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Production of $\beta$-Mannanase and $\beta$-Mannosidase from Sporolactobacillus sp. M201. (Sporolactobacillus sp. M201 균주에 의한 $\beta$-Mannanase와 $\beta$-Mannosidase의 생산)

  • 박원식;김화영;최용진
    • Microbiology and Biotechnology Letters
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    • v.26 no.3
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    • pp.232-237
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    • 1998
  • A bacterial strain producing high levels of an extracellular ${eta}$-mannanase and intracellular ${eta}$-mannosidase and ${alpha}$-galactosidase was isolated from soil. The strain isolated was identified as a strain of Sporolactobacillus sp. and designated as Sporolactobacillus sp. M20l. Synthesis of ${eta}$-mannanase by Sporolactobacillus sp. M20l was induced by sucrose, maltose, or locust bean gum. The highest induction rate was obtained with 2% locust bean gum added to the culture medium as a sole carbon source. On the other hand, induction of ${eta}$-mannosidase was observed only with locust bean gum. The optimal media for the enzyme production were established as follows: for ${eta}$-mannanase; 2% locust bean gum, 0.5% peptone, 0.2% KH$_2$PO$_4$, 80 mg/l MgSO$_4$, and 8 mg/l ZnSO$_4$ (pH 6.0), and for ${eta}$-mannosidase; 2% locust bean gum, 0.5% yeast extract, 0.2% KH$_2$PO$_4$, 80 mg/l MgSO$_4$, and 8 mg/l ZnSO$_4$ (pH 5.0). The optimal culture temperatures for production of ${eta}$-mannanase and ${eta}$-mannosidase were found to be 37$^{\circ}C$ and 3$0^{\circ}C$, respectively. Under the optimal culture conditions, the production of ${eta}$-mannanase and ${eta}$-mannosidase reached the highest levels of 10.6 units/ml and 1.35 units/ml after 30 h and 24 h cultivation, respectively.

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Comparative Analysis of Aniline Dioxygenase Genes from Aniline Degrading Bacteria, Burkholderia sp. HY1 and Delftia sp. HY99. (Aniline 분해균주 Burkholderia sp. HY1과 Delftia sp. HY99에서 유래된 Aniline Dioxygenases 유전자의 비교 분석)

  • Kahng, Hyung-Yeel;Oh, Kye-Heon
    • Microbiology and Biotechnology Letters
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    • v.35 no.2
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    • pp.104-111
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    • 2007
  • In this study, aniline dioxygenase genes responsible for initial catabolism of aniline in Burkholderia sp. HY1 and Delftia sp. HY99 were cloned and the amino acid sequences were comparatively analyzed, which already have been reported as bacteria utilizing aniline as a sole source of carbon and nitrogen, B. sp. HY1 was found to have at least a plasmid, and the plasmld-cured strain, B. sp. HY1-PC obtained using mitomycin C was tested with wild type strain to investigate whether the former maintained the degradability for aniline. This proved that the aniline oxygenase gene from B. sp. HY1 was located in chromosomal DNA, not in plasmid DNA. Aniline dioxygenase small subunits from B. sp. HY1 and D. sp. HY99 were found, based on 146 amino acids, to share 79% similarity. Notably, ado2 genes from B. sp. HY1 and D. sp. HY99 which were found to be terminal dioxygenase of aniline dioxygenase small subunit showed 99% similarity in the deduced amino acid sequences with tdnA2 of Frateuria sp. ANA-18 and danA2 of D. sp. AN3, respectively. Besides, enzyme assay and amino acid sequence analysis of catechol dioxygenase supported the previous report that B. sp. HY1 might occupy ortho-cleavage pathway using catechol 1,2-dioxygenase, while D. sp. HY99 might occupy catechol 2,3-dioxygenase for meta-cleavage pathway.

Analysis of Species Variety and Physiological Characteristics of Denitrifying Oligotrophic Bacteria Isolated from the Specific Environment in Korea (국내 특수 생태환경의 탈질 저영양 세균의 종 다양성 및 생리적 특성 분석)

  • Lee, Chang-Muk;Weon, Hang-Yeon;Kwon, Soon-Wo;Kang, Han-Chul;Koo, Bon-Sung;Yoon, Sang-Hong
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.210-217
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    • 2011
  • In an effort to isolate novel bacteria for the bioremediation of over-fertilized soils, we identified 135 denitrifying cells out of 3,471 oligotrophic bacteria pools (3.9%) using a denitrification medium supplemented with potassium nitrate as the sole nitrogen source. Soil samples were taken from ecologically well-conserved areas, including a mountain swamp around the demilitarized zone (Yongneup), two ecoparks (Upo and the Mujechi bog), and ten representative islands around the Korean peninsula (Jejudo, Daecheongdo, Socheongdo, Baekryeongdo, Ulrungdo, Dokdo, Geomundo, Hongdo, Huksando and Yeonpyeongdo). All of the 135 bacteria produced nitrogen gas from the denitrification medium, and were proved to be nitrate reductase positive by API-BioLog tests. Phylogenetic analysis using 16S rDNA sequences revealed that the 135 bacteria consisted of 44 different genera. Along with the most prominent, Proteobacteria (87.4%), we identified denitrifying bacteria from Firmicutes (9.4%), Actinobacteria (2.4%), and Bacteroidetes (0.8%). Physiological analyses of the 44 representative denitrifying bacteria, under various pH levels, growth temperatures and salt stresses, revealed 12 favorable denitrifying strains for soil bioremediation.

Isolation and Characterization of a Diesel-Degrading Bacterium, Gordonia sp. SD8 (디젤 분해 세균 Gordonia sp. SD8 분리 및 특성)

  • Hong, Sun-Hwa;Kim, Ji-Young;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.38 no.3
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    • pp.335-339
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    • 2010
  • A diesel-degrading bacterium, Gordonia sp. SD8, was isolated from soil contaminated with petroleum, and its diesel degradation was characterized in a soil as well as a liquid culture system. SD8 could grow in the mineral salt medium supplemented with diesel as a sole carbon and energy source. The maximum specific growth rate ($0.67{\pm}0.05\;d^{-1}$) and diesel degradation rate ($1,727{\pm}145$ mg-TPH $L^{-1}\;d^{-1}$) of SD8 showed at 20,000 mg-TPH $L^{-1}$ and $30^{\circ}C$, and then this bacterium could degrade high strength of diesel of 40,000 mg-TPH $L^{-1}$. The residual diesel concentration in the inoculated soil with SD8 was 3,724 mg-TPH kg-dry $soil^{-1}$ after 17 days, whereas the diesel concentration in the non-inoculated soil was $8,150{\pm}755$ mg-TPH kg-dry $soil^{-1}$. These results indicate that Gordonia sp. SD8 can serve as a promising microbial resource for the bioremediaion of contaminated soil with petroleum hydrocarbons including diesel.

Isolation and Characterization of Marine Bacterial Strain SH-1 Producing Agar-Degrading Enzymes (한천 분해효소를 생산하는 해양 미생물 SH-1의 분리 및 특성 분석)

  • Lee, Jae-Hag;Lee, Soon-Youl
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.324-330
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    • 2014
  • A marine bacterial strain producing agar-degrading enzymes was isolated from a mud flat in Jeboo-do (Korea) using a selective artificial sea water (ASW) agar plate containing agar as the sole carbon source. The isolate, designated as SH-1, was gram-negative, aerobic, and motile with single polar flagellum. 16S rRNA gene sequence similarity analysis showed the isolate SH-1 had the highest homology (96.5%) to marine bacterium Neiella marina J221. Cells could grow at $28-37^{\circ}C$ but not at $42^{\circ}C$, and the agarase activity of the cell culture supernatant was higher when grown at $28^{\circ}C$ than when grown at $37^{\circ}C$. Cells could grow when concentrations of 1-5% (w/v) NaCl were added to the growth media with the best growth observed at 3% NaCl, and the agardegrading enzyme activity of the cell culture supernatant was best when grown at 3% NaCl-containing growth media under the conditions we examined. The crude enzyme prepared from 48-h culture broth of strain SH-1 exhibited an optimum pH and temperature for agar-degrading activity at 7.0 and $40^{\circ}C$, respectively. Zymogram analysis of the crude supernatant and cell extract showed that strain SH-1 produced at least 3 agar-degrading enzymes with molecular weights of 15, 35, and 52 KD. Thinlayer chromatography (TLC) analysis also suggested that HS-1 produces ${\beta}$-agarase to degrade agarose to neoagarooligosaccharides.

BTEX Biodegradation in Contaminated Soil Samples Using Pure Isolates and Changes in the Mixed Microbial Community Structure (순수 분리 미생물을 이용한 오염 토양에서의 BTEX 생분해 특성과 미생물 군집 변화)

  • Chung, Kyung-Mi;Choi, Yong-Su;Hong, Seok-Won;Lee, Soo-Jin;Lee, Sang-Hyup
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.7
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    • pp.757-763
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    • 2006
  • In our previous studies, we have isolated bacteria from BTEX-contaminated sediment, which utilized BTEX as a sole carbon source and $NO_3$-N as an electron acceptor. For the possibility of field application, we have applied co-culture of those isolates in the BTEX-contaminated soil and evaluated their biodegradation efficiencies. To investigate the relationship between the isolates and indigenous microorganism in soil, changes of microbial community structure in soil samples with respect to time were monitored. To examine this, soil samples were artificially contaminated with benzene, toluene, ethylbenzene and o-xylene. BTEX-degrading bacteria such as Pseudomonas stutzeri strain 15(DQ 202712), Klebsiells sp. strain 20(DQ 202715) and Citrobacter sp. strain A(DQ 202713) were injected into the soil samples in the ratio of 2:1:1. Our results showed that the highest BTEX biodegradation efficiency was achieved when both BTEX and $NO_3-N$ existed simultaneously. The change in soil microbial community structure was characterized by PCR-DGGE analysis comparing the relative DGGE band intensities. The band intensities of indigenous microorganisms in the soil were reduced by injecting co-culture of the three isolates. On the contrary, the relative band intensities of the isolates were increased. Among the three isolates, Pseudomonas stutzeri strain 15 rendered the highest band intensity. This indicates that the Pseudomonas stutzeri was the dominant microbial species found in the soil samples.

Gibberellin Production by Newly Isolated Strain Leifsonia soli SE134 and Its Potential to Promote Plant Growth

  • Kang, Sang-Mo;Khan, Abdul Latif;You, Young-Hyun;Kim, Jong-Guk;Kamran, Muhammad;Lee, In-Jung
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.106-112
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    • 2014
  • Very few plant growth-promoting rhizobacteria (PGPR) are known to produce gibberellins (GAs). The current study aimed to isolate a phytohormone-producing PGP rhizobacterium from soil and assess its potential to enhance plant growth. The newly isolated bacterium was identified as Leifsonia soli sp. SE134 on the basis of partial 16S ribosomal RNA gene sequence. Application of L. soli culture filtrate significantly increased the biomass, hypocotyl, and root lengths of cucumber seeds as compared with non-inoculated sole medium and distilled water treated controls. Furthermore, the PGPR culture was applied to the GA-deficient mutant rice cultivar Waito-C. Treatment with L. soli SE134 significantly increased the growth of Waito-C rice seedlings as compared with controls. Upon chromatographic analysis of L. soli culture, we isolated, detected and quantified different GAs; namely, $GA_1$ ($0.61{\pm}0.15$), $GA_4$ ($1.58{\pm}0.26$), $GA_7$ ($0.54{\pm}0.18$), $GA_8$ ($0.98{\pm}0.15$), $GA_9$ ($0.45{\pm}0.17$), $GA_{12}$ ($0.64{\pm}0.21$), $GA_{19}$ ($0.18{\pm}0.09$), $GA_{20}$ ($0.78{\pm}0.15$), $GA_{24}$ ($0.38{\pm}0.09$), $GA_{34}$ ($0.35{\pm}0.10$), and $GA_{53}$ ($0.17{\pm}0.05$). Plant growth promotion in cucumber, tomato, and young radish plants further evidenced the potential of this strain as a PGP bacterium. The results suggest that GA secretion by L. soli SE134 might prove advantageous for its ameliorative role in crop growth. These findings can be extended for improving the productivity of different crops under diverse environmental conditions.

Structural and Kinetic Characteristics of 1,4-Dioxane-Degrading Bacterial Consortia Containing the Phylum TM7

  • Nam, Ji-Hyun;Ventura, Jey-R S.;Yeom, Ick Tae;Lee, Yongwoo;Jahng, Deokjin
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1951-1964
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    • 2016
  • 1,4-Dioxane-degrading bacterial consortia were enriched from forest soil (FS) and activated sludge (AS) using a defined medium containing 1,4-dioxane as the sole carbon source. These two enrichments cultures appeared to have inducible tetrahydrofuran/dioxane and propane degradation enzymes. According to qPCR results on the 16S rRNA and soluble di-iron monooxygenase genes, the relative abundances of 1,4-dioxane-degrading bacteria to total bacteria in FS and AS were 29.4% and 57.8%, respectively. For FS, the cell growth yields (Y), maximum specific degradation rate ($V_{max}$), and half-saturation concentration ($K_m$) were 0.58 mg-protein/mg-dioxane, $0.037mg-dioxane/mg-protein{\cdot}h$, and 93.9 mg/l, respectively. For AS, Y, $V_{max}$, and $K_m$ were 0.34 mg-protein/mg-dioxane, $0.078mg-dioxane/mg-protein{\cdot}h$, and 181.3 mg/l, respectively. These kinetics data of FS and AS were similar to previously reported values. Based on bacterial community analysis on 16S rRNA gene sequences of the two enrichment cultures, the FS consortium was identified to contain 38.3% of Mycobacterium and 10.6% of Afipia, similar to previously reported literature. Meanwhile, 49.5% of the AS consortium belonged to the candidate division TM7, which has never been reported to be involved in 1,4-dioxane biodegradation. However, recent studies suggested that TM7 bacteria were associated with degradation of non-biodegradable and hazardous materials. Therefore, our results showed that previously unknown 1,4-dioxane-degrading bacteria might play an important role in enriched AS. Although the metabolic capability and ecophysiological significance of the predominant TM7 bacteria in AS enrichment culture remain unclear, our data reveal hidden characteristics of the TM7 phylum and provide a perspective for studying this previously uncultured phylotype.

Activity Screening of the Proteolytic Enzymes Responsible for Post-mortem Degradation of Fish Tissues (어류의 사후 변화에 관여하는 단백질분해효소의 검색)

  • PYEUN Jae-Hyeung;LEE Dong-Soo;KIM Doo-Sang;HEU Min-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.3
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    • pp.296-308
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    • 1996
  • Proteolytic enzymes responsible for post-mortem degradation of the fish tissues have been studied in regard with screening the proteases distributed in the fish body by reacting with the specific synthesized substrates. Activities of cathepsin L, B, H, G, and D like enzymes were detected in the muscle crude protease from the both kind of fish, dark fleshed fish (anchovy, Engraulis japonica, and gizzard-shad, Clupanodo punctatus) and white fleshed fish (seabass, Lateolabrax japonicus, and sole, Pleuronichthys cornutus), however, those of chymotrypsin, trypsin, pepsin, and peptidase like enzymes were observed 3n the viscera crude pretense from the fish. Proteolytic activities of the muscle crude protease at pH 6.0 were similar to those of the viscera crude protease at pH 8.0, but, those of the viscera crude protease at pH 8.0 were about 2 times higher than those at pH 6.0. The muscle and viscera crude protease from anchovy showed the strongest proteolytic activity among the four fish crude proteases and the proteolytic activity of the viscera crude protease was approximately 100 times higher than that of the muscle crude protease, which suggest that viscera proteases were more contributed on the development of post-mortem changes than muscle proteases. With the degradation patterns on SDS-polyacrylamide gel electrophoresis against yellowtail myofibrillar proteins, the muscle and viscera crude protease of the four fishes were primary responsible for the degradation of myosin heavy chain, and myosin light chain and actin, respectively.

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Effect of Health Education Method for Korean Patients with Essential Hypertension on Their Compliance with Health Behaviors (보건교육방법이 본태성 고혈압 환자의 건강행위 이행에 미치는 효과)

  • 손경욱;유왕근
    • Korean Journal of Health Education and Promotion
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    • v.21 no.2
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    • pp.215-231
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    • 2004
  • The purpose of this study was to examine what factors affected patients who suffered from essential hypertension compliance with health behaviors, to help build a successful strategy to step up their compliance with health behaviors, and to seek effective ways to implement health education programs for patients with chronic disease. The subjects in this study were 60 people selected from among the patients who were diagnosed by physicians as having essential hypertension in S General Hospital in the city of P from April 10 through July 30, 2000, after health education was provided four times a month. The quasi- experimental design based on a control group pretest-posttest design was employed. The subjects were divided into three groups of 20 patients each: one was an experimental group to receive education in one-to-one interview, another was an experimental group to receive education as a group, and the third was a control group. The two experimental groups learned the same material through different methods, and the control group was given the same teaching materials and asked to comply with health behaviors on their own without instruction. After the three-week education was implemented in different ways, their compliance with health behaviors was measured. Collected data was analyzed by t-test, paired test, one-way analysis of variance, correlation analysis and regression analysis procedures. The findings of this study were as follows: 1. Concerning the effective type of health education, the group education produced the best results, followed by the one-to-one interviews and the sole use of print media. 2. Regarding the effect of compliance with health behaviors, the group- educated group got the highest score in compliance with health behaviors, but blood pressure lowered more significantly in the individual interview group. And the compliance with health behaviors had a significant negative correlational relationship with both systolic and diastolic blood pressure. 3. Parameter that had most significant correlational relationship with compliance with health behaviors was health locus of control, followed by self-efficacy and health perception. But there was no significant correlational relationship between compliance with health behaviors and knowledge of hypertension. 4. As a result of analyzing the impact of knowledge of hypertension, health locus of control, self-efficacy and health perception on compliance with health behaviors, self-efficacy was found to exercise most influence. Above-mentioned findings suggested that group education or one- to-one discussion would be more effective for health care for hypertension in koreans, as they could serve to have patients realize their own responsibility for health and to motivate their compliance with health behaviors, and there was a need to more positively utilize educational intervention for patients with chronic diseases, which could elevate not only compliance with health behaviors but self-efficacy.