해양암석 분말과 곰팡이 배양액에 의한 적조생물 편조류의 구제효과

현성희,신현웅,Hyun, Sung-Hee,Shin, Hyun-Woung 한국미생물학회 2006 미생물학회지 Vol.42 No.1

조류의 효과적인 제어방법 개발의 일환으로 해양암석분말의 효과를 보기위하여 해양암석분말과 곰팡이 배양 상등액을 이용하여 해양조류 제거율을 조사하였다. 편조류인 H. akashiwo와 P.minimum는 해양암석 분말을 30g/1의 농도로 살포한 실험군에서 높은 구제효과를 나타내었고, P. oxalicum (HCLE-34)배양상등액 살포는 5 mg/l 살포군에서도 구제효과를 나타내었다. 혼합 살포군에서도 배양 상등액 단독 살포군에서와 동등한 정도의 구제효과를 나타내었고, 완충용액 (pH 2.8)은 15 mg/l 살포군에서 60분 경과 후 75%의 구제효과를 나타내었으나, 전자현미경적 관찰에서는 암석분말과 배양 상등액 혼합 살포군, 배양 상등액 살포군, 암석분말 살포군 및 완충용액 살포군 순으로 세포의 파괴정도가 관찰되었다. 이러한 결과는 암석분말과 곰팡이 배양상등액이 해양에서 발생되는 편조류의 대발생을 조절할 수 있는 물질로 작용함을 시사한다. To see effect of marine rock powder and fungal culture supernatant, we analyzed the biodegradation rates of harmful marine dinoflagellate, Heterosigma akashiwo and Prorocentrum minimum for developing the effective control methodology of algal bloom. Relatively low removal rates were observed in the treatment of marine rock powder or buffer solution alone. However, the lysis of H. akashiwo and P. minimum was enhanced in the combined treatments of marine rock powder with fungal supernatant. The effective concentration and exposure time of fungal supernatant for the lysis of H. akashiwo and P. minimum were 5 ml/l and 30 minutes, respectively. These results suggest that the fungal supernatant may be a biocontrol agent for the control of algal blooms in seawater.

한국형 Streptococcus mutans의 분리 및 동정

현성희,장성렬,최영길 한국미생물학회 1989 미생물학회지 Vol.27 No.3

한국사람으로부터 치마우식증의 원인규능로 알려진 S. mutans를 분리, 동정하였고, 이들과 기존의 실험실균주간의 생리 및 생화학적 특성과 이들이 합성하는 다당류의 구성당을 비교하였다. 분리배지인 MS. MST와 선택배지인 MSP. MSPT. MSB 그리고 MSBT에서 모두 생장하고, mannitol dehydrogenase를 분비하는 균주 108, 110 및 120이 S. mutans로 분리, 동정되었다. 분리균주와 기존의 실험실균주간의 생리 및 생화학적 특성을 비교한 결과, 본 연구에서 분리한 3균주 모두 기존의 실험실균주와는 달리 hippuraterktnqnsgogyth를 가지고 있었다. 또한 균주 108은 lactose 발효 능력이, 균주 110은 sorbitol, lactose, inulin, melibiose, raffinose 발효능력이 , 균주 120은 inulin. raffinose 발효능력이 없는 것으로 나타나 기존의 실험 실균주와는 물론 이들 각각도 다른 생리 및 생화학적 특성을 나타내었다. 상기의 결과와 같이 분리균주 108, 110 그리고 120은 생리, 생화학적 특성 및 자당으로부터 합성되는 glucan의 양에 있어서도 기존의 실험실균주와 다르게 나타나 이들을 각각 한국형 S. mutans KHC108, KHCH10, KHC120이라 명명하였다. S. mutans known as a causative causative agent of dental caries was isolated from a carious lesion of Korean in the present study. The physiological, biochemical characteristics and polysaccharide pattern of these isolates were compated with those of four laboratory strains ; AHT(a), FA-1F(b), LM7(e), and OMZ65(g). One hundred strains of oral streptococci were isolated from dental caries sites of Korean (one male and one female). Among these, 3 strains were identified as S. mutans. These strains were able to grow on selective media MS, MST, MSP, MSP1, MSB, MSBT and were stained dark pink when sprayed with solutions of mannitol and TTC. So, these strains were called strain 108, 110, and 120, respectively. Strain 108, 110, and 120 were bacitracin resistnt. As these strains contained particularly hippurate hydrolysis enzyme, they were distinguished from laboratory strains. Apart from laboratory strains, the strain 108 was not capable of fermenting lactose, the strain 110 was not able to ferment sorbitol, inulin, melibiose, raffinose and the strain120 was incapable of fermenting inulin, raffinose. All fractions of extracellular and ecll bound polysaccharide of the strain108, 110, and 120 were consisted of more glucan than fructan. Aside from laboratory strains, the isolated strains were composed of more water-insoluble glucans related adherence on solid surface than water-soluble. According to these results, the strain108, 110, and 120 had native characteristecs of S. mutans, but they were different from laboratory strains in some characteristics. Therefore, each of them was given a name to S. mutans KHC108, KHC110, and KHC120, respcetively.

Penicillium oxalicum(HCLF-34)으로부터 분비되는 Anabaena cylindrica 세포벽 분해효소의 특성

현성희,최영길 한국미생물학회 1999 미생물학회지 Vol.35 No.3

Penicillium oxalicum 으로부터 세포외로 분비되는 Anabaena cylindrica 분해효소의 분자량은 renaturation SDS-PAGE에서 약 22kDa 으로 확인되었으며, 분해 효소의 농축은 ultrafiltration cut off fraction 중 30-10 kDa 구간에서 수획하였다. 최적 활성조건의 측정 결과 적정 pH는 3.5-4.0, 적정반응 온도는 $20^{\circ}C$, 그리고 온도 안정성은 $4^{\circ}C$ 이하에서 100% 이상, 20-$90^{\circ}C$ 범위에서는 50% 이상의 활성을 나타내었다. 금속이온 및 효소안정제의 영향에서는 $Na^+$,$K^+$, $Ba^(2+)$, $Mg^(2+)$, $Mn^(2+)$의 양이온과 BSA는 효소의 활성을 촉진시키는 반면, $Ca^(2+)$, $Cu^(2+)$의 양이온과 EDTA, PMSF 는 효소의 활성을 억제하는 작용을 하였다. 이러한 금속이온과 안정제의 영향에서 1가, 2가 양이온에 의해 활성이 증가하고, $Fe^(3+)$, $Ca^(2+)$, $Cu^(2+)$의 양이온에 의해서는 활성이 감소하는 결과는 대부분의 세포벽 분해효소가 갖는 특성과 유사한 결과였다. 분해효소는 A. cylindrica 과 Micrococcus. luteus 의 세포벽을 기질로 사용한 효소의 활성 반응에서 반응 시작 후 1시간에서 5시간 사이에 반응 산물로 환원당의 양이 급격히 증가하였다. The fuugus(Penicil1ium oralicum; HCLF-34) secreted the cyanobacteria lytic enzyme which had a molecular weight of about 22 kDa, a optimum temperature of $20^{\circ}C$, a optimum pH of 3.5, and a temperature-stable up to $50^{\circ}C$. The chemical ions such as sodium, potassium, barium, magnesium. and mangan ions appeared positive activity. but calcium, iron, copper ions, EDTA, and PMSF displayed negative activity: this results were the same as the characterilics of other cell wall lytic enzymes. This extracellular enzyme showed lytic aclivily against SDS-insoluble peptidoglycan of Anabaenrr cylinrlrica. The cell wall lylic enzyme of Penicilliurn oxalicum(HCLF-34) seemed to be glycosidase-like enzyme in the fact that ihe concentration of rcducing sugar was increased when the peptidoglycan of Anabaena qlinrlricn md Micrococcus luteus reacted with this enzyme

버섯배지로부터 분리한 고온성 곰팡이의 RAPD-PCR 분석

현성희,이호용,민봉희 대구대학교 기초과학연구소 2000 基礎科學硏究 Vol.17 No.2

Seven strains of thermophilic fungi were isolated from high temperature composting process for Pleurotus ostreatus. These isolates were identified the genera Trichophyton (S-1) and Sepedonium (S-2, S-3, S-5, S-6, S-7 and S-10) by morphological characters. Genetic similarity analysis of RAPD-PCR method by using the random primers showed the highest similarity of 66.98% between S-5 and S-7. The similarity between S-6 and S-10 strains was about 57.12%_ S-1 and the rest of isolates had below 10% similarity, it was revealed scarcely identity each other. The similarity between strain S-2, 3 and S-5, 6, 7, 10 was about 21%. Thus these two groups were different in morphological characters and RAPD-RCR method.

Penicillium oxalicum ( HCLF-34 ) 의 세포외 분비 효소에 의한 Anabaena cylindrica 의 분해

현성희,최영길 ( Soung Hee Hyun,Yong Keel Choi ) 한국환경생물학회 1998 환경생물 : 환경생물학회지 Vol.16 No.2

Penicillium oxalicum (HCLF-34) that showed excellent lytic characters of Anabaena cylindrica, was isolated by author. Algal lytic substance from this fungus was exchanged color of Anabaena cylindrica broth culture from dark green to pale reddish-brown color. After three hours in the mixed culture Anabaena cylindrica with lytic substance, the host was segmented, formed cluster, appeared speroblast and the sheath was lysed on the microscopy. In addition, the cell wall surface of the host was manifested the pore, lysed cell to cell junction region and gushed out cytoplasmic materials after one hour in the mixed culture on transmission electron microscopy.

Penicillium oxalicum(HCLF-34)으로부터 남조세균 (Anabaena cylindrica) 분해효소의 분리 및 동정

현성희,이호용,최영길 한국미생물학회 2000 미생물학회지 Vol.36 No.1

Penicillium oxalicum(HCLF-34)의 세포외 분비효소로부터 ultrafilration, gel filtration chromatograph와 anion exchange chromatography법을 이용하여 남조세균(Anabaena cylindrica) 분해효소를 분리하였다. 이 효소의 분자량은 약 22 kDa이며, renaturation SDS-PAGE에서 monomer로서 남조세균 분해 활성을 갖는다. 아미노산 서열은 N-말단부터$NH_(2)$-Glu-Ser-Tyr-Ser-Ser-Asn-Ala-Ala-Gly-Ala-Val-Leu-Ile---, 13개의 아미노산을 분석하였으며, 분석된 아미노산의 homology를 조사한 결과 aspergillopepsin II precursor(acid protease A)와 13개의 아미노산 중 11개(84%)의 유사도를 나타내었고, acid proteinase EapC precursor과 13개 중 10개(81%)의 유사도를 나타내었다. Algal lytic enzyme, an extracellular enzyme, was purified from the culture filtrate of Penicillium oxalicum(HCLF-34) by ultrafiltration, gel filtration chromatography, and anion exchange chromatography. The enzyme has a molecular mass of approximately 22 kDa, an it is a monomer by renaturation SDS-PAGE. The amino acid sequences of the enzyme was revealed to be NH2-Glu-Ser-Tyr-Ser-Ser-Asn-Ala-Ala-Gly-Ala-Val-Leu-Ile---, had about 84% identity with the mature light chain of aspergillopepsin II precursor and 81% identity with the mature protein of the acid proteinase EapC precursor.

해양암석 분말과 곰팡이 배양액에 의한 적조생물 Heterosigman akashiwo의 구제효과 /

현성희,민봉희,이호용 大邱大學校 基礎科學硏究所, 2003 基礎科學硏究 Vol.20 No.1

Penicillium oxalicum HCLF-34로부터 Acid Proteinase의 부분유전자 Cloning 및 Sequencing

현성희,천재순,강상순,김진규 한국미생물학회 2004 미생물학회지 Vol.40 No.1

Acid proteinase는Aspergillus niger (acid pretense A)와 Cryphonectria parasitica (acid proteinase EapC)에서 분비하는 단백질로서 치즈를 제조할 때 이용하는 단백질이다. 본 연구에서는 Penicillium oxalicum HCLF-34로부터 acid proteinase의 부분유전자를 기존에 밝혀진 acid proteinase의 homology 정보로부터 degenerate primer를 제작하여 PCR방법을 이용하여 cloning하였다. Cloning된 유전자로부터 438 bp 염기서열을 분석하였으며, 이 염기서 열을 146개의 아미노산 정보로 변환하여 acid proteinase family와 homology를 비교한 결과 acid protease A와 71% 아미노산 서열의 homology를 나타내었고, EapC와 67%의 아미노산 서열 homology가 확인되었다. Acid proteinase has been discovered in Aspergillus niger (acid protease A) and Cryphonectria parasitica (acid proteinase EapC) and it plays major roles in cheese formation from milk. In this study, a partial gene encoding acid proteinase in Penicillium oxalicum HCLF-34 was cloned by using PCR with degenerate primers corresponding to highly conserved regions of the acid proteinase. The partial acid proteinase gene in P. oxalicum HCLF-34 contains an open reading frame of 438 base pairs and encodes an acid proteinase protein of 146 amino aicds. The predicted amino acid sequences showed 71 % homology with acid protease A and 67% homology with EapC.

한국인의 혈청의 요산, 복부비만, 허리둘레 사이들의 관계

현성희,김인식,유세철,윤진석,안태호,이정화 대한임상검사과학회 2012 대한임상검사과학회지(KJCLS) Vol.44 No.2

The purpose of this study was to determine the association between serum uric acid, abdominal obesity, and waist circumference (WC) in Korean. A total of 208 healthy examinees were examined at Gimcheon Medical Center from July 2011 to December 2011. WC and body mass index (BMI) of subjects were measured as well as blood pressure, fasting blood sugar level, High Density Lipoprotein (HDL) and Low Density Lipoprotein (LDL) cholesterol, C-reactive protein (CRP), triglycerides, and serum uric acid (SUA). In both men (p<0.05) and women (p<0.01), BMI and SUA were significantly correlated with WC. In men, triglycerides (r=0.501, p<0.01) had a positive correlation with WC, and CRP (r=0.442, p<0.01) had a positive correlation with BMI. In women, triglycerides (r=0.470, p<0.01) and CRP (r=0.366, p<0.01) had a positive correlation with WC. Therefore, WC and SUA may be the most effective anthropometric indices of obesity for screening Korean population. .

시안세균 생장 억제 곰팡이의 분리 및 동정

현성희,성주연,최영길 ( Soung Hee Hyun,Ju Youn Soung,Yong Keel Choi ) 한국환경생물학회 1997 환경생물 : 환경생물학회지 Vol.15 No.2

Cyanobacteria (Anabaena cylindrica) lytic fungi were isolated from 4 reservoirs located in Kyonggi-Do, Korea. These isolates from Wangsong reservoir shown in eutrophication, were identified the genera Acremonium sp., and Penicillium sp. by morphological characters. The Penicillium sp. generally showed less algal lytic activity than the Acremonium sp. on the Anabaena cylindrica agar lawn, but the cultured supernatant of Penicillium sp. exhibited greater activity than the Acremonium sp. on Micrococcus luteus. In the mixed culture of Anabaena cylindrica and HCLF-34, the algae was segmented and lysed in 3 days. On the other hand, Anabaena cylindrica was aggregated, segmented, and lysed within 3 hours in the mixture of Anabaena cylindrica and the supernatant of HCLF-34. In addition, Anabaena cylindrica was segmented and the width sheath of Anabaena cylindrica was lysed on Scanning Electron Microscopy, in mixed culture of the algae and the fungi on agar plate.