• 한국식품영양과학회지
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• 제43권8호
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• pp.1289-1295
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• 2014

본 연구에서는 전통발효식품 13종과 천연물 4종에서 27균주를 분리한 후 생육속도가 빠르고 pH 저하 능력이 우수한 3종의 균주를 1차 선발하였으며, 분자생물학적 방법을 이용하여 동정한 결과 Staphylococcus warneri, Sta. epidermidis, Lactobacillus plantarum과 99% 상동성을 보였다. 선발된 3균주의 발효소시지 스타터로서 이용가능성을 알아보기 위해 발효소시지의 환경과 유사한 model-system에서 배양하며 pH 저하 능력, 총산 생성 능력, 생육 능력, 아질산염 소거 능력을 발효소시지의 스타터로 많이 사용되고 있는 5균주 및 상업용 2균주와 비교하였다. Modelsystem에서 pH 저하능과 총산 생성능, 생육능은 관련성이 있었으며, 상업용 균주보다 분리한 Sta. epidermidis DO 10-1, Lac. plantarum MLK 14-2가 우수한 결과를 나타내었다. 아질산염 소거능의 경우에도 분리한 3균주가 상업용 균주보다 상대적으로 빠른 속도를 보였다. 분리한 3균주는 발효소지지 스타터로서의 발효 소거 능력은 우수할 것으로 보이지만 Staphylococcus는 잠재적인 위험성이 제기되는 균주이므로 Lac. plantarum이 발효소지지 제조에 가장 적합할 것으로 사료된다.

• The Plant Pathology Journal
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• 제34권1호
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• pp.1-10
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• 2018

A previously unreported bacterial disease on chili pepper (Capsicum annuum L.) seedlings affecting as many as 4% of seedlings was observed in greenhouses in Chihuahua, Mexico (Delicias and Meoqui counties). Initial lesions appeared as irregular small spots on leaves and brown necrosis at margins tips were observed. Later, the spots became necrotic with a chlorotic halo. Advanced disease was associated with defoliation. A Gram negative, rod-shaped bacterium was isolated from diseased chili pepper seedlings. Three inoculation methods revealed that isolated strains produce foliage symptoms, similar to those observed in naturally infected seedlings. Pathogenic strains that caused symptoms in inoculated seedlings were re-isolated and identified to fulfill koch's postulate. Polyphasic approaches for identification including biochemical assays (API 20E and 50CH), carbon source utilization profiling (Biolog) and 16S rDNA, hsp60 and rpoB sequence analysis were done. Enterobacter cloacae was identified as the causal agent of this outbreak on chili pepper seedlings.

• 한국미생물·생명공학회지
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• 제48권3호
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• pp.358-365
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• 2020

An organic solvent- and bleach-stable protease-producing strain was isolated from a polluted river water sample and identified as Aeromonas veronii OB3 on the basis of biochemical properties (API 20E) and 16S rRNA sequence analysis. The strain was found to hyper-produce alkaline protease when cultivated on fish waste powder-based medium (HVSP, 4080 U/ml). The biochemical properties and compatibility of OB3 with several detergents and additives were studied. Maximum activity was observed at pH 9.0 and 60℃. The crude protease displayed outstanding stability to the investigated surfactants and oxidants, such as Tween 80, Triton X-100, and H₂O₂, and almost 36% residual activity when incubated with 1% SDS. Remarkably, the enzyme demonstrated considerable compatibility with commercial detergents, retaining more than 100% of its activity with Ariel and Tide (1 h, 40℃). Moreover, washing performance of Tide significantly improved by the supplementation of small amounts of OB3 crude protease. These properties suggest the potential use of this alkaline protease as a bio-additive in the detergent industry and other biotechnological processes such as peptide synthesis.

• KSBB Journal
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• 제26권1호
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• pp.62-68
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• 2011

To isolate Lactic acid bacteria for animals, we have screened from Kim-chi, swine intestine, swine feces, and dairy products by random selection and anti-viral, antipathogenic bacteria test. Among them, CLP-1 shown that inhibitory effect against rotavirus, porcine epidemic diarrhea (PED) virus, Salmonella sp, and E.coli. By examining biological property, API-ZYM and identified Lactobacillus plantarum by 16S rDNAgene sequence. CLP-1 determined resistance to low pH and bile salt. Futhermore, the cell body of CLP-1 adhered to the intestinal epithelium tissue of swine and Caco-2 cell. CLP-1 was examined on cell immune system modulating activity in vitro. The whole cell and cell culture supernatant was increasing of interferon-${\beta}$ activity. And then, CLP-1 increased prevention effect by Salmonella enteritidis infection in SPF chickens. And we determined similar result in pigs.

• Journal of Species Research
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• 제9권3호
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• pp.198-203
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• 2020

The goal of this study was to isolate and identify wild yeasts from soil samples. The 15 wild yeast strains were isolated from the soil samples collected in Pocheon city, Gyeonggi Province, Korea. Among them, four yeast stains were unrecorded, and 11 yeast stains were previously recorded in Korea. To identify wild yeasts, microbiological characteristics were observed by API 20C AUX kit. Pairwise sequence comparisons of the D1/D2 domain of the 26S rRNA were performed using Basic Local Alignment Search Tool(BLAST). Cell morphology of yeast strains was examined by phase contrast microscope. All strains were oval-shaped and polar budding and positive for assimilation of glucose, 2-keto-ᴅ-gluconate, N-acetyl-ᴅ-glucosamine, ᴅ-maltose and ᴅ-saccharose (sucrose). There is no official report that describes these four yeast species: one strain of the genus Kodamaea in the family Metschnikowiaceae and three strains of the Hannaella in the family Bulleribasidiaceae. Kodamaea ohmeri YI7, Hannaella kunmingensis YP355, Hannaella luteola YP230 and Hannaella oryzae YP366 were recorded in Korea, for the first time.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1985년도 워크샵 및 심포지엄 북한산국립공원의 식생
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• pp.73-81
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• 1985

We have previously isolated OsMADS4 gene that is a member of the class B MADS box genes from rice. In this study, another member of the class B MADS box genes was isolated from rice flower by the yeast two-hybrid screening method using OsMADS4 as bait. RNA blot analyses revealed that the clone, OsMADS16, was expressed in the second and third whorls, whereas the OsMADS4 transcripts were present in the second, third, and fourth whorls. These expression patterns of the OsMADS16 and OsMADS4 genes are very similar with those of AP3 and PI, the class B genes of Arabidopsis, respectively. In the yeast two-hybrid system, OsMADS4 interacted only with OsMADS16 among several rice MADS genes investigated, suggesting that OsMADS4 and OsMADS16 function as a heterodimer in specifying sepal and petal identities. We have also isolated OsMADS6 gene using OsMADS1 as a probe. Both are members of the AGL2 MADS family. Various MADS genes that encode for protein-protein interaction partners of the OsMADS6 protein were isolated by the yeast two-hybrid screening method. A majority of these genes belong to the AGL2 family. Sequence Homology, expression pattern, and ectopic expression phenotypes indicated that one of the interaction partners, OsMADS14, appears to be homologous to API, the class A MADS gene of Arabidopsis.

• 한국균학회지
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• 제49권4호
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• pp.545-553
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• 2021

In 2021, seven yeast strains were isolated from earthworm (Eisenia andrei) gut samples collected from the Nanji Water Regeneration Center in Goyang City, Gyeonggi Province, Korea. A total of seven yeasts were isolated, of which three strains have not been previously reported in Korea. To identify the yeasts, pairwise sequence comparisons of large subunit (LSU) rDNA sequences were performed using the basic local alignment search tool (BLAST). Assimilation test and cell morphology analysis were performed using the API 20C AUX kit and phase contrast microscope, respectively. Five of the seven strains were assigned to the genus Candida of the order Saccharomycetales of the class Saccharomycetes, and two to the genus Apiotrichum of the order Trichosporonales of the class Tremellomycetes. The yeast strain Candida sojae E2 belongs to the family Debaryomycetaceae, and Apiotrichum laibachii E8 and A. laibachii E9 belong to the family Trichosporonaceae. All strains were cultured in yeast mold agar for three days and showed different colony forms. C. sojae E2 was round and entire shaped, while A. laibachii E8 and A. laibachii E9 was round and convex shaped. This study focuses on the description of the three yeast strains that have not been officially reported in Korea.

• Journal of Species Research
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• 제10권4호
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• pp.350-355
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• 2021

In 2020, 11 Basidiomycetous yeast strains were isolated from soil samples collected from the forests of Namhansanseong in Korea. Among them, seven species were reported, but four species were unreported in Korea. To identify wild yeasts, pairwise sequence comparisons of D1/D2 domain of the 26S rRNA were performed using Basic Local Alignment Search Tool (BLAST). The cell morphologies and assimilation test are observed by phase contrast microscope and API 20C AUX kit, respectively. The 11 strains were assigned to the genera Rhodotorula (4 strains) of the order Sporidiobolales of the class Microbotryomycetes; and Cryptococcus(2 strains), Goffeauzyma (1 strains), Naganishia (2 strains) of the order Filobasidiales and Saitozyma (2 strains) of the order Tremellales of the class Tremellomycetes in the phylum Basidiomycota. The unreported yeast strains Cryptococcus gastricus 20n5-2, Goffeauzyma gilvescens 20n2-7, Naganishia adeliensis 20n8-1, and Naganishia friedmannii 20n24-1 belong to the family Filobasidiaceae. All strains had oval shaped cells and cream-colored colonies cultured on on YM agar for 3 days. In this study, we focus on the description of four unreported yeast species in Korea.

• 대한소아치과학회지
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• 제32권2호
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• pp.344-356
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• 2005

우리 인체에는 의학적으로 유용한 기능을 가진 세균들이 정상적으로 존재하며 구강내에도 독특한 기능을 나타내는 세균들이 상재한다. 본 연구에서는 치아우식증이 없는 소아의 타액에서 분리한 유산균 2주가 Streptococcus mutans에 의한 인공치태 형성과 혐기성 세균에 의한 휘발성 유황화합물 생성을 억제하는 것을 확인하고, API 50 CHL medium kit를 이용한 생화학적 검사와 16S rDNA sequencing으로 동정하여 다음과 같은 결과를 얻었다. 1. 분리균주는 2주 모두 그람양성 간균으로 과산화수소를 생성하였다. 2. 인공치태의 무게는 Streptococcus mutans의 단독 배양시 $124.4{\pm}30.4mg$이었으나, 분리균주와 병합 배양시에는 각각 $5.2{\pm}2.0mg,\;10.6{\pm}6.6mg$으로 현저하게 감소하였다(p<0.05). 3. Streptococcus mutans의 생균수는 단독 배양시 ml당 $3.4{\times}10^9$이었으나, 분리균주와 병합 배양시에는 각각 ml당 $4.6{\times}10^8$과 $2.4{\times}10^8$으로 감소하였다. 4. Fusobacterium nucleatum을 30분간 진탕한 후 측정한 상청액의 흡광도는 1.286이었으나, Fusobacterium nucleatum과 분리균주를 병합으로 30분간 진탕한 후 측정한 상청액의 흡광도는 각각 0.628과 0.497로 감소하였으며, 상호 결합 정도는 29.4%와 57.8%이었다. 5. Fusobacterium nucleatum의 단독 배양시 cysteine과 $FeSO_4$를 첨가한 배지를 가한 후 측정한 침전물의 배지 흡광도는 1.794이었으나, 분리균주와 병합 배양시 측정한 침전물의 배지 흡광도는 각각 1.144와 0.915로 감소하였으며, Porphyromonas gingivalis 단독 배양시 침전물의 배지 흡광도는 1.932이었으나 분리균주와 병합 배양시에는 침전물의 배지 흡광도가 각각 1.170과 1.266으로 감소하였다. 6. 분리균주를 API 50 CHL medium kit로 탄수화물 발효검사를 시행한 결과, 분리균주 1주는 Lactobacillus salivarius로, 다른 분리균주는 Lactobacillus delbrueckii subsp. lactis로 동정되었다. 7 분리균주를 16S rDNA partial sequencing으로 동정한 결과, 2주 모두 Lactobacillus salivarius subsp. salicinius와 유전자 유사치가 99.60%, 99.73%를 보여 Lactobacillus salivarius subsp. salicinius로 동정되었다. 이상의 결과를 종합하면 치아우식증이 없는 소아의 타액에서 분리된 유산균 중 과산화수소를 분비하여 인공치태 형성과 휘발성 유황화합물 생성을 억제하는 분리균주는 Lactobacillus salivarius subsp. salicinius로 동정되었다.

• 생명과학회지
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• 제22권7호
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• pp.912-919
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• 2012

Carboxymethylcellulose (CM-cellulose)와 Beechwood xylan을 각각 기질로 사용하여 trypan blue를 첨가하여 제작한 Agar-LB 배지 상에서 명확한 활성환을 형성하는 균주를 cellulase와 xylanase 생산 균주로 단리하였다. 단리한 균주 유래의 16S rRNA 유전자 및 API 50 kit를 분석한 결과 Bacillus subtilis와 약 99.5%의 높은 상동성을 보였기에 본 균주를 Bacillus subtilis로 동정하여 B. subtilis NC1로 명명하였다. B. subtilis NC1 유래 cellulase와 xylanase는 CM-cellulose와 Beechwood xylan에 대하여 각각 높은 효소 활성을 보였으며, 두 효소 모두 pH 5.0과 $50^{\circ}C$의 조건하에서 가장 높은 효소 활성을 보였다. B. subtilis NC1 균주 유래 cellulase와 xylanase 유전자를 cloning하기 위하여 shot-gun cloning 방법을 이용하여 B. subtilis NC1 염색체 DNA로부터 효소 유전자를 cloning하여 유전자 배열을 규명한 결과 cellulase 유전자는 아미노산 499개를 암호화하는 1,500 bp의 open reading frame (ORF)으로 이루어져 있었으며, 아미노산 배열로부터 추정되는 분자량은 55,251 Da 이었다. 그리고, xylanase에 대한 유전자는 아미노산 422개를 암호화하는 1,269 bp의 ORF로 이루어져 있었으며 유전자 유래 아미노산 배열로부터 추정되는 단백질 분자량은 47,423 Da 이었다. 두 효소의 아미노산 배열을 이용하여 상동성을 검토한 결과 cellulase는 glycoside hydrolase family (GH) 5에 속하는 cellulase와 xylanase는 GH30에 속하는 xylanase와 높은 상동성을 나타내었다.