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[Modified method for determining water pollution by using a Serratia marcescens culture].

The method for determining the degree of water pollution with the aid of a bacterial tracer has been improved by using: (1) a technique for maintaining the culture of Serratia marcescens; (2) a medium whose composition provided for a high level of prodigiosin in the bacterial culture; and (3) an express method for assaying the degree of pigmentation during growth of Ser. marcescens on solid media.

Culture Media↗
PubMed1975

Mutants of Serratia marcescens lacking cyclic nucleotide phosphodiesterase activity and requiring cyclic 3',5'-AMP for the utilization of various carbohydrates.

Adenine requiring mutants of Serratia marcescens SM-6-F'lac+ have been found to grow well in minimal-glucose medium solely supplemented with cAMP. From one of these ade strains double mutants (called ade cpd) were isolated which could no longer utilize cAMP but which still grew on 5'AMP. Dialyzed cell extracts (soluble fraction) of the double mutants, assayed for cAMP phosphodiesterase, were unable to hydrolyze cAMP whereas cell extracts of the parental strains yielded 5'AMP at a rate of 1.6-2.0 mumoles min-1 mg-1 protein. The loss of the phosphodiesterase activity in S. marcescens cpd W 1181 did not cause an accumulation of large amounts of cAMP as was found for the diesterase-negative mutant AB257pc-1 of Escherichia coli. The induced synthesis of beta-galactosidase in mutant cpd W 1181 showed about the same sensitivity to transient and permanent catabolite (glucose) repression as the corresponding cpd+ strain. Starting from S. marcescens cpd W 1182 three independent double mutants (called cpd cya) were isolated which required exogenous cAMP for utilizing various carbohydrates as carbon source, for motility and for the formation of extracellular lipase and the red pigment prodigiosine. The intracellular concentration of cAMP in these mutants, grown in nutrient broth, was 40-60% of that of the parental strain which is about 4 x 10(-4) M. However, the adenylate cyclase in cell extracts of the mutants W 1237 and W 1270 was like that of the corresponding cya+ strain (about 2 x 10(-2) mumoles min-1 mg-1 protein).

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1991

Lipopolysaccharide is the receptor for kappa phage in Serratia marcescens.

Kappa phage active on Serratia marcescens can form plaques on white and red strains with identical efficiencies. To identify the kappa phage receptor, the inactivation of the phage was studied after incubation with several bacterial subcellular fractions. The experiments demonstrated that kappa phage adsorbs to outer membrane fractions of susceptible cells. Proteinase K did not affect the rate of inactivation. Lipopolysaccharide proved to be the primary receptor for kappa phage. Prodigiosin content of the lipopolysaccharide fraction was low.

Adsorption↗
PubMed2006

N-acylhomoserine lactone-dependent cell-to-cell communication and social behavior in the genus Serratia.

Members of the genus Serratia are increasingly responsible for nosocomial infections, the treatment of which may be complicated by the appearance of multi-antibiotic-resistant strains. Some but not all Serratia strains and species produce N-acylhomoserine lactones (AHLs), and possess luxR and luxI homologous genes. Phylogenetic comparisons have provided evidence for the lateral transfer of these quorum-sensing systems, and in at least one strain of S. marcescens, transfer via a complex transposon has been experimentally demonstrated. AHL-dependent quorum sensing in Serratia controls population surface migration, biofilm development, the biosynthesis of a carbapenem antibiotic and production of the red pigment, prodigiosin. Serratia also possesses LuxS and produces autoinducer-2 (AI-2) which appears to function as a second quorum-sensing system controlling many of the same phenotypes as the LuxR/AHL systems.

4-Butyrolactone↗
PubMed2006

An outbreak of Serratia marcescens on the neonatal unit: a tale of two clones.

Serratia spp. are an important cause of hospital-acquired infections and outbreaks in high-risk settings. Twenty-one patients were infected or colonized over a nine-month period during 2001-2002 on a neonatal unit. Twenty-two isolates collected were examined for antibiotic susceptibility, beta-lactamase production and genotype. Random-amplified polymorphic DNA polymerase chain reaction and pulsed-field gel electrophoresis revealed that two clones were present. The first clone caused invasive clinical infection in four babies, and was subsequently replaced by a non-invasive clone that affected 14 babies. Phenotypically, the two strains also differed in their prodigiosin production; the first strain was non-pigmented whereas the second strain displayed pink-red pigmentation. Clinical features suggested a difference in their pathogenicity. No environmental source was found. The outbreak terminated following enhanced compliance with infection control measures and a change of antibiotic policy. Although S. marcescens continued to be isolated occasionally for another five months of follow-up, these were sporadic isolates with distinct molecular typing patterns.

Clone Cells↗
PubMed2000

Induction of apoptosis of activated murine splenic T cells by cycloprodigiosin hydrochloride, a novel immunosuppressant.

Two types of immunosuppressants, cycloprodigiosin hydrochloride (cPrG) and L-leucyl-L-leucine methyl ester (LeuLeuOMe), both have the ability to selectively inhibit the lysosomal function, and a related compound to cPrG, prodigiosin 25-C, and LeuLeuOMe have been reported to selectively inhibit the T cell function in vitro. We therefore examined the cell-type specificity of cPrG and LeuLeuOMe using murine splenocytes. Concanavalin A (Con A)- and lentil lectin-induced proliferation was suppressed by cPrG more profoundly than lipopolysaccharide-induced proliferation. At the optimal concentration, Con A induced the proliferation of both CD4+ and CD8+ cells, whereas at a supra-optimal concentration Con A induced rather selective proliferation of CD8+ cells. Irrespective of the dose of Con A, CD4+ and CD8+ cells were equally affected by cPrG. In contrast, LeuLeuOMe induced the selective loss of CD8+ cells. cPrG enhanced the apoptosis of murine splenocytes and nylon fiber column-purified T cells cultured in the presence of Con A, as shown by the decrease in cell size and/or DNA fragmentation. Overall, this study revealed that the cell-type specificity of cPrG is different from that of LeuLeuOMe, and that the immunosuppression by cPrG is associated with apoptosis.

Animals↗
PubMed1988

Serratia marcescens infection associated with early abortion in cows and buffaloes.

Serratia marcescens was isolated in pure culture from cases of septic abortion in 4 cows on one farm and 10 buffaloes on two other farms. A reddish vaginal discharge was observed after abortion in all animals and in the internal organs of the aborted fetuses. All but two of the isolates produced prodigiosin, and two of the isolates from buffaloes were atypical in that they fermented raffinose. O-serological, bacteriophage and bacteriocin typing revealed four different strains. All cows were infected by the same strain, and this strain was also isolated from the semen of a breeding bull on the same farm. In another farm a strain of serotype O 14 was isolated from 6 of 10 buffaloes, and two other distinct strains were isolated from the remainder. The strain from the cattle was sensitive to gentamicin and so were two of the buffalo isolates. The infected cows were treated with intra-uterine gentamicin and the organism disappeared from cervical mucus after 3 days. Each animal after abortion showed a raised titre of agglutinating antibody to their respective isolate. A survey of 1172 healthy buffaloes and cattle gave an incidence of 1.8% with raised titres towards S. marcescens.

Abortion, Septic↗
PubMed2006

Facile detection of acyl and peptidyl intermediates on thiotemplate carrier domains via phosphopantetheinyl elimination reactions during tandem mass spectrometry.

With the emergence of drug resistance and the genomic revolution, there has been a renewed interest in the genes that are responsible for the generation of bioactive natural products. Secondary metabolites of one major class are biosynthesized at one or more sites by ultralarge enzymes that carry covalent intermediates on phosphopantetheine arms. Because such intermediates are difficult to characterize in vitro, we have developed a new approach for streamlined detection of substrates, intermediates, and products attached to a phosphopantetheinyl arm of the carrier site. During vibrational activation of gas-phase carrier domains, facile elimination occurs in benchtop and Fourier-transform mass spectrometers alike. Phosphopantetheinyl ejections quickly reduce >100 kDa megaenzymes to <1000 Da ions for structural assignment of intermediates at <0.007 Da mass accuracy without proteolytic digestion. This "top down" approach quickly illuminated diverse acyl intermediates on the carrier domains of the nonribosomal peptide synthetases (NRPSs) or polyketide synthases (PKSs) found in the biosynthetic pathways of prodigiosin, pyoluteorin, mycosubtilin, nikkomycin, enterobactin, gramicidin, and several proteins from the orphan pksX gene cluster from Bacillus subtilis. By focusing on just those regions undergoing covalent chemistry, the method delivered clean proof for the reversible dehydration of hydroxymethylglutaryl-S-PksL via incorporation of 2H or 18O from the buffer. The facile nature of this revised assay will allow diverse laboratories to spearhead their NRPS-PKS projects with benchtop mass spectrometers.

Acylation↗
PubMed1999

Total Synthesis and Structural Refinement of the Cyclic Tripyrrole Pigment Nonylprodigiosin.

The first total synthesis of the cyclic prodigiosin derivative 4 is described, which constitutes a potential lead compound for the development of immunosuppressive agents. The key steps of this approach comprise a palladium-catalyzed Suzuki cross coupling reaction of the rather unstable pyrrole boronic acid derivative 17 with the electron rich pyrrolyl triflate 15 followed by a ring-closing metathesis reaction (RCM) of the resulting diene to form the macrocyclic ring of the target molecule. This transformation is best achieved by using the ruthenium indenylidene complex 21 as precatalyst. X-ray data of product 18.HCl thus formed suggest that the tautomeric form B properly describes the electron distribution within the heteroaromatic segment of this alkaloid, in which the central ring constitutes the azafulvene unit of the pyrrolylpyrromethene chromophore.

Journal Article↗
PubMed1999

Flexible Synthesis of Metacycloprodigiosin and Functional Derivatives Thereof.

A conceptually new approach to m-pyrrolophane derivatives is outlined providing ready access to compound 23 which can be elaborated into the immunosuppressive alkaloid metacycloprodigiosin 2 according to literature procedures. The key steps of this sequence involve a palladium-catalyzed macrocyclization reaction of vinyl epoxide 10, the conversion of the alpha-pyrone derivative 14 into the pyrrole targets, and the attachment of the side chain via a Wittig (or Peterson) olefination followed by hydrogenation of the alkene formed over Crabtree's catalyst. The flexibility of this route is demonstrated by the synthesis of several analogues of the parent compound 23 which may help to assess the structure/activity profile of the prodigiosin family of natural products in more detail. The unusual pyrone structure 14 used to encode the meta-bridged pyrrolophane units was characterized by X-ray crystallography.

Journal Article↗
PubMed2006

4-alkoxy- and 4-amino-2,2'-bipyrrole synthesis.

[Structure: see text] 4-alkoxy-2,2'-bipyrroles and the first examples of 4-amino-2,2'-bipyrroles have been synthesized by a diversity-oriented strategy from 4-hydroxyproline. The bipyrrole products offer interesting potential as building blocks for making pyrrole products, as demonstrated by the first synthesis of an amino prodigiosin analogue.

Chromatography, Liquid↗
PubMed1998

Pseudoalteromonas bacteriolytica sp. nov., a marine bacterium that is the causative agent of red spot disease of Laminaria japonica.

An aerobic, polarly flagellated marine bacterium that produces a prodigiosin-like pigment was isolated from the red-spotted culture beds of Laminaria japonica. Five isolates had unique bacteriolytic activity for both Gram-positive and -negative bacteria, which had never been observed among Alteromonas or related species. The isolates were identified as the causative agent of red spot disease of L. japonica seeds. The phenotypic features of the isolates were similar to these of Pseudoalteromonas rubra ATCC 29570T, but they could be differentiated using 10 traits (growth at 37 degrees C, requirement for organic growth factors, bacteriolytic activity, utilization of sucrose, N-acetylglucosamine, fumarate, succinate, D-galactose, L-proline and acetate). The G+C content of DNAs from the isolates was 44-46 mol%. The isolates constitute a new species, distinct from the other Alteromonas and Pseudoalteromonas species, as shown by DNA-DNA hybridization experiments and phylogenetic clustering of 16S rRNA gene sequences, for which the name Pseudoalteromonas bacteriolytica sp. nov. (type strain = IAM 14595T) is proposed. A set of phenotypic features which differentiate this new species from closely related Pseudoalteromonas and Alteromonas species is provided.

Base Sequence↗
PubMed1976

Facultative wood-digesting bacteria from the hind-gut of the termite Reticulitermes hesperus.

Among the facultative bacteria capable of growth on mesquite wood which were isolated from the asceptically dissected hind-gut of the termite Reticulitermes hesperus were two strains of Bacillus cereus, one strain each of Arthrobacter, Alcaligenes and Serratia, and a very small Gram-negative fermentative rod. The B. cereus strains, the Serratia marcescens strain and the Arthrobacter sp. grew well on a mineral salts alpha-cellusose agar. One of the Bacillus cereus strains and Serratia marcescens hydrolysed gels of carboxymethylcellulose. All isolates grew well with mesquite wood as the carbon source. The Serratia marcescens isolate produced prodigiosin but differed from a typed strain both in size and in some physiological characteristics.

Alcaligenes↗
PubMed1977

Taxonomy of the genus Serratia.

One hundred and fifty-six strains of Serratia and related bacteria including representatives of Enterobacter liquefaciens, Enterobacter cloacae, Enterobacter aerogenes, Erwinia carotovora, Erwinia chrysanthemi, Erwinia herbicola and Erwinia nimipressuralis were studied using 223 morphological, physiological, biochemical and carbon source utilization tests. The results were subjected to computer analysis. At the 80% similarity level all strains, except two, grouped into eight phenons representing: (A) Serratia marcescens with the neotype CCM303 (ATCCI3880); (B) S. marinorubra with the monotype NCTC10912 (ATCC27614); (CI) S. liquefaciens with the type ATCCI4460; (C2) S. plymuthica with the monotype CCM640 (ATCC183); (D) Erwinia herbicola with the neotype of Enterobacter agglomerans NCTC9381; (E) Enterobacter cloacae with the neotype NCTCI0005 and Erwinia nimipressuralis; (F) Erwinia carotovora with the type ATCC495, Erwinia atroseptica and Erwinia chrysanthemi; (G) Klebsiella mobilis with the neotype NCTCI0006. At the 70% similarity level the phenons formed two groups: (A, B, CI, C2) and (D, E, F, G). The following conclusions were drawn. (I) There are three species of enterobacteria producing prodigiosin: S. marcescens, S. plymuthica and S. marinorubra. (2) There are four species of Serratia, one colourless (S. liquefaciens). (3) Subphenons (biovars) are described within the four species of Serratia. (4) Non-pigmented wild-type strains of S. marcescens can generally be differentiated from pigmented strains by characters other than pigmentation, because subphenons are homogeneous with respect to pigmentation. This survey raised some problems of nomenclature because old descriptions could be found that could loosely fit the present phenons. Comparison with an authentic culture was considered to be the most objective way of identifying these phenons with earlier named species.

Bacteriological Techniques↗
PubMed1997

Serratia marcescens.

Over the last 30 years, Serratia marcescens has become an important cause of nosocomial infection. There have been many reports concerning the identification, antibiotic susceptibility, pathogenicity, epidemiological investigations and typing of this organism. Accurate identification is important in defining outbreaks. The API 20E system has been used widely, but is not individually satisfactory. The growth of S. marcescens in the environment has been investigated in relation to water, disinfectants and plastics such as blood bags. Certain extracellular products are unique to S. marcescens. Pigment (prodigiosin) biosynthesis by S. marcescens has been investigated fully since the emergence of the organism as a cause of infection. Many other aspects of the pathogenicity and virulence of S. marcescens have been studied, including adherence and hydrophobicity, lipopolysaccharide (LPS) and extracellular products. Two modes of adhesion to host epithelial surfaces have been suggested. These are mannose-resistant (MR) pili and mannose-sensitive (MS) pili. LPS, which is responsible for the biological activity of endotoxin, has been investigated fully and 24 somatic antigens have been described. The production of different enzymes by S. marcescens as virulence factors has also been reported, including chitinase, lipase, chloroperoxidase and an extracellular protein, HasA. Antibiotics used to treat serratia infection include beta-lactam agents, aminoglycosides and fluoroquinolones and a variety of different resistance mechanisms have been demonstrated. Typing methods used to study the epidemiology of S. marcescens include biotyping, bacteriocin typing, phage typing, plasmid analysis, polymerase chain reaction amplification of enterobacterial repetitive intergenic consensus sequences (ERIC-PCR) and ribotyping. Serological typing has also been used and this method seems to be a suitable first-line typing method for S. marcescens, although some strains remain untypable. RAPD-PCR has also been applied to a small number of isolates and seems to be a promising method, especially for rapid monitoring of an outbreak and tracing the source of initial infection.

Aminoglycosides↗
PubMed2003

Spore-forming Serratia marcescens subsp. sakuensis subsp. nov., isolated from a domestic wastewater treatment tank.

A strain (KREDT) that formed endospores and produced the pigment prodigiosin was isolated from activated sludge. The presence of spores in cells of strain KREDT was evident upon electron microscopy examination, heat treatment and the detection of dipicolinic acid in the cells. Biochemical characteristics, and 16S rDNA sequence and DNA-DNA homology data identified strain KREDT as Serratia marcescens. The major respiratory quinone of strain KREDT was found to be ubiquinone Q-8. The formation of endospores by Gram-negative bacteria has not been observed previously, and has never been reported in any species of Serratia. Here, it is shown that strain KREDT (JCM 11315T = CIP 107489T) represents a novel subspecies of S. marcescens, for which the name Serratia marcescens subsp. sakuensis is proposed.

DNA, Bacterial↗
PubMed2003

Vibrio ruber sp. nov., a red, facultatively anaerobic, marine bacterium isolated from sea water.

A red, heterotrophic, marine bacterium, designated strain VR1T, was isolated from a sea-water sample collected in the shallow coastal region of Keelung, Taiwan. Cells of the novel strain were facultatively anaerobic, Gram-negative rods that were motile by means of a polar flagellum. The strain grew optimally at 25-30 degrees C and pH 6-7. Growth required the presence of NaCl, the optimal concentration being about 2%. The red pigment produced by the cells was identified as prodigiosin. Strain VR1T grew anaerobically by fermenting glucose and other carbohydrates and producing acids and gases. The strain did not require either vitamins or other organic growth factors for growth. It contained 2-OH-16 : 0 and 3-OH-14 : 0 as the major cellular fatty acids. The DNA G + C content was 45.8 mol%. Phenotypic and chemotaxonomic characterization indicated that strain VR1T represents a novel species in the genus Vibrio. Strain VR1T is phenotypically similar to Vibrio gazogenes. However, the reduction of nitrate to nitrite, the ability to utilize D-arabinose, melibiose and L-glycine as sole carbon sources, the inability to utilize sorbitol as a sole carbon source, resistance to O/129 and susceptibility to erythromycin and novobiocin allow differentiation between V. gazogenes and strain VR1T. The name Vibrio ruber sp. nov. is proposed for the novel species, with strain VR1T (=CCRC 17186T =JCM 11486T) as the type strain.

Anaerobiosis↗
PubMed2026

Genome mining of alkaliphilic cyanobacterial consortia: identification of biosynthetic gene clusters in Sodalinema and associated heterotrophs.

Alkaline soda lakes are high-pH environments that host specialized microbial communities with potential for biotechnology and natural product discovery. We characterized three Sodalinema-dominated cyanobacterial consortia enriched from Canadian soda lakes over 510 days. Using hybrid metagenomic sequencing and metatranscriptomics across pH, alkalinity, and temperature gradients, we reconstructed high-quality metagenome-assembled genomes and assessed functional activity. All consortia converged toward cyanobacteria dominance and exhibited temperature optima between 21&#xb0;C and 30&#xb0;C. Phylogenetic analysis placed Sodalinema genomes within a distinct clade affiliated with Candidatus Sodalinema alkaliphilum. Genomic analysis indicated complete biosynthetic pathways for vitamin B5, vitamin B7, and the molybdenum cofactor, but incomplete pathways for vitamins B1, B9, and B12, consistent with patterns observed in Sodalinema yuhuli. Metatranscriptomic profiles showed increased expression of genes involved in phycocyanin and carotenoid biosynthesis at pH 10.2 relative to pH 8.5. Biosynthetic gene cluster analysis revealed that most secondary metabolic potential resided in heterotrophic community members. Roseinatronobacter encoded pathways for N-acyl homoserine lactones, osmoprotectants, betalactones, and prodigiosin, while Alkalimonas, Wenzhouxiangella, and members of the Kiloniellales encoded clusters for lanthipeptides, cyclodipeptides, hydrogen cyanide, and pyrroloquinoline quinone. These findings indicate functional partitioning within the consortia and highlight the contribution of heterotrophs to secondary metabolism.IMPORTANCEAlkaline soda lakes contain microbial communities adapted to high pH that remain underexplored for biotechnology. This study focuses on Sodalinema, a filamentous cyanobacterium that dominates enriched consortia from Canadian soda lakes, and its associated heterotrophic partners. We show that while Sodalinema drives primary productivity, heterotrophic bacteria encode most of the pathways for antimicrobial and signaling compounds. These interactions may support community stability and defense against competing microorganisms. By linking genomic potential with gene expression, this work identifies alkaline cyanobacterial consortia as a source of bioactive compounds and provides a framework for exploring extremophilic microbial communities for natural product discovery.

Sodalinema↗