• Title/Summary/Keyword: Wild strain

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The Attenuation Mechanism and Live Vaccine Potential of a Low-Virulence Edwardsiella ictaluri Strain Obtained by Rifampicin Passaging Culture

  • Shuyi Wang;Jingwen Hao;Jicheng Yang;Qianqian Zhang;Aihua Li
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.167-179
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    • 2023
  • The rifampicin-resistant strain E9-302 of Edwardsiella ictaluri strain 669 (WT) was generated by continuous passage on BHI agar plates containing increasing concentrations of rifampicin. E9-302 was attenuated significantly by 119 times to zebrafish Danio rerio compared to WT in terms of the 50% lethal dose (LD50). Zebrafish vaccinated with E9-302 via intraperitoneal (IP) injection at a dose of 1 × 103 CFU/fish had relative percentage survival (RPS) rates of 85.7% when challenged with wild-type E. ictaluri via IP 14 days post-vaccination (dpv). After 14 days of primary vaccination with E9-302 via immersion (IM) at a dose of 4 × 107 CFU/ml, a booster IM vaccination with E9-302 at a dose of 2 × 107 CFU/ml exhibited 65.2% RPS against challenge with wild-type E. ictaluri via IP 7 days later. These results indicated that the rifampicin-resistant attenuated strain E9-302 had potential as a live vaccine against E. ictaluri infection. A previously unreported amino acid site change at position 142 of the RNA polymerase (RNAP) β subunit encoded by the gene rpoB associated with rifampicin resistance was identified. Analysis of the whole-genome sequencing results revealed multiple missense mutations in the virulence-related genes esrB and sspH2 in E9-302 compared with WT, and a 189 bp mismatch in one gene, whose coding product was highly homologous to glycosyltransferase family 39 protein. This study preliminarily explored the molecular mechanism underlying the virulence attenuation of rifampicin-resistant strain E9-302 and provided a new target for the subsequent study of the pathogenic mechanism of E. ictaluri.

Studies on the m-Toluate Degradating Plasmid in Pseudomonas (m-Tluate를 분해하는 Peudomonas의 분리 및 Dgradative Pasmid와의 연관성에 관하여)

  • 박순희;하영칠;홍순우
    • Korean Journal of Microbiology
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    • v.17 no.1
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    • pp.25-41
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    • 1979
  • A strain able to grow up m-toluate minimal medium has been isolated after selective enrichment and given the name T81X, which was later identified as pseudomonase putida according to its morphological and biochemical characteristics. After treatment with plasmied specific curing agent, mitomycin C, followed by replica plating on m-toluate and xylene minimal agar plate, T81Xstrain has been shown to harbour a curable plasmid relating to the m-toluate and xylene metabolism. Spontaneous curing frquency of this plasmid was also greatly enhanced by growing on benzoate minimal medium. After then, it was also xylene metabogrowing on benzoate minimal medium. After then, it was found to be conjugally nontransmissible. From the comparative investigation of catechol 1,2-oxygenase and catechol 2,3-oxygenase activities in wild type and cured strain on various growth substrate, it appeared that T81X strain has both of these two enzymes while cured strain has catechol 1,2-oxygenase only. Growing on m-toluate minimal medium T81X strain should carry the genetic information necessary for coding the catechol 1,2-oxygense induced by m-toluate or benzoate, on that curable plasmid.

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Studies on the Cellulase of Penicillium sp. Isolated from Soils. -(1) Isolation of Penicillium sp. C8-14 strain from Kwangneung soil- (토양에서 분리한 Penicillum sp. 가 생산하는 Cellulase에 관한 연구(제1보) -광능지역 토양으로부터 Penicillum sp. C8-14주(株)의 분리-)

  • Kim, Yong-Bae;Yi, Pyung-Kuk;Choi, Seung-Ho
    • The Korean Journal of Mycology
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    • v.1 no.1
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    • pp.23-28
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    • 1973
  • The soil of Kwangneung area(Kyeunggi-Do) was inoculated directly into wheat-bran-media and after $3{\sim}4$ days of incubation, a Penicillium species whose cellulase activity was 1011u/g was isolated. With the treatment of mutagenic agents an improved strain(cellulase activity: 1303u/g) was obtained. This strain was screened again by mono-spore isolation method. Finally a strain C8-14 (cellulase activity: 2351u/g) which had lesser spores than the wild strain was obtained.

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Fungal Strain Improvement for Cellulase Production Using Repeated and Sequential Mutagenesis

  • Vu, Van-Hanh;Pham, Tuan-Anh;Kim, Keun
    • Mycobiology
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    • v.37 no.4
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    • pp.267-271
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    • 2009
  • A fungal strain producing a high level of cellulase was selected from 320 fungal isolates and identified as Aspergillus sp. This strain was further improved for cellulase production by sequential treatments by two repeated rounds of $\gamma$-irradiation of $Co^{60}$, ultraviolet treatment and four repeated rounds of treatment with N-methyl-N'-nitro-N-nitrosoguanidine. The best mutant strain, Aspergillus sp. XTG-4, was selected after screening and the activities of carboxymethyl cellulase, filter paper cellulase and $\beta$-glucosidase of the cellulase were improved by 2.03-, 3.20-, and 1.80-fold, respectively, when compared to the wild type strain. After being subcultured 19 times, the enzyme production of the mutant Aspergillus sp. XTG-4s was stable.

Site-specific Disruption of Glyoxylate Bypass and Its Effect in Lysine-producing Corynebacterium lactofermentum Strain

  • Kim, Youn-Hee;Lee, Heung-Shick
    • Journal of Microbiology and Biotechnology
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    • v.6 no.5
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    • pp.315-320
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    • 1996
  • The role of glyoxylate bypass in a lysine-producing Corynebacterium lactofermentum strain was analyzed. Unlike the wild type, the strain expressed enzymes of glyoxylate bypass during growth in the fermentation broth containing glucose as the carbon source. To evaluate the importance of glyoxylate bypass in the strain, we disrupted chromosomal aceA by using a cloned fragment of the gene. Site-specific disruption of aceA which codes for the isocitrate lyase, the first enzyme of the bypass, was confirmed by Southern blot analysis. The aceA mutant strain completely lost isocitrate lyase activity and ability to grow in a minimal medium containing acetate as the sole carbon source. The mutant strain was similar to its parental strain in growth characteristics and produced comparable amounts of lysine in shake flasks containing glucose as the carbon source. The amount of oxaloacetate accumulated in the fermentation medium was similar for both strains, suggesting that expression of glyoxylate bypass does not necessarily lead to the increase in intracellular oxaloacetate. These data clearly demonstrate that glyoxylate bypass does not function as one of the routes of carbon supply for lysine production in the strain. It appears that the leakiness of the glyoxylate bypass in the strain might be the result of a secondary mutation which arose during previous strain development by random mutagenesis.

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Screening of γ-Aminobutyric Acid (GABA)-Producing Wild Yeasts and their Microbiological Characteristics

  • Han, Sang-Min;Jeon, Sun-Jeong;Lee, Hyang-Burm;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.44 no.2
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    • pp.87-93
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    • 2016
  • From 182 non-pathogenic wild yeast isolates from flowers, Pichia silvicola UL6-1 and Sporobolomyces carnicolor 402-JB-1 were selected for potent ${\gamma}$-aminobutyric acid production and microbiological characteristics were investigated. Pichia silvicola UL6-1 formed ascospores and pseudomycelia. The strain was also halotolerant, growing well in 5% NaCl-containing yeast extract-peptone-dextrose (YPD) medium. Sporobolomyces carnicolor 402-JB-1 did not form ascospores or pseudomycelia and grew well on 10% glucose-yeast extract-peptone medium.

A Study of Useful Wild Mushrooms by Segregation and Identification Native in Middle Area (중부지방에서 자생하는 유용 야생 버섯의 분류 및 동정)

  • Noh, Jae-Goan;Park, Jae-Seong;Choi, Jae-Sun;Song, In-Gyu;Yun, Tae;Min, Kyeong-Beom
    • Journal of Mushroom
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    • v.7 no.2
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    • pp.49-56
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    • 2009
  • This study was conducted to preserve of mushroom resources and utility useful wild mushrooms by segregation and identification from 2005 to 2007. The mushroom strains were collected a center of native mushroom wild growth place of Chungbuk Province. The obtained results from this study were summarized as follows ; We collected 79 wild mushroom strains, and the collected wild mushrooms were composition of 32 strains of edible mushrooms, 3 strains of medicinal use mushrooms, 15 strains of poisonous mushrooms, and 29 strains indistinct mushrooms. The 28 strains were segregated and identified from 32 strains of edible mushrooms. The present preservation strains are 15 strains, and other 13 strains were damaged in tissue culture and preservation. We made specimen of wild mushroom by alcohol, and have preserved perennial mushrooms by drying. We photographed 79 strains of wild mushrooms.

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Agronomic Characteristics and Aromatic Compositions of Korean Wild Codonopsis lanceolata Collections Cultivated in Field (한국 야생더덕 수집종의 노지 재배시 생육 특성과 향기성분 조성)

  • 이승필;김상국;민기군;조지형;최부술;이상철;김길웅
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.2
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    • pp.188-199
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    • 1996
  • The native ecological environment and aromatic constituents of Korean wild Codonopsis lanceolata and one Japanese strain were investigated to find Codonopsis lanceolata strains showing high aromatics, and to know regional differences among these strains. The results were as follows : There were no remarkable differences among the Korean wild C. lanceolata strains in ecological environments. Recovery yield of essential oils was highest in Togyusan strain with 0.009%. Difference in protein band patterns among these strains was not recognized, and peroxidase and esterase pattern changes were appeared in different collected regions at the leaf and root tissues. Major aromatic constituents were 11 kinds of aliphatic alcohols such as trans-2-hexenal, 1-hexanol, cis-3-hexanol, and trans-2-hexanol. And Togyusan strain, Sobaeksan strain, and Kayasan strain have the highest aliphatic alcohols of plant essential oils. In particular, BHT(butylated hydroxytoluene), one of the antioxidants, was detected in Chirisan strains.

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Lysine Production by Thialysine Resistant Mutant of Candida utilis ( I ) - Isolation of High Lysine Excreting Mutant of Candida utilis - (Candida utilis의 Thialysine 내성맥리주에 의한 Lysine생산 ( I ) -Candida utilis의 Lysine을 생산하는 Thialysine 내성맥리주의 분리-)

  • Bang, Byung-Ho;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.11 no.3
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    • pp.175-180
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    • 1983
  • Thialysine significantly inhibited the growth of wild type strain Gondida utilis NCYC-359. In the absence of thialysine, the culture reached stationary phase after 24hr, however, in the presence of 0.5% thialysine, the culture reached stationary phase after 40hr, respectively. Effect of amino acid or vitamin was investigated on recovery of the growth of wild type strain from thialysine inhibition. Glycine, methionine, arginine and tryptophan recovered growth inhibition by thialyslne to some extent. However, vitamins were inert. Especially, lysine at one eighth concentration of thialysine recovered almost fully the growth inhibition. Thialysine resistant mutants were induced from the parent strain of Condida utilis NCYC-359 by NTG treatment. Colonies of thialysine resistant mutants were obtained on agar minimal medium supplemented with 0.1-0.5% thialysine. The frequency of thialysine resistant mutants induced by the first mutation was the highest at 0.1% The wild strain produced no appreciable lysine extracellularly. However, almost thialysine resistant mutants excreted appreciably. Lysine excretion increased after repeated mutation. Finally, of the thialysine resistant mutants induced by NTG, Condida utilis TRN-4006 was obtained. This strain excreted lysine (400$\mu\textrm{g}$/$m\ell$) into the medium with a concomitant decrease of lysine in the intracellular pool.

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Bacterial Cellulose 생산균 KJ-1으로 부터 uv와 NTG mutant들의 cellulose 생산수율의 증가

  • Kim, Hye-Eun;Son, Chang-Jin;Jeong, Seon-Yong;Kim, Seong-Jun
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.723-726
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    • 2001
  • This study was performed to improve the yield of bacterial cellulose(BC) by UV and NTG mutagenesis of strain KJ-1 which produced largely BC. some mutants showed high BC productivity with twice elevation compared to that the wild strain KJ-1. A difference was found in production and bioconversion phase of synthesized organic acid, such as gluconic acid, 2-keto gluconic acid, and 5-keto gluconic acid between mutants and strain KJ-1 in the static culture. The organic acid produced in secondary metabolism phase, were more rapidly consumed in the culture with the mutants than that the parent strain after glucose in the broth was conversed to a limiting substrate. Therefore, we suggested the reason for increasing of BC production that the mutant strain consumed more efficiently synthesized acids as substrates than that of the parent strain.

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