• Journal of Microbiology and Biotechnology
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• 제27권9호
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• pp.1593-1601
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• 2017

Vibrio species are generally recognized as pathogens predominant in seafood along coastal areas. The food industry has sought to develop efficient microbial detection methods. Owing to the limits of conventional methods, this study aimed to establish a rapid identification method for Vibrio isolated from Korea, based on matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Four different preparation procedures were compared to determine the appropriate means to pretreat Vibrio species, using 17 isolates and five reference strains. Extended direct transfer and full formic acid extraction methods using bacterial colonies on agar plates revealed very low identification rates. Formic acid and trifluoroacetic acid (TFA) extractions using bacterial broth cultures were also performed. All Vibrio isolates and reference strains prepared by TFA extraction were successfully identified to the species level (17/22, 77.3%) and to the genus level (5/22, 22.7%). Thus, TFA extraction was considered the most appropriate method to pretreat Vibrio species for MALDI-TOF MS. The remaining 33 isolates and two reference strains were prepared by TFA extraction and analyzed by MALDI-TOF MS. Overall, 50 isolates were identified to the species level (40/50, 80%) and to the genus level (10/50, 20%). All isolates were identified as 43 V. alginolyticus, six V. parahaemolyticus, and one V. vulnificus species. V. alginolyticus and V. parahaemolyticus were isolated from fish offal (87.5% and 12.5%, respectively), seawater (91.3%, 8.7%), and shellfish (62.5%, 37.5%), whereas V. alginolyticus and V. vulnificus were isolated from sediment (90.9% and 9.1%, respectively). This study established a reliable system of MALDI-TOF MS preparation and analysis for Vibrio identification.

• 한국응용과학기술학회지
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• 제32권4호
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• pp.669-684
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• 2015

본 연구는 웰빙(well-being)으로 각광받고 있는 천연물 관련 연구에 무화과 추출물의 활용 가능성을 확인하고자 다음의 연구를 수행하였다. 무화과를 세척 건조하여 알코올추출 방법으로 추출한 후, Column chromatography, G-Mass, IR, $^1H$-nmr 방법에 의하여 무화과 추출물의 이화학적 성질을 분석하였으며, 무화과 추출물을 두피 제품에 적용하기 위하여 항균, 항진균에 대한 항균성을 조사하였다. 무화과 추출물의 이화학적 분석은 열매와 잎으로 나누어 분석하였으며, 두 부분 모두 비듬과 피부병 치료 효능을 가진 항균 및 항진균 효능이 있음을 알 수 있었다. 또한 무화과 추출물에 대한 항균성은 액체배지 희석법에 의해 분석하였다. 그리고 소비자의 구매 욕구를 충족시키고, 무화과의 산업화 타당성을 검토하기 위하여 무화과 추출물 함유 수제 투명 및 고형 비누를 제조하여 타당성을 검토하였다.

• 한국미생물·생명공학회지
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• 제9권3호
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• pp.159-164
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• 1981

부분적 항상성변이주인 Bacillus megaterium (KFCC 10029)가 생산하는 페니실린 아미다제를 예로하여 강화된 $Ca^{++}$-alginate gel에 의한 포괄방법을 이용하는 효소 고정화 방법을 제시하였다. 발효액으로 부터 celite 흡착법에 의해 효소를 분리한 후 alginate의 gellatin용액에 혼합하고 $Ca^{++}$ 용액에서 응고시키고 glutaraldehyde로 처리하여 성형하였다. 이렇게 하여 얻은 고정화효소의 최적 pH 및 온도는 각각 8.0과 6$0^{\circ}C$였다. Km value 와 6-APA 및 페닐초산에 의한 저해 상수는 각각 2.6mM, 7.4mM, 21.2mM이었다. Gel의 증가된 물리적 강도 때문에 반응조 조작중 흡착효소의 유실을 성공적으로 없앨수 있었다. 관형식 반응조에서의 고정화 효소의 반감기는 4$0^{\circ}C$와 3$0^{\circ}C$에서 각각 6일 및 30일이었으며, 이것은 흡착효소와 비교해 볼 때 6-8배의 증가치이다. 결론적으로 alginate gel 포괄방법에 의한 효소고정화 방법에 있어, 본 연구에서 개발된 개량된 방법을 사용함으로써 고정화효소의 물리적 강도 및 안정도를 크게 증가시킬 수 있었다.

• 대한본초학회지
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• 제32권2호
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• pp.87-95
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• 2017

Objectives : Infectious diseases by Methicillin-Resistant Staphylococcus aureus (MRSA) are a growing problem worldwide. Characteristic of MRSA is endlessly mutation to resist antibiotics. Daehwanggo (DHG) is one of the oriental medicine prescriptions contained in Principles and Practice of Eastern Medicine. Daehwanggo was mainly used for external preparation from old times. The purpose of this study is to confirm possibility as supplementary drug of DHG about antibiotics through observation of synergy effect between DHG and commercial antibiotics and to observe restriction on growth of MRSA on any pathway through observation of mechanism. Methods : The minimum inhibitory concentration (MIC) of DHG against MRSA is $500{\sim}2000{\mu}g/m{\ell}$ by broth dilution method. In the checkerboard method, the combinations of DHG with antibiotics has partial synergistic effect or synergy effect and DHG markedly reduced the MICs of the antibiotics oxacillin (OX), gentamicin (GT) against MRSA. In the inhibition of resistance mechanism of DHG against MRSA, the expression of resistance gene and protein about ${\beta}-lactam$ antibiotic was reduced. Also, we observed the effect of DHG about cell membrane permeability against MRSA, and confirmed that DHG suppressed growth of strains by increasing cell membrane permeability. Results : Basis on the result, we speculate that DHG increase antibacterial activity of antibiotics against MRSA by changing the structure of cell wall of MRSA. Conclusions : These data suggest that Daehwanggo possesses possibility as supplementary drug about antibiotics against MRSA.

• The Plant Pathology Journal
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• 제33권3호
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• pp.337-344
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• 2017

To develop a commercial product using the mycoparasitic fungus Simplicillium lamellicola BCP, the scale-up of conidia production from a 5-l jar to a 5,000-l pilot bioreactor, optimization of the freeze-drying of the fermentation broth, and preparation of a wettable powder-type formulation were performed. Then, its disease control efficacy was evaluated against gray mold diseases of tomato and ginseng plants in field conditions. The final conidial yields of S. lamellicola BCP were $3.3{\times}10^9conidia/ml$ for a 5-l jar, $3.5{\times}10^9conidia/ml$ for a 500-l pilot vessel, and $3.1{\times}10^9conidia/ml$ for a 5,000-l pilot bioreactor. The conidial yield in the 5,000-l pilot bioreactor was comparable to that in the 5-l jar and 500-l pilot vessel. On the other hand, the highest conidial viability of 86% was obtained by the freeze-drying method using an additive combination of lactose, trehalose, soybean meal, and glycerin. Using the freeze-dried sample, a wettable powder-type formulation (active ingredient 10%; BCP-WP10) was prepared. A conidial viability of more than 50% was maintained in BCP-WP10 until 22 weeks for storage at $40^{\circ}C$. BCP-WP10 effectively suppressed the development of gray mold disease on tomato with control efficacies of 64.7% and 82.6% at 500- and 250-fold dilutions, respectively. It also reduced the incidence of gray mold on ginseng by 65.6% and 81.3% at 500- and 250-fold dilutions, respectively. The results indicated that the new microbial fungicide BCP-WP10 can be used widely to control gray mold diseases of various crops including tomato and ginseng.

• 한국버섯학회지
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• 제11권2호
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• pp.53-62
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• 2013

A total of 35 phosphate solubilizing bacterial strains were isolated from waste mushroom bed of Agaricus bisporus in Buyeo-Gun, Chungnam and screened for the production of indole acetic acid (IAA). The best IAA producing strain was identified as Pantoea rodasii using 16S rRNA analysis. In addition to the IAA production, this strain could act as an efficient phosphate solubilizer (1100 ${\mu}g$ $ml^{-1}$ after 5 days of incubation) also. The selected strain was cultured under different conditions in order to assess the optimum conditions for maximum IAA production. The nutrient broth (NB) medium was recorded as the best medium, where the maximum IAA production (229 ${\mu}g$ $ml^{-1}$) was recorded at the start of stationary phase (12 hours after inoculation) of the bacteria growth. The performance of the strain was found to be maximum at the temperature of $30^{\circ}C$ followed by $25^{\circ}C$. IAA production was found to be increased with increasing tryptophan concentration (from 0.1 to 0.6%), however beyond this limit, a slight reduction in IAA production was observed. The strains' ability to produce IAA was further confirmed by extraction of crude IAA and subsequent TLC analysis. A specific spot from the extracted IAA preparation was found corresponding with the standard spot of IAA with same $R_f$ value. The results of HPLC analysis conducted in identifying and quantifying the IAA production more precisely, are in agreement with the results of the assessment done with colorimetric method. As revealed by the results of the pot experiment, the isolated strain could significantly enhance the growth (as measured by shoot and root growth) of mung bean plants compared to that of non-inoculated plants. Therefore it can be concluded that the present strain, Pantoea rodasii has great potential to be used as bio-inoculants.

• Applied Biological Chemistry
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• 제47권4호
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• pp.379-383
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• 2004

Bacillus sp. 유래 ${\beta}-mannanase$의 brown copra meal에 대한 기질 특이성을 규명하기 위하여 정제효소 150 ml를 0.5% 기질에 $50^{\circ}C$, 24 hrs 가수분해후 TLC, FACE로 비교 검토한 결과 2종류의 galactosyl mannooligosaccharide로 구성되어 1차 activated carbon column chromatography을 이용해 250 ml/hr 유속으로 tube당 50ml씩 ethanol $0{\sim}30%$ linear gradient로 당을 분리하였다. Activated carbon column chromatography에 의한 당용액 0.2 ml와 5% phenol 0.2 ml를 가하여 혼합 후 $Conc.H_2SO_4$ 1 ml를 가하여 혼합한 후 20분간 방치하여 490 nm로 흡광도를 측정하여 TLC로 pattern을 검토한 후 $Gal^3Man_4$ 및 DP 7의 galactosyl manooligosaccharide의 fraction을 회수하여 2차 activated carbon column chromatography를 이용해 1차와 동일한 조전에서 분리 회수하여 중합도 5는 $Gal^3Man_4(63-mono-{\alpha}-D-galactopyranosyl-{\beta}-mannotetraose)$로 동정되었고, 중합도 7은 현재 동정중에 있다. Bifidobacterium속 균주(B. longum, B. bifidum)에 대한 생육활성을 비교 검토하기 위하여 MRS media에서 탄소원을 dextrose대신에 조제된 $Gal^3Man_4$를 첨가후 측정한 결과 $Gal^3Man_4$이 첨가되지 않은 MRS broth에 비해 생육촉진 활성을 보였다. 특히 B. longum에 대해서는 $Gal^3Man_4$를 dextrose대체 탄소원으로 처리시 10배의 생육활성이 증가하였다.