scholarly journals A Repeating Sulfated Galactan Motif Resuscitates Dormant Micrococcus luteus Bacteria

2018 ◽  
Vol 84 (13) ◽  
Author(s):  
Thomas Böttcher ◽  
Dávid Szamosvári ◽  
Jon Clardy
Keyword(s):  
Micrococcus Luteus ◽  
Bacterial Species ◽  
Model Organism ◽  
Growth Media ◽  
Specific Chemical ◽  
Content Type ◽  
Growth Initiation ◽  
Molecular Signal ◽  
Polysaccharide Composition ◽  
Growing Conditions

ABSTRACT Only a small fraction of bacteria can autonomously initiate growth on agar plates. Nongrowing bacteria typically enter a metabolically inactive dormant state and require specific chemical trigger factors or signals to exit this state and to resume growth. Micrococcus luteus has become a model organism for this important yet poorly understood phenomenon. Only a few resuscitation signals have been described to date, and all of them are produced endogenously by bacterial species. We report the discovery of a novel type of resuscitation signal that allows M. luteus to grow on agar but not agarose plates. Fractionation of the agar polysaccharide complex and sulfation of agarose allowed us to identify the signal as highly sulfated saccharides found in agar or carrageenans. Purification of hydrolyzed κ-carrageenan ultimately led to the identification of the signal as a small fragment of a large linear polysaccharide, i.e., an oligosaccharide of five or more sugars with a repeating disaccharide motif containing d -galactose-4-sulfate (G4S) 1,4-linked to 3,6-anhydro-α- d -galactose (DA), G4S-(DA-G4S) n ≥2 . IMPORTANCE Most environmental bacteria cannot initiate growth on agar plates, but they can flourish on the same plates once growth is initiated. While there are a number of names for and manifestations of this phenomenon, the underlying cause appears to be the requirement for a molecular signal indicating safe growing conditions. Micrococcus luteus has become a model organism for studying this growth initiation process, often called resuscitation, because of its apparent connection with the persistent or dormant form of Mycobacterium tuberculosis , an important human pathogen. In this report, we identify a highly sulfated saccharide from agar or carrageenans that robustly resuscitates dormant M. luteus on agarose plates. We identified and characterized the signal as a small repeating disaccharide motif. Our results indicate that signals inherent in or absent from the polysaccharide composition of solid growth media can have major effects on bacterial growth.

scholarly journals Osmoadaptation Strategy of the Most Halophilic Fungus, Wallemia ichthyophaga, Growing Optimally at Salinities above 15% NaCl

2013 ◽  
Vol 80 (1) ◽  
pp. 247-256 ◽  
Author(s):  
Janja Zajc ◽  
Tina Kogej ◽  
Erwin A. Galinski ◽  
José Ramos ◽  
Nina Gunde-Cimerman
Keyword(s):  
Biomass Production ◽  
Osmotic Shock ◽  
Model Organism ◽  
Salinity Range ◽  
Growth Media ◽  
Optimal Salinity ◽  
Content Type ◽  
Batch Cultures ◽  
Specific Growth Rates ◽  
Optimum Salinity

ABSTRACTWallemia ichthyophagais a fungus from the ancient basidiomycetous genusWallemia(Wallemiales, Wallemiomycetes) that grows only at salinities between 10% (wt/vol) NaCl and saturated NaCl solution. This obligate halophily is unique among fungi. The main goal of this study was to determine the optimal salinity range for growth of the halophilicW. ichthyophagaand to unravel its osmoadaptation strategy. Our results showed that growth on solid growth media was extremely slow and resulted in small colonies. On the other hand, in the liquid batch cultures, the specific growth rates ofW. ichthyophagawere higher, and the biomass production increased with increasing salinities. The optimum salinity range for growth ofW. ichthyophagawas between 15 and 20% (wt/vol) NaCl. At 10% NaCl, the biomass production and the growth rate were by far the lowest among all tested salinities. Furthermore, the cell wall content in the dry biomass was extremely high at salinities above 10%. Our results also showed that glycerol was the major osmotically regulated solute, since its accumulation increased with salinity and was diminished by hypo-osmotic shock. Besides glycerol, smaller amounts of arabitol and trace amounts of mannitol were also detected. In addition,W. ichthyophagamaintained relatively small intracellular amounts of potassium and sodium at constant salinities, but during hyperosmotic shock, the amounts of both cations increased significantly. Given our results and the recent availability of the genome sequence,W. ichthyophagashould become well established as a novel model organism for studies of halophily in eukaryotes.

scholarly journals Impact of growth media and pressure on the diversity and antimicrobial activity of isolates from two species of hexactinellid sponge

Microbiology ◽  
2021 ◽  
Vol 167 (12) ◽  
Author(s):  
Matthew J. Koch ◽  
Poppy J. Hesketh-Best ◽  
Gary Smerdon ◽  
Philip J. Warburton ◽  
Kerry Howell ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Deep Sea ◽  
Micrococcus Luteus ◽  
Industrial Applications ◽  
Sponge Species ◽  
Growth Media ◽  
Rrna Gene ◽  
Content Type ◽  
Link Type ◽  
Increased Pressure

Access to deep-sea sponges brings with it the potential to discover novel antimicrobial candidates, as well as novel cold- and pressure-adapted bacteria with further potential clinical or industrial applications. In this study, we implemented a combination of different growth media, increased pressure and high-throughput techniques to optimize recovery of isolates from two deep-sea hexactinellid sponges, Pheronema carpenteri and Hertwigia sp., in the first culture-based microbial analysis of these two sponges. Using 16S rRNA gene sequencing for isolate identification, we found a similar number of cultivable taxa from each sponge species, as well as improved recovery of morphotypes from P. carpenteri at 22–25 °C compared to other temperatures, which allows a greater potential for screening for novel antimicrobial compounds. Bacteria recovered under conditions of increased pressure were from the phyla Proteobacteria , Actinobacteria and Firmicutes , except at 4 %O2/5 bar, when the phylum Firmicutes was not observed. Cultured isolates from both sponge species displayed antimicrobial activity against Micrococcus luteus, Staphylococcus aureus and Escherichia coli .

scholarly journals Adaptations Accumulated under Prolonged Resource Exhaustion Are Highly Transient

mSphere ◽  
2020 ◽  
Vol 5 (4) ◽  
Author(s):  
Sarit Avrani ◽  
Sophia Katz ◽  
Ruth Hershberg
Keyword(s):  
Bacterial Species ◽  
Growth Media ◽  
Bacterial Survival ◽  
Content Type ◽  
Prolonged Starvation ◽  
E Coli ◽  
Resource Exhaustion ◽  
Shed Light ◽  
External Growth

ABSTRACT Many nonsporulating bacterial species can survive for years within exhausted growth media in a state termed long-term stationary phase (LTSP). We have been carrying out evolutionary experiments aimed at elucidating the dynamics of genetic adaptation under LTSP. We showed that Escherichia coli adapts to prolonged resource exhaustion through the highly convergent acquisition of mutations. In the most striking example of such convergent adaptation, we observed that across all independently evolving LTSP populations, over 90% of E. coli cells carry mutations to one of three specific sites of the RNA polymerase core enzyme (RNAPC). These LTSP adaptations reduce the ability of the cells carrying them to grow once fresh resources are again provided. Here, we examine how LTSP populations recover from costs associated with their adaptation once resources are again provided to them. We demonstrate that due to the ability of LTSP populations to maintain high levels of standing genetic variation during adaptation, costly adaptations are very rapidly purged from the population once they are provided with fresh resources. We further demonstrate that recovery from costs acquired during adaptation under LTSP occurs more rapidly than would be possible if LTSP adaptations had fixed during the time populations spent under resource exhaustion. Finally, we previously reported that under LTSP, some clones develop a mutator phenotype, greatly increasing their mutation accumulation rates. Here, we show that the mechanisms by which populations recover from costs associated with fixed adaptations may depend on mutator status. IMPORTANCE Many bacterial species can survive for decades under starvation, following the exhaustion of external growth resources. We have previously shown that bacteria genetically adapt under these conditions in a manner that reduces their ability to grow once resources again become available. Here, we study how populations that have been subject to very prolonged resource exhaustion recover from costs associated with their adaptation. We demonstrate that rapid adaptations acquired under prolonged starvation tend to be highly transient, rapidly reducing in frequency once bacteria are no longer starved. Our results shed light on the longer-term consequences of bacterial survival under prolonged starvation. More generally, these results may also be applicable to understanding longer-term consequences of rapid adaptation to additional conditions as well.

scholarly journals Coordinated Assembly of theBacillus anthracisCoat and Exosporium during Bacterial Spore Outer Layer Formation

mBio ◽  
2018 ◽  
Vol 9 (6) ◽  
Author(s):  
Tyler J. Boone ◽  
Michael Mallozzi ◽  
Alex Nelson ◽  
Brian Thompson ◽  
Mark Khemmani ◽  
...  
Keyword(s):  
Protein Interaction ◽  
Outer Layer ◽  
Protein Interaction Network ◽  
Bacterial Species ◽  
Model Organism ◽  
Interaction Network ◽  
Subcellular Location ◽  
Content Type ◽  
Outermost Layer

ABSTRACTBacterial spores produced by theBacillalesare composed of concentric shells, each of which contributes to spore function. Spores from all species possess a cortex and coat, but spores from many species possess additional outer layers. The outermost layer ofBacillus anthracisspores, the exosporium, is separated from the coat by a gap known as the interspace. Exosporium and interspace assembly remains largely mysterious. As a result, we have a poor understanding of the overarching mechanisms driving the assembly of one of the most ubiquitous cell types in nature. To elucidate the mechanisms directing exosporium assembly, we generated strains bearing mutations in candidate exosporium-controlling genes and analyzed the effect on exosporium formation. Biochemical and cell biological analyses argue that CotE directs the assembly of CotO into the spore and that CotO might be located at or close to the interior side of the cap. Taken together with data showing that CotE and CotO interact directlyin vitro, we propose a model in which CotE and CotO are important components of a protein interaction network that connects the exosporium to the forespore during cap formation and exosporium elongation. Our data also suggest that the cap interferes with coat assembly at one pole of the spore, altering the pattern of coat deposition compared to the model organismBacillus subtilis. We propose that the difference in coat assembly patterns between these two species is due to an inherent flexibility in coat assembly, which may facilitate the evolution of spore outer layer complexity.IMPORTANCEThis work dramatically improves our understanding of the assembly of the outermost layer of theB. anthracisspore, the exosporium, a layer that encases spores from many bacterial species and likely plays important roles in the spore’s interactions with the environment, including host tissues. Nonetheless, the mechanisms directing exosporium assembly into a shell surrounding the spore are still very poorly understood. In this study, we clarify these mechanisms by the identification of a novel protein interaction network that directs assembly to initiate at a specific subcellular location in the developing cell. Our results further suggest that the presence or absence of an exosporium has a major impact on the assembly of other more interior spore layers, thereby potentially explaining long-noted differences in spore assembly betweenB. anthracisand the model organismB. subtilis.

scholarly journals Bacterial Longevity Requires Protein Synthesis and a Stringent Response

mBio ◽  
2019 ◽  
Vol 10 (5) ◽  
Author(s):  
Liang Yin ◽  
Hongyu Ma ◽  
Ernesto S. Nakayasu ◽  
Samuel H. Payne ◽  
David R. Morris ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Bacterial Species ◽  
Rhodopseudomonas Palustris ◽  
Growth Arrest ◽  
Model Organism ◽  
Stringent Response ◽  
Homogeneous Population ◽  
Gram Negative ◽  
Content Type ◽  
Specific Proteins

ABSTRACT Gram-negative bacteria in infections, biofilms, and industrial settings often stop growing due to nutrient depletion, immune responses, or environmental stresses. Bacteria in this state tend to be tolerant to antibiotics and are often referred to as dormant. Rhodopseudomonas palustris, a phototrophic alphaproteobacterium, can remain fully viable for more than 4 months when its growth is arrested. Here, we show that protein synthesis, specific proteins involved in translation, and a stringent response are required for this remarkable longevity. Because it can generate ATP from light during growth arrest, R. palustris is an extreme example of a bacterial species that will stay alive for long periods of time as a relatively homogeneous population of cells and it is thus an excellent model organism for studies of bacterial longevity. There is evidence that other Gram-negative species also continue to synthesize proteins during growth arrest and that a stringent response is required for their longevity as well. Our observations challenge the notion that growth-arrested cells are necessarily dormant and metabolically inactive and suggest that such bacteria may have a level of metabolic activity that is higher than many would have assumed. Our results also expand our mechanistic understanding of a crucial but understudied phase of the bacterial life cycle. IMPORTANCE We are surrounded by bacteria, but they do not completely dominate our planet despite the ability of many to grow extremely rapidly in the laboratory. This has been interpreted to mean that bacteria in nature are often in a dormant state. We investigated life in growth arrest of Rhodopseudomonas palustris, a proteobacterium that stays alive for months when it is not growing. We found that cells were metabolically active, and they continued to synthesize proteins and mounted a stringent response, both of which were required for their longevity. Our results suggest that long-lived bacteria are not necessarily inactive but have an active metabolism that is well adjusted to life without growth.

scholarly journals CdrA Interactions within thePseudomonas aeruginosaBiofilm Matrix Safeguard It from Proteolysis and Promote Cellular Packing

mBio ◽  
2018 ◽  
Vol 9 (5) ◽  
Author(s):  
Courtney Reichhardt ◽  
Cynthis Wong ◽  
Daniel Passos da Silva ◽  
Daniel J. Wozniak ◽  
Matthew R. Parsek
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Matrix Protein ◽  
Bacterial Species ◽  
Model Organism ◽  
Biofilm Growth ◽  
Content Type ◽  
Multicellular Aggregates ◽  
The Matrix ◽  
Matrix Stability ◽  
First Time

ABSTRACTBiofilms are robust multicellular aggregates of bacteria that are encased in an extracellular matrix. Different bacterial species have been shown to use a range of biopolymers to build their matrices.Pseudomonas aeruginosais a model organism for the laboratory study of biofilms, and past work has suggested that exopolysaccharides are a required matrix component. However, we found that expression of the matrix protein CdrA, in the absence of biofilm exopolysaccharides, allowed biofilm formation through the production of a CdrA-rich proteinaceous matrix. This represents a novel function for CdrA. Similar observations have been made for other species such asEscherichia coliandStaphylococcus aureus, which can utilize protein-dominant biofilm matrices. However, we found that these CdrA-containing matrices were susceptible to both exogenous and self-produced proteases. We previously reported that CdrA directly binds the biofilm matrix exopolysaccharide Psl. Now we have found that when CdrA bound to Psl, it was protected from proteolysis. Together, these results support the idea of the importance of multibiomolecular components in matrix stability and led us to propose a model in which CdrA-CdrA interactions can enhance cell-cell packing in an aggregate that is resistant to physical shear, while Psl-CdrA interactions enhance aggregate integrity in the presence of self-produced and exogenous proteases.IMPORTANCEPseudomonas aeruginosaforms multicellular aggregates or biofilms using both exopolysaccharides and the CdrA matrix adhesin. We showed for the first time thatP. aeruginosacan use CdrA to build biofilms that do not require known matrix exopolysaccharides. It is appreciated that biofilm growth is protective against environmental assaults. However, little is known about how the interactions between individual matrix components aid in this protection. We found that interactions between CdrA and the exopolysaccharide Psl fortify the matrix by preventing CdrA proteolysis. When both components—CdrA and Psl—are part of the matrix, robust aggregates form that are tightly packed and protease resistant. These findings provide insight into how biofilms persist in protease-rich host environments.

scholarly journals Utilization of induction furnace steel slag based iron oxide nanocomposites for antibacterial studies

2021 ◽  
Vol 3 (3) ◽  
Author(s):  
J. Baalamurugan ◽  
V. Ganesh Kumar ◽  
T. Stalin Dhas ◽  
S. Taran ◽  
S. Nalini ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Iron Oxide ◽  
Building Materials ◽  
Induction Furnace ◽  
Micrococcus Luteus ◽  
Bacterial Species ◽  
Steel Slag ◽  
If Steel ◽  
Growth Inhibitory ◽  
Microplate Reader

AbstractMetals and metal oxide-based nanocomposites play a significant role over the control of microbes. In this study, antibacterial activity of iron oxide (Fe2O3) nanocomposites based on induction furnace (IF) steel slag has been carried out. IF steel slag is an industrial by-product generated from secondary steel manufacturing process and has various metal oxides which includes Al2O3 (7.89%), MnO (5.06), CaO (1.49%) and specifically Fe2O3 (14.30%) in higher content along with metalloid SiO2 (66.42). Antibacterial activity of iron oxide nanocomposites has been revealed on bacterial species such as Micrococcus luteus, Bacillus subtilis and Staphylococcus aureus. Micrococcus luteus has undergone maximum zone of inhibition (ZOI) of 12 mm for 10 mg/mL concentration of steel slag iron oxide nanocomposite. Growth inhibitory kinetics of bacterial species has been studied using ELISA microplate reader at 660 nm by varying the concentration of steel slag iron oxide nanocomposites. The results illustrate that IF steel slag is a potential material and can be utilized in building materials to increase the resistance against biodeterioration. Graphic abstract

scholarly journals Enhancing Terpene Yield from Sugars via Novel Routes to 1-Deoxy-d-Xylulose 5-Phosphate

2014 ◽  
Vol 81 (1) ◽  
pp. 130-138 ◽  
Author(s):  
James Kirby ◽  
Minobu Nishimoto ◽  
Ruthie W. N. Chow ◽  
Edward E. K. Baidoo ◽  
George Wang ◽  
...  
Keyword(s):  
Bacterial Species ◽  
Xylose Reductase ◽  
Pentose Phosphate ◽  
Terpene Biosynthesis ◽  
Content Type ◽  
E Coli ◽  
Pathway Expression ◽  
Terpene Synthesis ◽  
Selection For

ABSTRACTTerpene synthesis in the majority of bacterial species, together with plant plastids, takes place via the 1-deoxy-d-xylulose 5-phosphate (DXP) pathway. The first step of this pathway involves the condensation of pyruvate and glyceraldehyde 3-phosphate by DXP synthase (Dxs), with one-sixth of the carbon lost as CO2. A hypothetical novel route from a pentose phosphate to DXP (nDXP) could enable a more direct pathway from C5sugars to terpenes and also circumvent regulatory mechanisms that control Dxs, but there is no enzyme known that can convert a sugar into its 1-deoxy equivalent. Employing a selection for complementation of adxsdeletion inEscherichia coligrown on xylose as the sole carbon source, we uncovered two candidate nDXP genes. Complementation was achieved either via overexpression of the wild-typeE. coliyajOgene, annotated as a putative xylose reductase, or via various mutations in the nativeribBgene.In vitroanalysis performed with purified YajO and mutant RibB proteins revealed that DXP was synthesized in both cases from ribulose 5-phosphate (Ru5P). We demonstrate the utility of these genes for microbial terpene biosynthesis by engineering the DXP pathway inE. colifor production of the sesquiterpene bisabolene, a candidate biodiesel. To further improve flux into the pathway from Ru5P, nDXP enzymes were expressed as fusions to DXP reductase (Dxr), the second enzyme in the DXP pathway. Expression of a Dxr-RibB(G108S) fusion improved bisabolene titers more than 4-fold and alleviated accumulation of intracellular DXP.

scholarly journals Evaluation of Commercial Disinfectants against Staphylococcus lentus and Micrococcus spp. of Poultry Origin

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Otun Saha ◽  
Nadira Naznin Rakhi ◽  
Arif Istiaq ◽  
Israt Islam ◽  
Munawar Sultana ◽  
...  
Keyword(s):  
Micrococcus Luteus ◽  
Bacterial Species ◽  
16S Rrna Gene Sequencing ◽  
Average Diameter ◽  
Diffusion Method ◽  
Dilution Method ◽  
Rrna Gene ◽  
Poultry Farms ◽  
Rrna Gene Sequencing ◽  
Selection Of

Introduction. Effective sanitation strategies for poultry farms require an appropriate selection of the disinfectant based on the contaminants present and their sensitivity to the disinfectants. Aim. The current study investigated the prevalence of streptococci/micrococci in poultry farms of Bangladesh and the efficacy of commercial disinfectants (Savlon, Lysol, Quatovet, Virkon S, and Virocid) along with alcohol against these pathogens to adopt appropriate strategies. Materials and Methods. Conventional approaches and the 16S rRNA gene sequencing were performed to confirm the isolates at the species level along with microtiter biofilm assay to determine their biofilm-forming ability. Efficacy of the disinfectants was tested against those isolates using agar well diffusion and minimum inhibitory concentration (MIC) test by broth dilution method using different dilutions of the disinfectants. Results. Staphylococcus lentus (n = 32), Micrococcus luteus (n = 7), and Micrococcus aloeverae (n = 4) were confirmed among 102 presumptively screened streptococci/micrococci isolates from 43 samples. No single disinfectant showed equally high efficacy against all three bacterial species in agar well diffusion test, although Virocid showed the lowest MIC against all three of them. Lysol was least effective among the commercial disinfectants by both MIC and diffusion method, although each commercial disinfectant was more effective than alcohol. Considering both the average diameter of the inhibition zones and the MIC values, efficacy can be interpreted as Virocid > Quatovet > Savlon > Virkon S > Lysol. Although the efficacy decreased with decreasing concentration, the disinfectants retained a satisfactory level of efficacy at 50% concentration. Among test pathogens, M. aloeverae was the most sensitive to the disinfectants and the weakest biofilm producers, whereas 4/14 S. lentus and 1/5 M. luteus were strong biofilm producers, which may cause more reduction in the efficacy in environmental conditions. Conclusion. As no ideal disinfectant was found in the study, the efficacy of the disinfectants should be routinely evaluated and validated to ensure the sanitation standards in the poultry sector.

scholarly journals Improved Antimicrobial Activities of Synthetic-Hybrid Bacteriocins Designed from Enterocin E50-52 and Pediocin PA-1

2014 ◽  
Vol 81 (5) ◽  
pp. 1661-1667 ◽  
Author(s):  
Santosh Kumar Tiwari ◽  
Katia Sutyak Noll ◽  
Veronica L. Cavera ◽  
Michael L. Chikindas
Keyword(s):  
Micrococcus Luteus ◽  
Electrical Potential ◽  
Gram Negative Bacteria ◽  
Additional Advantage ◽  
Gram Negative ◽  
Content Type ◽  
Intracellular Atp ◽  
Hybrid Peptides ◽  
C Terminus ◽  
N Terminus

ABSTRACTTwo hybrid bacteriocins, enterocin E50-52/pediocin PA-1 (EP) and pediocin PA-1/enterocin E50-52 (PE), were designed by combining the N terminus of enterocin E50-52 and the C terminus of pediocin PA-1 and by combining the C terminus of pediocin PA-1 and the N terminus of enterocin E50-52, respectively. Both hybrid bacteriocins showed reduced MICs compared to those of their natural counterparts. The MICs of hybrid PE and EP were 64- and 32-fold lower, respectively, than the MIC of pediocin PA-1 and 8- and 4-fold lower, respectively, than the MIC of enterocin E50-52. In this study, the effect of hybrid as well as wild-type (WT) bacteriocins on the transmembrane electrical potential (ΔΨ) and their ability to induce the efflux of intracellular ATP were investigated. Enterocin E50-52, pediocin PA-1, and hybrid bacteriocin PE were able to dissipate ΔΨ, but EP was unable to deplete this component. Both hybrid bacteriocins caused a loss of the intracellular concentration of ATP. EP, however, caused a faster efflux than PE and enterocin E50-52. Enterocin E50-52 and hybrids PE and EP were active against the Gram-positive and Gram-negative bacteria tested, such asMicrococcus luteus,Salmonella entericaserovar Enteritidis 20E1090, andEscherichia coliO157:H7. The hybrid bacteriocins designed and described herein are antimicrobial peptides with MICs lower those of their natural counterparts. Both hybrid peptides induce the loss of intracellular ATP and are capable of inhibiting Gram-negative bacteria, and PE dissipates the electrical potential. In this study, the MIC of hybrid bacteriocin PE decreased 64-fold compared to the MIC of its natural peptide counterpart, pediocin PA-1. Inhibition of Gram-negative pathogens confers an additional advantage for the application of these peptides in therapeutics.

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