• Title/Summary/Keyword: strain SK1

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Purification, Characterization, and Cloning of Trimethylamine Dehydrogenase from Methylophaga sp. Strain SK1

  • Kim, Hee-Gon;Kim, Yan;Lim, Heon-Man;Shin, Hyun-Jae;Kim, Si-Wouk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.4
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    • pp.337-343
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    • 2006
  • Trimethylamine dehydrogenase (TMADH, EC 1.5.99.7), an iron-sulfur flavoprotein that catalyzes the oxidative demethylation of trimethylamine to form dimethylamine and formaldehyde, was purified from Methylophaga sp. strain SK1. The active TMADH was purified 12.3-fold through three purification steps. The optimal pH and temperature for enzyme activity was determined to be 8.5 and $55^{\circ}C$, respectively. The $V_{max}\;and\;K_m$ values were 7.9 nmol/min/mg protein and 1.5 mM. A genomic DNA of 2,983 bp from Methylophaga sp. strain SK1 was cloned, and DNA sequencing revealed the open reading frame (ORF) of the gene coding for TMADH. The ORF contained 728 amino acids with extensive identity (82%) to that of Methylophilus methylotrophus $W_3A_1$.

Degradation Properties of n-Alkane Assimilating Pseudomonas putida 3SK Carrying $CAM::TOL^{*}$ Plasmid and NAH Plasmid

  • Chun, Hyo-Kon;Cho, Kyung-Yun;Kho, Yung-Hee
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.270-273
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    • 1994
  • Pseudomonas putida 3SK, which was constructed by the conjugal transfet of the $CAM::TOL^{*}$ plasmid of Pseudomonas putida CSnA and the NAH plasmid of Pseudomonas putida KCTC 2403 into n-alkane assimilating Pseudomonas putida KCTC 2405, showed a broad degradation spectrum and floc-forming ability. This strain degraded m-toluic acid, naphthalene, camphor and decane simultaneously. $Hg^{2+}$ at the concentration of 1 ppm in the minimal medium could not inhibit the growth of this strain. The degradation of m-toluic acid by Pseudomonas putida 3SK was not repressed by the easily utilizable compounds, such as glucose and succinate. But, the addition of formalin inhibited the growth of Pseudomonas putida 3SK. After the cultivation of this strain on the artificial wastewater containing m-toluic acid, naphthalene, camphor and decane for 24 hr, the initial COD value (1500) of the artificial wastewater was declined to 300.

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InAs 양자점 크기에 따른 광학적 특성 평가

  • Han, Im-Sik;Park, Dong-U;No, Sam-Gyu;Kim, Jong-Su;Kim, Jin-Su
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.187-187
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    • 2013
  • 양자점(Quuantum dot, QD)은 0차원 특성을 가지는 구조로 양자 구속 효과로 인하여 bulk와 는 다른 구조적, 광학적, 전기적 특성을 가지고 있다. InAs QD는 size와 barrier의 bandgap 조절을 이용하여 쉽게 bandgap을 바꿀 수 있는 장점이 있어 solar cell, semiconductor laser diode, infrared photodetector 등으로 많은 연구가 이루어지고 있다. 일반적으로 Stranski-Krastanov (SK) mode로 성장한 InAs QD는 보통 GaAs epilayer와의 lattice mismatch (7%)를 이용하여 성장을 하고 이로 인하여 strain을 가지고 있고 QD의 density와 stack이 높을수록 strain이 커진다. 하지만 sub-monolayer (SML) QD 같은 경우 wetting layer가 생기는 지점인 1.7 ML이하에서 성장되는 성장 방식으로 SK-QD보다는 작은 strain을 가지게 된다. 또 QD의 size가 작아 SK-QD보다 큰 bandgap을 가지고 있다. 본 연구에서는 분자선 에피택시(molecular beam epitaxy, MBE)를 이용하여 semi-insulating GaAs substrate 위에 InAs QD를 0.5/1/1.5/1.7/2/2.5 monolayer로 성장을 하였다. GaAs과 InAs의 성장온도와 성장속도는 각각 $590^{\circ}C$, 0.8 ML/s와 $480^{\circ}C$, 0.2 ML/s로 성장을 하였으며 적층사이의 interruption 시간은 10초로 고정하였고 10주기를 성장하였다. Photoluminescence (PL)측정 결과 SML-QD는 size에 따라서 energy가 1.328에서 1.314 eV로 약간 red shift를 하였고 SK-QD의 경우 1.2 eV의 energy정도로 0.1 eV이상 red shift 하였다. 이는 QD size에 의하여 energy shift가 있다고 사료된다. 또 wetting layer의 경우 1.41 eV의 energy를 가지는 것으로 확인 하였다. SML-QD는 SK-QD 보다 반치폭(full width at half maximum, FWHM)이 작은 것은 확인을 하였고 strain field의 감소로 해석된다. 하지만 SML-QD의 경우 SK-QD보다 상대적으로 작은 PL intensity를 가지고 있었다. 이를 개선하기 위해서는 보다 높은 QD density를 요구하게 되는데 growth temperature, V/III ratio, growth rate 등을 변화주어서 연구할 계획이다.

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Characterization of Methylophaga sp. strain SK1 Cytochrome $c_L$ Expressed in Escherichia coli

  • Kim, Hee-Gon;Phan, Trongnhat;Jang, Tae-Sa;Koh, Moon-Joo;Kim, Si-Wouk
    • Journal of Microbiology
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    • v.43 no.6
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    • pp.499-502
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    • 2005
  • Methylophaga sp. strain SK1 is a new restricted facultative methanol-oxidizing bacterium that was isolated from seawater. The aim of this study was to characterize the electron carriers involved in the methanol oxidation process in Methylophaga sp. strain SK1. The gene encoding cytochrome $c_L$ (mxaG) was cloned and the recombinant gene was expressed in Escherichia coli $DH5\alpha$ under strict anaerobic conditions. The recombinant cytochrome $c_L$ had the same molecular weight and absorption spectra as the wild-type cytochrome $c_L$ both in the reduced and oxidized forms. The electron flow rate from methanol dehydrogenase (MDH) to the recombinant cytochrome $c_L$ was similar to that from MDH to the wild-type cytochrome $c_L$. These results suggest that recombinant cytochrome $c_L$ acts as a physiological primary electron acceptor for MDH.

Biological Treatment of Piggery Liquid Manure by Malodor Reducing Bacteria (악취 저감용 세균에 의한 돈분뇨의 생물학적 처리)

  • Quan, Xiao-Tian;Shin, Jae-Hyeong;Wang, Yan-Qing;Choi, Min-Gyung;Kim, Sang-Min;Kim, Soo-Ki
    • Journal of Life Science
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    • v.32 no.12
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    • pp.971-978
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    • 2022
  • Sulfur-oxidizing, ammonium-oxidizing, and nitrogen-oxidizing media were used to isolate bacteria to degrade malodor gas effectively in piggery manure or soil. Twelve different strains were isolated: Paenibacillus amylolyticus, Rhodococcus jostii, Rhodococcus qingshengii, Rhodococcus opacus, Alcaligenes faecalis, Alcaligenes faecalis, Kastia adipate, Kastia adipata, Microbacterium oxydans, Halomonas campisalis, Acinetobacter oleivorans, and Micrococcus luteus. By inoculating each strain in the piggery liquid manure by 1%, the pH in most strain treatments was maintained at 8.0. Total bacterial counts were maintained at 7.3~7.9 log CFU/ml until 15 days, and then they dropped dramatically down to 5.1~5.5 log CFU/ml. On the 30th day, the treatment group inoculated with Rhodococcus opacus SK2659 showed a relatively high level of ammonium nitrogen removal, which was 39% of that of the control group. When Rhodococcus opacus SK2659 was inoculated, H2S concentration after 100 days was 3.23% compared with the control (no inoculation), suggesting that Rhodococcus opacus SK2659 is an excellent strain for removing malodor gas. The gas production of the treatments was lower than that of the control. The total accumulated amount of gas production in most strain treatments was a quarter of the gas production compared to the control throughout the experimental periods. Acinetobacter oleivorans SK2675 showed the lowest level at 12.39% compared to the control in gas production. In conclusion, the use of mixture strains, such as Rhodococcus opacus SK2659 and Acinetobacter oleivorans SK2675 isolated in this study could increase the efficacy of malodor gas reduction in the biological treatment of piggery manure.

Development of an ${\alpha}-amylase-hyperproducing$ mutant of Bacillus licheniformis and its characteristics (${\alpha}-Amylase$ 고생산성 Bacillus licheniformis 변이주의 개발과 특성 분석)

  • Jeong, Heo-Jin;Jung, Kyung-Hwa;Chang, Jong-Soo;Yoon, Ki-Hong;Park, Seung-Hwan;Kim, Hoon
    • Applied Biological Chemistry
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    • v.41 no.1
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    • pp.18-22
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    • 1998
  • A mutant strain which hyperproduced thermostable ${\alpha}-amylase$ was obtained by chemical mutagenesis of Bacillus licheniformis. The mutant strain, SK-5, produced the enzyme about 50 times higher than the original strain. The mutant was longer and slimmer in shape, slower in growth compared to the original strain. Nucleotide sequence analysis of the SK-5 ${\alpha}-amylase$ gene revealed no changes in the the structural gene. The changes found in the promoter region might be responsible for the hyperproduction of the enzyme by the mutant. No structural changes in the enzyme structure could be observed when the secreted enzymes at various culture times were analyzed by Western blot.

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Production and fermentation characteristics of seafood kimchi started with Leuconostoc mesenteriodes SK-1 isolated from octopus baechu kimchi (문어 배추김치에서 분리한 Leuconostoc mesenteroides SK-1을 이용한 수산물김치의 제조 및 발효 특성)

  • Jang, Mi-Soon;Jung, Ko-Eun;Yun, Jae-Ung;Nam, Ki-Ho
    • Food Science and Preservation
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    • v.23 no.7
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    • pp.1050-1057
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    • 2016
  • This study was carried out to investigate the effectiveness of Leuconostoc mesenteroides isolated from octopus baechu kimchi as a potential starter for seafood kimchi. L. mesenteroides is lactic acid bacterium currently used as a starter for kimchi production. We selected the most effective L. mesenteroides strain from the 7 strains isolated from octopus baechu kimchi and, based on biochemical properties and 16S rRNA sequencing, identified the selected strain as L. mesenteroides SK-1. The SK-1 strain exhibited acid-tolerance, good survival capacity, and excellent dextran productivity. We investigated the effects the SK-1 of starter on seafood kimchi fermentation. Octopus baechu kimchi was fermented with L. mesenteroides SK-1 at $4^{\circ}C$ for 35 d. The decrease in pH and increase in acidity in octopus baechu kimchi fermented with the SK-1 starter occurred more quickly than that in the control kimchi indicating that. Octopus baechu kimchi with SK-1 starter has a relatively slow rate of increase in lactic acid production. As a result, octopus baechu kimchi prepared with L. mesenteroides SK-1 can be maintained at a suitable ripening degree over an extended period of time compared to that of the control kimchi, Moreover, the octopus baechu kimchi started with L. mesenteroides SK-1 has excellent sensory properties, including a refreshing taste, and a weak sour odor.

Characterization of an Iron- and Manganese-containing Superoxide Dismutase from Methylobacillus Sp. Strain SK1 DSM 8269

  • Seo, Sung Nam;Lee, Jae Ho;Kim, Young Min
    • Molecules and Cells
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    • v.23 no.3
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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.

Purification and Some Properties of Soluble Cytochrome c from Methylobacillus sp. Strain SK1 (Methylobacillus sp. Strain SK1에 존재하는 Soluble cytochrome c의 정제 및 특성)

  • 김시욱;노영태;김영민
    • Korean Journal of Microbiology
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    • v.29 no.6
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    • pp.380-386
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    • 1991
  • Three types of soluble cytochrome c were purified to homogeneity from Methylobacillus sp. strain SK1 which grows only on methanol. Cytochrome c-I was purified 58.5-fold in seven steps. Cytochrome c-II and c-III were purified 57.3- and 122.1-fold in eight steps, respectively. The molecular weights of the cytochrome c-I was determined to be 12,500, while those of the cytochrome c-II and c-III were 16,000. The isoelectric points of the c-I, c-II and c-III were found to be 8.8, 6.6, and 6.6 respectively. The spectrum of reduced cytochrome c-I showed .alpha.-, .betha.-, .gamma.-peaks at 551.4, 522.2, and 416.6nm. The peaks for c-II were found at 551.0, 521.6, and 416.5nm, while those for c-III were shown at 551.2, 521.8, and 416.0 nm. The spectra of oxidized cytochrome c-I, c-II, and c-III showed .gamma.-peak at 411.8, 409.0, and 410.2 nm, respectively. The absorption coefficients of .alpha.- and .gamma.-peak for c-I in the reduced state were determined as 47 and 197 $mM^{-1}$ $cm^{-1}$ , respectively. The coefficients of .alpha.- and .gamma.-peak for c-II were determined to be 43 and 137 $mM^{-1}$ $cm^{-1}$ , while those for c-III were 41 and 172 $mM^{-1}$ $cm^{-1}$ , respectively. The c-I and c-III were found to bind carbon monoxide.

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Phase Diagrams and Stable Structures of Stranski-Krastanov Structure Mode for III-V Ternary Quantum Dots

  • Nakaima, Kazuno;Ujihara, Toru;Miyashita, Satoru;Sazaki, Gen
    • Proceedings of the Korea Association of Crystal Growth Conference
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    • 1999.06a
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    • pp.81-114
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    • 1999
  • The strain, surface and inerfacial energies of III-V ternary systems were calculated for three kinds of structure modes: the Frank-van der Merwe (FM) mode, the Stranski-Krastanov (SK) mode an the Volmer-Wever (VW) mode. The free energy for each mode was estimated as functions of the thickness and composition or lattice misfit. Through comparison of the free energy of each mode, it was found that the thickness-composition phase diagrams of III-V ternary systems can be determined only by considering the balance of the free energy and three kinds of structure modes appear in the phase diagrams. The SK mode appears only when the lattice misfit is large and/or the lattice layer is thick. The most stable structure of the SK mode is a cluster with four lattice layers or minimum thickness on a wetting layer of increasing lattice layers. The VW mode appears when the lattice misfit is large and the lattice layer is thin and only in the InPSb/InP and GaPSb/GaP systems which have the largest lattice misfit of III-V ternary systems. The stable region of the SK mode in the GaPSb/GaP and InPSb/InP phase diagrams is largest of all because the composition dependence of the strain energy of these systems is stronger than that of the other systems. The critical number of lattice layers below which tow-dimensional (2D) layers precede the three-dimensional (3D) nucleation in the SK mode at x=1.0 depnds on the lattice misfit.

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