PURPOSE: To report a case of corneal ulcer due to Alcaligenes faecalis in a patient with a preexisting corneal ulcer. CASE SUMMARY: A 58-year-old male patient presented with a corneal ulcer without a history of any trauma. The patient had a history of corneal ulcer 9 months earlier. The patient had previously been diagnosed with diabetic retinopathy and neovascular glaucoma, and his visual acuity was no light perception. Corneal scraping and culture yielded Alcaligenes faecalis susceptible to most antibiotics in the antibiotic susceptibility test. After treatment with empirical systemic antibiotics and eyedrops, his eye improved with a remaining corneal scar. CONCLUSIONS: Alcaligenes faecalis should be considered as a causal pathogen of corneal ulcer in patients with suspicious compromised ocular surface, such as previous corneal ulcer.
Alcaligenes ; Alcaligenes faecalis ; Anti-Bacterial Agents ; Cicatrix ; Corneal Ulcer ; Diabetic Retinopathy ; Eye ; Glaucoma, Neovascular ; Humans ; Light ; Male ; Middle Aged ; Ophthalmic Solutions ; Visual Acuity

Curdlan is a water insoluble exopolysaccharide produced by Alcaligenes faecalis under nitrogen-limiting conditions. After excretion, the polysaccharide is attached the cell wall. Thus enhancement of biomass production during the cell growth phase is important to curdlan production. A strategy of increasing nitrogen source to improve biomass production was adopted for curdlan production by Alcaligenes faecalis (ATCC 31749). In the batch fermentation of curdlan, a relatively higher NH4Cl level of 3.6 g/L with continuous glucose feeding increased the cell density leading to improvement of curdlan production. However, excessive NH4Cl would inhibit curdlan production and biomass production was not improved significantly. In addition, feeding of ammonia water at the initial phase replaced NaOH solution to control pH at 7.0. Subsequently, feeding of NaOH solution was resumed to control pH at 5.6 for curdlan production after ammonia was consumed. As a result, biomass production and curdlan yield were both enhanced remarkably. Feeding of ammonia water during the first 24 h led to biomass production of 18.8 g/L. However, higher cell density did not lead to increase in curdlan production. The maximum curdlan production (72 g/L) was obtained by feeding ammonia water for the first 14 h, during which the cell density was about 11.9 g/L.
Alcaligenes faecalis ; cytology ; metabolism ; Ammonium Chloride ; pharmacology ; Cell Culture Techniques ; methods ; Cell Proliferation ; Fermentation ; beta-Glucans ; metabolism

Considering Alcaligenes faecalis pencillin G acylase(AfPGA), which possesses the attractive characteristics for beta-lactam antibiotics conversions, the gene of PGA was cloned into an expressing vector pKKFPGA. The recombinant plasmid contained multicopy replicon(COLE 1), trc promoter, AfPGA gene, rrnB transcript terminator and ampicillin marker transformed Escherichia coli DH5alpha. As both the recombinant plasmid and the host DH5alpha had no laclq gene, the trc promoter was always active and the AfPGA could be constitutively expressed without IPTG induction in the host DH5alpha. In the shaking flask, the recombinant cell was inoculated into the fermentation medium (tryptone 10g/L, yeast extract 5g/L, MgSO4 x 7 H2O 1g, KH2 PO4 2g/L, K2HPO4 x 3H2O 5g/L, Na2HPO4 x 12H2O 7g/L, (NH4)2SO4 1.2g/L, NH4Cl 0.2 g/L, NaCl 0.1g/L, dextrin 30g/L) and cultured at 28 degrees C for 20h. The production of AfPGA reached 2,590u/L(NIPAB method), with a cell-density-specific activity of more than 300(u/L)/A600, this yield increased 432 fold higher than the native expression of Alcaligenes faecalis . Without ammonium sulphate fractionation and dialysis, the supernatant of crude extract was directly loaded on DEAE-Sepharose CL 6B column equilibrated by phosphate buffer (50mmol/L, pH7.8), and the enzyme fraction was not absorbed on the column but impurities were absorbed. Subsequently the effluent was added ammonium sulphate to 1mol/L and loaded on Butyl-Sepharose CL 4B column equilibrated by 50mmol/L phosphate buffer pH7.8-1mol/L ammonium sulphate. The enzyme was eluted as concentration of ammonium sulphate in phosphate buffer decreased to 0, PGA was eluted. After these two column chromatography, the enzyme was enriched 20 times with a 91% activity recovery. The purified enzyme had a specific activity of 68.6u/mg protein. However, the overproduction of PGA was often limited by translocation and/or periplasmic processing steps, subsequently resulted in intracellular accumulation of various types of PGA precursors and then formed inclusion bodies in the cytoplasm and/or periplasm. In this study, 5% PGA precursors formed as inclusion bodies in the cytoplasm while no inclusion bodies formed in the periplasm. It suggested most PGA precursors were transported to the periplasm and matured to active PGA and also explained why PGA gene was highly expressed in the host DH5alpha. On the other hand, inclusion bodies in the cytoplasm indicated that the maturation of PGA in the host DHSalpha was limited by the translocation step.
Alcaligenes faecalis ; enzymology ; Blotting, Western ; Escherichia coli ; genetics ; Penicillin Amidase ; genetics ; isolation & purification ; Recombinant Proteins ; biosynthesis ; isolation & purification

The effect of initial ammonium chloride level on production of curdlan in Alcaligenesfaecalis was investigated. It was found that ammonium chloride was the limiting substrate for cell growth during the batch fermentation process. However, the cell growth and curdlan production could not be enhanced by solely increasing the initial ammonium chloride level. The pH drop in the broth due to the consumption of ammonium chloride also effected the cell growth and curdlan production. By simultaneously increasing the initial ammonium chloride concentration and implementing an optimal pH control strategy, which is to control pH at 7.0 in the growth phase, and then shift to 5.6 in the production phase, the biomass and curdlan production in batch fermentation were increased markedly. If the initial ammonium chloride concentration was increased from 1.1 g/L to 3.6 g/L, biomass concentration of 7.2 g/L was obtained, and the final curdlan concentration reached 30.5 g/L, which was 51.7% higher than that of the former case. As the cell growth was improved due to the increase of the initial ammonium chloride concentration, the agitation speed and aeration rates must be enhanced to suit the higher oxygen uptake requirement. However, as curdlan molecules is subject to the structural breakage due to the high shear stress at higher agitation speed, an overall optimal condition for both productivity and quality of curdlan should be considered comprehensively.
Alcaligenes faecalis ; drug effects ; growth & development ; metabolism ; Ammonium Chloride ; pharmacology ; Culture Media ; Dose-Response Relationship, Drug ; Fermentation ; Hydrogen-Ion Concentration ; beta-Glucans ; metabolism

A two-stage pH control method was employed in batch fermentation of curdlan by Alcaligenes faecalis WX-C12 where cell-growth stage was constantly controlled at pH 7.0 and stationary stage was controlled at a constant pH as well. The influence of pH control on the curdlan production was investigated. The optimal pH control of batch process for curdlan production was obtained when cell-growth stage was controlled at pH 7.0 and stationary stage was constantly controlled at pH 5.6. Production and productivity of curdlan, QP and YP/S reached 28.19 g/L, 291 mg/(L.h), 132.27 mg/(L.h.g) and 0.659, an improvement of 20.4%, 38.1%, 38.1% and 29.5% compared to a pH uncontrolled operation respectively.
Alcaligenes ; growth & development ; metabolism ; Fermentation ; Glucans ; biosynthesis ; Hydrogen-Ion Concentration ; beta-Glucans

Agitator is one of the essential factors to realize high efficient fermentation for high aerobic and viscous microorganisms, and the influence of different impeller combination on the fermentation process is very important. Welan gum is a microbial exopolysaccharide produced by Alcaligenes sp. under high aerobic and high viscos conditions. Computational fluid dynamics (CFD) numerical simulation was used for analyzing the distribution of velocity, shear rate and gas holdup in the welan fermentation reactor under six different impeller combinations. The best three combinations of impellers were applied to the fermentation of welan. By analyzing the fermentation performance, the MB-4-6 combination had better effect on dissolved oxygen and velocity. The content of welan was increased by 13%. Furthermore, the viscosity of production were also increased.
Alcaligenes ; metabolism ; Fermentation ; Hydrodynamics ; Industrial Microbiology ; methods ; Oxygen ; Polysaccharides, Bacterial ; biosynthesis ; Viscosity

Positive bacterial culture was obtained in 33 out of 70 cultures of the tip of used Foley catheter, performed during the period of 20 months from March l st, 1977 to October 31st, 1978 and the following results were obtained. l. Almost twice much positive bacterial culture was obtained from the culture of used catheter tip (45. 7%) compared with the urine culture taken from the same patient at the same time (24. 3%). 2. Positive culture rate of the catheter tip in the group who was catheterized at operating room (32.7%) was much lower than in the group catheterized at ward (76.2%). Very few positive culture was obtained from the catheter tip used in a patient who had normal urine findings previous to catheterization if this indwelling catheter removed within 72 hours. 4. The most common organism on the culture of the catheter tip was proteus sp. (9 cases), and less commonly, pseudomonas (3 cases), enterococci (3 cases), yeast (3 cases), E. coli (2 cases), klebsiella (2 cases), alcaligenes (2 cases), enterobact.(2 cases) and citrobact.(2 cases) were found.
Alcaligenes ; Catheterization ; Catheters* ; Catheters, Indwelling ; Humans ; Klebsiella ; Operating Rooms ; Proteus ; Pseudomonas ; Yeasts

PURPOSE: To report a case of Alcaligenes xylosoxidans endophthalmitis following cataract extraction and IOL implantation. In Korea, Alcaligenes xylosoxidans endophthalmitis has not been reported. METHODS: The patient was transferred for endophthalmitis at 6 days after cataract extraction and IOL implantation. Alcaligenes xylosoxidans was founded in culture. We performed pars plana vitrectomy with removal of IOL and lens capsule, and then we supplied systemic IV antibiotics with fortified topical antibiotics. RESULTS: Sixty-four days after vitrectomy, corrected visual acuity was 1.0 and anterior segment showed no inflammatory cell.
Alcaligenes* ; Anti-Bacterial Agents ; Cataract Extraction* ; Cataract* ; Endophthalmitis* ; Humans ; Korea ; Visual Acuity ; Vitrectomy

In this research a strain of isolated Pseudomonas alcaligenes which causes degradation of dexamethasone was acclimated further and its proteins of every position in the bacterium were separated by the osmotic shock method. The separated intracellular proteins which had the highest enzyme activity were extracted by the salting out with ammonium sulfate and were purified with the cation exchange chromatography and gel chromatography. The purified proteins which was active to cause degradation of dexamethasone had been detected were cut with enzyme and were analyzed with mass spectrometry. The results showed that the degradation rate to dexamethasone by acclimated Pseudomonas alcaligenes were increased from 23.63% to 52.84%. The degrading enzymes were located mainly in the intracellular of the bacteria and its molecular weight was about 41 kD. The specific activity of the purified degrading enzymes were achieved to 1.02 U x mg(-1). Its 5-peptide amino acid sequences were consistent with some sequences of the isovaleryl-CoA dehydrogenase. The protein enzyme may be a new kind degrading enzyme of steroidal compounds. Our experimental results provided new strategies for cleanup of dexamethasone in water environment with microbial bioremediation technique.
Amino Acid Sequence ; Bacterial Proteins ; chemistry ; Chromatography ; Dexamethasone ; chemistry ; Molecular Weight ; Pseudomonas alcaligenes ; enzymology

Background: A clinician reported unusually high incidence of A. xylosoxidans isolation from aspirated tissues in outpatient clinic. Methods: A. xylosoxidans isolates from January 2002 to June 2002 were investigated. The infection control nurse reviewed medical records and observed the procedures of tissue aspiration and culture at the clinical microbiology laboratory. Specimens were obtained for investigational cultures from dye, aspiration gun, slide alcohol sponge, tray, sink. water of sink, buffer solution, microscope, computer, and telephone. Results: A. xyloxosidans was isolated from twenty-four patients during 6 months. None of 24 cases had any typical signs or symptoms of infections by A. xylosoxidans. Observation of tissue aspiration and culture procedure revealed that buffer solution was used for prevention of specimen drying after tissue aspiration. Culture of the buffer solution yielded a heavy growth of A. xylosoxidans from four out of ten specimens. A. xylosoxidans was not isolated from any other investigational specimens. Conclusions: This was supposed to represent pseudoepidemic. Contaminated buffer solution was documented as the cause of this pseudoepidemic. The usage of buffer solution was stopped. During the follow-up period of 2 months, no additional A. Xylosoxidans was cultured from aspirated tissues.
Alcaligenes* ; Ambulatory Care Facilities ; Follow-Up Studies ; Humans ; Incidence ; Infection Control ; Medical Records ; Methods ; Porifera ; Telephone ; Water