• Journal of Applied Biological Chemistry
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• v.56 no.2
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• pp.89-93
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• 2013

Gamma-aminobutyric acid (GABA) aminotransferase (gabT) and succinic semialdehyde dehydrogenase (gabD) genes from Pseudomonas fluorescens KCCM 12537 were cloned into a single pETDuet-1 vector and co-expressed in Escherichia coli BL21(DE3) simultaneously. The mixture of both enzymes, called GABase, is the key enzyme for the enzymatic analysis of GABA. The molecular mass of the GABA aminotransferase and succinic semialdehyde dehydrogenase were determined to be 52.8 and 46.7 kDa following computations performed with the pI/Mw program, respectively. The GABase activity between pH 6.0 and 9.0 for 24 h at $4^{\circ}C$ remained over 75%, but under pH 6.0 decreased rapidly. The GABase activity between 25 and $35^{\circ}C$ by the treatment at pH 8.6 for 30 min remained over 80%, but over $35^{\circ}C$ decreased rapidly. When the activity against GABA was defined as 100%, the purified GABase activity against 5-aminovaleric acid having a similar structure to GABA showed 47.7% and GABase activity against ${\beta}$-alanine, ${\varepsilon}$-amino-n-caproic acid, $_L$-ornithine, $_L$-lysine, and $_L$-aspartic acid showed between 0.3 to 2.3%. The GABA content was analyzed with this co-expressed GABase, compared with the other GABase which was available commercially. As a result, the content of GABA extracted from brown rice, dark brown rice, and black rice were $26.4{\pm}3.5$, $40.5{\pm}4.7$ and $94.7{\pm}9.3{\mu}g/g$, which were similar data of other GABase in the error ranges.

• Korean Journal Plant Pathology
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• v.12 no.1
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• pp.99-108
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• 1996

3종의 빙핵세균 Peudomonas syringae 8401, Pseudomonas fuorescens 8701, Erwinia herbicola 8701의 세포 외막으로부터 아무런 변성제도 사용치 않고 sucrose density gradient centrifugation, Sephacryl gel filtration chromatography, DEAE-cellulose ion exchange chromatography, non-denaturing buffer를 이용한 PAGE, electroelution, SDS-PAGE를 통해 빙핵활성 단백질을 고도로 정제할 수 있었다. P. suringae와 P. fluorescens에서는 각각 3종류(155 kD, 75 kD, 50 kD)의 빙핵활성 단백질이, E. herbicola에서는 155 kD를 제외한 2종류(75 kD, 50 kD)의 빙핵활성 단백질은 이 연구를 통해 처음 밝혀진 것으로 , 지금까지 보고된 빙핵활성 단백질(150 KD 이상)보다는 훨씬 작은 것이다. 이는 빙핵활성을 나타내는 단백질의 기본단위는 이 실험의 결과만에 의하면 최대 50 kD임을 시사한다. 이들 단백질은 그 유래된 세균의 종류나 또는 단백질 분자량의 크기에 관계없이 모두 -5.5~7.5$^{\circ}C$에서 물을 동결시키는 높은 빙핵활성을 갖고 있었다. 이는 지금까지 보고된 어느 정제단백질의 빙핵활성보다 높은 것이다. 정제된 단백질의 빙핵활성은 trypsin 처리에 의해 상실되었고, pH6~8범위에서는 안정하였으며, pH5이하, pH9이상에서는 활성을 상실하였다. 보존온도에 대한 영향은 3$0^{\circ}C$이상이 되면 점차 활성이 감소하는 경향을 보이다 37$^{\circ}C$이상에서는 활성이 완전히 상실되었다. 금속이온으로서 Hg\ulcorner이온과 SDS에 의해 활성이 상실되었으나 phosphatidylinositol의 첨가에 의해서는 활성이 약간 증가(-1$^{\circ}C$)하였다.

• Journal of Environmental Health Sciences
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• v.33 no.5
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• pp.462-469
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• 2007

Isolation and application of oil-degradation microbes from the oil-contaminated soil and the determination of optimal operation conditions about the peat moss, the addition for the oil-biodegradation. After all experiments, we have acquired three important conclusions: First, we found out the 4 microbes, Pseudomonas fluorescens, Pseudomonas aeruinosa, Kurtia sp., Bacillus ceres, with excellent capability for the oil-degradation; Second, the optimal operating conditions of the peat moss for TPH treatment were pH $7{\sim}8$, temperature $25{\sim}30^{\circ}C$, water content 20%, mixing 2 times/ day, addition volume 2%; Third, in case of the application to the oil-contaminated soil with 4 mixed microbes, the removal efficiency of TPH was increased from 54% to 83% in oil-contaminated soil and from 65% to 85% in oil-contaminated soil with the peat moss.

• Journal of Environmental Health Sciences
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• v.32 no.5 s.92
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• pp.478-484
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• 2006

There was no significant difference in the CADR (Clean Air Delivery Rate) between physical aerosols, NaCl and smoke, and biological aerosols, airborne MS2 virus and P. fluorescens, which implicate that the hybrid-type of air purifier, applying the unipolar ion emission and the radiant catalytic ionization, imposed identical reduction effect on both physical aerosol and bioaerosol. Ventilation decreases the efficiency of air cleaning by unipolar ionization because high ventilation diminishes the particle concentration reduction effect. The particle removal efficiency decreases with increase in the chamber volume because of the augmented ion diffusion and higher ion wall loss rate. Particle size affects the efficiency of air ionization. The efficiency is high for particles with very small diameter because reduction of charge increases with particle size. If there is no increasing supply of ions, the efficiency of air cleaning by unipolar ionization increases with respect to initial concentration of particles because of the large space charge effect at high particle concentration and amplified electric field.

• Korean Journal Plant Pathology
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• v.12 no.4
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• pp.381-388
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• 1996

맥류세균성 줄무늬병균의 선택배양기(KM-1)를 개발하여 이병식물체 및 토양으로부터 Xanthomonas campestris pv. translucens를 선택적으로 분리할 수 있는 효율성을 검토하였다. KM-1배양기의 구성성분은 증류수 1 L당 lactose 10 g, D(+)trehalose 4.0 g, thiobarbituric acid 0.2 g, K\ulcornerHPO\ulcorner 및 KH\ulcornerPO\ulcorner 각각 0.8 g, yeast extract 30 mg, NH\ulcornerCl 1 g, cycloheximide 100 mg, tobramycin 8.0 mg, ampicillin 1.0 mg 및 Bacto agar 15 g이며 1 N NaOH로 pH 6.6으로 조절하였다. X. c. t.의 균주별 KM-1의 배양효율은 비선택성 농후배지인 Wilbrinks agar에 비하여 1.30정도였으며, 기타 토양전염성식물병원세균 Agrobacterium tumefaciens, Agrobacterium rhizogenes, Erwinia carotovora var. atroseptica, Erwinia carotovora var. carotovora, Corynebacterium insidiosum, 및 기타 토양생존 부생세균 Bacillus cereus, Pseudomonas aeruginosa, Pseudomonas fluorescens, and Pseudomonas putida 등의 생장을 완벽하게 억제하였다. KM-1의 저장기간(shelf-life)도 5$^{\circ}C$에서 2개월 동안 선택성을 유지하였다. 따라서 본 병원균의 전염원의 생존 등 발생생태연구에 활용될 수 있는 가치가 충분히 인정되었다.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.5
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• pp.888-892
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• 2000

Antimicrobial activity of Caesalpina sappan L. extract (CS extract) against 6 kinds of food spoilage and pathogenic organisms was studied. The growth of Listeria monocytogenes Brie 1, Escherichis coli ATCC 11775, Staphylococcus aureus ATCC 11775, and Pseudomonas fluorescens ATCC 11775 was inhibited about 4 to 5 $log_{10}$ cycle in Tryptic soy Broth(TSB) containing 1% CS extract. Bacillus subtilis KCTC 102 and Vibrio parahaemolyticus ACTT 17802 did not show apparent growth in the same medium. Effect of CS extract on preservation of ground meat was also investigated. The range of pH change was 5.0~5.2 in CS extract added ground meat, 5.2~6.0 in CS extract not added ground meat (control) during storage at 4$^{\circ}C$ for 30 days. Number of total bacteria after 15 days storage was $10^{6}$/g in CS extract added ground meat, 10$^3$/g in control. Redness of ground meats was improved significantly by addition of 1% CS extract during storage at 4$^{\circ}C$ for 30 days. The sensory quality of 1% CS extract added hamburger patty was similar to that of the control in taste, flavor, and overall acceptability.

• Journal of Microbiology and Biotechnology
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• v.17 no.8
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• pp.1300-1307
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• 2007

This study assessed the possible role of different traits in selected plant growth-promoting rhizobacteria (PGPR) for improving wheat growth and yield under natural conditions. Rhizobacteria exhibiting 1-aminocyclopropane-1-carboxylate (ACC)-deaminase activity were isolated and screened for their growth-promoting activity in wheat under axenic conditions. Five isolates belonging to Pseudomonas and one Burkholderia caryophylli isolate that showed promising performances under axenic conditions were selected and characterized for in vitro ACC-deaminase activity, chitinase activity, auxin production, P solubilization, and root colonization. These isolates were then used as inocula for wheat cultivated under natural conditions in pot and/or field trials. Significant increases in root elongation, root weight, tillers per pot, 1,000-grain weight, and grain and straw yields were observed in response to inoculation with PGPR in the pot trials. Inoculation with these PGPR was also effective under field conditions and increased the wheat growth and yield significantly. However, the efficacy of the strains was inconsistent under the axenic, pot, and field conditions. Pseudomonas fluorescens ($ACC_{50}$), which exhibited a relatively high in vitro ACC-deaminase activity, chitinase activity, auxin production, and P solubilization and more intensive root colonization, was the most efficient isolate under the field conditions. Therefore, these results demonstrated that ACC-deaminase activity is an efficient parameter for the selection of promising PGPR under axenic conditions. However, additional traits of PGPR, including auxin production, chitinase activity, P solubilization, and root colonization, are also important for selecting PGPR as biofertilizers.

• Journal of Microbiology and Biotechnology
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• v.25 no.8
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• pp.1349-1360
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• 2015

The rhizospheric zone abutting plant roots usually clutches a wealth of microbes. In the recent past, enormous genetic resources have been excavated with potential applications in host plant interaction and ancillary aspects. Two Pseudomonas strains were isolated and identified through 16S rRNA and rpoD sequence analyses as P. fluorescens QAU67 and P. putida QAU90. Initial biochemical characterization and their root-colonizing traits indicated their potential role in plant growth promotion. Such aerobic systems, involved in gluconic acid production and phosphate solubilization, essentially require the pyrroloquinoline quinine (PQQ)-dependent glucose dehydrogenase (GDH) in the genome. The PCR screening and amplification of GDH and PQQ and subsequent induction of mutagenesis characterized their possible role as antioxidants as well as in growth promotion, as probed in vitro in lettuce and in vivo in rice, bean, and tomato plants. The results showed significant differences (p ≤ 0.05) in parameters of plant height, fresh weight, and dry weight, etc., deciphering a clear and in fact complementary role of GDH and PQQ in plant growth promotion. Our study not only provides direct evidence of the in vivo role of GDH and PQQ in host plants but also reveals their functional inadequacy in the event of mutation at either of these loci.

• Research in Plant Disease
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• v.22 no.1
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• pp.1-8
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• 2016

Elsinoe fawcettii causing citrus scab was suppressed by rhizobacterial strains such as Burkholderia gladioli MRL408-3, TRH423-3 and Pseudomonas fluorescens THJ609-3, TRH415-2 having antifungal activity. The leaf surface of Satsuma mandarin, which was pre-treated with the rhizobacterial strains, was observed by a scanning electron microscope (SEM) after inoculation with E. fawcettii. The number of lesions was reduced on the leaves pre-treated with the rhizobacterial strains compared to those of untreated leaves. Especially, the lesions numbers was apparently reduced on the leaves pre-treated with B. gladioli MRL408-3. The observation by SEM revealed that not only the germination rate but also the length of germ tube of the pathogen were decreased on the rhizobacterial strains pre-treated leaves. These inhibition of the fungal growth was more strongly expressed on the leaves pre-treated with commercial fungicide imibenconazole, by which the lesions was rarely found on the leaves. Based on these results, it was suggested that rhizobacterial strains may inhibit the germination and growth of the E. fawcettii on the surface of citrus leaves, resulting in decrease of disease severity.

• Journal of Food Hygiene and Safety
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• v.22 no.2
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• pp.77-81
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• 2007

Antimicrobial activity of both fresh Prunus mume and Prunus mume liqueur byproduct (PLB), generated after producing Prunus mume liqueur were examined against various pathogeinc bacteria such as Listeria monocytogenes Scott A, Listeria monocytogenes ATCC 19115, Bacillus cereus KCCM 11341, Staphylococcus aureus KCCM 12255, Pseudomonas fluorescens ATCC 21541, Escherichia coli O157:H7, Salmonella Typhimurium ATCC 14028, and Shigella sonnei. PLB showed strong antibacterial effects against tested pathogenic bacteria.L. monocytogenes ATCC 19115, B. cereus KCCM 11341, S. sonnei, and E. coli O157:H7 were not detected in trytpic soy broth containing 1% of prunus mume or PLB after 24-hour incubation at $37^{\circ}C$, respectively. Prunus mume showed higher antimicrobial activities than that of PLB against tested pathogens.