• KSBB Journal
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• 제25권1호
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• pp.79-84
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• 2010

In the present study, we isolated a new bacterial strain producing invertase (EC 3.2.1.26) and determined optimized culture condition in flask culture. The strain was identified as Bacilus flexus determined by the 16S rDNA sequencing method. The invertase was produced only in the sucrose medium as the sole carbon source. Potassium nitrate was an adequate nitrogen source for enzyme production, whereas meat peptone showed the highest bacterial growth. Enzyme production was increased about 2-fold when $MgSO_4\cdot7H_2O$ was supplemented to the growth media. The optimum temperature was found to be $30^{\circ}C$ for both enzyme production and bacterial growth. Invertase exhibited pH optima in the range 5.0-6.0 and have a temperature optimum at $40^{\circ}C$, similarly to other invertases found from different microbial sources. Several mineral ions (K and Fe) stimulated the invertase activity, whereas some bioelements (Ag, Mg, and Mn) inhibited enzyme activity. Under the optimized culture condition, the maximum enzyme production (over 250 units/mL) was achieved at 20 h. To the best of our knowledge, this is the first time to report on invertase production by Bacilus flexus.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.430-434
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• 2004

Three extracellular protease-deficient Bacillus subtilis strains were transformed with the plasmid pCK98 containing the endo-$\beta$-1,4-glucanase (Eng) gene of B. subtilis BSE616. The three transformants, B. subtilis DB104 (pCK98), WB600 (pCK98) and WB700 (pCK98), produced the same high level of enzyme activity and showed similar patterns of cell growth and enzyme production. When B. subtilis DB 104 (pCK98), a two-extracellular protease deficient strain, was cultured for 22 h, almost all the secreted enzyme was found to be in the completely cleaved form by both activity staining and Western blotting studies. B. subtilis WB600 (pCK98), a six-extracellular protease-deficient strain, produced a partially cleaved form in addition to the intact form of the enzyme, although the degree of internal cleavage of the enzyme was greatly reduced. With B. subtilis WB700 (pCK98), a seven-extracellular protease-deficient strain, almost all the enzyme was produced as the intact uncleaved form. This study illustrates that a role of the V pr protease is to degrade foreign proteins produced in B. subtilis and WB700 is a suitable expression system for producing the intact form of the Eng and other foreign proteins that may lose at least part of their efficacy due to internal proteolytic cleavage.

• 한국미생물·생명공학회지
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• 제45권1호
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• pp.12-18
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• 2017

전국에서 수집한 발효식품으로부터 단백질 분해 효소활성을 보유한 균주를 분리하였다. Skim milk를 첨가한 TSA 평판배지에서 투명환을 형성한 8점의 균주 중 김치에서 분리된 MBE/L865 균주가 대조구인 KCTC13112 균주에 비하여 약 2.6배의 효소활성을 나타내었으며 Bacillus subtilis로 동정하여 한국미생물보존센터에 KCCM43059 균주로 기탁하였다. B. subtilis KCCM43059 균주의 최적 배양조건은 $37^{\circ}C$, pH 8으로 대조구인 KCTC13112 균주의 비성장속도보다 우수하였다. 반응조건 $60^{\circ}C$, pH 6에서 $429.37{\pm}18.65U/mg$ protein의 최적 효소활성을 나타내었으며 동일조건에서 대조구 균주보다 효소활성이 우수하였다. 또한 B. subtilis KCCM43059 균주는 대조구 균주에 비하여 NaCl에 상대적으로 높은 내성을 나타내었다.

• 한국식품영양과학회지
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• 제27권2호
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• pp.252-258
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• 1998

Biosurfactant를 생산하는 미생물을 토양으로부터 분리하였다. 그 중에서 표면장력 및 계면장력 감소능에서 가장 우수한 L-417주를 순수분리하여 동정한 결과, No-Cardia속으로 판명되었다. Biosurfactant 생산을 위한 최적 배지조성은 3% n-hexadecane, 0.1% $NaNO_3$, 0.02% $K_2HPO_4$, 0.01% $KH_2PO_4$, 0.01% $MgSO_4 \;.\;7H_2O$, 0.01% $CaCl_2$ 0.02% yeast extract였으며, 최적 온도와 pH는 각각 $30^{\circ}C$와 6.0이였다. 이러한 조건에서 500ml용 shaking flask에 최적 배지 50ml를 넣어 배양했을 경우 대수증식기 말기인 4일째에 균의 증식과 유화활성이 가장 높게 나타남에 따라 Nocardia sp. L-417에 의한 bio-surfactant의 생산은 균의 생육과 밀접한 관련이 있는 것으로 판단된다. 이계면활성제는 산업적으로 널리 사용되는 bunker A, paraffin, corn oil 및 oilve oil 등에 대해서도 비교적 높은 유화활성을 나타내였다.

• 환경생물
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• 제22권3호
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• pp.394-399
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• 2004

S. marcescens LSY4가 충전된 BOD센서를 제작하여 표준오염물질을 포함한 수용액의 BOD를 측정하였다. 배양시간이 9∼16시간일 때에는 배양시간에 따른 BOD센서의 감응도에 큰 차이를 나타내지 않았으며, 균체량도 0.22~0.75mg $cm^{-2}$의 범위에서는 거의 동일한 센서의 감응도를 나타내었다. 수용액의 pH가 4~9사이로 변화할 경우 센서의 감응도가 가역적으로 변화하였으나 수용액이 더 산성이거나 더 염기성이 되면 센서의 감응도가 비가역적으로 저하되었다. 수용액에 중금속이온이 첨가되면 센서의 감응도가 감소하였으며, $Zn^{2+}$나 $Cd^{2+}$보다 $Cu^{2+}$혹은 $Ag^+$가 첨가되었을 때 센서의 감응도가 더 급격히 감소하였다. 중금속에 대한 내성이 유도된 균체를 충전하였을 때 중금속이온의 첨가에 따른 센서의 감응도 감소가 크게 완화되었으며, 이런 효과는 $Cd^{2+}$내성주보다 $Cu^{2+}$내성주에서 더 현저하였다.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 추계학술대회
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• pp.105-105
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• 2019

Much of bacteria inhabit intestine and affect health. To elucidate the composition of intestinal bacteria and biological activities of plant materials on the bacteria, bacterial strains are need to be isolated and identified. In previous study, we isolated 41 fecal bacteria of BALB/c mice and the strains were identified as 11 species including Lactobacillus murinus and not classified bacterium. To expand the bacterial resources, we tried to isolate more bacteria from C57BL/6 mice. Fresh feces was suspended and serially diluted in distilled water. The aliquots were inoculated on GAM agar plate and incubated anaerobically at $37^{\circ}C$ for 48 h. Each of colony formed was picked up and incubated again on GAM agar plate for stock and sampling. The bacteria gained were analyzed and identified by 16S rRNA gene. The bacterial strain were listed up. Major strain was Lactobacillus murinus which was observed as an abundant strain of BALB/c mice. The resources could be used for experiments of biological activities of plant materials and microbial composition of intestinal contents of experimental animals.

• Journal of Microbiology and Biotechnology
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• 제32권3호
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• pp.378-386
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• 2022

Scenedesmus obliquus ABC-009 is a microalgal strain that accumulates large amounts of lutein, particularly when subjected to growth-limiting conditions. Here, the performance of this strain was evaluated for the simultaneous production of lutein and biofuels under three different modes of cultivation - photoautotrophic mode using BG-11 medium with air or 2% CO₂ and heterotrophic mode using YM medium. While it was found that the highest fatty acid methyl ester (FAME) level and lutein content per biomass (%) were achieved in BG-11 medium with CO₂ and air, respectively, heterotrophic cultivation resulted in much higher biomass productivity. While the cell concentrations of the cultures grown under BG-11 and CO₂ were largely similar to those grown in YM medium, the disparity in the biomass yield was largely attributed to the larger cell volume in heterotrophically cultivated cells. Post-cultivation light treatment was found to further enhance the biomass productivity in all three cases and lutein content in heterotrophic conditions. Consequently, the maximum biomass (757.14 ± 20.20 mg/l/d), FAME (92.78 ± 0.08 mg/l/d), and lutein (1.006 ± 0.23 mg/l/d) productivities were obtained under heterotrophic cultivation. Next, large-scale lutein production using microalgae was demonstrated using a 1-ton open raceway pond cultivation system and a low-cost fertilizer (Eco-Sol). The overall biomass yields were similar in both media, while slightly higher lutein content was obtained using the fertilizer owing to the higher nitrogen content.

• Journal of Microbiology and Biotechnology
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• 제32권4호
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• pp.437-446
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• 2022

In this study, to obtain icaritin with high pharmacological activities from icariin, which has a content ratio of over 58% in the total flavonoids of Epimedium herb, a special Epimedium flavonoid-glycosidase was produced, purified and characterized from Aspergillus sp.y848 strain. The optimal enzyme production was gained in a medium containing 5% (w/v) wheat bran extract and 0.7% (w/v) Epimedium leaf powder as the enzyme inducer, and strain culture at 30℃ for 6-7 days. The molecular weight of the enzyme was approximately 73.2 kDa; the optimal pH and temperature were 5.0 and 40℃. The enzyme Km and Vmax values for icariin were 15.63 mM and 55.56 mM/h. Moreover, the enzyme hydrolyzed the 7-O-glucosides of icariin into icariside II, and finally hydrolyzed 3-Orhamnoside of icariside II into icaritin. The enzyme also hydrolyzed 7-O-glucosides of epimedin B to sagittatoside B, and then further hydrolyzed terminal 3-O-xyloside of sagittatoside B to icarisiede II, before finally hydrolyzing 3-O-rhamnoside of icarisiede II into icaritin. The enzyme only hydrolyzed 7-O-glucoside of epimedin A or epimedin C into sagittatoside A or sagittatoside C. It is possible to prepare icaritin from the high-content icariin in Epimedium herb using this enzyme. When 2.5% icariin was reacted at 40℃ for 18-20 h by the low-cost crude enzyme, 5.04 g icaritin with 98% purity was obtained from 10 g icariin. Also, the icaritin molar yield was 92.5%. Our results showed icaritin was successfully produced via cost-effective and relatively simple methods from icariin by crude enzyme. Our results should be very useful for the development of medicines from Epimedium herb.

• Journal of Microbiology and Biotechnology
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• 제32권7호
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• pp.877-884
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• 2022

Probiotics are microorganisms that can benefit host health when ingested in a live state, and lactic acid bacteria are the most common type. Among fungi, Saccharomyces boulardii (SB) is the only strain known to have a probiotic function with beneficial effects on colitis; however, information on other probiotic yeast strains is limited. Therefore, this study aimed to discover yeast strains expressing intestinal anti-inflammatory activities by exhibiting probiotic properties in dextran sodium sulfate (DSS)-induced colitis mice model. Nuruk (Korean traditional fermentation starter) containing various microbial strains was used as a source for yeast strains, and S. cerevisiae 28-7 (SC28-7) strain was selected with in vitro and in vivo characteristics to enable survival in the intestines. After 14 days of pretreatment with the yeast strains, DSS was co-administered for six days to induce colitis in mice. The results revealed that the disease activity index score was lowered by SC28-7 treatment compared to the DSS group, and the colon length and weight/length ratio were recovered in a pattern similar to that of the normal group. SC28-7 administration significantly reduced the secretion of pro-inflammatory cytokines in the serum and modified the mRNA expression of inflammatory cytokines (interleukin-1β, transforming growth factor-β, and interferon-γ) and proteins involved in gut barrier functions (mucin 2, mucin 3, zonula occludens-1, and occludin) in colon tissues. These results indicate that SC28-7 attenuates DSS-induced colon damage and inflammation, supporting its future use as a probiotic yeast for treating and preventing intestinal inflammatory diseases such as inflammatory bowel disease.

• 한국유기농업학회지
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• 제30권3호
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• pp.409-422
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• 2022

본 연구에서는 흰비단병에 대한 길항 미생물과 유기농업자재를 선발하여 혼합 처리 시 방제 효과를 확인하고자 하였다. 토양에서 분리한 39균주 중 균사 생장 억제력이 가장 높은 JJ2-01 균주를 길항 미생물로 선발하였다. 또한, 유기농업자재의 흰비단병 발병도를 확인하여 병 억제 효과가 있는 유기농업자재로 마늘 오일을 선발하였다. 선발 미생물과 마늘 오일을 혼합하거나 마늘 오일을 단독으로 사용할 경우 병 억제 효과는 유사하게 나타났지만, 마늘 오일을 단독으로 사용할 경우 토양의 acid phosphatase의 활성이 감소하였다. 반면, 마늘 오일과 선발 미생물 JJ2-01 균주를 혼합 처리하였을 때 urease 활성이 증가하였다. 본 실험의 결과를 종합하면, 선발 미생물 JJ2-01 균주와 마늘 오일을 혼합하여 사용하면 고추 흰비단병을 효과적으로 억제하고 토양의 질과 건전성을 유지 또는 향상하는데 도움이 될 수 있을 것이라 판단된다.