• Journal of Microbiology
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• 제39권3호
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• pp.195-201
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• 2001

The effect of moisture content an the reductive dechlorination of polychlorinated biphenyls and population dynamics of dechlorinating microorganisms was investigated in sediments spiked with Aroclor 1248. In sediment slurry with an overlying water layer, dechlorination ensued after a 4-week lag period and reduced the average number of chlorines per biphenyl from 3.91 to 3.15 after 48 weeks. In the sediments of reduced moisture content, however, dechlorination occurred after a lag period of 12 weeks and decreased the average number of chlorines per biphenyl to only 3.62, and the dechlorination rate was also slower. When the population size of dechlorinators, methanogens, and sulfate-reducing bacteria was determined by the most probable number techniques, however, no difference was found between the slurry and the low-moisture sediments, except for methanogens. The growth of dechlorinating populations coincided with the end of the lag period and they then increased by 3 orders of magnitude in two conditions. Specific growth rate of dechlorinators showed little difference between the slurry and the low-moisture sediments; however, growth yield was high in the sediments of reduced moisture content. The reduction of sediment moisture decreased the dechlorination rate and extent of PCBs but did not inhibit the growth of PCB dechlorinators.

• Journal of the Korean Wood Science and Technology
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• 제50권3호
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• pp.159-166
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• 2022

CO₂ emissions are the primary reason for global warming; hence, biological and chemical technologies for converting CO₂ into useful compounds are being actively studied. Biological methods using enzymes can convert CO₂ under mild conditions. Formate dehydrogenase (FDH) is a representative CO₂ conversion enzyme. Its function was revealed after isolation from bacteria, yeast, and plants. In this study, we evaluated the CO₂ conversion potential of FDH isolated from wood-rotting fungi. After isolating the FDH gene (TvFDH) from Trametes versicolor, we cloned the full-length FDH from T. versicolor and expressed it in a cell-free expression system. The gene encoding TvFDH was identified as 1,200 bp open reading frame (ORF) and the expected molecular weight of the protein was approximately 42 kDa. Overexpression of the recombinant crude protein including TvFDH was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Enzyme activities and metabolite analyses confirmed the efficiency of TvFDH for CO₂ reduction.

• Journal of Microbiology and Biotechnology
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• 제25권8호
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• pp.1299-1306
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• 2015

This study aims to investigate the mechanism of action of the cinnamon bark essential oil (CB), when used singly and also in combination with piperacillin, for its antimicrobial and synergistic activity against beta-lactamase TEM-1 plasmid-conferred Escherichia coli J53 R1. Viable count of bacteria for this combination of essential oil and antibiotic showed a complete killing profile at 20 h and further confirmed its synergistic effect by reducing the bacteria cell numbers. Analysis on the stability of treated cultures for cell membrane permeability by CB when tested against sodium dodecyl sulfate revealed that the bacterial cell membrane was disrupted by the essential oil. Scanning electron microscopy observation and bacterial surface charge measurement also revealed that CB causes irreversible membrane damage and reduces the bacterial surface charge. In addition, bioluminescence expression of Escherichia coli [pSB1075] and E. coli [pSB401] by CB showed reduction, indicating the possibility of the presence of quorum sensing (QS) inhibitors. Gas-chromatography and mass spectrometry of the essential oil of Cinnamomum verum showed that trans-cinnamaldehyde (72.81%), benzyl alcohol (12.5%), and eugenol (6.57%) were the major components in the essential oil. From this study, CB has the potential to reverse E. coli J53 R1 resistance to piperacillin through two pathways; modification in the permeability of the outer membrane or bacterial QS inhibition.

• 약학회지
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• 제32권5호
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• pp.295-303
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• 1988

For microbial production of acetaminophen, a popular analgesic-antipyretic from aniline, we screened various fungi and bacteria. And we succeeded to some extents in acetaminophen production by successful protoplast fusion between S. lividans and S. globisporus and also between S. rimosus and S. aureofaciens. However, more fertile results might be brought via performing the cloning of acetanilide p-hydroxylation genes of Streptomyces in yeast. This study was initiated to determine whether the acetanilide p-hydroxylase of Streptomyces is cytochrome P-450 species or non-heme iron protein species. The p-hydroxylationactivity on acetanilide in S. aureofaciens ATCC 10762 was found to be unstable on exposing to the air. However, 100,000xg supernatant of the cell free extracts which were prepared in $N_2$ atmosphere showed the p-hydroxylation activity. Characteristic absorption peak of cytochrome P-450 after reduction with dithionite and addition of CO was not observed in the region of 450nm. Moreover, metyrapone and 2, 6-dichloroindophenol did not affect this enzyme activity, but sodium azide, sodium cyanide, cupric sulfate, cadmium chloride, ${\alpha}$, ${\alpha}'-dipyridyl$, and o-phenanthroline reduced p-hydroxylase activity considerably. S. fradiae NRRL 2702 was shown to have strong p-hydroxylation activity in intact cells. This activity disappeared in its cell free extracts. In its 100,000xg supernatant, however, characteristic absorption peak of cytochrome P-450 after reduction with dithionite and addition of CO was observed at 446nm. Thus, the results herein presented suggest that acetanilide p-hydroxylase of Streptomyces aureofaciens is not related to cytochrome P-450 and may include non-heme iron protein for its activity. However, it is not clear whether acetanilide p-hydroxylase in S. fradiae belongs to the same category of S. aureofaciens.

• 대한환경공학회지
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• 제28권4호
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• pp.438-446
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• 2006

팔당호 경안천 하류 정체수역에서 퇴적물 인 용출 저감을 위한 capping 소재를 개발하고자 실험실 규모의 batch test를 수행하였다. 조사지역 퇴적물 평균 입도는 7.7 ${\phi}$로 매우 세립하고, 유기탄소 함량은 2.4%로 매우 높게 나타났다. 최적의 capping 소재 선정을 위한 인 용출 실험을 위해 150 L 반응기 5개에 모래층, powder-gypsum($CaSO_4{\cdot}2H_2O$), granule-gypsum, 복합층(gypsum+sand)로 capping을 한 경우와 control을 45일간 비교 평가하였다. 모래로 capping할 경우, 퇴적물로부터 용출되는 인을 약 50% 차단 가능한 것으로 나타났다. 그러나 모래층 1 cm로는 $CH_4$ gas 발생으로 인한 퇴적물의 재부유와 유기물 분해에 의한 수층 용존산소 감소(3 mg/L 이하)를 초래하는 것으로 나타났다. 따라서 모래로 capping을 할 경우에는 모래층이 더 높아져야 할 것으로 판단된다. powder-Gypsum과 granule-Gypsum은 80% 이상의 인 용출 저감 효과를 보였으나 수층 ${SO_4}^{2-}$ 농도가 증가되어 상수원보호구역에 적용하기 어려운 것으로 나타났다. 따라서 ${SO_4}^{2-}$의 용해도를 감소시키기 위해 Fe-Gypsum, $SiO_2$-gypsum 소재를 개발하였다. powder-Gypsum은 물과 결합시 경화되어 퇴적물 층위에 공극이 전혀 없는 차단막을 만들기 때문에 gypsum 층에 crack이 생길 경우, $CH_4$ gas 발생으로 인한 인 용출이 한꺼번에 일어날 수 있다. 복합층(granule-Gypsum+sand(1 cm))으로 capping을 할 경우, 인 용출 차단 효과가 높을 뿐만 아니라, ${SO_4}^{2-}$ 농도 역시 powder-Gypsum, granule-Gypsum 보다 수층으로 용출이 적어 상수원보호구역에 적합한 것으로 나타났다. Gypsum으로 capping을 할 경우, 빠른 초기속성작용(early diagenesis)과 ${SO_4}^{2-}$ 농도가 퇴적물에 충분히 공급되어 SRB(sulfate reducing bacteria)의 활성이 높아져 methanogensis의 진행을 저하시킬 수 있는 것으로 나타났다.

• 대한환경공학회지
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• 제28권5호
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• pp.563-572
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• 2006

정체수역 호소 퇴적물로부터 인 용출 저감을 위한 최적의 In-situ 처리 방법을 제시하고자 실험실 규모의 batch test를 수행하였다. 실험에 사용된 퇴적물의 성상은 평균입도 $7.7{\phi}$(mud)로 매우 세립하고, 유기탄소 함량은 2.4%로 매우 높다. 인 용출 실험은 수층 미생물의 영향을 고려한 경우와 미생물의 영향을 배제한 호소수와 증류수로 구분하여 총 12개 컬럼을 비교 평가하였다. In-situ capping 소재로는 모래와 황토를 사용하였으며, in-situ chemical treatment로는 Fe-Gypsum, $SiO_2$-Gypsum의 산화제를 적용하였다. 또한 산화제와 모래층을 결합시킨 복합층에 대해서도 비교 평가하였다. 미생물의 영향을 고려한 호소수의 경우, 실험 초기에는 인의 농도가 급격히 감소하다 10일 이후부터 서서히 증가되는 것을 확인할 수 있었다. 이는 수층의 미생물에 의해 인이 섭취(uptake)되었다가 다시 용출되는데 시간이 소요되는 것으로 나타났다. 30일간 퇴적물로부터 용출된 인의 flux는 control의 경우 $3.0mg/m^2{\cdot}d$로 매우 높고, 모래층(5 cm), 황토층(5 cm)으로 rapping을 한 경우 역시 충분한 압밀이 이루어지지 않아 $2.5mg/m^2{\cdot}d,\;1.8mg/m^2{\cdot}d$로 인 저감 효율이 낮았다. 화학적 소재를 적용한 Fe-gypsum와 $SiO_2$-gypsum은 $1.4mg/m^2{\cdot}d$로 control과 비교시 인 용출 저감 효율이 약 40% 이상으로 나타났다. 복합층으로 rapping을 한 경우는 $1.0mg/m^2{\cdot}d$로 60% 이상의 높은 인 저감 효율을 보였다. 즉, 오염퇴적물에 산화제로서 gypsum($CaSO_4{\cdot}2H_2O$) 적용은 인의 용출을 감소시키며, 퇴적물내 황산염이 서서히 용해되어 SRB(sulfate reducing bacteria)의 활성과 유기물의 광물화를 촉진시킬 수 있다.

• 미생물학회지
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• 제53권2호
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• pp.83-96
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• 2017

부산시 영도구의 혁신지구의 인공해수천은 높아진 하상과 조류의 특성으로 인해 물이 순환되지 않고 더구나 주위의 오수가 유입되고 있어서 수질이 나빠지고 악취를 발생하고 있다. 이 문제를 해결하기 위한 방안으로 가장 오염되고 조류이동을 감안한 하천의 지점에 생물증강법을 적용하여 친환경적, 효율적으로 하천을 정화하고자 하였다. 현장에서 활성화된 복합미생물을 가장 오염된 지점(Site 2)에 7~10일 간격으로 투입하여, 수질의 pH, 온도, DO, ORP, SS, COD, T-N, 및 T-P를 측정하였고 또한 하상퇴적토의 COD 및 미생물군집을 분석하였다. 13개월 후 Site 2의 수질 SS, COD, T-N 및 COD (퇴적토)의 처리효율은 각각 51%, 58%, 27% 및 35%으로 나타났으나 T-P는 오히려 47% 증가를 보였다. 대부분의 측정지점에서 황산염환원세균(sulfate reducing bacteria)그룹 (Desulfobacteraceae_uc_s, Desulfobacterales_uc_s, Desulfuromonadaceae_uc_s, Desulfuromonas_g1_uc and Desulfobacter postgatei)과 Anaerolinaeles의 밀도는 대체적으로 생물증강에 의한 정화가 진행될수록 감소하였으며, 반면에 Gamma-proteobacteria (NOR5-6B_s and NOR5-6A_s), Bacteroidales_uc_s, 및 Flavobacteriales_uc_s의 밀도는 증가하는 경향이었다. 상대적으로 COD가 낮고 DO가 높은 청정지점인 St. 4에서는 호기성미생물인 Flavobacteriaceae_uc_s가 우점하였다. 이러한 미생물군은 하천의 정화과정을 추적할 수 있는 지표미생물로 활용될 수 있을 것으로 판단되었다. 생물증강 시행 후의 대표적 시점 퇴적토시료의 미생물군집 alpha diversity 지수(OTUs, Chao1 및 Shannon 지수)는 시행 전에 비해 감소하는 경향을 보였으며, 또한 beta diversity 분석기법(fast unifrac 분석)으로 분석한 결과 정화 전이나 초기에 비해서 정화가 진행될수록 전반적으로 시료에 무관하게 미생물군집의 유사성을 보여 생물증강이 현장 토착 미생물의 군집구조를 변화시키고 있음을 확인하였다. 이러한 사실로 보아 본 복합미생물에 의한 현장 생물증강법은 brine water 및 담수로 이루어진 오염된 하천을 환경친화적, 경제적으로 정화할 수 있는 대안으로 판단이 되었다.

• 한국축산식품학회지
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• 제35권2호
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• pp.211-215
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• 2015

From the previous study, Enterococcus faecium SH01 was isolated from mukeunji, an over-ripened kimchi, and it produced bacteriocin SH01. Bacteriocin SH01 showed an inhibitory effect against Listeria monocytogenes ATCC 19111, a bacterial strain causing human listeriosis. Crude bacteriocin SH01 was purified by ammonium sulfate precipitation and its inhibitory activity at two concentrations (500 and 1,000 AU/g) against Listeria monocytogenes ATCC 19111 was investigated in ground beef at increasing temperatures (5, 10, 15, and 20℃) for 8 d. The number of Listeria monocytogenes ATCC 19111 significantly decreased (p<0.05) as the concentration of bacteriocin increased from 500 to 1,000 AU/g. Intrinsic crude protease activities in ground beef were examined and increased as the temperature increased. Experiments varying both the concentrations of added bacteriocin SH01 and temperature demonstrated a maximum inhibition (2.33 log reduction of bacteria) in samples containing 1,000 AU/g of bacteriocin SH01 incubated at 20℃. When the crude bacteriocin SH01 solution (1,280 AU/mL) was incubated with crude protease solutions at different temperatures, its activity decreased by only half (640 AU/mL), as assessed in an agar well diffusion assay. The finding that the antilisterial activity of bacteriocin SH01 increased with temperature can be explained by the fact that higher temperatures increase bacterial membrane fluidity, thereby promoting the cellular penetration of bacteriocin SH01 into L. monocytogenes. Bacteriocin SH01 may be an excellent candidate as a biopreservative for controlling L. monocytogenes growth in ground beef.

• The Plant Pathology Journal
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• 제29권4호
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• pp.374-385
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• 2013

Environmental stresses induce several plant pathogenic bacteria into a viable but nonculturable (VBNC) state, but the basis for VBNC is largely uncharacterized. We investigated the physiology and morphology of the copper-induced VBNC state in the plant pathogen Ralstonia solanacearum in liquid microcosm. Supplementation of $200{\mu}M$ copper sulfate to the liquid microcosm completely suppressed bacterial colony formation on culture media; however, LIVE/DEAD BacLight bacterial viability staining showed that the bacterial cells maintained viability, and that the viable cells contain higher level of DNA. Based on electron microscopic observations, the bacterial cells in the VBNC state were unchanged in size, but heavily aggregated and surrounded by an unknown extracellular material. Cellular ribosome contents, however, were less, resulting in a reduction of the total RNA in VBNC cells. Proteome comparison and reverse transcription PCR analysis showed that the Dps protein production was up-regulated at the transcriptional level and that 2 catalases/peroxidases were present at lower level in VBNC cells. Cell aggregation and elevated levels of Dps protein are typical oxidative stress responses. $H_2O_2$ levels also increased in VBNC cells, which could result if catalase/peroxidase levels are reduced. Some of phenotypic changes in VBNC cells of R. solanacearum could be an oxidative stress response due to $H_2O_2$ accumulation. This report is the first of the distinct phenotypic changes in cells of R. solanacearum in the VBNC state.

• Molecules and Cells
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• 제23권3호
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• pp.370-378
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• 2007

A superoxide dismutase was purified 62-fold in seven steps to homogeneity from Methylobacillus sp. strain SK1, an obligate methanol-oxidizing bacterium, with a yield of 9.6%. The final specific activity was 4,831 units per milligram protein as determined by an assay based on a 50% decrease in the rate of cytochrome c reduction. The molecular weight of the native enzyme was estimated to be 44,000. Sodium dodecyl sulfate gel electrophoresis revealed two identical subunits of molecular weight 23,100. The isoelectric point of the purified enzyme was found to be 4.4. Maximum activity of the enzyme was measured at pH 8. The enzyme was stable at pH range from 6 to 8 and at high temperature. The enzyme showed an absorption peak at 280 nm with a shoulder at 292 nm. Hydrogen peroxide and sodium azide, but not sodium cyanide, was found to inhibit the purified enzyme. The enzyme activity in cell-free extracts prepared from cells grown in manganese-rich medium, however, was not inhibited by hydrogen peroxide but inhibited by sodium azide. The activity in cell extracts from cells grown in iron-rich medium was found to be highly sensitive to hydrogen peroxide and sodium azide. One mol of native enzyme was found to contain 1.1 g-atom of iron and 0.7 g-atom of manganese. The N-terminal amino acid sequence of the purified enzyme was Ala-Tyr-Thr-Leu-Pro-Pro-Leu-Asn-Tyr-Ala-Tyr. The superoxide dismutase of Methylobacillus sp. strain SK1 was found to have antigenic sites identical to those of Methylobacillus glycogenes enzyme. The enzyme, however, shared no antigenic sites with Mycobacterium sp. strain JC1, Methylovorus sp. strain SS1, Methylobacterium sp. strain SY1, and Methylosinus trichosproium enzymes.