人体健康产生有益影响。令人惊讶的是,我们发现这9个物种都有相似的孔隙形成毒素和药物出口,可能在发病机制中发挥作用。这些物种有氨基酸、碳水化合物和蛋白质的转运体,这对它们的有机生活方式和适应各自的生态位至关重要。然而,严格意义上的益生菌物种两歧双歧杆菌含有较少的这种转运体,因此表明与宿主细胞和其他肠道微生物的相互作用有限。本研究结果与我们之前对多种拟杆菌、大肠杆菌/沙门氏菌和乳酸杆菌的转运体的研究结果进行了比较。总的来说,双歧杆菌的转运体(基于总蛋白的百分比)比先前研究的细菌种类群更大,并且优先于初级主动转运系统而不是次级载体。综上所述,这些结果为这些益生菌的生理和致病潜力提供了有用的信息,这些益生菌的转运体反映了它们的生理和致病潜力。
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引用次数: 1
Proteogenomic Insights into the Physiology of Marine, Sulfate-Reducing, Filamentous Desulfonema limicola and Desulfonema magnum. 对海洋硫酸盐还原丝状脱硫浮游动物(Desulfonema limicola)和脱硫浮游动物(Desulfonema magnum)生理学的蛋白质基因组学见解。
IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2021-02-19 DOI: 10.1159/000513383
Vanessa Schnaars, Lars Wöhlbrand, Sabine Scheve, Christina Hinrichs, Richard Reinhardt, Ralf Rabus

The genus Desulfonema belongs to the deltaproteobacterial family Desulfobacteraceae and comprises marine, sulfate-reducing bacteria that form filaments and move by gliding. This study reports on the complete, manually annotated genomes of Dn. limicola 5ac10T (6.91 Mbp; 6,207 CDS) and Dn. magnum 4be13T (8.03 Mbp; 9,970 CDS), integrated with substrate-specific proteome profiles (8 vs. 11). The richness in mobile genetic elements is shared with other Desulfobacteraceae members, corroborating horizontal gene transfer as major driver in shaping the genomes of this family. The catabolic networks of Dn. limicola and Dn. magnum have the following general characteristics: 98 versus 145 genes assigned (having genomic shares of 1.7 vs. 2.2%), 92.5 versus 89.7% proteomic coverage, and scattered gene clusters for substrate degradation and energy metabolism. The Dn. magnum typifying capacity for aromatic compound degradation (e.g., p-cresol, 3-phenylpropionate) requires 48 genes organized in operon-like structures (87.7% proteomic coverage; no homologs in Dn. limicola). The protein complements for aliphatic compound degradation, central pathways, and energy metabolism are highly similar between both genomes and were identified to a large extent (69-96%). The differential protein profiles revealed a high degree of substrate-specificity for peripheral reaction sequences (forming central intermediates), agreeing with the high number of sensory/regulatory proteins predicted for both strains. By contrast, central pathways and modules of the energy metabolism were constitutively formed under the tested substrate conditions. In accord with their natural habitats that are subject to fluctuating changes of physicochemical parameters, both Desulfonema strains are well equipped to cope with various stress conditions. Next to superoxide dismutase and catalase also desulfoferredoxin and rubredoxin oxidoreductase are formed to counter exposure to molecular oxygen. A variety of proteases and chaperones were detected that function in maintaining cellular homeostasis upon heat or cold shock. Furthermore, glycine betaine/proline betaine transport systems can respond to hyperosmotic stress. Gliding movement probably relies on twitching motility via type-IV pili or adventurous motility. Taken together, this proteogenomic study demonstrates the adaptability of Dn. limicola and Dn. magnum to its dynamic habitats by means of flexible catabolism and extensive stress response capacities.

Desulfonema属属于脱硫杆菌科(Desulfobacteraceae)脱硫杆菌属(deltaproteobacterial),由海洋硫酸盐还原细菌组成,它们能形成菌丝并通过滑行移动。本研究报告了 Dn. limicola 5ac10T(6.91 Mbp;6 207 CDS)和 Dn. magnum 4be13T(8.03 Mbp;9 970 CDS)的完整人工注释基因组,并整合了底物特异性蛋白质组图谱(8 对 11)。移动遗传元件的丰富程度与其他脱硫杆菌科成员相同,这证实了水平基因转移是形成该家族基因组的主要驱动力。Dn. limicola 和 Dn. magnum 的分解代谢网络具有以下一般特征:分配的基因数分别为 98 个和 145 个(基因组份额分别为 1.7% 和 2.2%),蛋白质组覆盖率分别为 92.5% 和 89.7%,底物降解和能量代谢基因群分散。Dn. magnum 的芳香化合物降解能力(如对甲酚、3-苯基丙酸酯)需要 48 个基因,这些基因组成了操作子样结构(蛋白质组覆盖率为 87.7%;Dn. limicola 中没有同源基因)。两个基因组中脂肪族化合物降解、中心途径和能量代谢的蛋白质互补高度相似,并在很大程度上(69-96%)得到了鉴定。差异蛋白图谱显示,外围反应序列(形成中心中间体)具有高度的底物特异性,这与预测的两种菌株的大量感觉/调节蛋白相吻合。相比之下,在测试的底物条件下,能量代谢的中心途径和模块是组成性形成的。由于其自然栖息地的理化参数会发生波动性变化,因此这两种 Desulfonema 菌株都能很好地应对各种压力条件。除了超氧化物歧化酶和过氧化氢酶之外,还形成了脱硫铁氧还蛋白和红宝石还蛋白氧化还原酶,以抵御分子氧的侵袭。还检测到多种蛋白酶和伴侣蛋白,它们在热或冷休克时起着维持细胞平衡的作用。此外,甘氨酸甜菜碱/脯氨酸甜菜碱运输系统可对高渗透压做出反应。滑行运动可能依赖于通过 IV 型纤毛器的抽动运动或冒险运动。总之,这项蛋白质基因组研究表明,Dn. limicola和Dn. magnum能通过灵活的分解代谢和广泛的应激反应能力来适应其动态栖息地。
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引用次数: 0
Polyhydroxybutyrate: A Useful Product of Chlorotic Cyanobacteria. 聚羟基丁酸盐:一种有益的绿蓝藻产物。
IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2021-01-01 Epub Date: 2021-05-12 DOI: 10.1159/000515617
Moritz Koch, Karl Forchhammer

Polyhydroxybutyrate (PHB) is a carbon polymer with diverse functions, varying greatly on the organism producing it. This microreview describes the current knowledge about PHB metabolism, structure, and different physiological roles with a special focus on cyanobacteria. Despite the physiological function of PHB in the cyanobacterial phylum still being unknown, these organisms provide the unique opportunity to directly convert atmospheric CO2 into bioplastic using a solar-based process. Recent research on PHB metabolism in the cyanobacterial model organism Synechocystis revealed a sophisticated control of PHB granule formation. Novel insights about the metabolic background of PHB synthesis resulted in the engineering of the first cyanobacterial superproducer strain.

聚羟基丁酸酯(PHB)是一种具有多种功能的碳聚合物,在产生它的生物体中变化很大。这篇微评论描述了目前关于PHB代谢,结构和不同生理作用的知识,特别关注蓝藻。尽管蓝藻门中PHB的生理功能尚不清楚,但这些生物提供了利用太阳能将大气中的二氧化碳直接转化为生物塑料的独特机会。最近对蓝藻模式生物聚囊藻中PHB代谢的研究揭示了PHB颗粒形成的复杂控制。关于PHB合成的代谢背景的新见解导致了第一个蓝藻超级生产者菌株的工程。
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引用次数: 13
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Microbial Physiology
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