• Journal of Microbiology
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• 제42권3호
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• pp.188-193
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• 2004

Xanthobacter flavus strain UE15 was isolated in wastewater obtained from the Ulsan industrial complex, Korea. This strain functions as a 1,2-dichloroethane (1,2-DCA) degrader, via a mechanism of hydrolytic dechlorination, under aerobic conditions. The UE15 strain was also capable of dechlorinating other chloroaliphatics such as 2-chloroacetic acid and 2-chloropropionic acid. The dhlA gene encoding 1,2-DCA dechlorinase was cloned from the genomic DNA of the UE15 strain, and its nucleotide sequence was determined to consist of 933 base pairs. The deduced amino acid sequence of the DhlA dechlorinase exhibited 100％ homology with the corresponding enzyme from X. autotrophicus GJ10, but only 27 to 29％ homology with the corresponding enzymes from Rhodococcus rhodochrous, Pseudomonas pavonaceae, and Mycobacterium sp. strain GP1, which all dechlorinate haloalkane compounds. The UE15 strain has an ORF1 (1,356 bp) downstream from the dhlA gene. The OFR1 shows 99％ amino acid sequence homology with the transposase reported from X. autotrophicus GJ10. The transposase gene was not found in the vicinity of the dhlA in the GJ10 strain, but rather beside the dhlB gene coding for haloacid dechlorinase. The dhlA and dhlB genes were confirmed to be located at separate chromosomal loci in the Xanthobacter flavus UE15 strain as well as in X. autotrophicus GJ10. The dhlA and transposase the UE15 strain were found to be parenthesized by a pair of insertion sequences, 181247, which were also found on both sides of the transposase gene in the GJ10 strain. This unique structure of the dhlA gene organization in X. flavus strain UE15 suggested that the dechlorinase gene, dhlA, is transferred with the help of the transposase gene.

• Journal of Microbiology
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• 제42권3호
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• pp.216-222
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• 2004

In a previous paper, the ogdH gene that encodes 2-oxoglutarat dehydrogenase was isolated from Salmonella typhimurium. The catalytic N-terminal region in the enzyme was found to be very specific for the Salmonella species. Therefore, the aim of the present study was to detect S. typhimurium in food sources using primers designed for OGDH-l and OGDH-2 which were based on the salmonella-specific region of the ogdH gene. A simple polymerase chain reaction (PCR) detection method was developed to detect low numbers of S. typhimurium in a chicken meat microbial consortium. Using the ogdH-specific primers under stringent amplification conditions and for gene probe analysis, fewer than 100 colony-forming units (CFUs) were detectable when pure cultures were employed. When the PCR assay was run on S. typhimurium-contaminated meat contents, only the positive meat samples containing as few as 200 CFUs reacted to the assay. The method employed for sample processing is simple and it was determined to provide a sensitive means of detecting trace amounts of S. typhimurium-specific sequences in the presence of mixed meat microbial populations. When compared with six representative intestinal gram-negative bacterial strains in foods, including Vibrio parahaemolyticus, V. vulnificus, Enterobacter cloacae, E. coli O157:H7, Pseudomonas aeruginosa, and Proteus sp., S. typhimurium had a unique and distinct PCR product (796 bp). In conclusion, the two OGDH primers were found to be rapid and sensitive detectors of Salmonella spp for the PCR method.

• Journal of Microbiology and Biotechnology
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• 제17권1호
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• pp.52-57
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• 2007

Growth promotion of wild plants by some plant growth-promoting rhizobacteria (PGPR) was examined in the microcosms composed of soils collected separately from a grass-covered site and a nongrass-covered site in a lakeside barren area at Lake Paro, Korea. After sowing the seeds of eight kinds of wild plants and inoculation of several strains of PGPR, the total bacterial number and microbial activity were measured during 5 months of study period, and the plant biomasses grown were compared at the end of the study. Acridine orange direct counts in the inoculated microcosms, $1.3-9.8{\times}10^9\;cells{\cdot}g\;soil^{-1}$ in the soil from the grass-covered area and $0.9-7.2{\times}10^9\;cells{\cdot}g\;soil^{-1}$ in the soil from the nongrass-covered site, were almost twice higher than those in the uninoculated microcosms. The number of Pseudomonas sp., well-known bacteria as PGPR, and the soil dehydrogenase activity were also higher in the inoculated soils than the uninoculated soils. The first germination of sowed seeds in the inoculated microcosm was 5 days earlier than the uninoculated microcosm. Average lengths of all plants grown during the study period were 26% and 29% longer in the inoculated microcosms starting with the grass-covered soil and the nongrass-covered soil, respectively, compared with those in the uninoculated microcosms. Dry weights of whole plants grown were 67-82% higher in the inoculated microcosms than the uninoculated microcosms. Microbial population and activity and growth promoting effect by PGPR were all higher in the soils collected from the grass-covered area than in the nongrass-covered area. The growth enhancement of wild plants seemed to occur by the activities of inoculated microorganisms, and this capability of PGPR may be utilized for rapid revegetation of some barren lands.

• Journal of Microbiology and Biotechnology
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• 제19권1호
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• pp.42-50
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• 2009

Fifty-four Pectobacterium carotovorum subsp. carotovorum strains isolated in Korea were characterized by a spectrum of antibacterial activities against 7 indicator strains chosen to represent various regions and host plants. All P. carotovorum subsp. carotovorum isolates tested could be grouped into 4 classes depending on the pattern of antibacterial substance production. All tested strains had DNA fragment(s) homologous to the genes encoding carotovoricin and 21 of them had genes homologous to DNA invertase. Sixteen strains had genes homologous to the genes encoding carocin S1. Several isolates produced antibacterial substances active against strains in Brenneria, Pantoea, and Pectobacterium genera that belonged formerly to the genus Erwinia. Strains in Pseudomonas or Xanthomonas sp. were not sensitive to the antibacterial substances produced by P. carotovorum subsp. carotovorum, except for X. albilineans that was sensitive to antibacterial substances produced by most strains in P. carotovorum subsp. carotovorum and P. betavasculorum KACC10056. These results demonstrated the diverse patterns of antibacterial substance production and the possibility of the existence of new antibacterial substance(s) produced by P. carotovorum subsp. carotovorum isolated in Korea.

• Journal of Microbiology and Biotechnology
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• 제24권1호
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• pp.97-105
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• 2014

Quorum quenching (QQ) with a microbial vessel has recently been reported as an economically feasible biofouling control platform in a membrane bioreactor (MBR) for wastewater treatment. In this study, a quorum quenching MBR with a ceramic microbial vessel (CMV) was designed to overcome the extremely low F/M ratio inside a microbial vessel. The CMV was prepared with a monolithic ceramic microporous membrane and AHL-degrading QQ bacteria, Pseudomonas sp. 1A1. The "inner flow feeding mode" was introduced, under which fresh feed was supplied to the MBR only through the center lumen in the CMV. The inner flow feeding mode facilitated nutrient transport to QQ bacteria in the CMV and thus enabled relatively long-term maintenance of cell viability. The quorum quenching effect of the CMV on controlling membrane biofouling in the MBR was more pronounced with the inner flow feeding mode, which was identified by the slower increase in the transmembrane pressure as well as by the visual observation of a biocake that formed on the used membrane surface. In the QQ MBR with the CMV, the concentrations of extracellular polymeric substances were substantially decreased in the biocake on the membrane surface compared with those in the conventional MBR. The CMV also showed its potential with effective biofouling control over long-term operation of the QQ MBR.

• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1880-1893
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• 2015

In this study, Pseudomonas R0-14, which was isolated from Arctic soil samples, showed a clear halo when grown on M9 medium agarose plates containing olive oil-rhodamine B as substrate, suggesting that it expressed putative lipase(s). A putative lipase gene, lipR, was cloned from R0-14 by genome walking and Touchdown PCR. lipR encodes a 562-amino-acid polypeptide showing a typical α/β hydrolase structure with a catalytic triad consisting of Ser₁₅₃-Asp₂₀₂-His₂₆₀ and one α-helical lid (residues 103-113). A phylogenetic analysis revealed that LipR belongs to the lipase subfamily I.3. LipR was successfully expressed in Escherichia coli, purified, and biochemically characterized. Recombinant LipR exhibited its maximum activity towards p-nitrophenyl butyrate at pH 8.5 and 60℃ with a K_m of 0.37 mM and a k_cat of 6.42 s^-1. It retained over 90% of its original activity after incubation at 50℃ for 12 h. In addition, LipR was activated by Ca²⁺, Mg²⁺, Ba²⁺, and Sr²⁺, while strongly inhibited by Cu²⁺, Zn²⁺, Mn²⁺, and ethylenediaminetetraacetic acid. Moreover, it showed a certain tolerance to organic solvents, including acetonitrile, isopropanol, acetone, methanol, and tert-butanol. When algal oil was hydrolyzed by LipR for 24 h, there was an enrichment of n-3 long-chain polyunsaturated fatty acids, including eicosapentaenoic acid (1.22%, 1.65-fold), docosapentaenoic acid (21.24%, 2.04-fold), and docosahexaenoic acid (36.98%, 1.33-fold), and even a certain amount of diacylglycerols was also produced. As a result, LipR has great prospect in industrial applications, especially in food and/or cosmetics applications.

• 한국수산과학회지
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• 제10권1호
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• pp.31-36
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• 1977

우리나라 충무연안의 해양 미생물의 월별 분포를 파악하고 이에 따라 효과적인 양식업 대책수립은 물론, 어패류로 인한 식중독 발생 예방 재료를 얻고자 1976년 4월부터 1977년 3월까지 매월 해수, 이토, 어류 및 패류등을 수집하여 조사한 결과를 요약하면 다음과 같다. 1. 해수, 이토, 어류 및 패류 732 시료에서 1,426 균주를 분리 하였다. 분리된 균종은 Pseudomonas fluorescens가 450균주, Achromobacter liquifacience 422 균주 Vibrio Parahaemolyticus가 72균주, V. alginolyticus 234균주, Proteus vulgaris가 248 균주였다. 2. V. parohaemolyticus는 732 시료중 72 균주로서 $9.84\%$ 였으며 이들중에도 해수, 이토에서 $52\%$이상을 차지하였다. 또 넓게 분포되어 있음을 알았다. 3. 양식장(뗏목)의 이토를 채취 했을 때 이토속에서 많은 Gas가 발생함을 알았고 이것은 대부분 패류의 분비물 퇴적으로 추정되었다.. (부패가 일어남) 또한 이러할 곳은 용존산소량이 적을뿐 아니라, 패류에 산소가 부족되면 신진대사가 잘 안되므로 비만도(肥滿度)가 저하한다. 이러한 해역에서 폐사된 패류를 많이 볼 수 있었다. 4. 조사지역별 분포를 보면 Station 10, 9, 8, 11, 1의 순이였다. (Fig. 1)특히 St. 10은 여객선이 많이 출입하는 충무항이며 Station 9는 주위 매축공사로 인하여 분포율이 컸고 St. 8은 분뇨처리 탱크가 가까이 있기 때문에 영향이 크다고 느껴진다. 본 연구는 1976년도 문교부 학술연구 조성비로 이루어졌다. 통영수전 한학수 교장님과 문교부에 감사를 드리며 본 실험을 도운 이정태 조교님, 재료 학명등에 협조하여 주신 김무상 교수님, 가공과 신영호, 강숙희양께 사의를 표한다.

• 한국토양비료학회지
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• 제37권2호
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• pp.116-123
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• 2004

유기재배를 위한 화학비료 대체 혼합발효 유기질비료를 개발하기 위하여 깻묵, 어박, 쌀겨, 골분 등의 유기질 자재와 일라이트, 패화석 등을 배합하여 제조하였으며 70일간의 제조과정 중 이화학적 특성 및 미생물 밀도 변화를 조사하였다. 유기질비료의 주요 성분인 깻묵과 쌀겨는 각각 7.6%와 2.6%의 질소와 3.6%와 4.6%의 인산, 1.4%와 2.2%의 칼륨을 함유하였다. 골분은 29.2%의 인산을 함유하였으며 일라이트의 칼륨 농도는 3.8%이였다. 유기질비료의 온도는 혼합 2일후 $50^{\circ}C$ 이상으로 상승하였으며 발효 40일 경과 후 상온과 비슷한 온도로 내려갔다. 발효 초기 수분 함량은 36.3%였으며 1개월 후 16.0%로 감소하였고 그 후 변화가 거의 없었다. 발효 초기에 유기물 함량에 비해 질소의 손실이 커서 탄질비가 증가하였다가 발효 10일 경과 후 서서히 감소하였다. 암모니아태 질소 함량은 혼합 시 $1,504mg\;kg^{-1}$이었으며 10일 후 최고 농도인 $5,530mg\;kg^{-1}$까지 증가하였다가 서서히 감소하였다. 그 반면에 질산태질소 함량은 발효 전 기간 동안 저농도로 유지되었는데 이러한 경향은 높은 pH와 관련이 있다고 판단되었다. 약 2개월간의 발효 후 유기질비료의 질소, 인산, 칼리 및 유기물 함량은 각각 2.7%, 2.8%, 1.8% 및 35.9%이었다. 발효과정 중 유기질비료내 생존하는 호기성세균, Bacillus sp. 및 방선균의 총균수는 각각 $12.5{\times}10^{10}$, $45.5{\times}10^5$와 $13.6{\times}10^5cfu\;g^{-1}$이었다. Pseudomoaas sp.는 발효초기에 $71.9{\times}10^7cfu\;g^{-1}$이었으나 발효 20일 후에는 온도 상승으로 급격히 감소하여 거의 나타나지 않았다. 균류 중 효모는 발효 초기에 많이 검출되었으며 사상균수은 발효 후기에 많았다.

• 한국미생물·생명공학회지
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• 제34권3호
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• pp.196-203
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• 2006

김치로부터 3종의 EPS 생성 유산균을 분리하였다. 분리균주는 Leuconostoc kimchii, Leuconostoc citreum, Leuconostoc mesenteroides로 동정되었으며, 각각 Leuconostoc kimchii GJ2, Leuconostoc citreum C3, Leuconostoc mesenteroides C11로 명명하였다. EPS생성 김치유산균의 장내 정착성 여부 확인 결과 pH 3.0의 0.05 M sodium phosphate buffer와 인공위액에서 2시간 처리한 후에도 3균주 모두 초기균수($10^8\;CFU/ml$)를 유지하였으며 인공담즙에서 24시간 처리한 후에도 3균주 모두 초기균수($10^8\;CFU/ml$)를 유지하며 3종의 EPS생성 김치유산균 Leu. kimchii GJ2, Leu. citreum C3, Leu. mesenteroides C11이 장내 정착하면서 probiotics로서 작용할 수 있을 것임을 확인하였다. 반면, 대조군으로서 분리균주가 EPS를 생성하지 않았을 때의 경우에는 내산성, 인공위액, 인공담즙에서 처리 후 생균수가 $10^{1-2}\;CFU/ml$ 감소현상을 보여 균체의 EPS생성 여부가 probiotics로서 기능할 수 있는데 결정적 요인임을 보여주었다. 특히 Leu. kimchii GJ2의 경우 E. coli, Pseudomonas, Listeria, Micrococcus, Salmonella, Staphylococcus, Streptococcus속 등의 유해 균주에서 항균활성을 나타내었으며, 생산 항균물질은 단백질성 물질로 확인되었다. 김치유산균 Leu. kimchii GJ2, Leu. citreum C3, Leu. mesenteroides C11로부터의 EPS 생성량은 sucrose(5%) 배지에서 각각 21.49 g/l, 16.46 g/l, 22.98 g/l 였으며, 정제 시에도 각각 14.61 g/l, 7.73 g/l, 4.77 g/l로 기존의 EPS 생성 유산균에서의 생산량에 비해 10배 이상의 높은 생산량을 나타내었다. TLC 및 HPLC를 이용한 EPS 구성 당 확인 결과 3균주 모두 glucose로만 구성된 homopolysaccharides로 확인되었다. EPS 생성량이 가장 좋은 Leu. kimchii GJ2의 평균 분자량은 360,606 Da이었으며, 나머지 두 균주에 대해서는 생성 EPS 형태와 점도의 차이로 미루어 보아 생성 EPS의 분자구조와 분자량이 서로 다른 것으로 판단하였다.

• 한국유기농업학회지
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• 제17권1호
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• pp.83-94
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• 2009

양송이버섯 재배 후 문제가 되고 있는 폐상퇴비는 다음 버섯 재배의 촉박성 때문에 폐상하여 재배사 주변에 방치하는 경우가 많으며, 이들은 병해충 및 수질오염의 원인이 됨으로 신속한 처리가 필요하다. 폐상된 양송이재배 부산물에는 형광성 Pseudomonas속, 방선균 등 다양한 미생물이 분포하였고, 양송이 재배농가별로 폐상퇴비 중의 미생물상 차이가 많았다. 폐상퇴비에서 분리한 세균들은 토마토 시들음병균. 풋마름병균에 대해 길항성을 보였다. 농가별 양송이 폐상퇴비의 이화학성은 pH는 $5.8{\sim}6.7$이었고, EC는 $316{\sim}528\;dS\;m^{-1}$로 농가마다 많은 차이를 보였고, T-N의 농도는 $151{\sim}1.99%$, $P_2O_5$은 $0.27{\sim}0.55%$, $K_2O$은 $1.71{\sim}3.57%$, CaO는 $1.79{\sim}5.85%$, MgO는 $0.88{\sim}1.33%$였다. 양송이 폐상퇴비의 농가실증 시험결과 시기별 토마토의 생육과 당도는 농과관행퇴비 처리구와 뚜렷한 차이가 없었고 생육시기별 토양이화학성은 농가관행에 비해 EC와 K, Ca는 증가하였지만, 인산은 감소하였다. 처리별 토양미생물상의 변화에는 차이가 없었으며, 수량은 농가관행구에 비해 약간 감소하는 경향을 보였다. 풋마름병 발병률은 무처리에 비해 폐상퇴비 처리구에서 상대적으로 낮았다.