• 한국미생물·생명공학회지
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• 제16권3호
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• pp.199-204
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• 1988

Naphthalene을 유일한 탄소원으로 이용하는 균을 분식 배양과 연속식 배양에 의해서 토양과 폐수로부터 분리하였다. 이 균은 Pseudomonas putida로 동정되었으며, 최적 pH와 온도는 각각 7.0과 3$0^{\circ}C$ 이었다. 분리된 균은 1,5-dihydroxynaphthalene을 naphthalene보다 더욱 잘 이용하였으며 benzoate와 salicylate도 이용하였다. 또한 catechol dl meta-분해경로를 통해서 분해되었으며, ampicillin, chloramphenicol, kanamycin, streptomycin에 대해서 강한 저항성을 지니고 있었으며, naphthalene의 분해에 관여하는 약 110kb 크기의 plasmid를 1개 지니고 있었다.

• 한국지하수토양환경학회지:지하수토양환경
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• 제16권5호
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• pp.74-81
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• 2011

The fluorene-degrading strain Sphingobacterium sp. KM-02 was isolated from PAHs-contaminated soil near a mineimpacted area by selective enrichment techniques. Fluorene added to the Sphingobacterium sp. KM-02 culture as sole carbon source was 78.4% removed within 120 h. A fluorene degradation pathway is tentatively proposed based on identification of the metabolic intermediates 9-fluorenone, 4-hydroxy-9-fluorenone, and 8-hydroxy-3,4-benzocoumarin. Further the ability of Sphingobacterium sp. KM-02 to bioremediate 100 mg/kg fluorene in soil matrix was examined by composting under laboratory conditions. Treatment of microcosm soil with the strain KM-02 for 20 days resulted in a 65.6% reduction in total amounts. These results demonstrate that Sphingobacterium sp. KM-02 could potentially be used in the bioremediation of fluorene from contaminated soil.

• Journal of Microbiology and Biotechnology
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• 제7권1호
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• pp.43-48
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• 1997

Three strains capable of degrading a chlorophenol were isolated by selective enrichment from soils contaminated with industrial wastewater. A Pseudomonas solanacearum TCP114 could use 2,4,6-trichlorophenol (TCP) as sole carbon and energy source, while two strains of Pseudomonas testosteroni CPW301 and Arthrobacter ureafaciens CPR706 could use 4-CP. All isolates also grew well on phenol. The degradation of one component by a pure strain was strongly affected by the presence of other compounds in the medium, CPW301 and CPR706 entirely lost the ability to degrade 4-CP and phenol in the presence of TCP. TCP114 also lost the ability to degrade phenol when 4-CP was added to the culture medium. These restrictions on the degradability could be overcome by employing defined mixed cultures (TCP114 and one strain of 4-CP degrading strains). All three components were successfully degraded by defined mixed cultures through their cooperative activities. It was also demonstrated that defined mixed cultures could be immobilized by using calcium alginate for the semi-continuous degradation of the three component mixture. Immobilization could not only accelerate the degradation rate, but also allowed the reuse of the cell mass several times without loss of the cells' degrading capabilities.

• Bulletin of the Korean Chemical Society
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• 제35권2호
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• pp.383-391
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• 2014

Biotin-S-S-Phosphine was designed and synthesized as a potential tool for a proteomic study of O-GlcNAcmodified proteins. This reagent features a disulfide linker between a triarylphosphine moiety, which allows selective conjugation to azide-containing proteins, and a biotin moiety that can allow easy isolation through its strong affinity toward avidin-coated solid beads. The disulfide linkage within this reagent can allow the easy release of the bound molecules of interest, which is difficult to achieve when a biotin:avidin pair is used alone, by reducing the disulfide bond of the reagent with DTT. Preliminary in vitro biological assays with azidelabeled and unlabeled cell lysates and a pure protein Nup62 showed that the Biotin-S-S-Phosphine reagent is highly reactive toward the free thiol groups of proteins. When a molecular tool with a disulfide linker is applied to the enrichment of the molecules of interest from other species, it is important to block the free-thiols of the sample using exhaustive alkylation prior to the Staudinger ligation reactions to restore the bioorthogonal nature of this reaction.

• Journal of Microbiology and Biotechnology
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• 제24권7호
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• pp.936-942
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• 2014

Using racemic (R,S)-2-(4-chlorophenyl)-3-methylbutyramide, an intermediate for the chiral pyrethroid insecticide Esfenvalerate, as a sole nitrogen source in a minimal medium, several strains with high enatioselectivity (${\geq}98%$) were isolated by enrichment techniques. One of the strains, LG 31-3, was identified as Burkholderia multivorans, based on physiological and morphological tests by a standardized Biolog station for carbon source utilization. A novel amidase was purified from B. mutivorans LG 31-3 and characterized. The enzyme exhibited (S)-selective amidase activity on racemic (R,S)-2-(4-chlorophenyl)-3-methylbutyramide. Addition of the racemic amide induced the production of the enantioselective amidase. The molecular mass of the amidase on SDS-PAGE analysis was shown to be 50 kDa. The purified amidase was subjected to proteolytic digestion with a modified trypsin. The N-terminal and internal amino acid sequences of the purified amidase showed a high sequence homology with those deduced from a gene named YP_366732.1 encoding indole acetimide hydrolase from Burkholderia sp. 383.

• 미생물학회지
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• 제16권3호
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• pp.111-121
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• 1978

About 40 temperature-sensitive mutants have been isolated as a preliminary step to study the spore germination, the cell cycle, and the control of macromolecular synthesis in Aspergillus nidulans. To obtain temperature-sensitive mutants rapidly and effectively, the selective enrichment method using antifungal antibiotic nystatin was developed. Based on the data which had applied to the concentration of auxotrophic mutants by the earlier investigators, the optimal concentration and the time of treatment at the nonpermissive temeprature were determined as 50 to 100 units per ml and 4.5 hr., respectively. Out of 41 ts mutants assigned to the strain symbol PK, thirteen that seemed to be arrested at the earlystage of spore germination were subjected to the further cytological and genetic analysis. Elght of these mutants are able to form germ tube and five not. Staining with acid fuchsin for the 5PK strains shows that one irreversible mutant, PK6 strain able to form germ tube, accumulate mitotic spindle, being arrested in mitosis. Another PK15 and PK23 strain have more than one intact nucleolus without germ tube formation at the restrictive temperature. the temperature-senstive mutation in PK12 strain, the onlystrain which is able occurred in certain gene specific for the germination of spore. All of the ts markers are recessive and complement each other in heterokaryon between two different ts markers at the restrictive temperature.

• 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 Industrial and Engineering Chemistry
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• 제67권
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• pp.461-468
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• 2018

A highly sensitive, selective and rapid method for the determination of lead based on the reaction of lead (II) with 5-(4'-chlorophenylazo)-6-hydroxypyrimidine-2,4-dione (CPAHPD) and the solid phase extraction of the Pb(II)-CPAHPD complex with Amberlite XAD-2000 was developed, in the presence of pH 5.6 buffer solution and Triton X-114 medium. CPAHPD reacts with lead to form a violet complex with a molar ratio of 2:1 (CPAHPD to lead). This complex was enriched by the solid phase extraction with Amberlite XAD-2000. An enrichment factor of 500 was obtained by elution of the complex from the resin with a minimal amount of isopentyl alcohol(0.2 mL). In isopentyl alcohol medium,the molar absorptivity of the complex is $1.13{\times}10^6L\;mol^{-1}cm^{-1}$ at 647 nm. Beer's law is obeyed in the range of $5.0-160ng\;mL^{-1}$ in the measured solution. The relative standard deviation for 10 replicate samples of $50ng\;mL^{-1}$ level is 1.26%. The detection and quantification limits reaches 1.5 and $4.7ng\;mL^{-1}$ in the original samples. The presented procedure was successfully applied for determination of lead content in real samples such as vegetables, waters, biological and soil samples with satisfactory results.

• The Plant Pathology Journal
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• 제40권3호
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• pp.282-289
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• 2024

Fire blight is a bacterial disease caused by Erwinia amylovora. In Korea, fire blight was first reported in 2015 in an orchard. If the infection is confirmed, all trees in the orchard must be removed and the orchard must remain closed for 3 years. Since 2020, if the number of trees infected with fire blight is less than 5% of the total trees in the orchard, only the infected tree and adjacent trees are removed in Korea. Three years after removal, the trees can be replanted after confirming that the orchard soil is free from E. amylovora. In this study, a protocol was established for detecting E. amylovora in soil via selective enrichment, using tryptic soy broth with 0.05% bile salts and 50 ㎍/ml cycloheximide, and real-time polymerase chain reaction. This protocol resulted in a 1,000-times improved detection limit for E. amylovora in soil samples compared to that in unenriched samples. Soil monitoring was performed for orchards where fire blight-infected trees had been removed 3-27 months prior; the selected orchards were monitored every 3 months. Monitoring confirmed that E. amylovora was not present in the soil at any site in any of the orchards. A new detection protocol facilitates the monitoring of E. amylovora in soil and could help permit the replanting of trees in orchards. Also monitoring results provide evidence that trees can be planted earlier.

• 한국식품위생안전성학회지
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• 제31권6호
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• pp.439-444
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• 2016

두 종류의 Cronobacter 선택배지(DFI agar, R&F agar)의 분유 및 건조호박 내 Cronobacter의 선택 분리능을 real-time PCR법과 함께 비교하였다. 분유에서의 Cronobacter 검출률은 세 가지 방법에서 유의적인 차이를 나타내지 않았으나(p < 0.05), 건조호박의 경우 R&F배지와 real-time PCR법이 DFI에서보다 유의적으로 높은 검출률을 보였다(p < 0.05). 배지 간 선택성에 있어서도, R&F 선택배지는 건조호박에서 DFI에 비해 유의적으로 높은 선택성을 나타냈다(p < 0.05). Real-time PCR 및 R&F배지의 사용은 분유뿐만 아니라, 건조 호박 등의 높은 경쟁세균총을 갖는 영유아식의 원료로 사용될 수 있는 식품군에서도 Cronobacter를 효과적으로 검출할 수 있는 방법으로 사료된다.