• Title/Summary/Keyword: 25S rDNA

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Screening and Characterization of Oleaginous Microalgal Species from Northern Xinjiang

  • Wu, Lei;Xu, Liangliang;Hu, Chunxiang
    • Journal of Microbiology and Biotechnology
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    • v.25 no.6
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    • pp.910-917
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    • 2015
  • A total of 646 strains, including green algae and diatoms, were isolated from 220 samples to screen microalgae with high lipid productivity (LP). The samples were obtained from nine habitats in Northern Xinjiang, China in June 2013. This study initially identified eight lipidrich strains, namely, Desmodesmus intermedius XJ-498, D. intermedius XJ-145, D. intermedius XJ-99, Monoraphidium pusillum XJ-489, M. dybowskii XJ-435, M. dybowskii XJ-151, Mychonastes homosphaera XJ-488, and Podohedriella falcata XJ-176, based on 18S rDNA sequencing. The strains were cultured in a photobioreactor for the same period. Results showed that the specific growth rate (day-1) of M. pusillum XJ-489 was the highest (1.14 ± 0.06), and the biomass concentration (g/l) of D. intermedius XJ-99 was the highest (2.84 ± 0.3). Futhermore, the lipid content (%) of M. dybowskii XJ-151 was the highest (33.5 ± 4.38), and the lipid productivity (mg l-1 day-1) of My. homosphaera XJ-488 was the highest (86.41 ± 9.04). C16 to C18 accounted for 86% to 98% of the total lipid, and the biodiesel qualities of the selected algae corresponded to international standards. This study suggests that My. homosphaera XJ-488, D. intermedius XJ-99, and M. dybowskii XJ-151 are the most potential strains for biodiesel production among all the isolated strains.

Characterization of Diesel Oil-Degrading Bacteria (디젤유 분해균주의 특성 및 토양배양)

  • 안민정;한윤전;임현섭;최기현;권오범;정병철
    • Korean Journal of Microbiology
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    • v.39 no.2
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    • pp.108-113
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    • 2003
  • Diesel oil-degrading bacterial strains were isolated from diesel oil contaminated soil and called HS series (HS1, HS2 and HS3). These strains were identified as Acinetobacter sp. (HS1) and Pseudomonas sp. (HS2 and HS3) based on Biolog test, cellular fatty acid composition, and 16S rDNA sequence analysis. These strains were coltivated in liquid minimal media containing 2% diesel oil, and diesel oil-degrading activity was measured. As result, all strains degraded over 70% of total diesel oil. But PAH (polycyclic aromatic hydrocarbon)- and pris- tane-degrading rate of these strain was below 20% of total PAH and pristane. The HS 1 strain showed highest hydrophobicity and low emulsifying activity among the experimental strains and high diesel oil-degrading activity. From the above-mentioned result, microcosm experiment was performed with the HS1 strain. The HS1 strain showed a degrading activity of over 80% of total diesel oil in microcosm test. And microbial activity was correlated to diesel oil-degrading activity. Therefore, it is suggested that the HS1 strains could be effectively used for the bioremediation for diesel oil.

Characterization and Methanol Biosynthesis of a Methane-Oxidizing Bacterium, Methylomonas sp. SM4, Isolated from Rice Paddy Field Soil (논에서 분리한 메탄산화세균 Methylomonas sp. SM4의 특성과 메탄올 생합성)

  • Park, Sung Min;Madhavaraj, Lavanya;Kim, Si Wouk
    • KSBB Journal
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    • v.32 no.2
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    • pp.124-132
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    • 2017
  • A methane-oxidizing bacterium was isolated from rice paddy field soil around Jeollanam-do province, Korea, and characterized. The isolate was gram-negative, orange pigmented and short rod ($1.1-1.2{\times}1.6-1.9{\mu}m$). It was catalase and urease-negative but oxidase-positive. The strain utilized methane and methanol as sole carbon and energy sources. It had an ability to grow with an optimum pH 7.0 and an optimum growth temperature $30^{\circ}C$. The strain was resistant to antibiotic polymyxin B but sensitive to streptomycin, kanamycin, ampicillin, chloramphenicol and rifampicin. The isolate required copper for their growth with concentration range of $2-25{\mu}M$, with an optimum of $10{\mu}M$. Under optimal culture condition, specific cell growth rate and generation time were found to be $0.046hr^{-1}$ and 15.13 hr, respectively. Phylogenetic analysis based on 16S rDNA sequences indicated that the strain formed a tight phylogenetic lineage with Methylomonas koyamae with a value of 99.4% gene sequence homology. So, we named the isolate as Methylomonas sp. SM4. 8.6 mM methanol was accumulated in the reaction mixture containing 70 mM sodium formate and 40 mM $MgCl_2$ (MDH inhibitor) under atmosphere of methane:air (40:60) mixture for 24 hr at $30^{\circ}C$.

Morphological and Molecular Characteristics of Anisakis typica Larvae in Two Species of Threadfin Bream, Nemipterus hexodon and N. japonicus, from the Gulf of Thailand

  • Tunya, Rattanachai;Wongsawad, Chalobol;Wongsawad, Pheravut;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.58 no.1
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    • pp.15-25
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    • 2020
  • The third stage larvae (L3) of Anisakis typica were detected in 2 species of threadfin bream, Nemipterus hexodon and N. japonicus, from the Gulf of Thailand, and were morphologically and molecularly characterized. Total 100 threadfin breams, 50 Nemipterus hexodon and 50 N. japonicus, were examined with naked eyes after the opening of abdominal cavity with scissors. Almost all infected larvae remained alive and active even the fish were transported for 1-2 days. Anisakid larvae were exclusively distributed in the body cavity and rarely in the liver. The prevalence of A. typica L3 were 68.0% and 60.0% in N. hexodon and N. japonicus and their infection intensities were 3.5 and 4.2 per fish infected each. Morphological and morphometric analysis were performed by viewing specimens under both a light microscope and a scanning electron microscope. Interestingly, the protruded mucron of Anisakis typica under SEM showed a distinct cylindrical shape that differed from the cone shape of A. simplex. The protruded mucron could be used to identify A. typica L3 larvae in the future. A comparison of the ITS1-5.8S-ITS2 rDNA nucleotide sequences of these species revealed high blast scores with A. typica. Conclusively, it was confirmed that A. typica L3 are prevalent in threadfin breams from the Gulf of Thailand, and their morphological and molecular characters are something different from those of other anisakid larvae, including A. simplex and A. pegreffii.

Characterization of Fusarium oxysporum Isolated from Paprika in Korea

  • Cha, Sang-Do;Jeon, Young-Jae;Ahn, Geum-Ran;Han, Jae-In;Han, Kap-Hoon;Kim, Seong-Hwan
    • Mycobiology
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    • v.35 no.2
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    • pp.91-96
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    • 2007
  • In the present study we first report in Korea the identification and characterization of Fusarium oxysporum isolated from rotten stems and roots of paprika (Capsicum annuum var. grossum) at Masan, Kyungsangnamdo in 2006. The fungal species produced white aerial mycelia accompanying with dark violet pigment on PDA. The optimal temperature and pH for the growth of the species was $25^{\circ}C$ and pH 7, respectively. Microscopic observation of one of isolates of the species shows that its conidiophores are unbranched and monophialides, its microconidia have oval-ellipsoidal shape with no septate and are of $3.0{\sim}11{\times}1.5{\sim}3.5\;{\mu}m$ sizes, its macroconidia are of $15{\sim}20{\times}2.0{\sim}3.5\;{\mu}m$ sizes and have slightly curved or slender shape with $2{\sim}3$ septate. The results of molecular analysis show that the ITS rDNA of F. oxysporum from paprika shares 100% sequence identity with that of known F. oxysporum isolates. The identified species proved it's pathogenicity by causing rotting symptom when it was inoculated on paprika fruits. The growth of F. oxysporum from paprika was suppressed on PDA by agrochemicals such as benomyl, tebuconazole and azoxystrobin. The identified species has the ability of producing extracelluar enzymes that degrade cellobiose and pectin.

Lactic acid bacteria strains selected from fermented total mixed rations improve ensiling and in vitro rumen fermentation characteristics of corn stover silage

  • Huang, Kailang;Chen, Hongwei;Liu, Yalu;Hong, Qihua;Yang, Bin;Wang, Jiakun
    • Animal Bioscience
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    • v.35 no.9
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    • pp.1379-1389
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    • 2022
  • Objective: This study identified the major lactic acid bacteria (LAB) strains from different fermented total mixed rations (FTMRs) via metataxonomic analysis and evaluated the ability of their standard strain as ensiling inoculants for corn stover silage. Methods: The bacterial composition of eight FTMRs were analyzed by 16S rDNA sequencing. Corn stover was ensiled without LAB inoculation (control) or with 1×106 cfu/g LAB standard strain (Lactobacillus vaginalis, Lactobacillus reuteri, Lactobacillus helveticus, or Lactobacillus paralimentarius) selected from the FTMRs or 10 g/t commercial silage inoculant (CSI) around 25℃ for 56 days. For each inoculation, a portion of the silage was sampled to analyze ensiling characteristics at time intervals of 0, 1, 3, 7, 14, 28, and 56 days, gas production (GP), microbial crude protein and volatile fatty acids as the measurements of rumen fermentation characteristics were evaluated in vitro with the silages of 56 days after 72 h incubation. Results: Lactobacillus covered >85% relative abundance of all FTMRs, in which L. pontis, L. vaginalis, L. reuteri, L. helveticus, and L. paralimentarius showed >4% in specific FTMRs. CSI, L. helveticus, and L. paralimentarius accelerated the decline of silage pH. Silage inoculated with L. paralimentarius and CSI produced more lactic acid the early 14 days. Silage inoculated with L. paralimentarius produced less acetic acid and butyric acid. For the in vitro rumen fermentation, silage inoculated with CSI produced more potential GP, isobutyric acid, and isovaleric acid; silage inoculated with L. helveticus produced more potential GP and isovaleric acid, silage inoculated with L. paralimentarius or L. reuteri produced more potential GP only. Conclusion: The standard strain L. paralimentarius (DSM 13238) is a promising ensiling inoculant for corn stover silage. The findings provide clues on strategies to select LAB to improve the quality of silage.

Cultural Characteristics of Rhizoctonia cerealis Isolated from Diseased Wheat Fields and Evaluation of the Resistance of Korean Winter Cereal Crops (밀 잎집눈무늬병원균(Rhizoctonia cerealis)의 배양적 특성과 국내육성 맥류 품종의 저항성 평가)

  • Lee, Eun-Sook;Lee, Wang-Hyu;Kang, Chun-Sik;Kim, Mi-Jung;Kim, Tae-Soo;Park, Jong-Chul
    • Research in Plant Disease
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    • v.17 no.1
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    • pp.19-24
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    • 2011
  • It was identified as a sharp eyespot (Rhizoctonia cerealis) that the isolates from abnormal symptoms in wheat that showed yellowing leaves, necrotic spot on stem base and dead tillers. These isolates have slower growth property and fewer mycelia than Rhizoctonia solani AG-1(1A) (KACC 40106). They showed binuclear cell, same media cultural and DNA characteristics to R. cerealis. They caused same symptoms on leaves and stem base appeared in artificial inoculation test, comparing to diseased wheat fields and also affect to maturing of kernels. They have optimal growth temperature and acidity on the artificial media as $20{\sim}25^{\circ}C$ and pH 5~7, respectively. In the investigation of varietal resistance of Korean winter cereal crops to sharp eyespot, there was no resistant in wheat cultivars that all materials infected over 20% diseased ratio. 12 cultivars including 'Anbaekmil', however, considered to moderate resistance with 20 to 30% infection ratio. The others crops using in feeding, whole crop barley, oat, rye and triticale were resistant below 15% diseased degree except the rye that showed over 50% infection rate. It was the first evaluation to sharp eyespot resistance for the Korean feeding crop cultivars. Most tested Korean barley cultivars for malting and food were moderate and susceptible to the sharp eyespot. Only 3 hulled barley, 'Tapgolbori', 'Albori' and 'Seodunchalbori', showed resistance with less than 10% diseased ratio. All tested naked barley cultivars showed susceptible response to the disease.

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.

Characteristics of isolated lactic acid bacteria and their effects on the silage quality

  • Wang, Siran;Yuan, Xianjun;Dong, Zhihao;Li, Junfeng;Guo, Gang;Bai, Yunfeng;Zhang, Junyu;Shao, Tao
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.6
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    • pp.819-827
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    • 2017
  • Objective: Four lactic acid bacteria (LAB) strains isolated from common vetch, tall fescue and perennial ryegrass on the Tibetan Plateau were characterized, and their effects on the fermentation quality of Italian ryegrass (Lolium multiflorum Lam.) silage were studied. Methods: The four isolated strains and one commercial inoculant (G, Lactobacillus plantarum MTD-1) were evaluated using the acid production ability test, morphological observation, Gram staining, physiological, biochemical and acid tolerance tests. The five LAB strains were added to Italian ryegrass for ensiling at three different temperatures ($10^{\circ}C$, $15^{\circ}C$, and $25^{\circ}C$). Results: All isolated strains (LCG3, LTG7, I5, and LI3) could grow at $5^{\circ}C$ to $20^{\circ}C$, pH 3.0 to 8.0 and NaCl (3.0%, 6.5%). Strains LCG3, LTG7, I5, and LI3 were identified as Lactobacillus plantarum, Pediococcus acidilactici, Lactobacillus paraplantarum, and Lactobacillus casei by sequencing 16S rDNA, respectively. All LAB inoculants significantly (p<0.05) increased lactic acid (LA) contents and ratios of lactic acid to acetic acid, and reduced pH and ammonia nitrogen/total nitrogen (AN/TN) compared with uninoculated silages at various temperatures ($10^{\circ}C$, $15^{\circ}C$, and $25^{\circ}C$). Compared to the commercial inoculant G, I5, and LI3 showed similar effects on improving the silage quality of Italian ryegrass at $10^{\circ}C$ and $15^{\circ}C$, indicated by similar pH, LA content and AN/TN. Conclusion: All inoculants could improve the silage fermentation quality at various temperatures ($10^{\circ}C$, $15^{\circ}C$, and $25^{\circ}C$). At the temperature of $10^{\circ}C$ and $15^{\circ}C$, strain I5 and LI3 had similar effects with the commercial inoculant G on improving the silage quality of Italian ryegrass.

Production and Characterization of Alkaline Protease of Micrococcus sp. PS-1 Isolated from Seawater (해수에서 분리한 Micrococcus sp. PS-1이 생산하는 단백질 분해효소의 생산과 효소학적 특성)

  • Jin, Young-Rang;Yu, Sun-Nyoung;Kim, Kwang-Youn;Kim, Sang-Hun;Park, Seul-Ki;Kim, Hyeun-Kyeung;Lee, Yong-Seok;Choi, Yong-Lark;Ji, Jae Hoon;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.23 no.2
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    • pp.273-281
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    • 2013
  • The purpose of this research was to investigate the production and characterization of alkaline protease from Micrococcus sp. PS-1 newly isolated from seawater. Micrococcus sp. PS-1 was grown in Luria-Bertani (LB) medium. Its optimal temperature and pH for growth were $30^{\circ}C$ and 7.0, respectively. The effect of nitrogen sources was investigated on optimal enzyme production. A high level of alkaline protease production occurred in LB broth containing 2% skimmed milk. The protease was purified in a 3-step procedure involving ultrafiltration, acetone precipitation, and dialysis. The procedure yielded a 16.43-purification fold, with a yield of 54.25%. SDS-PAGE showed that the enzyme had molecular weights of 35.0 and 37.5 kDa. Its maximum protease activity was exhibited at pH 9.0 and $37^{\circ}C$, and its activity was stable at pH 8.0-11.0 and $25-37^{\circ}C$. The protease activity was strongly inhibited by PMSF, EDTA, and EGTA. Taken together, the results demonstrate that the protease enzyme from Micrococcus sp. PS-1 probably belongs to a subclass of alkaline metallo-serine proteases.