• 미생물학회지
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• 제26권2호
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• pp.129-136
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• 1988

To investigate the influence of cosubstrate supplement and ammonium sulfate on lignin degradation by Pseudomonas diminuta KM-4-2, isolated in the laboratory, the strain was cultured on the lignin media which contained lignin as a source of carbon and the culture filtrate was analyzed by Sephadex G-75 column chromatography. It was found that polymerization was not appeared unlike wood-rot fungi. When the carbohydrates were added, the peak of lignin at 280nm by UV scanning spectra of the filtrate, was significantly increased. In order to determine the effect of ammonium sulfate on the ligninolytic activity, the isolated strain was incubated in the media containing 0.1%, 0.25% and 0.5% of nitrogen concentration in the Warburg flask and the rate of oxygen uptake was esitmated by Warbuge Respirometer. As a result, the activity was maximum at 0.1% of nitrogen concentration and thereafter decreased in parallel with nitrogen concentration.

• Journal of Microbiology
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• 제37권4호
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• pp.200-205
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• 1999

7-Aminocephalosporanic acid (7-ACA) is the initial compound in preparation of cephalosporin antibiotics widely used in clinical treatment. Bacteria producing glutaryl 7-ACA acylase, which convert cephalosporin C to 7-ACA, has been screened in soil samples. A bacterial strain exhibiting high glutaryl 7-ACA acylase activity, designated KAC-1, was isolated and identified as a strain of Pseudomonas diminuta by characterizing its morphological and physiological properties. The screening procedures include culturing on enrichment media containing glutaric acid, glutamate, and glutaryl 7-aminocephalosporanic acid as selective carbon sources. To enhance enzyme production, optimal cultivation conditions were investigated. This strain grew optimally at pH 7 to 9 and in temperatures of 20 to 40 C, but acylase production was higher when the strain was grown at 25 C. Glutaric acid, glutamate and glucos also acted as inducers for acylase production. In a jar fermenter culture, P. diminuta KAC-1 produce acylase in a growth-associated manner. The substrate specificity of KAC-1 acylase by cell extract showed that this enzyme had specificity toward glutaryl 7-ACA, glutaryl 7-ADCA, but not cephalosporin C.

• Journal of Microbiology and Biotechnology
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• 제11권2호
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• pp.326-328
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• 2001

Pseudomonas diminuta (ATCC 19146) has been typically used in the bacterial challenge test for validation of the sterilizing filtration process. Cell size is critical for determining the retention characteristics of membrane filters with pore-size of $0.2{\mu}m$. The changes of cell sizes after osmotic shocks at 150, 260, 500, and 700 mosM were measured by a particle size analyzer and the changes of their buoyant densities were analyzed with a Percoll gradient. The results indicated that there were no significant differences when cells were cultured in 260 mosM medium and osmotically shocked at 500 and 700 mosM. However, the osmotically shocked cells at 150 mosM showed a 38% increase of the cell size compared to the cells at 260 mosM. From these study, we concluded that the worst case condition for validation of a sterilizing filter would be 500 mosM, not because of changes in the cell size, but due to decrease in cell viability under those conditions.

• Journal of Microbiology and Biotechnology
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• 제11권3호
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• pp.452-457
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• 2001

The glutaryl 7-aminocephalosporanic acid (glutaryl 7-ACA) acylase was purified from Pseudomonas diminuta KAC-1 cells isolated from soil, and characterized. The acylase was purified by procedures including ammonium sulfate fractionation and column chromatographies on DEAE-Sepharose, Phenyl-Sepharose, Q-Sepharose, and Superose 12H/R. The negative acylase was found to be composed of two subunits with molecular masses of approximately 55 kDa and 17 kDa, respectively. The isoelectric point of the enzyme was 4.0. The specific activities of the purified acylase were 8.0 and 7.0 U/mg on glutaryl 7-ACA and glutaryl 7-aminodesacetoxy cephalosporanic acid (glutaryl 7-ADCA), respectively, and $K_m$ values were 0.45 mM for glutaryl 7-ADCA and 0.67 mM for glutaryl 7-ADCA. The enzyme had a pH optimum at 8.0 and a tmperature optimum at $40^{\circ}C$. The acylase catalyzed the synthesis of glutaryl 7-ACA from glutaric acid and 7-ACA as well as the hydrolysis of glutaryl 7-ADCA, although the reaction rate of the synthesis was slower than that of the hydrolysis. In addition, it was found that the enzyme had a glutaryl transferase activity, thereby transferring the glutaryl group from one cephalosporin nucleus to another.

• 한국식품영양과학회지
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• 제24권4호
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• pp.631-635
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• 1995

In order to develop effective manufacturing method and to improve quality of low-salt fermented squid(10% of table salt), we investigated the effects of temperature, salinity and pH on the growth of Staphylococcus xylosus, Micrococcus varians, Pseudomonas diminuta and Pseudomonas D2 isolated from of low-salt fermented squid and the growth characteristics of these bacteria during fermentation were elucidated. All bacteria showed good growth during the process of low-salt fermented squid(pH 6~7 ; concentration of NaCl, 7~10% ; temperature, 7~1$0^{\circ}C$) and their cell numbers increased as fermentation proceeded under the same fermentation condition.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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• pp.510-515
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• 2005

Pseudomonas cepacia BY21 was found to produce glutaryl acylase that is capable of deacylating glutaryl-7-aminocephalosporanic acid (glutaryl-7-ACA) to 7-aminocephalosporanic acid (7-ACA), which is a starting material for semi-synthetic cephalosporin antibiotics. Amino acids of the reported glutaryl acylases from various Pseudomonas sp. strains show a high similarity (>93% identity). Thus, with the known nucleotide sequences of Pseudomonas glutaryl acylases in GenBank, PCR primers were designed to clone a glutaryl acylase gene from P. cepacia BY21. The unknown -subunit gene of glutaryl acylase from chromosomal DNA of P. cepacia BY21 was cloned successfully by PCR. The -subunit amino acids of P. cepacia BY21 acylase (GenBank accession number AY948547) were similar to those of Pseudomonas diminuta KAC-1 acylase except that Asn408 of P. diuminuta KAC-1 acylase was changed to Leu408.

• 한국식품과학회지
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• 제31권6호
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• pp.1619-1627
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• 1999

염 농도와 감마선 조사 선량을 달리한 오징어젓갈의 미생물 균총 변화를 살펴보았다. 총균, Lactobacillus spp., Staphylococcus spp.는 발효기간에 따라 지속적으로 생장하여 발효 40일째에 염도 5%와 10%에서는 $10^8\;CFU/mL$까지, 염도 20%에서는 $10^7\;CFU/mL$까지 생장하였으며 5 kGy 이상 감마선 조사구는 초기 균수 감소로 발효 후기의 미생물 밀도가 낮아졌다. 염도가 증가할수록 감마선 조사가 미생물 생장에 미치는 영향이 컸으며, 감마선 조사에 의한 오징어젓갈 관련 발효 미생물의 생장억제는 염도 10%에 10 kGy의 감마선 조사 조건이 적합하였다. 발효미생물의 동정 결과, L. plantarum이 전 실험구에 걸쳐 가장 일반적인 발효균으로 분리되었으며 Lactobacillus sp. 2, Mic. varians, Streptococcus sp. 1 등은 염도 5%의 실험구에만, Mic. morrhuae 등은 20%의 염농도에서만 분리되어 염도에 따른 microflora의 차이를 나타내었다. 또한, L. brevis, Ped. halophilus, Pse. diminuta 등은 방사선 감수성이 큰 미생물군이었으며, L. plantarum, Mic. morrhuae, Pseudomonas sp. 3 등은 방사선에 대한 저항성이 높았다. 전체적으로 보아 염도 20% 실험구에서의 분리 미생물이 15종으로 가장 다양한 분포를 보였으며 염도가 낮아질수록, 감마선 조사선량이 높아질수록 종의 다양성은 낮았다.

• 한국산학기술학회논문지
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• 제13권10호
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• pp.4910-4916
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• 2012

세라믹담체에 적용한 4종의 해양박테리아 (Aeromonas hydrophila, Chryseomonas indologenes, Pseudomonas diminuta, Vibrio parahaemolyticus)의 저농도 질소 인 제거 효율의 변화를 분석하였다. 해양박테리아는 광양만에서 분리 동정하였다. 담체에 적용한 4종의 해양박테리아 모두 대조군에 보다 약 3배 정도의 성장률이 증가하였으며, 암모니아서 질소 제거효율도 30% 이상 증가하였다. 질산성 질소의 제거 효율은 A. hydrophila 균주가 가장 높았으며, 인의 제거는 P. diminuta 균주가 가장 높은 것으로 나타났다. 본 연구의 결과는 세라믹담체는 질소-인 제거 효율 증진에 좋은 재료이며, 분리된 해양박테리아는 현장의 저농도 질소-인 조절에 유용할 수 있음을 보여준다.

• 한국산학기술학회논문지
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• 제13권7호
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• pp.3267-3274
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• 2012

371 균주의 해양박테리아를 광양만에서 분리하였다. 우점종은 Pseudomonas aeruginosa, Aeromonas hydrophila, P. fluorescens, P. paucimobilis, Chryseomonas luteola, P. vescularis 등이었다. 영양염과 유기물을 제거할 수 있는 해양박테리아를 탐색하기 위하여 암모니아성 질소(100 mg/L), 질산성 질소(100 mg/L) 및 인(10 mg/L)이 각각 포함된 10 mL의 marine broth 2216 (DIFCO)에 해양박테리아를 접종(1.0%, v/v)하고 12시간 배양하였다. 스크리닝 테스트 결과 25% 이상 $COD_{Cr}$을 제거하는 해양박테리아는 16종, 15% 이상의 암모니아 질소를 제거하는 해양박테리아는 9종, 60% 이상의 질산 질소를 제거하는 해양박테리아는 11종 그리고 90% 이상의 인을 제거하는 해양박테리아는 13종이 분리되었다. Aeromonas hydrophila, Chryseomonas indologenes, Pseudomonas diminuta, Vibrio parahaemolyticus 균주가 유기물 및 영양염류 제거실험을 위해 선정되었다. 회분식 시험을 위해 4종의 해양박테리아를 $COD_{Cr}$ 250 mg/L, $NH_3-N$ 40 mg/L, ${NO_3}^{-}-N$ 40 mg/L, ${PO_4}^{3-}-P$ 10 mg/L이 각각 첨가되어 있는 변형 marine broth에 접종하고, 10시간 배양하면서 제거효율을 측정하였다.

• Journal of Microbiology and Biotechnology
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• 제23권2호
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• pp.195-204
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• 2013

While structured packing modules are known to be efficient for surface wetting and gas-liquid exchange in abiotic surface catalysis, this model study explores structured packing as a growth surface for catalytic biofilms. Microbial biofilms have been proposed as selfimmobilized and self-regenerating catalysts for the production of chemicals. A concern is that the complex and dynamic nature of biofilms may cause fluctuations in their catalytic performance over time or may affect process reproducibility. An aerated continuous trickle-bed biofilm reactor system was designed with a 3 L structured packing, liquid recycling and pH control. Pseudomonas diminuta established a biofilm on the stainless steel structured packing with a specific surface area of 500 $m^2m^{-3}$ and catalyzed the oxidation of ethylene glycol to glycolic acid for over two months of continuous operation. A steady-state productivity of up to 1.6 $gl^{-1}h^{-1}$ was achieved at a dilution rate of 0.33 $h^{-1}$. Process reproducibility between three independent runs was excellent, despite process interruptions and activity variations in cultures grown from biofilm effluent cells. The results demonstrate the robustness of a catalytic biofilm on structured packing, despite its dynamic nature. Implementation is recommended for whole-cell processes that require efficient gas-liquid exchange, catalyst retention for continuous operation, or improved catalyst stability.