This study was performed to analyze 6 day-term variations in bacterial communities contaminating the floor of two dry saunas that were operated at 64degrees C (low temp) and 76degrees C (high temp). Bacteria were sampled daily from the saunas for 6 days from Monday to Saturday. Genomic DNA was isolated directly from bacteria-collected cotton swabs. The diversity of the bacterial communities collected from the saunas was analyzed using thermal gradient gel electrophoresis (TGGE). The total numbers of DNA bands separated by TGGE for bacteria collected from the low temp and high temp sauna were 20 and 18, respectively, during the 6 days. Seven of 20 bacteria in the low temp sauna and eight of 18 bacteria in the high temp sauna were detected more than three times over the 6 experimental days. Twelve of the 26 bacterial genera contaminating the saunas were cross detected. Bacteria belonging to the genera Moraxella and Acinetobacter were selectively detected in the low temp sauna, whereas those belonging to Aquaspirillum, Chromobacterium, Aquabacterium, Gulbenkiania, Pelomonas, and Aquitalea were selectively detected in the high temp sauna. Three species of bacteria contaminating both the low and high temp saunas were thermophile or thermoduric. The results indicate that the sauna-contaminating bacteria may have been transferred from outside the saunas by user traffic but did not inhabit the saunas.
Acinetobacter ; Bacteria ; Chromobacterium ; DNA ; Electrophoresis ; Floors and Floorcoverings ; Moraxella ; Steam Bath

Two initial models of cold-adapted lipase PsLip1 have been constructed, based on homology with the bacterial lipases Chromobacterium viscosum(CvLip) and Pseudomonas cepacia(PcLip), whose X-ray structures have been solved and refined to high resolution. The mature polypeptide chains of these lipases have 84% similarity. The models of Mod1 and Mod2 have been compared with the tertiary structures of CvLip and PcLip, respectively, and analyzed in terms of stabilizing interactions. Several structural aspects that are believed to contribute to protein stability have been compared: the number of conserved salt bridges, aromatic interactions, hydrogen bonds, helix capping, and disulfide bridges. The 3-dimensional structural model of PsLip1 has been constructed in order to elucidate the structural reasons for the decreased thermostability of the enzyme in comparison with its mesophilic counterparts.
Chromobacterium ; Hydrogen ; Lipase ; Models, Structural ; Protein Stability ; Pseudomonas

Child ; Chromobacterium ; Female ; Humans ; Sepsis ; drug therapy ; etiology ; microbiology ; mortality

Chromobacterium violaceum is a gram negative straight rod, 0.8-1.2 by 2.5 to 6.0 m, which is motile by one polar flagella and one to four lateral flagella. The organism inhabits soil and water and is often found in semitropical and tropical climates. Infections in humans are rare. We report a case of infection caused by strains of C. violaceum. A 38-year-old male patient was admitted to KyungHee University Hospital, Seoul, Korea on July 28th, 2003, after a car accident. The patient had multiple trauma and lacerations. He had an open wound in the left tibial area from which C. violaceum was isolated. The strain was resistant to ampicillin, tobramycin, ampicillin/sulbactam, ceftriaxone and cefepime, but was susceptible to amikacin, gentamicin, ciprofloxacin, levofloxacin, trimethoprim/sulfamethoxazole and piperacillin/tazobactam. The patient was treated successfully by debridement, cephapirin sodium and astromicine sulfate.
Wound Infection/*etiology ; Male ; Humans ; Gram-Negative Bacterial Infections/*etiology ; Chromobacterium/*isolation & purification ; Adult ; *Accidents, Traffic

Cylindrocarpon destructans is an ascomycete soil-borne pathogen that causes ginseng root rot. To identify effective biocontrol agents, we isolated several bacteria from ginseng cultivation soil and evaluated their antifungal activity. Among the isolated bacteria, one isolate (named JH7) was selected for its high antibiotic activity and was further examined for antagonism against fungal pathogens. Strain JH7 was identified as a Chromobacterium sp. using phylogenetic analysis based on 16S rRNA gene sequences. This strain was shown to produce antimicrobial molecules, including chitinases and proteases, but not cellulases. Additionally, the ability of JH7 to produce siderophore and solubilize insoluble phosphate supports its antagonistic and beneficial traits for plant growth. The JH7 strain suppressed the conidiation, conidial germination, and chlamydospore formation of C. destructans. Furthermore, the JH7 strain inhibited other plant pathogenic fungi. Thus, it provides a basis for developing a biocontrol agent for ginseng cultivation.
Ascomycota ; Bacteria ; Cellulases ; Chromobacterium* ; Fungi ; Genes, rRNA ; Germination ; Panax* ; Peptide Hydrolases ; Plants ; Soil

Chromobacterium violaceum (C. violaceum) is a gram negative, facultative anaerobe widely distributed in natural aquatic environments and is sensitive to temperature. Its infection has a predilection to tropical or subtropical regions that between latitude 35degrees N and 35degrees S. Infection due to C. violaceum is rare. But human infection with this organism can result in severe, systemic disease with a high fatality rate. C. violaceum infection may rapidly progress to sepsis with multiple organ abscesses, predominantly in lungs, liver, and spleen. The organism is usually resistant to most antibiotics commonly prescribed for gram-negative bacterial infection like aminoglycosides, extended spectrum beta-lactam antibiotics etc. In Korea, two cases of C. violaceum infections were reported in patients injured in a Guam airplane accident, and one case was reported local C. violaceum infection. Here, we report a case of fatal C. violaceum postoperative wound infection in Korea.
Abscess ; Aircraft ; Aminoglycosides ; Ankle Fractures* ; Ankle* ; Anti-Bacterial Agents ; Chromobacterium* ; Gram-Negative Bacterial Infections ; Guam ; Humans ; Korea ; Liver ; Lung ; Sepsis ; Spleen ; Surgical Wound Infection ; Wound Infection* ; Wounds and Injuries*

In proteins of thermophilic bacteria, Gly is tend to be replaced by Ala and Lys is tend to be replaced by Arg to adapt the high temperature. In order to improve the thermal stability of phenylalanine hydroxylase (PAH) from Chromobacterium violaceum, all the Gly on PAH were mutated to Ala and Lys to Arg. Positive mutant enzymes with improved thermal stability were selected, followed by combined mutation and characterization. The results revealed that half-lives of K94R and G221A mutants at 50 °C were 26.2 min and 16.8 min, which were increased by 1.9-times and 0.9-times than the parent enzyme (9.0 min). The residual activity of K94R/G221A mutant was improved to 65.6% after keeping at 50 °C for 1 h, which was 6.6 time higher than the parent enzyme (8.6%). Circular dichroism (CD) spectroscopy revealed that Tm values of the parent enzyme, K94R, G221A and K94R/G221A were 51.5 ℃, 53.8 ℃, 53.1 ℃ and 54.8 ℃, respectively. According to the protein structure simulation, the two mutations were located on flexible loop. In the K94R mutant, the mutated Arg94 on the surface of the enzyme formed an extra hydrogen bond with Ile95, which stabilized the located loop. In the G221A mutant, the mutated Ala221 formed hydrophobic interaction with Leu281, which could stabilize both the loop and flexible area of the C-terminus of G221A. The results not only provided a reference for protein modification on thermal stability, but also laid the foundation for application of phenylalanine hydroxylase in the field of functional foods.
Bacterial Proteins ; biosynthesis ; genetics ; Chromobacterium ; enzymology ; Enzyme Stability ; Hot Temperature ; Kinetics ; Mutagenesis, Site-Directed ; Mutation ; Phenylalanine Hydroxylase ; biosynthesis ; genetics ; Protein Engineering

Romipeptides A and B (1 and 2), two new romidepsin derivatives, and three known compounds, chromopeptide A (3), romidepsin (4) and valine-leucine dipeptide (5) were isolated from the fermentation broth of Chromobacterium violaceum No. 968. Their structures were elucidated by interpretation of their UV, HR-ESI-MS and NMR spectra. The absolute configuration of compound 1 and 2 were established by single crystal X-ray diffraction analysis. Compounds 1-5 were evaluated for their anti-proliferative activities against three human cancer cell lines, SW620, HL60, and A549. The results showed most of these compounds exhibited antitumor activities in vitro, in which compound 2 displayed potent cytotoxicity to SW620, HL60 and A549 cell lines, with IC of 12.5, 6.7 and 5.7 nmol·L, respectively.
Antineoplastic Agents ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Chemistry Techniques, Analytical ; Chromobacterium ; metabolism ; Depsipeptides ; chemistry ; pharmacology ; Dipeptides ; chemistry ; Drug Screening Assays, Antitumor ; Fermentation ; Humans ; Molecular Structure ; Peptides, Cyclic ; chemistry