• Title/Summary/Keyword: ITS rDNA

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Studies on Proteolytic and Fibrinolytic Activity of Bacillus subtilis JM-3 Isolated from Anchovy Sauce (멸치액젓으로부터 분리한 Bacillus subtilis JM-3의 단백질 분해활성과 혈전 용해 활성에 관한 연구)

  • Lee, Sang-Soo;Kim, Sang-Moo;Park, Uk-Yeon;Kim, Hee-Yun;Shin, Il-Shik
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.283-289
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    • 2002
  • This study was performed to search for potential microorganism that has rapid fermenting and physiological function from anchovy sauce. We isolated three bacterial strains, JM-1, JM-2, and JM-3 with proteolytic and fibrinolytic activity from anchovy sauce. Among the 3 bacterial strains, JM-3 showed the strongest proteolytic and fibrinolytic activity. Bacterial strain JM-3 was gram-positive rod, motile and formed endospore. The 16S rRNA of bacterial strain JM-3 was amplified by PCR and then its sequence was determined by ABI 310 genetic analyzer. The 16S rRNA sequence of bacterial strain JM-3 was compared to BLAST DNA database and identified to Bacillus subtilis with 99% of homology. The optimum temperature, pH and NaCl concentration for growth of B. subtilis JM-3 were $40^{\circ}C$, 5.0 and 0%, respectively. The optimum temperature, pH and NaCl concentration for proteolytic and fibrinolytic enzyme production of B. subtilis JM-3 were same as optimum conditions for growth. At 20% of NaCl concentration which is common NaCl concentration of fish sauce, B. subtilis JM-3 showed about 60% of proteolytic and fibrinolytic activity of 0% NaCl concentration. From above results, we found that B. subtilis JM-3 will be able to used for starter of functional fish sauce.

Antifungal activity against cheese fungi by lactic acid bacteria isolated from kimchi (김치 분리 유산균의 치즈 곰팡이 항진균 활성)

  • Choi, Ha Nuel;Oh, Hyun Hee;Yang, Hee Sun;Huh, Chang Ki;Bae, In Hyu;Lee, Jai Sung;Jeong, Yong Seob;Jeong, Eun Jeong;Jung, Hoo Kil
    • Food Science and Preservation
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    • v.20 no.5
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    • pp.727-734
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    • 2013
  • The antifungal activity against cheese fungi by lactic acid bacteria isolated from kimchi was investigated. Eight fungi were isolated from cheese in the cheese ripening room. Two of them were identified as Penicillium and Cladosporium via ITS-5.8S rDNA analysis. Twenty-two species of lactic acid bacteria with antifungal activity were isolated from kimchi. Two of them were identified as Lactobacillus and Pediococcus via 16S rRNA sequence analysis. Of the 22 lactic acid bacteria species, six were selected (L. sakei subsp. ALJ011, L. sakei subsp. ALI033, L. sakei subsp. ALGy039, P. pentosaceus ALJ015, P. pentosaceus ALJ024 and P. pentosaceus ALJ026) due to their higher activity against the eight fungi isolated from cheese in the cheese ripening room; and among the six species, the P. pentosaceus ALJ015 and P. pentosaceus ALJ024 isolates from the Jeonju area kimchi and the L. sakei subsp. ALI033 isolate from the Iimsil area kimchi had higher antifungal activity than the other lactic acid bacteria. The minimum inhibitory concentration (MIC) of L. sakei subsp. ALI033 against the eight fungi isolated from cheese in the cheese ripening room was $62.5{\mu}g/mL$.

Synthesis and Secretion of Mutant Mannose-Binding Lectin (돌연변이 Mannose-binding Lectin 합성과 세포 병리적 연구)

  • Jang, Ho-Jung;Chung, Kyung Tae
    • Journal of Life Science
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    • v.23 no.3
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    • pp.347-354
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    • 2013
  • Innate immunity is the ability to differentiate infectious agents from self. The innate immune system is comprised of a complicated network of recognition and effector molecules that act together to protect the host in the early stage of an infectious challenge. Mannose-binding lectin (MBL or mannose-binding protein, MBP) belongs to the family of $Ca^{2+}$-dependent lectins (C-type lectin with a collagen-like domain), which are considered an important component of innate immunity. While it is associated with increased risk and severity of infections and autoimmunity, the most frequent immuno-deficiency syndrome was reported to be low MBL level in blood. Deficiency of human MBL is caused by mutations in the coding region of the MBL gene. Rat homologue gene of human MBL gene was used to study functions of wild type and mutant MBL proteins. Although extensive studies have yielded the structural information of MBL, the functions of MBL, especially mutant MBL, still require investigation. We previously reported the cloning of rat wild-type MBL gene and the production of a truncated form of MBL protein and its antibody. Here, we present the cloning of mutant MBL cDNA in collagen-like domain (R40C, G42D, and G45E) using site-directed mutagenesis and differential behaviors of wild type and mutant MBL in cells. The major difference between wild type and mutant MBL was that while wild type MBL was secreted, mutant MBL was inhibited for secretion, retained in endoplasmic reticulum, and still functioned as a lectin.

Characterization of Starch-Utilizing Yeast Saccharomycopsis fibuligera Isolated from Nuruk (누룩으로부터 분리된 전분대사 효모 Saccharomycopsis fibuligera 균주의 생육특성)

  • Choi, Da-Hye;Park, Eun-Hee;Kim, Myoung-Dong
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.407-412
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    • 2014
  • A number of Saccharomycopsis fibuligera strains that can hydrolyse and utilize starch as a carbon source were isolated from nuruk, a traditional Korean starter for rice wine fermentation, and their specific growth rates on starch-containing medium were compared to choose the prominent strain. S. fibuligera strain MBY1320 showed a higher growth rate at $42^{\circ}C$ than that of strain S. fibuligera KCTC7806, indicating that S. fibuligera MBY1320 has more thermo-tolerant machinery for starch hydrolysis and utilization than KCTC7806. Although the activity of ${\alpha}$-amylase at $30^{\circ}C$ was significantly lower for S. fibuligera MBY1320 than KCTC7806 (3,812.5 U vs. 14,878.5 U), S. fibuligera MBY1320 showed a much higher glucoamylase activity at $42^{\circ}C$ than S. fibuligera KCTC7806 (5,048.9 U vs. 13,152.3 U). Thus, a new S. fibuligera strain, with a higher starch-hydrolysing activity at elevated temperatures than that of other types of strain, this study reports.

The Characteristics of Tetrachloroethylene (PCE) Degradation by Pseudomonas putida BJ10 (Pseudomonas putida BJ10의 Tetrachloroethylene (PCE) 분해 특성)

  • Choi, Myung-Hoon;Kim, Jai-Soo;Lee, Sang-Seob
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.311-316
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    • 2008
  • In this study, biological PCE degradation by using a BTEX degrading bacterium, named BJ10, under aerobic conditions in the presence of toluene was examined. According to morphological, physiological characteristics, 16S rDNA sequencing and fatty acid analysis, BJ10 was classified as Pseudomonas putida. As a result of biological PCE degradation at low PCE concentrations (5 mg/L), PCE removal efficiency by P. putida BJ10 was 52.8% for 10 days, and PCE removal rate was 5.9 nmol/hr (toluene concentration 50 mg/L, initial cell density 1.0 g (wet weight)/L, temperature 30, pH 7 and DO $3.0{\sim}4.2\;mg/L$. At high PCE concentration (100 mg/L), PCE removal efficiency by P. putida BJ10 was 20.3% for 10 days, and PCE removal rate was 46.0 nmol/hr under the same conditions. The effects of various toluene concentration (5, 25, 50, 100, 200 mg/L) on PCE degradation were examined under the same incubation conditions. The highest PCE removal efficiency of PCE was 57.0% in the initial PCE concentration of 10 mg/L in the presence of 200 mg/L toluene for 10 days. Furthermore, the additional injection of 5.5 mg/L PCE (total 7.6 mg/L) made 63.0% degradation for 8 days in the presence of 50 mg/L toluene under the same conditions. Its removal rate was 13.5 nmol/hr, which was better than the initial removal rate (8.1 nmol/hr).

Purification of Methioninase from Pseudomonas putida and Its Effect on the Uptake of ^11C-Methionine in Vivo. (Pseudomonas putida 유래 Methioninase의 정제 및 생체내 ^11C-Methionine 섭취에 미치는 영향)

  • 변상성;박귀근
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.377-382
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    • 2003
  • Purification of methioninase resulted in a yield of 69%, and SDS-PAGE analysis of the purified product revealed a single band of approximately 43 kDa in molecular weight. in vitro experiments with cancer cells incubated in methionine-free media demonstrated an increase in $^{11}$ C-methionine uptake to 25.8$\pm$1.1% at 6 hr, 31.8$\pm$0.8% at 24 hr, and 62.2$\pm$0.6% at 48hr, compared to controls. Treatment of the cancer cells with purified methioninase showed no decrease in survival after a 2 hr incubation with 0.01 U/ml, but survival of RR1022 cells decreased 30% after 24 to 48 hr incubation. SKOV-3 cells showed a 5% and 14% decrease in survival with 0.1 and 1 U/ml methioninase after 24 hr. After 48hr survival decreased 15% and 24% with 0.1 and 1 U/ml methioninase. Measurements of $^{11}$ C-methionine uptake in RR1022 cells demonstrated no change at 2 hr, but a 13.7$\pm$4.7% and 40.7$\pm$2.6% increase in uptake at 24 and 48 hr, respectively. SKOV-3 cells also showed no change at 2 hr, but had a 17.7$\pm$7.2% and 38.9$\pm$4.9% increase in $^{11}$ C-methionine uptake after 24 hr and 48 hr treatment with methioninase, respectively. $^{11}$ C-methionine PET imaging revealed clear visualization of both the tumors and contralateral infectious lesions. Administration of rMET appeared to result in a slight increase in tumor:nontumor contrast on $^{11}$ C-methionine PET images. Injection of purified methioninase also produced PET images where tumor uptake was higher than that of infectious lesions.

Isolation and Characterization of a Marine Bacterium Producing Thermotolerant Agarase (내열성 한천분해효소를 생산하는 해양세균의 분리 및 특성)

  • Park Ceun-Tae;Lee Dong-Ceun;Kim Nam Young;Lee Eo-Jin;Jung Jong-Ceun;Lee Jae-Hwa;Heo Moon-Soo;Lee Jung-Hyun;Kim Sang-Jin;Lee Sang-Hyeon
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.884-888
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    • 2005
  • An agar-degrading bacterium was isolated from north-eastern sea of Jeju island and cultured in marine agar 2216 media. Biochemical and morphologicl characteristics and 165 rRNA gene revealed that isolated strain was member of Agarivorans genus, and named Agarivorans sp. JA-1. Agarase was produced as growth-related and expressed regardless of agar presence. Optimal pH was 8 at 50 mM Clycine-NaOH buffer, and activity was maximum at $40^{\circ}C$E Enzymatic activity was maintained over $80\%$ at $60^{\circ}C$t and $70\%$ at $80^{\circ}C$ which is thermotolerant. Hence isolated novel Agarivorans sp. JA-1 strain and its beta-agarase could be used for the production of functional oligosaccharide from agar in solution state.

Characterization of Cellulase from Bacillus subtilis NSC Isolated from Soil (토양으로부터 단리한 Bacillus subtilis NSC 유래 Cellulase의 특성 규명)

  • Kim, Sang Jin;Park, Chang-Su
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.228-233
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    • 2018
  • We isolated microorganisms from soil, which is sampled at forest, Kyeonbuk, Korea, as cellulolytic microorganisms. The isolated strains were identified by analysis of 16S rRNA gene from the starins. The result, four kinds of Bacillus subtilis, one kind of Bacillus amyloliquefaciens, and one kind of Bacillus cereus were identified. Among these strains, Bacillus subtilis was selected due to its high cellulase activity and this strain was named as Bacillus subtilis CNS. The optimum pH and temperature of the cellulase from Bacillus subtilis CNS was pH 5.0 and $40^{\circ}C$, respectively. In the investigation of pH and temperature stability, the cellulase from Bacillus subtilis NSC stabled pH 4.0~6.0 range and until $40^{\circ}C$ for 30 min perfectly. In the enzyme activity for various cellulosic substrate, cellulase from Bacillus subtilis CNS showed the highest activity for CM-cellulose. And, the enzyme activities for alkali swollen cellulose, Alpha-cellulose, Sigmacell-cellulose, and Avicel were approximately 31%, 8%, 8% and 4% of activity for CM-cellulose, respectively. In the degradation of CM-cellulose, the 0.26 U/ml and 0.52 U/ml of cellulase showed 0.43 and 0.76 U/ml activity for CM-cellulose after the reaction of 120 min, respectively.

Lactobacillus plantarum (KACC 92189) as a Potential Probiotic Starter Culture for Quality Improvement of Fermented Sausages

  • Ba, Hoa Van;Seo, Hyun-Woo;Seong, Pil-Nam;Kang, Sun-Moon;Kim, Yoon-Seok;Cho, Soo-Hyun;Park, Beom-Young;Ham, Jun-Sang;Kim, Jin-Hyoung
    • Food Science of Animal Resources
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    • v.38 no.1
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    • pp.189-202
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    • 2018
  • This study was conducted to evaluate the effects of fermenting temperature on the applicability of Lactobacillus plantarum for production of fermented sausages as starter cultures, and its applicable efficiency was also compared with those inoculated with commercial starter culture or non-inoculated control. The L. plantarum isolated from a naturally-fermented meat, identified by 16S rDNA sequencing and again identified by de novo Assembly Analysis method was used as a starter culture. Six treatments: 3 with L. plantarum at different fermenting temperatures (20, 25 and $30^{\circ}C$), and other 3 treatments (1 with commercial starter culture, 1 with its mixture with L. plantarum and 1 non-inoculated control) fermented under the same conditions ($25^{\circ}C$) were prepared. Results revealed that the fermenting temperature considerably affected the pH change in samples added with L. plantarum; the highest pH drop rate (1.57 unit) was obtained on the samples fermented at $30^{\circ}C$, followed by those at $25^{\circ}C$ (1.3 unit) and $20^{\circ}C$ (0.99 unit) after 4 days fermentation. Increasing the temperature up to $30^{\circ}C$ resulted in significantly lower spoilage bacteria count (5.15 log CFU/g) and lipid oxidation level in the products inoculated with L. plantarum. The sensory analysis also showed that the samples added with L. plantarum at $30^{\circ}C$ had significantly higher odor, taste and acceptability scores than those fermented at lower temperatures. Under the same processing condition, although the L. plantarum showed slightly lower acidification than the commercial starter culture, however, it significantly improved the eating quality of the product.

Occurrence of Brown Rot on Apricot Caused by Monilinia fructicola in Korea (Monilinia fructicola 에 의한 살구 잿빛무늬병)

  • Choi, In-Young;Kim, Ju;Seo, Kyoung-Won;Oh, Hun-Tak;Cho, Chong-Hyeon;Kim, Jin-Ho;Song, Young-Ju
    • Research in Plant Disease
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    • v.22 no.2
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    • pp.122-126
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    • 2016
  • In June 2015, an exhibited typical signs and symptoms of brown rot was observed on fruit of Apricot cvs. Modern and Alexander at an incidence of 5% of fruit in Jeonju, Korea. Early symptoms on fruit showed small, circular, light brown spots that eventually destroyed the entire fruit. Small sporodochia appeared on the fruit surface. Fruit susceptibility to brown rot increases during the 1 to 2 weeks period prior to harvest. The conidia were one-celled, hyaline, lemon-shaped, $14.6-18.0{\times}8.5-11{\mu}m$, and borne in branched monilioid chains. Based on the morphological characteristics and phylogenetic analysis of internal transcribed spacer (ITS), the fungus was identified as Monilinia fructicola. A BLAST search revealed that sequences of the fungus shared 100% identity to those of M. fructicola. Pathogenicity of a representative isolate was proved by artificial inoculation, fulfilling Koch's postulates. To our knowledge, this is the first confirmed report on the occurrence of M. fructicola on apricot in Korea.