• Title/Summary/Keyword: 25S rDNA

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Characterization and Organic Hydrocarbons Degradation Potential of Euryhaline Marine Microorganism, Bacillus sp. EBW4 Isolated from Polychaete (Perinereis aibuhitensis) (갯지렁이(Perinereis aibuhitensis)에서 분리한 광염성 해양 미생물 Bacillus sp. EBW4의 특성 및 유기물 분해능 분석)

  • Shin, Seyeon;Yundendorj, Khorloo;Lee, Sang-Suk;Lee, Dong-Heon;Kang, Kyoung-Ho;Kahng, Hyung-Yeel
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.38-45
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    • 2013
  • In this study, euryhaline marine microorganism, Bacillus sp. strain EBW4 isolated from polychaete (Perinereis aibuhitensis) of Suncheon Bay was physiologically, biochemically and genetically characterized. Based on 16S rRNA sequence, EBW14 was found to share 98.25% similarity with Bacillus hemicentroti $JSM076093^T$, 97.96% similarity with Bacillus hwajinponensis SW-$72^T$ and 96.28% similarity with B. algicoa $KMM3737^T$, respectively. The temperature range for the growth of strain EBW4 was $4-40^{\circ}C$, NaCl concentration range 0-17% and pH range pH 5-9, revealing that EBW4 was euryhaline bacterium. Major fatty acids in strain EBW4 were composed of anteiso $C_{15:0}$ (48.2%), iso $C_{16:0}$ (12.1%), anteiso $C_{17:0}$ (11.6%) and iso $C_{14:0}$ (9.4%). EBW4 was found to have DNase, amylase, protease and lipase for the degradation of macromolecules such as DNA, carbohydrates, proteins, lipids, etc. The enzyme activities of alkaline phosphatase, esterase (C4), leucine arylamidase and ${\alpha}$-chymotrypsin were also found in strain EBW4. Analysis of the biodegradation ability of EBW4 for organic hydrocarbons under different salinity conditions using synthetic water waste revealed that EBW4 exhibited the ability to degrade organic hydrocarbons very quickly, suggesting strain EBW4 may be a good candidate for the application to various industries.

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).

Manufacture of Cutting-Gouda Cheese using Lactobacillus rhamnosus_p1 and the Physicochemical Properties of Gouda Cheese during Ripening Periods (Lactobacillus rhamnosus_p1을 이용한 절단형 고다치즈 제조방법 및 숙성 중 품질특성)

  • Park, Jong-Hyuk;Jung, Hoo-Kil;Moon, Hye-Jung;Oh, Jeon-Hui;Lee, Joo-Hee;Kim, Myung-Kon;Na, Sang-Eon;Kim, Youn-Jeong;Hwang, Young-Tae
    • Journal of Dairy Science and Biotechnology
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    • v.32 no.2
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    • pp.131-139
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    • 2014
  • The aim of this study was to manufacture Cutting-Gouda cheese and to investigate the change in physicochemical properties of Cutting-Gouda cheese made with Lactobacillus rhamnosus_p1. Lactic acid bacteria were isolated from Gouda cheese ripened for more than 1 year. They were identified as 2 strains of L. rhamnosus, Lactobacillus casei, Lactobacillus curvatus, and Staphylococcus saprophyticus by 16S rDNA sequencing and named L. rhamnosus_p1, L. casei_p2, L. curvatus_p3, L. rhamnosus_p4 and S. saprophyticus_p5. The proteolytic activities of isolated strains against casein were measured using prepared skim milk agar plates. L. rhamnosus_p1 showed the highest proteolytic activity. Cutting-Gouda cheese was made with L. rhamnosus_p1, and its physicochemical properties (moisture, protein, fat, ash and free amino acid content) were measured during ripening periods. Because of the modified atmosphere packaging ($N_2{^-}$), there was no change in moisture, protein, fat, and ash in the experimental group. The total amount of free amino acids in the control and experimental group gradually increased during ripening periods. The sensory evaluation showed that the experimental group was preferable to the control group. This result suggests that L. rhamnosus_p1 has potential to be developed as a new starter for Gouda cheese.

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Clinical Findings of Phenylketonuria Patients in Korea (페닐케톤뇨증의 임상적 고찰)

  • Shin, Ik Soon;Lee, Dong Hwan
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.12 no.1
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    • pp.14-22
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    • 2012
  • Objectives: This study was performed to review clinical manifestations of hyperphenylalaninemia patients in Korean. Methods: 178 cases of hyperphenyalaninemia were diagnosed at department of pediatrics, Soonchunhyang University Hospital from January, 1983 to August. We reviewed DNA analysis, MR imaging, EEG, radiography of the left hand and wrist, bone densitometry (BMD), IQ test of hyperphenyalaninemia patients. Results: Out of 178 cases, 161 cases were diagnosed classic phenylketonuria and 17 cases were diagnosed BH4 deficiency. 122 cases performed DNA analysis. R243Q (10.3%), Y204C (9.9%), and IVS4-1G>A (8.1%) mutations were predominant. 22 cases underwent MR imaging. Varying degrees of symmetrical high signal intensity were noted on T2-weighted sequences in the periventricular deep white matter of 15 cases. 23 cases were performed EEG. 12 cases (52.3%) showed abnormal pattern. EEG abnormalities showed in 11 cases. On lumbar BMD four of 11 cases (36%) showed reduced bone density of more than 1 S.D. in four of 11 cases, bone age was less than chronological age by at least one year. 18 cases were performed IQ test. Mean IQ scores was $84{\pm}21.6$. Among older than 15 years (9 cases), Mean IQ scores was $72{\pm}21.2$. PTPS deficiency was 14 cases, DHPR deficiency was 2 cases, and GTPCH deficiency was 1 case. Conclusion: We confirmed there were varieties of DNA mutations. And MR imaging and EEG were nonspecific in PKU patients. Older children showed lower IQ score. Low phenylalanine diet prevents brain damage in PKU patient. Not only first few years of life but also lifetime, Keeping low phenylalanine diet is important.

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Quality Evaluation by the Addition of Pine Needle and Artemisia princeps Extracts in Vinegared Kochujang (솔잎, 약쑥추출물을 첨가한 초고추장의 품질평가)

  • Kim, Eun-Lyang;Kang, Sun-Chul
    • Applied Biological Chemistry
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    • v.50 no.3
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    • pp.167-177
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    • 2007
  • This study was carried out to develop natural vinegared kochujang. We first formulated vinegared kochujang containing pine needle (PN) and Artemisia princeps (AP) extracts. A dominant strain was isolated from the vinegared kochujang and identified as Bacillus amyloliquefaciens ER282 based on the 16S rDNA sequence. The plant extracts of PN and AP were also investigated for their antibacterial activity and showed enough good activities to use for vinegared kochujang as a natural preservative. Quality characteristics of vinegared kochujang were evaluated, based on total cell numbers, pH, acidity, sugar content, color and sensory evaluation during storage at $25^{\circ}C$ and $37^{\circ}C$, respectively, for 3 weeks. No significant differences were found in the pHs and acidities of vinegared kochujang added with the plant extracts of PN and AP during their storage compared to starting time; however, total cell numbers were gradually decreased during the storage at $25^{\circ}C$ and $37^{\circ}C$, as the storage period was increased. For sensory evaluation, overall preference continuously declined depending on storage period, but the addition of 3% extracts of PN could delay the quality loss of vinegared kochujang during storage at $25^{\circ}C$ and $37^{\circ}C$ for 3 weeks.

First Report of Powdery Mildew Caused by Erysiphe cruciferarum on Arabidopsis thaliana in Korea

  • Choi, Hyong-Woo;Choi, Young-Jun;Kim, Dae-Sung;Hwang, In-Sun;Choi, Du-Seok;Kim, Nak-Hyun;Lee, Dong-Hyuk;Shin, Hyeon-Dong;Nam, Jae-Sung;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.25 no.1
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    • pp.86-90
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    • 2009
  • In November 2008, typical powdery mildew symptoms were observed on leaves of Arabidopsis thaliana ecotype Col-0 plants in a growth room under controlled laboratory conditions at Korea University, Seoul. The disease was characterized by the appearance of white powder-like fungal growth on the surface of infected leaves. As the disease progressed, infected leaves exhibited chlorotic or necrotic brown lesions, and leaf distortion and senescence. Conidiophores of the causal fungus were hyaline, unbranched, 3-4 celled, cylindrical, and $80-115{\times}6-9{\mu}m$ in size. Singly produced conidia (pseudoidium type) were hyaline, oblong to cylindrical or oval in shape, and $26-55{\times}15-20{\mu}m$ in size with a length/width ratio of average 3, angular/rectangular wrinkling of outer wall and no distinct fibrosin bodies. Appressoria on the hyphae were multi-lobed. These structures are typical of the powdery mildew Oidium subgenus Pseudoidium, anamorph of the genus Erysiphe. The measurements of the fungal structures coincided with those of Erysiphe cruciferarum. The phylogenetic analysis using ITS rDNA sequences revealed that the causal fungus Erysiphe sp. KUS-F23994 is identical to E. cruciferarum. The isolated fungus incited powdery mildew symptoms on the inoculated Arabidopsis leaves, which proved Koch's postulates. Taken all data together, we first report the occurrence of powdery mildew disease of A. thaliana caused by Erysiphe cruciferarum in Korea.

Polymorphism of Bovine Lymphocyte Antigen DRB3.2 Alleles in Iranian Native Sarabi Cows

  • Pashmi, M.;Ghorashi, S.A.;Salehi, A.R.;Moini, M.;Javanmard, A.;Qanbari, S.;Yadranji-Aghdam, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.6
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    • pp.775-778
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    • 2006
  • Sarabi cows (n = 136) from the Sarabi Breeding Station were genotyped at bovine lymphocyte antigen (BoLA)-DRB3.2 locus by a genotyping system that used the polymerase chain reaction and restriction fragment length polymorphism. Genomic DNA was extracted from whole blood samples. A two-step polymerase chain reaction was carried out in order to amplify a 284 base-pair fragment of target gene. Nested-PCR products were digested with three restriction endonuclease enzymes RsaI, BstYI and HaeIII. Digested fragments were analyzed by polyacrylamide gel electrophoresis. Twenty-six BoLA-DRB3.2 alleles were identified with frequencies ranging from 0.4 to 15.1%. Six new allele types observed in this study have not been reported previously. Identified alleles include: BoLA-DRB3.$2^*1$, $^*2$, $^*4$, $^*6$, $^*8$, $^*12$, $^*13$, $^*14$, $^*15$, $^*16$, $^*17$, $^*23$, $^*24$, $^*25$, $^*28$, $^*32$, $^*34$, $^*35$, $^*36$, $^*37$, $^*42$, $^*46$, $^*51$, $^*kba$, $^*laa$ and $^*vaa$. Their frequencies were found to be 0.4, 0.4, 0.7, 11.4, 1.1, 1.8, 2.9, 2.2, 4.4, 9.6, 1.1, 13.6, 0.4, 0.4, 1.1, 0.7, 0.4, 6.2, 2.2, 3.7, 1.1, 7.7, 1.5, 15.1, 2.6 and 7.3% respectively. The six most frequent alleles (DRB3.2 $^*6$, $^*16$, $^*23$, $^*46$, $^*kba$ and $^*vaa$) accounted for 64.7% of the alleles in the population of this herd. Numerous studies on this locus, covering different breeds, has revealed the existence of various alleles in this locus, and new investigations have introduced novel alleles. With respect to the high number of the observed alleles in this survey and the novelty of some alleles with no previous record of reporting, it is plausible to conclude that the BoLA-DRB3.2 locus is highly polymorphic in Iranian native Sarabi cows.

Characterization of a heat-resistant antimicrobial peptide secreted by Bacillus subtilis A405 (Bacillus subtilis A405 균주가 생성하는 내열성 항균 peptide의 특성 검정)

  • Koo, Bon-Sung;Lee, Seung-Bum;Yoon, Sang-Hong;Song, Gae-Kyung;Chung, Dae-Sung;Byun, Myung-Ok;Ryu, Jin-Chang
    • The Korean Journal of Pesticide Science
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    • v.2 no.3
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    • pp.28-35
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    • 1998
  • An antimicrobial peptide producing bacterium, Bacillus subtilis A405, was screened and identified among 700 of antagonistic bacteria. The heat-resistant antimicrobial peptide, AMP-405, was purified from the broth culture of B. subtilis A405 through $20{\sim}40%$ ammonium sulfate precipitation and ultrafiltration. The AMP-405 exhibited strong antimicrobial activities against Botrytis cinerea, Cercospora sp., Fusarium oxysporum, Penicillium digitatum, Celletotrichum gloeosporioides, Rhizoctonia solani, Pythium ultimum, Pyricularia oryzae, Escherichia coli, Pseudomonas spp. and Candida albicans. The molecular weight of the peptide was about 3.0 kDa determined by SDS-PAGE, Native-PAGE and Tris-Tricine gradient electrophoresis, and composed of 9 kinds of amino acid such as aspartic acid, glycine, serine, glutamine, valine, leucine, isoleucine, proline, tyrocine. To determine the efficiency of AMP-405 as a potential maintenance of fruits freshness, cherry tomato was srored at $25^{\circ}C$ for 2 weeks after treatment with $50{\mu}g/ml$ of AMP-405 and $10^{5}$ spores/ml of Botrytis cinerea simultaneously. Treatment with AMP-405 resulted in significantly less infection by Botrytis cinerea, than the treatment with tap water as a control.

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Identification and Characterization of Lactobacillus salivarius subsp. salivarius from Korean Feces

  • Bae, Hyoung-Churl
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2004.05a
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    • pp.89-119
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    • 2004
  • This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

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Optimization of γ-Aminobutyric Acid Production Using Lactobacillus brevis spp. in Darae Sap (Lactobacillus brevis 균주를 이용한 다래 수액에서의 감마아미노뷰티르산 (γ-Aminobutyric Acid) 생산 최적화)

  • Jeong, Myeong-Kyo;Jeong, Ji-Hee;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.48 no.3
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    • pp.214-222
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    • 2016
  • This study was performed to increase the production of ${\gamma}$-aminobutyric acid (GABA) by lactic acid bacteria (Lactobacillus brevis CFM11) and manufacture an optimum medium using the sap from Darae (Actinidia arguta). The concentration of GABA in the fermented sap was determined using GABase enzymatic assay. The isolated L. brevis CFM11 produced $605.67{\mu}g/mL$ GABA after incubation for 24 hours at $37^{\circ}C$ in broth. The sap was fermented by L. brevis CFM11 under optimum conditions of $32^{\circ}C$ for 48 hours with 40% rice bran extract, 1.0% sucrose, 3.0% soytone, 0.2% magnesium sulfate, and 0.2% MSG. The fermented sap produced a concentration of $1366.13{\mu}g/mL$ GABA. These results demonstrate that fermenting Darae sap using L. brevis CFM11 can produce a fermented sap beverage with increased GABA content.