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PubMed2000

Effect of some antibiotics on pigmentation in Serratia marcescens.

Serratia marcescens is characterized by its ability to produce a red pigment called prodigiosin. It is well known that there are some substances affecting pigmentation in bacteria. Cefoxitin, erythromycin, tobramycin, co-trimoxazole, imipenem and nitrofurantoin were found to have an inhibitory effect on pigmentation in a S. marcescens strain isolated from urine. It was also shown that the LD50 dose determined by inoculation of eight-week-old BALB/c mice with serial dilutions of overnight cultures of pigmented and nonpigmented variants was lower (LD50 = 300 x 10(3.94)) in the nonpigmented variant than in the pigmented one (LD50 = 300 x 10(5.58)). In addition, the Sereny test showed that in contrast to the pigmented variant, the nonpigmented variant caused keratitis in guinea pig eye.

Animals↗
PubMed1982

Pigments and antibiogram of transconjugants from nonpigmented mutants of Serratia marcescens.

Serratia marcescens produces a characteristic red pigment, prodigiosin, which is formed by the enzymatic coupling of 4-methoxy-2,2'-bipyrrole-5-bipyrrole-5-carboxaldehyde (MBC) and 2-methyl-3-amylpyrrole (MAP). Many clinical isolates which are resistant to multiple antibiotics are nonpigmented. However, the relationship of pigmentation (or nonpigmentation) to drug resistance of the strains has not yet been established. In this study we demonstrated the pigment synthesizing capability in the transconjugants obtained from nonpigmented mutants WF and 9-3-3 of S. marcescens under the condition of cell-to-cell contact. Mutant WF produces MAP while mutant 9-3-3 synthesized only MBC. After genetic transfer, the color of the recombinant colonies was red indicating the successful transfer of the pigment synthesizing capability. The antibiogram of the transconjugants indicated that they inherited the resistance characteristics to polymyxin B and chloramphenicol from their parent strains. further supportive evidence was obtained by spectroscopic and high performance liquid chromatographic analysis of the resulting pigments extracted from the pigmented transconjugants. The pigments produced by the transconjugants were similar, if not identical, to those produced by the wild type strain 08 and those synthesizes syntrophically. The possibility of simultaneous transfer of pigment synthesizing capability and drug resistance remains to be explored .

Conjugation, Genetic↗
PubMed1988

Cloning and sequence of the gene encoding the major structural component of mannose-resistant fimbriae of Serratia marcescens.

Serratia marcescens US46, a human urinary tract isolate, exhibits mannose-resistant hemagglutination and agglutinates yeast cells, thereby indicating that it has two types of adhesins. We constructed a cosmid library for the DNA of this organism and isolated DNA clones carrying genes for mannose-sensitive (MS) and mannose-resistant (MR) fimbriae. On introduction of the cloned genes into Escherichia coli K-12, MS and MR fimbriae were formed. These fimbriae were functionally and morphologically indistinguishable from those of S. marcescens. Subcloning of these gene clusters revealed that the genes encoding MS fimbriae reside on a 9-kilobase (kb) DNA fragment, while those encoding MR fimbriae are present on a 12-kb fragment. Transposon insertion and maxicell analyses revealed that formation of MR fimbriae is controlled by several genes which reside on the 9-kb fragment. The nucleotide sequence of smfA, the gene encoding the major structural component of MR fimbriae, revealed that this gene encodes a 174-amino-acid polypeptide with a typical procaryotic signal peptide. The primary structure of the smfA product showed significant homology with the primary structure of the E. coli fimbrial subunit.

Amino Acid Sequence↗
PubMed2004

Identification and characterization of the pswP gene required for the parallel production of prodigiosin and serrawettin W1 in Serratia marcescens.

Serratia marcescens mutants defective in production of the red pigment prodigiosin and the biosurfactant serrawettin W1 in parallel were isolated by transposon mutagenesis of strain 274. Cloning of the DNA fragment required for production of these secondary metabolites with different chemical structures pointed out a novel open reading frame (ORF) named pswP. The putative product PswP (230 aa) has the distinct signature sequence consensus among members of phosphopantetheinyl transferase (PPTase) which phosphopantetheinylates peptidyl carrier protein (PCP) mostly integrated in the nonribosomal peptide synthetases (NRPSs) system. Since serrawettin W1 belongs to the cyclodepsipeptides, which are biosynthesized through the NRPSs system, and one pyrrole ring in prodigiosin has been reported as a derivative of L -proline tethered to phosphopantetheinylated PCP, the mutation in the single gene pswP seems responsible for parallel failure in production of prodigiosin and serrawettin W1.

Bacterial Proteins↗
PubMed2005

Cloning, sequencing, and characterization of the SdeAB multidrug efflux pump of Serratia marcescens.

Serratia marcescens is an important nosocomial agent known for causing various infections in immunocompromised individuals. Resistance of this organism to a broad spectrum of antibiotics makes the treatment of infections very difficult. This study was undertaken to identify multidrug resistance efflux pumps in S. marcescens. Three mutant strains of S. marcescens were isolated in vitro by the serial passaging of a wild-type strain in culture medium supplemented with ciprofloxacin, norfloxacin, or ofloxacin. Fluoroquinolone accumulation assays were performed to detect the presence of a proton gradient-dependent efflux mechanism. Two of the mutant strains were found to be effluxing norfloxacin, ciprofloxacin, and ofloxacin, while the third was found to efflux only ofloxacin. A genomic library of S. marcescens wild-type strain UOC-67 was constructed and screened for RND pump-encoding genes by using DNA probes for two putative RND pump-encoding genes. Two different loci were identified: sdeAB, encoding an MFP and an RND pump, and sdeCDE, encoding an MFP and two different RND pumps. Northern blot analysis revealed overexpression of sdeB in two mutant strains effluxing fluoroquinolones. Analysis of the sdeAB and sdeCDE loci in Escherichia coli strain AG102MB, deficient in the RND pump (AcrB), revealed that gene products of sdeAB are responsible for the efflux of a diverse range of substrates that includes ciprofloxacin, norfloxacin, ofloxacin, chloramphenicol, sodium dodecyl sulfate, ethidium bromide, and n-hexane, while those of sdeCDE did not result in any change in susceptibilities to any of these agents.

Anti-Bacterial Agents↗
PubMed1993

Improvement of culture conditions for L-proline production by a recombinant strain of Serratia marcescens.

Serratia marcescens SP511 was previously reported to be an L-proline-producing strain that harbors a recombinant plasmid carrying the mutant type of the proline operon. This strain produced 65 g/L of L-proline in a medium containing 22% sucrose and urea after 5 d of incubation under the conventional culture conditions. We searched for more suitable culture conditions for more abundant L-proline production by SP511. To improve the supply of a nitrogen source to cells, ammonium was used instead of urea and fed to a culture under control of the pH of the medium. The concentrations of MgSO4 and K2HPO4 were increased, and in addition, sucrose was continuously added to the culture at a final concentration of 32%. Under these conditions, the cell amount was increased twofold over that under the previous conditions and L-proline production reached a maximum of more than 100 g/L after 4 d of incubation.

Ammonia↗
PubMed1973

Effect of iron and salt on prodigiosin synthesis in Serratia marcescens.

Serratia marcescens wild-types ATCC 264 and Nima grew but did not synthesize prodigiosin in a glycerol-alanine medium containing 10 ng of Fe per ml. Wild-type 264 required the addition of 0.2 mug of Fe per ml for maximal growth and prodigiosin synthesis; Nima required 0.5 mug of Fe per ml. Three percent, but not 0.1%, sea salts inhibited prodigiosin synthesis in a complex medium containing up to 10 mug of Fe per ml. NaCl was the inhibitory sea salt component. The inhibition was not specific for NaCl; equimolar concentrations of Na(2)SO(4), KCl, and K(2)SO(4) also inhibited prodigiosin synthesis. Experiments with strains 264 and Nima and with mutant WF which cannot synthesize 4-methoxy-2-2'-bipyrrole-5-carboxyaldehyde (MBC), the bipyrrole moiety of prodigiosin, and with mutant 9-3-3 which cannot synthesize the monopyrrole moiety 2-methyl-3-amylpyrrole (MAP) showed that both MBC synthesis and the reaction condensing MAP and MBC to form prodigiosin were relatively more sensitive to NaCl inhibition than the MAP-synthesizing step. The capacity of whole cells to condense MAP and MBC was present, but inactive, in cells grown in NaCl; removal of the NaCl from non-proliferating salt-grown cells restored the activity. Other evidence suggests the existence of a common precursor to the MAP- and MBC-synthesizing pathways.

Aldehydes↗
PubMed1992

Improvement of nitrogen supply for L-threonine production by a recombinant strain of Serratia marcescens.

Serratia marcescens T-2000 was previously reported to be an L-threonine-producing strain that harbors the recombinant plasmid carrying the mutant-type threonine operon. This strain produced 55 g of L-threonine/L of the medium containing urea as a nitrogen source after 72 h of cultivation. In the urea-containing medium, transitory stop of the growth was observed during the early period of cultivation when the entire amount of ammonium ion formed from urea via heat decomposition disappeared in the medium. This indicated that the shortage of ammonium supply in cells might delay both the cell growth and the L-threonine production. The use of ammonia water as a nitrogen source for L-threonine production was therefore studied, because microbial cells generally assimilate this source more readily than urea. When ammonia water was automatically fed to the medium so as to maintain the pH of the medium at around 7, the growth was accelerated, and the L-threonine production reached a maximum of 65 g/L at 48 h. Under these conditions, sucrose, a carbon source, was continuously fed to the medium, resulting in the production of 100 g of L-threonine/L at 96 h. Thus, the L-threonine production of the recombinant L-threonine-producing strain could be increased by devising the method for supply of a nitrogen source.

Ammonia↗
PubMed1987

Reliability of the colistin disk test in identification of Serratia marcescens and Serratia liquefaciens.

Resistance to polymycin B or E (colistin) in the disk test, which is used as a means of identification, was tested in 43 strains of Serratia marcescens and 26 of Serratia liquefaciens. While all strains had MIC values greater than 32 mg/l for colistin, false susceptibility to both polymyxins was encountered in the 24 h disk test in 9% of Serratia marcescens and 12-96% of Serratia liquefaciens strains, depending on the kind of polymyxin, temperature and duration of incubation. Using colistin disks and incubation at 25 degrees C for 48 h, the percentage of false susceptible results could be minimized.

Colistin↗
PubMed2005

Purification, crystallization and preliminary X-ray analysis of the outer membrane complex HasA-HasR from Serratia marcescens.

Serratia marcescens is able to acquire iron using its haem-acquisition system (;has'), which contains an outer membrane receptor HasR and a soluble haemophore HasA. After secretion, HasA binds free haem in the extracellular medium or extracts it from haemoproteins and delivers it to the receptor. Here, the crystallization of a HasA-HasR complex is reported. HasA and HasR have been overexpressed in Escherichia coli and the complex formed and crystallized. Small platelets and bunches of needles of dimensions 0.01 x 0.1 x 1 mm were obtained. A native data set has been collected to 6.8 A.

Amino Acid Sequence↗
PubMed1999

A plasmid-mediated beta-lactamase with carbapenem-hydrolyzing activity from Serratia marcescens.

Serratia marcescens W-313/pSW313 was isolated from a clinical specimen that showed resistance to various beta-lactams. beta-Lactamase from this strain hydrolyzed oxyiminocephalosporins and carbapenems, but did not hydrolyze 2-carboxypenam and aztreonam. The kinetic parameters of this enzyme were similar to those of the carbapenem-hydrolyzing beta-lactamases mediated by R-plasmid from Pseudomonas aeruginosa and S. marcescens, whose properties were previously reported.

Journal Article↗
PubMed1978

Differentiation of Serratia marcescens and Serratia liquefaciens by tests for lipase and phospholipase production.

The production of lipase and phospolipase by certain members of the Enterobacteriaceae was examined by thin-layer chromatography of resting-cell suspensions incubated with triolein or lecithin. Most strains of Serratia marcescens produced both enzymes while most strains of Serratia liquefaciens exhibited strong lipase but only a minor phospholipase activity. Enterobacter spp. (25 strains), Klebsiella pneumoniae (20 strains), Escherichia coli (15 strains), Citrobacter freundii (7 strains) and Proteus spp. (20 strains) lacked both types of enzymic activity except for the following: three strains of Enterobacter cloacae, two of Proteus mirabilis and three of Proteus vulgaris possessed slight lipase activity; about one-half of the Enterobacter aerogenes and Enterobacter hafniae strains examined produced slight phospholipase activity. It is suggested that tests for lipase and phospholipase should be used in conjunction with those for DNAase production and sugar fermentation for the differentiation of S. marcescens and S. liquefaciens.

Arabinose↗
PubMed1970

Cation transport in Serratia marcescens and Serratia marinorubra.

The sodium, potassium, and magnesium ion contents of Serratia marcescens and those of its salt-tolerant relative, S. marinoruba, were determined by atomic-absorption spectrometry. The intracellular K(+) and Mg(2+) contents of both microorganisms were found to be dependent on the ionic strength of the growth or suspending medium. The Mg(2+) content of S. marinoruba was generally greater than that of S. marcescens. The Na(+) content of the cells was normally low and did not increase as the cells aged or when the cells were grown in media of high ionic strength. The transport of K(+) by resting cells suspended in hypertonic solution was studied by chemical and light-scattering techniques and was found to be more rapid in S. marcescens than in S. marinorubra. The slower rate of K(+) transport in S. marinorubra is probably related to the lower glycogen reserves found in resting cells of this microorganism. K(+) transport was found to have a pH optimum of 5.5 to 6.1 for S. marcescens, and the K(m) for K(+) was approximately 1.6 mm. Na(+) and Mg(2+) were not taken up by the cells, although the presence of Mg(2+) tended to decrease rates of K(+) uptake. Tris-(hydroxymethyl)aminomethane, routinely used for resuspending the cells, was apparently taken up by the cells at pH >7.

Absorption↗
PubMedDate not supplied

Antibiotic susceptibility of Serratia marcescens and Serratia liquefaciens.

BACKGROUND: Over a period of 20 years, a total of 1,603 Serratia isolates were recovered from clinical specimens and examined for susceptibility to 29 antimicrobial drugs using the Bauer-Kirby agar disk diffusion test. Serratia marcescens was recovered most frequently (n = 1,409), followed by S. liquefaciens (n = 172); other Serratia species were scarce. During the 2-decade observation period there occurred 35 putative episodes/clusters of nosocomial cross-infection and 1 pseudo-outbreak due to S. marcescens, but none due to S. liquefaciens. METHODS: The antimicrobial susceptibility data for S. marcescens and S. liquefaciens were subdivided into two observation periods: I = 1980-1993, and II = 1993-1999. The crude data (series A) obtained for S. marcescens were corrected in two ways: by the omission of repetitive patient isolates (series B) and the additional removal of outbreak isolates except for index case isolates (series C). RESULTS AND CONCLUSIONS: Comparison of data obtained in series IC and IIC disclosed an increase in the susceptibility of S. marcescens to ampicillin + sulbactam, cefotaxime, chloramphenicol, doxycycline, fosfomycin, gentamicin, piperacillin, piperacillin + tazobactam, timentin and tobramycin during observation period II. Conversely, there was a decrease in susceptibility to ciprofloxacin, nalidixic acid and trovafloxacin, and slightly diminished susceptibility to norfloxacin and ofloxacin during observation period II as compared with the previous period. The crude data obtained for S. liquefaciens required no correction, as there were only a few repeat isolates. There was an increase in susceptibility to ampicillin, ampicillin + sulbactam, cefuroxime, doxycycline, fosfomycin, nitrofurantoin and polymyxin B (clear inhibition zones). However, there was an inexplicable decrease in susceptibility to piperacillin + tazobactam. Cocarde growth around polymyxin B disks was noted with 55.8% of the S. marcescens isolates as compared with 6.8% of the S. liquefaciens isolates. Slime around fluoroquinolone inhibition zones was produced by 83.4% of the S. marcescens isolates. Slime production around carbapenem inhibition zones was noted with 52% of the S. liquefaciens isolates, but with only a single isolate of S. marcescens.

Anti-Bacterial Agents↗
PubMed2003

Spontaneous dermal abscesses and ulcers as a result of Serratia marcescens.

Serratia sp have only rarely been reported as isolates from leg ulcers. We describe the case of a middle-aged man with a medical history significant for alcohol-induced cirrhosis who presented with rapidly progressive skin ulcers initially starting as purple nodules. These skin ulcers and underlying dermal abscesses were found to be a result of S marcescens, with the presumed portal of entry being a toe-web infection.

Abscess↗
PubMed2001

Cloning of a Serratia marcescens DNA fragment that induces quinoprotein glucose dehydrogenase-mediated gluconic acid production in Escherichia coli in the presence of stationary phase Serratia marcescens.

Serratia marcescens ER2 was isolated from an endorhizosphere sample based on its high level of mineral phosphate solubilizing (MPS) activity. This phenotype was correlated with expression of the direct oxidation pathway. An ER2 plasmid library constructed in Escherichia coli strain DH5alpha was screened for MPS activity. A recombinant clone DH5alpha (pKG3791) was capable of gluconic acid (GA) production and tricalcium phosphate solubilization but only in the presence of stationary phase ER2 cells. GA production in DH5alpha (pKG3791) was apparently the result of the quinoprotein glucose dehydrogenase activity because AG121 (a Tn5 knockout of gcd) carrying pKG3791 did not produce GA under the same conditions. GA production by DH5alpha (pKG3791) was not observed when ER2 was replaced by another PQQ-producing strain bacterium. These data add to a growing body of evidence that E. coli contains some type of PQQ biosynthesis pathway distinct from those previously characterized in Gram-negative bacteria and that these genes may be induced under appropriate conditions.

Calcium Phosphates↗
PubMed1998

The Leu-3 residue of Serratia marcescens metalloprotease inhibitor is important in inhibitory activity and binding with Serratia marcescens metalloprotease.

Serratia marcescens metalloprotease inhibitor (SmaPI) is a proteinase inhibitor toward Serratia marcescens metalloprotease (SMP). In sequential deletion analysis of the N-terminal region of the SmaPI, SmaPIs starting at Ser-2 and Leu-3 residues, respectively, had nearly a full inhibitory activity toward SMP. However, SmaPI starting at Ala-4 residue showed severely decreased inhibitory activity. Furthermore, kinetic analysis demonstrated that SmaPI starting at the Ala-4 residue had an inhibition constant for SMP approximately fourfold higher than that of wild-type SmaPI. The interactions of Leu-3 with SMP contribute 0.73 kcal mol-1 to the overall stability of the SMP-SmaPI complex (8.44 kcal mol-1). To elucidate the detailed role of the Leu-3 residue in inhibitory activity of SmaPI, several site-directed mutations were introduced. The inhibitory activities of Leu-3 mutants in which the Leu-3 has been converted to Ala, Asp, Gly, Ile, Lys, Phe, or Pro were correlated with the hydrophobicities of substituted amino acids. About 0.3 kcal mol-1 is attributable to the side chain of the Leu-3 residue in the binding with SMP. From these results, it is suggested that (i) in contrast with the Erwinia chrysanthemi inhibitor, Gly-1 and Ser-2 of SmaPI are not critical and (ii) the hydrophobicity of Leu-3 may be important in its inhibitory activity and binding with SMP.

Amino Acid Sequence↗