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A preliminary morphological screening and isolation of bacterial colony from latex industrial wastewater was carried out. Bacteria colonies from latex processing wastewater were isolated from a local latex processing industry. It was found that 17 bacterial isolates had been purified grown on nutrient agar under 35˚C. The colonies were then purified and morphologically indicated via Gram staining and motility test. After morphological observation, it was identified that out of 17 isolates, 9 isolates were Gram positive and 8 isolates were Gram negative. There are 11 out of 17 colonies were rod-shaped bacterial colonies, while the other 6 colonies were cocci-shaped bacteria. There were 11 colonies of gliding bacteria, three colonies were non-motile bacteria and the other three colonies were flagellated bacteria. This study is only limited to morphological observation as the main aim of this study was to investigate the potential occurrence of viable growth in treated latex processing wastewater. The bacterial colonies were classified base on their morphological properties shown. This study has classified several genera such as Staphylococcus, Escherichia, Thiobacillus, Arthrobacter and other Genus. The growth curve of 17 isolates studied and the chemical oxygen demand were determined.

The Gram-negative gliding bacteria Lysobacter represent a new and rich source for bioactive natural products. In an effort to discover new antibiotics, we found a cryptic biosynthetic gene cluster (BGC) in Lysobacter sp. 3655 that shared a high similarity with the putative lysocin BGC identified in silico previously from Lysobacter sp. RH2180-5. Lysocins are cyclic lipodepsipeptides with potent activity against MRSA (methicillin-resistant Staphylococcus aureus) using a novel mode of action, but the lysocin BGC had not been experimentally verified so far. Using an activity-guided screening, we isolated the main antibiotic compound and confirmed it to be lysocin E. However, the putative lysocin BGC was barely transcribed in the wild type, in which lysocins were produced only in specific conditions and in a negligible amount. To activate the putative lysocin BGC, we screened for strongly transcribed housekeeping genes in strain 3655 and found several powerful promoters. Upon engineering the promoters into the BGC, the lysocin gene transcription was significantly enhanced and the lysocin yield was markedly increased. With readily detectable lysocins production in the engineered strains, we showed that lysocin production was abolished in the gene deletion mutant and then restored in the complementary strain, even when grown in conditions that did not support the wild type for lysocin production. Moreover, the engineered strain produced multiple new lysocin congeners. The determination of the lysocin BGC and the Lysobacter promoters will facilitate the ongoing efforts for yield improvement and new antibiotic biosynthesis using synthetic biology strategies.

Motile bacteria usually rely on external apparatus like flagella for swimming or pili for twitching. By contrast, gliding bacteria do not rely on obvious surface appendages to move on solid surfaces.Flavobacterium johnsoniaeand other bacteria in the Bacteroidetes phylum use adhesins whose movement on the cell surface supports motility. InF. johnsoniae, secretion and helicoidal motion of the main adhesin SprB are intimately linked and depend on the type IX secretion system (T9SS). Both processes necessitate the proton motive force (PMF), which is thought to fuel a molecular motor that comprises the GldL and GldM cytoplasmic membrane proteins. Here we show thatF. johnsoniaegliding motility is powered by the pH gradient component of the PMF. We further delineate the interaction network between the GldLM transmembrane helices (TMH) and show that conserved glutamate residues in GldL TMH are essential for gliding motility, although having distinct roles in SprB secretion and motion. We then demonstrate that the PMF and GldL trigger conformational changes in the GldM periplasmic domain. We finally show that multiple GldLM complexes are distributed in the membrane suggesting that a network of motors may be present to move SprB along a helical path on the cell surface. Altogether, our results provide evidence that GldL and GldM assemble dynamic membrane channels that use the proton gradient to power both T9SS-dependent secretion of SprB and its motion at the cell surface.

Myxobacteria are unicellular, Gram-negative, soil-dwelling, gliding bacteria that belong to class δ-proteobacteria and order Myxococcales. They grow and proliferate by transverse fission under normal conditions, but form fruiting bodies which contain myxospores during unfavorable conditions. In view of the escalating problem of antibiotic resistance among disease-causing pathogens, it becomes mandatory to search for new antibiotics effective against such pathogens from natural sources. Among the different approaches, Myxobacteria, having a rich armor of secondary metabolites, preferably derivatives of polyketide synthases (PKSs) along with non-ribosomal peptide synthases (NRPSs) and their hybrids, are currently being explored as producers of new antibiotics. The Myxobacterial species are functionally characterized to assess their ability to produce antibacterial, antifungal, anticancer, antimalarial, immunosuppressive, cytotoxic and antioxidative bioactive compounds. In our study, we have found their compounds to be effective against a wide range of pathogens associated with the concurrence of different infectious diseases.

Morphological mutations in bacterial cell make them the most miscellaneous microscopic group. Their non-flagellated species known as gliding bacteria exhibit self-powered motion and leave an adhesive trail of slime. The self-propelled motion in some gliding bacteria is achieved as a result of backward surface wave in the cell envelope. Motivated by this fact, an undulating surface on a layer of couple-stress fluid is used to model the motion of such gliding bacteria. Five different wave profiles, namely, sawtooth, sinusoidal, triangular, trapezoidal, and square profiles are used to model the waveform of the undulating wave in the outer cell surface. The inclination of the surface is also integrated into the model. The flow equations are set up under the lubrication assumption. Stream function is derived as an elementary function of an organism’s speed, undulation amplitude, and couple-stress parameter with its flow rate. Speed of the glider and flow rate (satisfying equilibrium conditions) are computed by employing modified Newton–Raphson method. These refined values are further utilized to compute the power dissipation. Effects of different waveforms, inclination angle, gravitational and couple-stress parameters on the speed of the microorganism and rate of energy expended are also quantified. Slime velocity is also plotted for fixed glider. In addition, making use of the obtained realistic set of values of the organism’s speed, flow rate, occlusion parameter, and couple-stress parameter, streamline patterns of the slime are plotted and discussed in detail.

Capnocytophaga species are commensal gliding bacteria that are found in human and animal oral microbiota and are involved in several inflammatory diseases, both in immunocompromised and immunocompetent subjects. This study contributes to increased knowledge of this genus by characterizing a novel species isolated from a dental plaque sample in a male with gingivitis. We investigated morphological and chemotaxonomic characteristics using different growth conditions, temperature, and pH. Cellular fatty acid methyl ester (FAME) analysis was employed with gas chromatography/mass spectrometry (GC/MS). Phylogenetic analysis based on 16S rRNA, orthologous average nucleotide identity (OrthoANI), and digital DNA–DNA hybridization (dDDH) relatedness were performed. The Marseille-Q4570T strain was found to be a facultative aerobic, Gram-negative, elongated, round-tipped bacterium that grew at 25–56 °C and tolerated a pH of 5.5 to 8.5 and an NaCl content ranging from 5 to 15 g/L. The most abundant fatty acid was the branched structure 13-methyl-tetradecanoic acid (76%), followed by hexadecanoic acid (6%) and 3-hydroxy-15-methyl-hexadecanoic acid (4%). A 16S rDNA-based similarity analysis showed that the Marseille-Q4570T strain was closely related to Capnocytophaga leadbetteri strain AHN8855T (97.24% sequence identity). The OrthoANI and dDDH values between these two strains were, respectively, 76.81% and 25.6%. Therefore, we conclude that the Marseille-Q4570T strain represents a novel species of the genus Capnocytophaga, for which the name Capnocytophaga bilenii sp. nov. is proposed (=CSUR Q4570).

Clostridium perfringens strains cause a wide variety of human and animal disease, including gas gangrene or myonecrosis. Production of toxins required for myonecrosis, PFO and CPA, is regulated by the C. perfringens Agr-like (CpAL) system via the VirSR two-component system. Myonecrosis begins at the site of infection from where bacteria migrate deep into the host tissue likely using a previously described gliding motility phenotype. We therefore assessed whether gliding motility was under the control of the CpAL/VirSR regulon. The migration rate of myonecrosis-causing C. perfringens strain 13 (S13) was investigated during a 96 h period, including an adaptation phase with bacterial migration (∼1.4 mm/day) followed by a gliding phase allowing bacteria faster migration (∼8.6 mm/day). Gliding required both an intact CpAL system, and signaling through VirSR. Mutants lacking ΔagrB, or ΔvirR, were impaired for onward gliding while a complemented strain S13ΔagrB/pTS1303 had the gliding phenotype restored. Gene expression studies revealed upregulated transcription of pili genes (pilA1, pilA2 and pilT) whose encoded proteins were previously found to be required for gliding motility and CpAL/VirSR-regulated pfoA and cpa toxin genes. Compared to S13, transcription of cpa and pfoA significantly decreased in S13ΔagrB, or S13ΔvirR, strains but not that of pili genes. Further experiments demonstrated that mutants S13ΔpfoA and S13Δcpa migrated at the same rate as S13 wt. We demonstrated that CpAL/VirSR regulates C. perfringens gliding motility and that gliding bacteria have an increased transcription of toxin genes involved in myonecrosis.

3D imaging of live bacteria with optical microscopy techniques is a challenge due to the small size of bacterial cells, meaning that previous studies have been limited to observing motility behavior in 2D. We introduce the application of confocal multiwavelength interference reflection microscopy to bacteria, which enables visualization of 3D motility behaviors in a single 2D image. Using the model organism Myxococcus xanthus, we identified novel motility behaviors that are not explained by current motility models, where gliding bacteria exhibit aperiodic changes in their adhesion to an underlying solid surface. We concluded that the 3D behavior was not linked to canonical motility mechanisms and that IRM could be applied to study a range of microbiological specimens with minimal adaptation to a commercial microscope., The deltaproteobacterium Myxococcus xanthus is a model for bacterial motility and has provided unprecedented insights into bacterial swarming behaviors. Fluorescence microscopy techniques have been invaluable in defining the mechanisms that are involved in gliding motility, but these have almost entirely been limited to two-dimensional (2D) studies, and there is currently no understanding of gliding motility in a three-dimensional (3D) context. We present here the first use of confocal interference reflection microscopy (IRM) to study gliding bacteria, revealing aperiodic oscillatory behavior with changes in the position of the basal membrane relative to the substrate on the order of 90 nm in vitro. First, we use a model planoconvex lens specimen to show how topological information can be obtained from the wavelength-dependent interference pattern in IRM. We then use IRM to observe gliding M. xanthus bacteria and show that cells undergo previously unobserved changes in their adhesion profile as they glide. We compare the wild type with mutants that have reduced motility, which also exhibit the same changes in the adhesion profile during gliding. We find that the general gliding behavior is independent of the proton motive force-generating complex AglRQS and suggest that the novel behavior that we present here may be a result of recoil and force transmission along the length of the cell body following firing of the type IV pili. IMPORTANCE 3D imaging of live bacteria with optical microscopy techniques is a challenge due to the small size of bacterial cells, meaning that previous studies have been limited to observing motility behavior in 2D. We introduce the application of confocal multiwavelength interference reflection microscopy to bacteria, which enables visualization of 3D motility behaviors in a single 2D image. Using the model organism Myxococcus xanthus, we identified novel motility behaviors that are not explained by current motility models, where gliding bacteria exhibit aperiodic changes in their adhesion to an underlying solid surface. We concluded that the 3D behavior was not linked to canonical motility mechanisms and that IRM could be applied to study a range of microbiological specimens with minimal adaptation to a commercial microscope.

Three filamentous gliding bacteria from the German Collection of Microorganisms and Cell Cultures, Hp g11, Hp g471 and Hp g472, were subjected to a phylogenetic analysis. These organisms had previously been classified as members of the genus Herpetosiphon based on their growth physiology and morphology. However, a taxonomic assignment at the species level had not been carried out. Analysis of 16S rRNA sequences now confirmed the close relationship of strain Hp g472 to Herpetosiphon aurantiacus DSM 785T (98.6 % nucleotide identity) and Herpetosiphon geysericola DSM 7119T (97.7 %). The results of DNA–DNA hybridization experiments further implied that strain Hp g472 should be classified as a distinct species. The DNA G+C content of strain Hp g472 was 49.9 mol%. The major quinone was MK-10 and the predominant cellular fatty acids were C18 : 1, C16 : 1 and C16 : 0. Based on phenotypic, chemotaxonomic and phylogenetic data it was concluded that strain Hp g472 represents a novel species of the genus Herpetosiphon , for which the name Herpetosiphon gulosus sp. nov. is proposed. The type strain is Hp g472T (=DSM 52871T=NBRC 112829T). In contrast to Hp g472T, the strains Hp g11 and Hp g471 exhibited closest 16S rRNA gene sequence similarity (>99 %) with ‘Herpetosiphon giganteus’ Hp a2. The distinctive genotypic and phenotypic properties of the latter supported the revival of the name as Herpetosiphon giganteus (ex Reichenbach & Golecki, 1975) sp. nov., nom. rev. We propose the previously deposited reference strain DSM 589T=NBRC 112828T as the type strain.

The Delta-proteobacterium,Myxococcus xanthus, has been used as a model for bacterial motility and to provide insights of bacterial swarming behaviours. Fluorescence microscopy techniques have shown that various mechanisms are involved in gliding motility, but these have almost entirely been limited to 2D studies and there is currently no understanding of gliding motility in a 3D context. We present here the first use of confocal interference reflection microscopy (IRM) to study gliding bacteria, and we reveal aperiodic oscillatory behaviour with changes in the position of the basal membrane relative to the coverglass on the order of 90 nmin vitro. Firstly, we use a model plano-convex lens specimen to show how topological information can be obtained from the wavelength-dependent interference pattern in IRM. We then use IRM to observe glidingM. xanthusand show that cells undergo previously unobserved changes in their height as they glide. We compare the wild-type with mutants of reduced motility, which also exhibit the same changes in adhesion profile during gliding. We find that the general gliding behaviour is independent of the proton motive force-generating complex, AglRQS, and suggest that the novel behaviour we present here may be a result of recoil and force transmission along the length of the cell body following firing of the Type IV pili.

Desulfonema ishimotonii strain Tokyo 01T is a filamentous sulfate-reducing bacterium isolated from a marine sediment. In this study, the genome of this strain was sequenced and analyzed with a focus on gene transfer from phylogenetically distant organisms. While the strain belongs to the class Deltaproteobacteria, hundreds of proteins encoded in the genome showed the highest sequence similarities to those of organisms outside of the class Deltaproteobacteria, suggesting that more than 20% of the genome is putatively of foreign origins. Many of these proteins had the highest sequence identities with proteins encoded in the genomes of filamentous bacteria, including giant sulfur oxidizers of the orders Thiotrichales, cyanobacteria of various genera, and uncultured bacteria of the candidate phylum KSB3. As mobile genetic elements transferred from phylogenetically distant organisms, putative inteins were identified in the GyrB and DnaE proteins encoded in the genome of strain Tokyo 01T. Genes involved in DNA recombination and repair were enriched in comparison to the closest relatives in the same family. Some of these genes were also related to those of organisms outside of the class Deltaproteobacteria, suggesting that they were acquired by horizontal gene transfer from diverse bacteria. The genomic data suggested significant genetic transfer among filamentous gliding bacteria in phylogenetically dispersed lineages including filamentous sulfate reducers. This study provides insights into the genomic evolution of filamentous bacteria belonging to diverse lineages, characterized by various physiological functions and different ecological roles.

For many bacteria, motility is essential for survival, growth, virulence, biofilm formation and intra/interspecies interactions. Since natural environments differ, bacteria have evolved remarkable motility systems to adapt, including swimming in aqueous media, and swarming, twitching and gliding on solid and semi-solid surfaces. Although tremendous advances have been achieved in understanding swimming and swarming motilities powered by flagella, and twitching motility powered by Type IV pili, little is known about gliding motility. Bacterial gliders are a heterogeneous group containing diverse bacteria that utilize surface motilities that do not depend on traditional flagella or pili, but are powered by mechanisms that are less well understood. Recently, advances in our understanding of the molecular machineries for several gliding bacteria revealed the roles of modified ion channels, secretion systems and unique machinery for surface movements. These novel mechanisms provide rich source materials for studying the function and evolution of complex microbial nanomachines. In this review, we summarize recent findings made on the gliding mechanisms of the myxobacteria, flavobacteria and mycoplasmas.

Gliding bacteria are an assorted group of rod-shaped prokaryotes that adhere to and glide on certain layers of ooze slime attached to a substratum. Due to the absence of organelles of motility, such as flagella, the gliding motion is caused by the waves moving down the outer surface of these rod-shaped cells. In the present study we employ an undulating surface model to investigate the motility of bacteria on a layer of non-Newtonian slime. The rheological behavior of the slime is characterized by an appropriate constitutive equation, namely the Carreau model. Employing the balances of mass and momentum conservation, the hydrodynamic undulating surface model is transformed into a fourth-order nonlinear differential equation in terms of a stream function under the long wavelength assumption. A perturbation approach is adopted to obtain closed form expressions for stream function, pressure rise per wavelength, forces generated by the organism and power required for propulsion. A numerical technique based on an implicit finite difference scheme is also employed to investigate various features of the model for large values of the rheological parameters of the slime. Verification of the numerical solutions is achieved with a variational finite element method (FEM). The computations demonstrate that the speed of the glider decreases as the rheology of the slime changes from shear-thinning (pseudo-plastic) to shear-thickening (dilatant). Moreover, the viscoelastic nature of the slime tends to increase the swimming speed for the shear-thinning case. The fluid flow in the pumping (generated where the organism is not free to move but instead generates a net fluid flow beneath it) is also investigated in detail. The study is relevant to marine anti-bacterial fouling and medical hygiene biophysics.

A group of rod-shaped, aerobic, Gram-stain-negative, gliding bacteria producing flexirubin-type pigment was isolated from environmental samples collected in Antarctica in 2009-2014. Phylogenetic analysis of the almost complete 16S rRNA gene sequences revealed two separated branches belonging to the genus Flavobacterium. All analyses confirmed that strains of group I and II represent two novel species of the genus Flavobacterium, for which the names Flavobacterium chryseum sp. nov. (type strain CCM 8826T=P3160T=LMG 30615T) and Flavobacterium psychroterrae sp. nov. (type strain CCM 8827T=P3922T=LMG 30616T) are proposed.

Lysobacter enzymogenes C3 is a predatory strain of gram-negative gliding bacteria that produces antifungal antibiotics by the polyketide synthetic pathway. Outer membrane vesicles (OMV) are formed as a stress response and can deliver virulence factors to host cells. The production of OMV by C3 and their role in antifungal activity are reported here. Vesicles in the range of 130-150 nm in diameter were discovered in the cell-free supernatants of C3 cultures. These OMV contain molecules characteristic of bacterial outer membranes, such as lipopolysaccharide and phospholipids. In addition, they contain chitinase activity and essentially all of the heat stable antifungal activity in cell supernatants. We show here that C3 OMV can directly inhibit growth of the yeast Saccharomyces cerevisiae as well as the filamentous fungus Fusarium subglutinans. The activity is dependent on physical contact between OMV and the cells. Furthermore, fluorescent lipid labeling of C3 OMV demonstrated transfer of the membrane-associated probe to yeast cells, suggesting the existence of a mechanism of delivery for membrane-associated molecules. Mass spectrometric analysis of C3 OMV extracts indicates the presence of molecules with molecular weights identical to some of the previously identified antifungal products of C3. These data together suggest that OMV act as an important remote mobile component of predation by Lysobacter.ImportanceThe data presented here suggest a newly discovered function of outer membrane vesicles (OMV) that are produced from the outer membrane of the bacterial species Lysobacter enzymogenes C3. We show that these OMV can be released from the surface of the cells to deliver antibiotics to target fungal organisms as a mechanism of killing or growth inhibition. Understanding the role of OMV in antibiotic delivery can generally lead to improved strategies for dealing with antibiotic-resistant organisms. These results also add to the evidence that some bacterially produced antibiotics can be discovered and purified using methods designed for isolation of nanoscale vesicles. Information on these systems can lead to better identification of active molecules or design of delivery vehicles for these molecules.

The human microbiome is an assemblage of diverse bacteria that interact with one another to form communities. Bacteria in a given community are arranged in a three-dimensional matrix with many degrees of freedom. Snapshots of the community display well-defined structures, but the steps required for their assembly are not understood. Here, we show that this construction is carried out with the help of gliding bacteria. Gliding is defined as the motion of cells over a solid or semi-solid surface without the necessity of growth or the aid of pili or flagella. Genomic analysis suggests that gliding bacteria are present in human microbial communities. We focus on Capnocytophaga gingivalis which is present in abundance in the human oral microbiome. Tracking of fluorescently-labeled single cells and of gas bubbles carried by fluid flow shows that swarms of C. gingivalis are layered, with cells in the upper layers moving more rapidly than those in the lower layers. Thus, cells also glide on top of one another. Cells of non-motile bacterial species attach to the surface of C. gingivalis and are propelled as cargo. The cargo cell moves along the length of a C. gingivalis cell, looping from one pole to the other. Multi-color fluorescent spectral imaging of cells of different live but non-motile bacterial species reveals their long-range transport in a polymicrobial community. A swarm of C. gingivalis transports some non-motile bacterial species more efficiently than others and helps shape the spatial organization of a polymicrobial community.SignificanceWe describe a situation in which bacteria typical of the human oral microbiome are organized spatially by gliding cells, species of Capnocytophaga, that move backwards and forwards over the substratum. The mobile adhesins that pull the cells over the substratum also attach to cells of non-motile bacterial species, which are carried up and down the motile cells as cargo. The synchronized transport of non-motile cargo bacteria helps shape a polymicrobial community.

Gliding bacteria are virtually everywhere. These organisms are phylogenetically diverse with their hundreds of types, different shapes and several modes of motility. One possible mode of gliding motility in the rod shaped bacteria is that they propel themselves by producing undulating waves in their body. Few bacteria glides near the solid surface over the slime without any aid of flagella so the classical Navier-Stokes equations are incapable of explaining the slime rheology at the microscopic level. Micropolar fluid dynamics however provides a solid framework for mimicking bacterial physical phenomena at both micro and nano-scales, and therefore we use the micropolar fluid to characterize the rheology of a thin layer of slime and its dominant microrotation effects. It is also assumed that there is a certain degree of slip between slime and bacterial undulating surface and also between slime and solid substrate. The flow equations are formulated under long wavelength and low Reynolds number assumptions. Exact expressions for stream function and pressure gradient are obtained. The speed of the gliding bacteria is numerically calculated by using a modified Newton-Raphson method. Slip effects and effects of non-Newtonian slime parameters on bacterial speed and power are also quantified. In addition, when the glider is fixed, the effects of slip and rheological properties of micropolar slime parameters on the velocity, micro-rotation (angular velocity) of spherical slime particles, pressure rise per wavelength, pumping and trapping phenomena are also shown graphically and discussed in detail. The study is relevant to emerging biofuel cell technologies and also bacterial biophysics. Keywords: Gliding bacteria, Undulating surface model, Micropolar fluid, Slip effects, Exact solution, Modified Newton-Raphson method

In recent decades, bacterial cell biology has seen great advances, and numerous model systems have been developed to study a wide variety of cellular processes, including cell division, motility, assembly of macromolecular structures, and biogenesis of cell polarity. Considerable attention has been given to these model organisms, which include Escherichia coli , Bacillus subtilis , Caulobacter crescentus , and Myxococcus xanthus . Studies of these processes in the pathogenic bacterium Mycoplasma pneumoniae and its close relatives have also been carried out on a smaller scale, but this work is often overlooked, in part due to this organism's reputation as minimalistic and simple. In this minireview, I discuss recent work on the role of the M. pneumoniae attachment organelle (AO), a structure required for adherence to host cells, in these processes. The AO is constructed from proteins that generally lack homology to those found in other organisms, and this construction occurs in coordination with cell cycle events. The proteins of the M. pneumoniae AO share compositional features with proteins with related roles in model organisms. Once constructed, the AO becomes activated for its role in a form of gliding motility whose underlying mechanism appears to be distinct from that of other gliding bacteria, including Mycoplasma mobile . Together with the FtsZ cytoskeletal protein, motility participates in the cell division process. My intention is to bring this deceptively complex organism into alignment with the better-known model systems.

Various phylogenetic categorized groups of rod-shaped bacteria exhibit the gliding mechanism over solid surfaces without any aid of flagella. It is postulated that such gliding bacteria push themselves by means of generating waves in their own surface and leave an adhesive trail of slime. In the subsequent analysis, two different non-Newtonian fluid models i.e., Bingham model and Carreau model are used to describe the complex rheology of the slime. We assumed that the solid substrate present beneath the bacterium is inclined at a certain angle to the horizontal. We derive equations governing the flow for each fluid model under long wavelength assumption. In order to calculate the bacterial gliding speed and slime flow rate we employ finite difference method combined with Broyden root finding algorithm. The obtained speed and energy dissipation are plotted for various inclination angles and rheological parameters. Moreover, by using the realistic (calculated) pairs of the glider’s speed and flow rate, the flow patterns are shown graphically and discussed in detail. The present work is an extension to our previous study Ali et al. (2016) in which a reduced constitutive equation of Carreau model was solved by perturbative approach to obtain the gliding speed on a straight substrate valid for small values of rheological parameters. The most significant outcome of the present numerical work is that the speed of gliding bacteria can be controlled by adjusting the slime rheology and inclination angle.

The phylum Bacteroidetes is large and diverse, with rapid gliding motility and the ability to digest macromolecules associated with many genera and species. Recently, a novel protein secretion system, the Por secretion system (PorSS), was identified in two members of the phylum, the gliding bacterium Flavobacterium johnsoniae and the nonmotile oral pathogen Porphyromonas gingivalis . The components of the PorSS are not similar in sequence to those of other well-studied bacterial secretion systems. The F. johnsoniae PorSS genes are a subset of the gliding motility genes, suggesting a role for the secretion system in motility. The F. johnsoniae PorSS is needed for assembly of the gliding motility apparatus and for secretion of a chitinase, and the P. gingivalis PorSS is involved in secretion of gingipain protease virulence factors. Comparative analysis of 37 genomes of members of the phylum Bacteroidetes revealed the widespread occurrence of gliding motility genes and PorSS genes. Genes associated with other bacterial protein secretion systems were less common. The results suggest that gliding motility is more common than previously reported. Microscopic observations confirmed that organisms previously described as nonmotile, including Croceibacter atlanticus , “ Gramella forsetii ,” Paludibacter propionicigenes , Riemerella anatipestifer , and Robiginitalea biformata , exhibit gliding motility. Three genes ( gldA , gldF , and gldG ) that encode an apparent ATP-binding cassette transporter required for F. johnsoniae gliding were absent from two related gliding bacteria, suggesting that the transporter may not be central to gliding motility.

Bacteria which do not have organelles of motility, such as flagella, adopt gliding as a mode of locomotion. In gliding motility bacterium moves under its own power by secreting a layer of slime on the substrate. The exact mechanism by which a glider achieves motility is yet in controversy but there are evidences which support the wave-like undulation on the surface of the organism, as a possible mechanism of motility. Based on this observation, a model of undulating sheet over a layer of slime is examined as a possible model of the gliding motion of a bacterium. Three different non-Newtonian constitutive equations namely, finite extendable nonlinear elastic-peterline (FENE-P), Simplified Phan-Thien-Tanner (SPTT) and Rabinowitsch equations are used to capture the rheological properties of the slime. It is found that the governing equation describing the fluid mechanics of the model under lubrication approximation is same for all the considered three constitutive equations. In fact, it involves a single non-Newtonian parameter which assumes different values for each of the considered constitutive relations. This differential equation is solved using both perturbation and semi-analytic procedure. The perturbation solution is exploited to get an estimate of the speed of the glider for different values of the non-Newtonian parameter. The solution obtained via semi-analytic procedure is used to investigate the important features of the flow field in the layer of the slime beneath the glider when the glider is held fixed. The expression of forces generated by the organism and power required for propulsion are also derived based on the perturbation analysis.

Among the bacteria that glide on substrate surfaces, Mycoplasma mobile is one of the fastest, exhibiting smooth movement with a speed of 2.0-4.5 μm s−1 with a cycle of attachment to and detachment from sialylated oligosaccharides1. To study the gliding mechanism at the molecular level, we applied an assay with a fluorescently-labeled and membrane-permeabilized ghost model, and investigated the motility by high precision colocalization microscopy2. In the presence of free sialyllactose in the solution, which reduced the number of functional gliding units, stepwise displacement was shown for the first time to occur in gliding bacteria. Regardless of the random orientation of the binding target on the substrate surface, ghosts exhibit clean steps with a size of 70 nm in the gliding direction in the presence of methylcellulose reducing diffusive motions. Although the number of gliding machines involved in forward movement during the unitary step was not conclusively determined in our assay, this might be a new characteristic not to seen in conventional linear molecular motors, which follow the unitary distances between binding sites arranged on a track of cytoskeletons.We also quantified the motility of halophylic archaea, Halobacterium salinarum, at the single cell level in a microscopic detail. Symmetrical intermittent pauses during rotation was detected, possibly indicating unitary steps consuming single ATPs. The energetic efficiency of the motor was estimated as ∼70% from the size of stepping angle.REFERENCES1Miyata, M. 2010. Annu. Rev. Microbiol. 64:519-537.2Kinosita, Y., D. Nakane, et al., T.Nishizaka. 2014. Proc. Natl. Acad. Sci. USA. 111:8601-8606.

In recent years the discovery of some most important antibiotic compounds obtained by fermenting environmental microbes has been reported, providing proof that isolation and fermentation of producer strains is a significant approach to decifer novel structural types of antibiotics. Whereas many microbial taxa and environments have been well investigated in the past (e.g. soil-borne actinomycetes), the high diversity of microbial populations in certain habitats, e.g. marine sediments, has to date only been exploited marginally. Myxobacteria, the most prominent class of gliding bacteria, are well known for their ability to produce structurally intriguing natural products; however, so far no myxobacterial antibiotic has been developed for clinical use. In our studies, the antibacterial activity of the myxobacterial metabolite corallopyronin A was further investigated. Feeding studies with labeled precursors allowed to deduce all building blocks for the formation of corallopyronin A, whereby its biosynthesis from two chains probably connected by a Claisen-type reaction and the incorporation of bicarbonate into the methyl carbamate functionality can be regarded as unusual characteristics. A trans-AT type mixed PKS/NRPS gene cluster containing a β-branching cassette was identified as the putative basis for corallopyronin A biosynthesis in Corallococcus coralloides. Our research also resulted in the cultivation of several unusual marine myxobacteria which produce antibiotically active molecules.

Bacterial gliding motility is the smooth movement of cells on solid surfaces unaided by flagella or pili. Many diverse groups of bacteria exhibit gliding, but the mechanism of gliding motility has remained a mystery since it was first observed more than a century ago. Recent studies on the motility of Myxococcus xanthus, a soil myxobacterium, suggest a likely mechanism for gliding in this organism. About forty M. xanthus genes were shown to be involved in gliding motility, and some of their protein products were labeled and localized within cells. These studies suggest that gliding motility in M. xanthus involves large multiprotein structural complexes, regulatory proteins, and cytoskeletal filaments. In this review, we summarize recent experiments that provide the basis for this emerging view of M. xanthus motility. We also discuss alternative models for gliding.

SUMMARY In bacteria, motility is important for a wide variety of biological functions such as virulence, fruiting body formation, and biofilm formation. While most bacteria move by using specialized appendages, usually external or periplasmic flagella, some bacteria use other mechanisms for their movements that are less well characterized. These mechanisms do not always exhibit obvious motility structures. Myxococcus xanthus is a motile bacterium that does not produce flagella but glides slowly over solid surfaces. How M. xanthus moves has remained a puzzle that has challenged microbiologists for over 50 years. Fortunately, recent advances in the analysis of motility mutants, bioinformatics, and protein localization have revealed likely mechanisms for the two M. xanthus motility systems. These results are summarized in this review.

Recent studies of the gliding bacteria Mycoplasma mobile have identified a family of proteins called the Gli family which was considered to be involved in this novel and yet fairly unknown motility system. The 349 kDa protein called Gli349 was successfully isolated and purified from the bacteria, and electron microscopy imaging and antibody experiments led to the hypothesis that it acts as the “leg” of M. mobile, responsible for attachment to the substrate as well as for gliding motility. However, more precise evidence of the molecular shape and function of this protein was required to asses this theory any further. In this study, an atomic force microscope (AFM) was used both as an imaging and a force measurement device to provide new information about Gli349 and its role in gliding motility. AFM images of the protein were obtained revealing a complex structure with both rigid and flexible parts, consistent with previous electron micrographs of the protein. Single-molecular force spectroscopy experiments were also performed, revealing that Gli349 is able to specifically bind to sialyllactose molecules and withstand unbinding forces around 70 pN. These findings strongly support the idea that Gli349 is the “leg” protein of M. mobile, responsible for binding and also most probably force generation during gliding motility.

Myxobacteria are gliding bacteria that belong to the delta-Proteobacteria and are known for their unique biosynthetic capabilities. Among myxobacteria, Nannocystis spp. are most closely related to marine myxobacteria and their secondary metabolism has hardly been investigated. Phenylnannolones A (1), B (2) and C (3) were obtained from a culture of Nannocystis exedens that was isolated from the intertidal region of Crete. Compound 1 had inhibitory activity toward the ABCB1 gene product P-glycoprotein and reversed daunorubicin resistance in cultured cancer cells. Phenylnannolone A has an unusual structural architecture; it is composed of an ethyl-substituted polyene chain linked to a pyrone moiety on one side and to a phenyl ring on the other. The investigation of the biosynthesis with labelled precursors revealed acetate, butyrate and phenylalanine as building blocks for 1. The labelling pattern suggested novel biochemical reactions for the biosynthesis of the starter unit.

Bacterial motility is essential for chemotaxis, virulence and complex social interactions leading to biofilm and fruiting body formation. Although bacterial swimming in liquids with a flagellum is well understood, little is known regarding bacterial movements across solid surfaces. Gliding motility, one such mode of locomotion, has remained largely mysterious because cells move smoothly along their long axis in the absence of any visible organelle. In this review, I discuss recent evidence that focal adhesion systems mediate gliding motility in the social bacterium Myxococcus xanthus and combine this evidence with previous work to suggest a new working hypothesis inspired from knowledge in apicomplexan parasites. I then propose experimental directions to test the model and compare it to other pre-existing models. Finally, evidence on gliding mechanisms of selected organisms are presented to ask whether some features of the model have precedents in other bacteria and whether this complex biological process could be explained by a single mechanism or involves multiple distinct mechanisms.

Gliding bacteria are a taxonomically heterogeneous group of rod-shaped prokaryotes that adhere to and translocate on certain substrata by mechanism(s) that are unknown. Organelles of motility, such as flagella, have not been observed on these bacteria. Furthermore, their rigidified cell walls preclude amoeboid type locomotion. The present investigation analyzes a motility model that involves an undulating cell surface which transmits stresses through a layer of exuded slime to the substratum. The non-Newtonian nature of the slime is taken into account by considering a specific fluid of the differential type, namely an incompressible homogenous fluid of grade three. The non-linear differential equation resulting from the balance of momentum and mass is solved analytically and numerically.

The antifungal and cytotoxic metabolites vioprolides A-D (1–4) were obtained from the myxobacterium Cystobacter violaceus strain Cb vi35 by fermentation in the presence of the adsorber resin XAD-1180. Peptolide structures 1–4 composed of eight amino acids and a glyceric acid were determined by spectroscopic methods and total hydrolysis. The absolute configurations of the building blocks were established by GC analysis on chiral stationary phases. Common building blocks of 1–4 are l-Ala, l-Cys, D-Leu, l-Thr, l-MeVal, (E)-dehydrobutyrine (E-Dhb), and l-glyceric acid (l-Gla). Two variable positions are occupied by l-Pro or l-Homopro and l-Pro or trans-(2S,4R)-4-methylazetidinecarboxylic acid [(2S,4R)-Maz]. Two defined isomers of 1 were detected in CDCl3 in a ratio of 3 : 2 (2D-1H NOESY) and assigned to rotational isomers at the amide bond of ThrMeVal.

The absolute configuration of tartrolon B (1), a boron-containing macrodiolide from Sorangium cellulosum, was established by X-ray structure analysis as (2R,3R,4R,7S,8S,11S). Compared to the related boron-containing macrodiolides boromycin (2), aplasmomycin (3), and borophycin (4), and identical substructure and configuration of the boron-binding site was found for all four compounds. The biosynthesis of 1 was investigated by feeding experiments with 13C-labeled precursors. According to NMR analyses, the carbon atoms from C-1 to C-18 are derived from acetate units, the two methyl groups at C-22 and C-23 are derived from methionine, and a three-carbon starter unit C-19 to C-21 is derived from glycerol. The biosynthesis of 1 is closely related to those of 2 and 3 but with respect to the origin of the starter unit it is more distant to that of 4.

As a result of the extension of our screening to strains of Chondromyces four new antifungal and highly cytostatic depsipeptides were isolated, the chondramides A (1) – D (4). Their macrocyclic structures comprise three amino acids and a polyketide, viz. alanine, N-methyltryptophan, α-methoxyl-β-tyrosine, and a 7-hydroxytrimethyloctenoic acid. Thus, they were found to be closely related to a small class of cyclodepsipeptides including jaspamide, the geodiamolides, and neosiphoniamolide which all have been isolated from marine sponges.

Two groups of 3-substituted indole 1, 6–9 as well as 4-substituted quinoline derivatives 2–5 and harman (10) were isolated from various species and genera of myxobacteria. They were identified by their spectroscopic data. Some of the metabolites show moderate antifungal activity. Their biosynthetic origin in myxobacteria was demonstrated for 1, 2, and 3 by incorporation of L-[1'-14C]tryptophan.

During the search for thermophilic myxobacteria in geothermal environments, four myxobacteria-like cultures that grew at temperatures of up to 50°C and optimally at 45–49°C were enriched from various hot springs in Japan. Three of the cultures were derived from freshwater hot springs and one from a coastal saline spring. Although all lacked fruiting structures, they were bacteriolytic and formed diffusive shallow crater-like colonies. Even after repeated enrichment procedures using Escherichia coli-prey agar media, the crater-like cultures were usually surrounded by thin, film-like, spreading colonies of other gliding bacteria that grew faster. Within the crater-like sunken area, rod-shaped bacterial cells were observed. 16S rRNA gene sequences from the myxobacterial cultures were amplified by nested PCR with myxobacteria-specific primers, subcloned, and phylogenetically analyzed. All of the myxobacterial clones obtained from the three cultures were assigned to the suborder Sorangineae. These clones were distantly positioned within this suborder, and mostly shown to represent new genera of myxobacteria. These results suggested that diverse species of moderately thermophilic myxobacteria, including many as yet undescribed, ubiquitously inhabit hot spring environments.

Chemical examination of the culture broth extracts of the marine gliding bacterium Saprospira grandis (ATCC 23116) has resulted in the isolation of four new diterpenoids of the neoverrucosane class. The structures of the new diterpenoids, compounds 1-4, were assigned by combined spectroscopic methods emphasizing 2D NMR experiments. The relative stereochemistry of 1 was determined by 2D ROESY NMR methods, while the absolute stereochemistry was assigned by application of the modified Mosher method. This study adds to the rare observation of terpene production by prokaryotic microorganisms and suggests that marine gliding bacteria may be a significant source for new terpenoid secondary metabolites.

A 5'-nuclease PCR assay, targeting 16S rDNA, was developed to detect a group of gliding bacteria that digest Skeletonema costatum cells and are phylogenetically close to Cytophaga latercula. The detection limit was 15 molecules of the target DNA in one reaction mixture. The assay is so strict that the probe did not hybridize to DNA fragments with one nucleotide mismatch, even though the amount of DNA fragments was increased to the order of 1010 molecules. The assay was applied to DNA extracted from natural seawater of Yoshimi Bay, Hibiki-nada Sea, Japan, during the period from August 1998 to February 1999. A positive result was obtained only for a seawater sample of September 10, 1998. Among 30 PCR clones obtained from the 5'-nuclease PCR product of the positive sample, 25 clones gave positive results and 4 clones negative results in the assay. The positive clones examined were identical in the structure of the probe region, whereas negative clones had one nucleotide mismatch or deletion. The results indicate that the assay detects only the target sequence even in natural seawater DNA.

Bull, C. T., Shetty, K. G., and Subbarao, K. V. 2002. Interactions between myxobacteria, plant pathogenic fungi, and biocontrol agents. Plant Dis. 86:889-896. Myxobacteria are soil dwelling gram-negative gliding bacteria that form fruiting bodies containing myxospores. Although myxobacteria produce a wide range of antibiotics and lytic enzymes that assist in their ability to prey on other microorganisms, their role in agriculture has received little attention. Myxococcus spp. were isolated from soils in organic and conventionally managed strawberry production and transplant fields in the absence of soil fumigation. Fumigation with methyl bromide and chloropicrin virtually eliminated these organisms from soil. However, soil fumigation had no effect on the frequency of isolation of Myxococcus spp. from strawberry roots. Six Myxococcus spp. were tested in vitro against eight soilborne plant pathogenic fungi (Cylindrocarpon spp., Fusarium oxysporum f. sp. apii, Phytophthora capsici, Pythium ultimum, Rhizoctonia spp., Sclerotinia minor, Verticillium albo-atrum, and V. dahliae) and against two fungal biological control agents (Gliocladium virens and Trichoderma viride). Phytophthora capsici, Pythium ultimum, Rhizoctonia spp., S. minor, and T. viride were completely inhibited by all of the Myxococcus spp. tested. F. oxysporum f. sp. apii was the least sensitive to the myxobacteria, and no inhibition occurred with some Myxococcus spp. Inhibition of the other fungi tested was variable. Myxococcus coralloides inhibited nearly all the fungi tested. The ability of bacterial biological control agents to produce antibiotics and other secondary metabolites determined whether or not they were lysed by myxobacteria. Secondary metabolite production regulated by gacS protected Pseudomonas fluorescens strain CHA0 from lysis by myxobacteria. More specifically, phenazine antibiotics produced by Pseudomonas aureofaciens strain 30-84 protected it from lysis.

Background Many microorganisms, including myxobacteria, cyanobacteria, and flexibacteria, move by gliding. Although gliding always describes a slow surface-associated translocation in the direction of the cell's long axis, it can result from two very different propulsion mechanisms: social (S) motility and adventurous (A) motility. The force for S motility is generated by retraction of type 4 pili. A motility may be associated with the extrusion of slime, but evidence has been lacking, and how force might be generated has remained an enigma. Recently, nozzle-like structures were discovered in cyanobacteria from which slime emanated at the same rate at which the bacteria moved. This strongly implicates slime extrusion as a propulsion mechanism for gliding. Results Here we show that similar but smaller nozzle-like structures are found in Myxococcus xanthus and that they are clustered at both cell poles, where one might expect propulsive organelles. Furthermore, light and electron microscopical observations show that slime is secreted in ribbons from the ends of cells. To test whether the slime propulsion hypothesis is physically reasonable, we construct a mathematical model of the slime nozzle to see if it can generate a force sufficient to propel M. xanthus at the observed velocities. The model assumes that the hydration of slime, a cationic polyelectrolyte, is the force-generating mechanism. Conclusions The discovery of nozzle-like organelles in various gliding bacteria suggests their role in prokaryotic gliding. Our calculations and our observations of slime trails demonstrate that slime extrusion from such nozzles can account for most of the observed properties of A motile gliding.

Gliding is a means of locomotion on rigid substrates utilized by a number of bacteria includingmyxobacteria and cyanobacteria. One of the hypotheses advanced to explain this motility mechanism hinges on the role played by the slime filaments continuously extruded from gliding bacteria. This paper solves in full a non-linear mechanical theory that treats as dissipative shocks both the point where the extruded slime filament comes in contact with the substrate, called the filament's foot, and the pore on the bacterium outer surface from where the filament is ejected. We prove that kinematic compatibility for shock propagation requires that the bacterium uniform gliding velocity (relative to the substrate) and the slime ejecting velocity (relative to the bacterium) must be equal, a coincidence that seems to have already been observed., Comment: arXiv admin note: text overlap with arXiv:1402.6360

Gliding bacteria are a taxonomically heterogeneous group of rod-shaped prokaryotes that adhere to and translocate (parallel to their longitudinal axis) on certain substrata by mechanism(s) that are unknown. Organelles of motility, such as flagella, have not been observed on these bacteria. Furthermore, their rigidified cell walls preclude amoeboid-type locomotion. Here, we examine a hydrodynamical model of motility involving an undulating cell surface which transmits stresses through a layer of exuded slime to the substratum. It is shown that this mechanism generates a force that can propel the glider at a realistic speed and which requires an output of power that is much less than the organism's metabolic rate of energy production. For the case of a slime that is Newtonian, we find that the lift force on the glider is zero. Therefore, there is one degree of freedom among the dimensionless variables formed from the quantities: speed of the glider; amplitude, wavelength and phase speed of the undulations, and the thickness and viscosity of the secreted slime. Optimization schemes obtain a relation among the dimensionless groups formed from these variables. However for non-Newtonian slime, represented by the model of the third-order approximation of a simple fluid, a lift force is generated due to the normal stresses. Also for this shear-thinning viscoelastic fluid, the power required for translation is reduced.

Microscopy of organic-rich, sulfidic sediment samples of marine and freshwater origin revealed filamentous, multicellular microorganisms with gliding motility. Many of these neither contained sulfur droplets such as the Beggiatoa species nor exhibited the autofluorescence of the chlorophyll-containing cyanobacteria. A frequently observed morphological type of filamentous microorganism was enriched under anoxic conditions in the dark with isobutyrate plus sulfate. Two strains of filamentous, gliding sulfate-reducing bacteria, Tokyo 01 and Jade 02, were isolated in pure cultures. Both isolates oxidized acetate and other aliphatic acids. Enzyme assays indicated that the terminal oxidation occurs via the anaerobic C(1) pathway (carbon monoxide dehydrogenase pathway). The 16S rRNA genes of the new isolates and of the two formerly described filamentous species of sulfate-reducing bacteria, Desulfonema limicola and Desulfonema magnum, were analyzed. All four strains were closely related to each other and affiliated with the delta-subclass of Proteobacteria. Another close relative was the unicellular Desulfococcus multivorans. Based on phylogenetic relationships and physiological properties, Strains Tokyo 01 and Jade 02 are assigned to a new species, Desulfonema ishimotoi. A new, fluorescently labeled oligonucleotide probe targeted against 16S rRNA was designed so that that it hybridized specifically with whole cells of Desulfonema species. Filamentous bacteria that hybridized with the same probe were detected in sediment samples and in association with the filamentous sulfur-oxidizing bacterium Thioploca in its natural habitat. We conclude that Desulfonema species constitute an ecologically significant fraction of the sulfate-reducing bacteria in organic-rich sediments and microbial mats.

Sequencing the 16S rDNA and phylogenetic analysis were performed on algicidal bacteria belong ing to the family Flavobacteriaceae, Chattonella-killing Cytophaga sp. strain J18/MOI, Heterocapsa killing Cytophaga sp. strains AA8-2 and AA8-3, and Gymnodinium growth-inhibiting Flavobacterium sp. strain 5N-3. These algicidal strains were clustered phylogenetically within the marine Cytophaga/Flavobacter/Bacteroides (CFB) group. Flavobacterium sp. 5N-3 was phylogenetically clustered with Psychroserpens burtonensis but not with the algicidal gliding bacteria. The sequence simi larity between strain 5N-3 and P. burtonensis was only 92%. The strains J18/M01 and AA8-2 (AA8-3) exhibited the high level of sequence similarity of 98.5%. Despite the differences of host algae and of lo cality and year of isolation, these algicidal bacteria were phylogenetically located on the same terminal branch. The PCR amplification of the 16S rDNA of the CFB strains using a single set of J18/MO1-specific primers resulted in specific detection of the J18/M01, indicating that the J18/M01-specific sequence in the 16S rDNA is useful for selective detection of the strain J18/M01.

Gliding and near-surface swimming of microorganisms are described as a mobile form of microbial adhesion that need not necessarily be reversible. It is argued that the reversibility of microbial adhesion depends on the depth of the secondary interaction minimum, calculated from the forces between an organism and a substratum acting in a direction perpendicular to the substratum surface. The mobility of adhering microorganisms depends on lateral interactions between the organisms. On ideally homogeneous and smooth model surfaces, only mobile adhesion occurs because the multibody, lateral interactions are weak compared with the thermal or Brownian motion energy of the organisms. Minor chemical or structural heterogeneities, which exist on all real-life surfaces, yield a lateral interaction on adhering microorganisms. This causes their immobilization, which helps to explain the physicochemical nature of microbial gliding or near-surface swimming. Moreover, these lateral interaction energies are one order of magnitude smaller than the Lifshitz-Van der Waals, electrostatic, and acid-base forces acting perpendicular to substratum surfaces that are responsible for adhesion.

Many bacteria glide over surfaces without the aid of flagella. Gliding is still somewhat mysterious, but recent studies show that it involves specialized secretory systems that assemble membrane-associated filaments, and the recognition of extracellular components that trigger movement via transmembrane transducers.

The myxobacterial metabolites chivosazole A (1) and its variants (2–6) were discovered in Sorangium cellulosum, strain So ce12, which simultaneously provides the broad spectrum antibiotic sorangicin as well as the sorangiolides and disorazoles. The antifungal and cytotoxic chivosazoles (1–6) are novel glycosides of 6-deoxyglucopyranose derivatives and an aglycon that includes an oxazole in its 31-membered macrolide ring. The aglycon itself, chivosazole F (7), was formed by strain So ce885 and showed similar activity antibiotic and cytotoxic. The biogenetic origin of the structural elements in chivosazole F (7) was studied by feeding experiments with [1-13C]-, [1,2-13C]acetate, [methyl-13C]methionine and [1-13C]serine. Accordingly, the aglycon 7 is a polyketide assembled by condensation of nine acetate units, one serine unit and a further seven acetate units. While C-1 of serine is part of the macrolide ring, the amido to hydroxyl part of the serine together with C-1 of the adjacent N-acetyl unit form the 1,3-oxazole ring. C- and O-methyl groups are derived from methionine.