• 미생물학회지
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• 제39권3호
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• pp.192-196
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• 2003

다양한 유산균 자원을 확보하기 위한 일환으로, 동치미에서 유산균을 분리하였다. 동치미에서 분리된 균주는, 당이용성과 16S rRNA 유전자분석을 통하여,Lactobacillus plantarum HB1으로 밝혀졌다. 이 균주는 Gram 양성균이었으며, catalase를 갖고 있지 않았다. 16S rRNA유전자의 염기배열 120 bp을 분석해 본 결과,1～88 bp 영역에서는 기존의 것과 거의 100%의 상동성을 보여 주었고,나머지 32 bp의 영역에서는 상당한 변이를 보여 주었다. 따라서 본 균주는 기존의 L. plantarum과는 다른 균주임을 확인할 수 있었다. 동치미의 무에 포함되어 있는 질산염은 체내에서 아질산염으로 바뀐다. 아질산염은 아민과 반응을 일으켜 니트로사민이 되는데, 이 성분은 위암 발생을 일으키는 주요인자중의 하나이다. 동치미에서 분리된 본 균주의 배양액에 400 ${\mu}M$의 아질산염을 첨가한 경우 1시간30분 후에 거의 소거되었다. 아질산염 소거작용에 관여하는 본 균주 배양액 중의 특정성분의 분리, 소거작용 기작 등에 관한 보다 깊이 있는 연구가 앞으로 필요하다.

• KSBB Journal
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• 제18권4호
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• pp.289-293
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• 2003

독소루비신 고 생산성 산업균주인 BR-Dox의 충분한 양의 세포를 얻어내기 위해 R2YE 배지를 사용하였으며, 방선균의 포자형성이 잘 일어나는 R2YE 배지의 각 성분별로 첨가한 결과, CaCO$_3$를 첨가하였을 때 인위적인 원형질체 유도에 적합한 양의 세포를 얻어낼 수 있었다. 독소루비신 생합성 능력이 향상된 균주를 선별하기 위해 BR-Dox를 배양하여 세포를 인위적으로 원형질체로 유도하여 세포벽을 재생시킨 결과, 특이적으로 독소루비신 색소로 추정되는 붉은 색을 많이 내는 콜로니를 선별하여 정성 및 정량분석을 수행하였다. 선별된 파생균주 BR-Dox4와 BR-Dox6의 경우 TLC 정성분석 및 HPLC 정량분석 결과, BR-Dox에 비해 각각 25.2%, 12.2%의 생산성이 향상되었다. 본 연구결과는 인위적 원형질체 도입과 세포벽 재생을 통한 새로운 개념의 방선균 균주개량 방법을 제시하고, 이를 이용한 독소루비신 고생산성 균주개발의 가능성을 제시하였다.

• 한국구조물진단유지관리공학회 논문집
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• 제20권3호
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• pp.111-121
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• 2016

본 연구에서는 철근의 인장시험에서 변형률에 대한 상세 측정을 위하여 이미지 프로세싱을 이용하고, 변형률 분포와 넥킹구간을 평가하였다. 이미지 프로세싱 방법으로는 기존의 회색조영상을 이용한 방법이 아닌 칼라영상의 색상정보를 분석하여, 원하는 타겟과 가장 일치하는 영역과 그 외의 영역으로 구분하여 이진영상으로 변환하는 방법을 사용하였다. 변환된 이진영상에서 개별 타겟들의 도심점을 산출한 후 각 도심점의 상대변위값을 변형률로 환산하였다. 인장실험은 ASTM A615 기준으로 제작된 grade 60 철근 중 D22와 D29에 대해서 시험을 수행하였다. 이미지 프로세싱을 이용하여 계측된 변형률 결과를 기존 변형률 게이지를 이용하여 계측한 결과와 비교하여, 본 연구에서 사용한 이미지 프로세싱 방법에 대해서 검증하였다. 이미지 프로세싱을 이용하여 초기 탄성구간의 변형률 뿐만 아니라 넥킹구간에서 발생한 0.5(50%) 이상의 변형률도 계측이 가능한 것을 확인하였다. 본 연구결과 이미지 프로세싱을 통하여 기존 변형률 게이지의 계측한계를 극복가능하고, 다양한 지점에서 자유롭게 계측할 수 있음을 알 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제10권4호
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• pp.496-501
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• 2000

Escherichia cole NZN111, which is known as a pfl ldhA double mutant strin, was metabolically engineered to produce succinic acid by overexpressing malic enzyme into the E. coli controlled by a trc promoter. Fermentation studies were carried out in a LB medium by first growing cells aerobically to an $OD_{600}$ of 5. At this point, 0.01 mM IPTG was added to induce the overexpression of malic enzyme and the agitation speed was gradually lowered. When the culture $OD_{600}$ reached 11, a complete anaerobic condition was achieved by flushing with a $CO_3-H_2$ gas mixture. When NZN111(pTrcML) was cultured at $37^{\circ}C$, the final succinic acid concentration of 2.8 g/l could be obtained after 30 h of anaerobic cultivation. The fermentation results were analyzed by the calculation of metabolic fluxes. Metaolic flux analysis showed that about 85% of phosphoenolpyruvate (PEP) was converted to pyruvate, and further converted to malic acid by malic enzyme.

• 한국미생물·생명공학회지
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• 제45권1호
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• pp.43-50
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• 2017

본 연구로부터 WJ-1 균주는 제주도에서 수집된 토양샘플로부터 동정되었는데, 형태분화관찰 및 16S rRNA 유전자 염기서열분석과 DNA-DNA hybridization 분석을 통하여 S. glaucescens의 신아종으로 분류되었다. 균주 WJ-1의 주요 cellular fatty acid와 게놈내 G+C 농도는 각각 $C_{15:0}$ anteiso (42.99%)와 74.73 mol%였다. 이 균은 배양액으로부터 준비된 조효소액의 xylanase 활성은 중성 pH 조건 및 $55^{\circ}C$에서 활성이 가장 높았다. S. glaucescens의 조효소액을 이용하여 xylan으로부터 xylotriose 및 xylotetraose를 포함하는 xylooligosaccharide를 제조할 수 있다. 본 연구는 S. glaucescens의 아종에 관한 최초의 보고이며, 관련 종에서 xylanase 활성에 관한 최초의 보고이다. 본 연구 결과로부터, WJ-1 균주는 lignocellulosic biomass의 이용 및 기능성 xylooligosacchade 생산에 유용하게 활용될 수 있을 것으로 기대된다.

• Journal of Microbiology and Biotechnology
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• 제16권6호
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• pp.870-879
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• 2006

The present study describes the formation of succinic acid by a nonvirulent, highly osmotolerant Klebsiella pneumoniae strain SAP (succinic acid producer), its profile of metabolites, and enzymes of the succinate production pathway. The strain produced succinate along with other metabolites such as lactate, acetate, and ethanol under aerobic as well as anaerobic growth conditions. The yield of succinate was higher in the presence of $MgCO_3$ under $N_2$ atmosphere as compared with that under $CO_2$ atmosphere. Analysis of intracellular metabolites showed the presence of a smaller PEP pool than that of pyruvate. Oxaloacetate, citrate, and $\alpha$-ketoglutarate pools were considerably larger than those of isocitrate and fumarate. In order to understand the synthesis of succinate, the enzymes involved in end-product formation were studied. Levels of phosphoenolpyruvate carboxykinase, fumarate reductase, pyruvate kinase, and acetate kinase were higher under anaerobic growth conditions. Based on the profiles of the metabolites and enzymes, it was concluded that the synthesis of succinate took place via oxaloacetate, malate, and fumarate in the strain under anaerobic growth conditions. The strain SAP showed potential for the bioconversion of fumarate to succinate under $N_2$ atmosphere in the presence of $MgCO_3$. At an initial fumarate concentration of 10 g/l, 7.1 g/l fumarate was converted to 7 g/l succinate with a molar conversion efficiency of 97.3%. The conversion efficiency and succinate yield were increased in the presence of glucose. Cells grown on fumarate contained an 18-fold higher fumarate reductase activity as compared with the activity obtained when grown on glucose.

• Environmental Engineering Research
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• 제20권1호
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• pp.79-87
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• 2015

To treat leather industry wastewater (LIW) containing high nitrogen concentration, eight aerobic denitrifiers were isolated from sludge existing in an LIW-treatment aeration tank. Among them, one strain named as KH8 had showed the great ability in denitrification under an aerobic condition, and it was identified as Pseudomonas aeruginosa R12. The aerobic denitrification ability of the strain KH8 was almost comparable to its anaerobic denitrification ability. In lab-scale aerobic denitrifications performed in 1-L five-neck flasks for 48 hr, denitrification efficiency was found to be much improved as the strain KH8 held a great majority in the seeded cells. From the nitrogen balance at the cell-combination ratio of 10:1 (the strain KH8 to the other seven isolates) within the seeded cells, the percentage of nitrogen loss during the aerobic denitrification process was estimated to be 58.4, which was presumed to be converted to $N_2$ gas. When these seeded cells with lactose were applied to plant-scale aeration tank for 56 day to treat high-strength nitrogen in LIW, the removal efficiencies of $COD_{Cr}$ and TN were achieved to be 97.0% and 89.8%, respectively. Under this treatment, the final water quality of the effluent leaving the treatment plant was good enough to meet the water-quality standards. Consequently, the isolated aerobic denitrifiers could be suitable for the additional requirement of nitrogen removal in a limited aeration-tank capacity. To the best of our knowledge, this is the first report of aerobic denitrifiers applied to plant-scale LIW treatment.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1217-1226
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• 2004

A psychrotrophic bacterial strain, Pseudomonas fluorescens BM07, synthesized unsaturated fatty acids (UFA) from fructose in response to lowering of growth temperature, and incorporated them into both polyhydroxyalkanoic acid (PHA) and membrane lipid. The blocking of PHA synthesis by adding 5 mM 2-bromooctanoic acid to the growth medium, containing 70 mM fructose, was found to be a useful means to profile the composition of membrane lipid by gas chromatography. As the growth temperature changed from 35 to $50^{\circ}C$, the total content of two UFA, 3-hydroxy-cis-5­dodecenoic acid ($C_{12:1}$) and 3-hydroxy-cis-7-tetradecenoic acid ($C_{14:1}$), in PHA increased from 31 to 44 $mol\%$. The growth at lower temperatures also led to an increase in the level of two major UFA, palmitoleic acid (C16:1 cis9) and cis-vaccenic acid (C18:1 cis11), in membrane lipid. A fraction of these membrane-lipid UFA was converted to their corresponding cyclopropane fatty acids (CFA). The CFA conversion was a function of culture time, exhibiting biphasic increase before and after entering the stationary phase. However, pH changes in growth media had no effect on the CFA conversion, which is contrary to the case of E. coli reported. The cells grown at $30^{\circ}C$ responded to a cold shock (lowering the medium temperature down to $10^{\circ}C$) by increasing the level of C16:1 cis9 and C 18: I cis II up to that of $10^{\circ}C$-grown control cells and concomitantly decreasing the relative level of cis-9,10­methylenehexadecanoic acid (the CFA converted from C16:1 cis9) from 14 to 8 $mol\%$, whereas the 10-grown cells exhibited little change in the lipid composition when exposed to a warmer environment of $30^{\circ}C$ for 12 h. Based on this one- way response, we suggest that this psychrotrophic strain responds more efficiently and sensitively to a cold shock than to a hot shock. It is also suggested that BM07 strain is a good producer of two unsaturated 3-hydroxyacids, $C_{12:1}\;and\;C_{141:1}$.

• 한국미생물·생명공학회지
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• 제25권5호
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• pp.520-526
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• 1997

Bacterial strains which degrade Arabian Light crude oil were isolated by enrichment culture from oil-spilled soil. The strain A54 was finally selected after testing emulsifying activity and oil conversion rate. Strain A54 was identified as a Acinetobacter sp. based on the morphological, biochemical and physiological characteristics. It appears to be highly specialized for growth on Arabian Light crude oil in minimal salts medium since it showed preference for oil or degradation products as substrates for growth. It was found that it could grow on at least fifteen different hydrocarbons. The optimum cultural and environmental conditions were as follows; 25$\circ$C for temperature, 7,5 for pH, 2.0% for NaCl concentration and 2.0% for crude oil concentration. Additionally, the optimal concentration of NH$_{4}$NO$_{3}$, and K$_{2}$HPO$_{4}$, were 12.5 mM and 0.057 mM, respectively. Cell growth and emulsifying activity as a function of time were also determined. Crude oil degradation and the reduction of product peaks were identified by the analysis of remnant oil by gas chromatography. Approximately 63% of crude oil were converted into a form no longer extractable by mixed organic solvents.

• KSBB Journal
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• 제26권6호
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• pp.557-560
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• 2011

It has been known that Δ-desaturase (TAD5) in the biosynthetic pathway of long chain polyunsaturated fatty acids of Thraustochytrium aureumis responsible for the conversion of di-homo-${\gamma}$-linolenic acid (C20：4) into arachidonic acid (C20：4). The genetic sequence analysis on TAD5 of Thraustochytrium aureum ATCC34304 used in this study showed that it has two amino acid changes when compared to that of Thraustochytrium aureum TAD5 first reported in 2003. Accordingly, Thraustochytrium aureum ATCC34304 TAD5 was named TAD5_1. TAD5_1-inserted methylotropic Pichia pastoris was prepared and then cultured with a precursor fatty acid, di-homo-${\gamma}$-linolenic acid. GC analysis confirmed that a certain amount of the precursor fatty acid was converted into arachidonic acid. In this study, not only a recombinant Pichia pastoris with the typical activity of ${\Delta}5$-desaturase which plays an essential role in the biosynthesis of LCPUFAs was successfully made but also the preparationpotential of a recombinant Pichia pastoris strain which may synthesize eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) that are important in maintaining and improving human's brain function was proposed.