• Title/Summary/Keyword: mutant strain

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Survival Factor Gene FgSvf1 Is Required for Normal Growth and Stress Resistance in Fusarium graminearum

  • Li, Taiying;Jung, Boknam;Park, Sook-Young;Lee, Jungkwan
    • The Plant Pathology Journal
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    • v.35 no.5
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    • pp.393-405
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    • 2019
  • Survival factor 1 (Svf1) is a protein involved in cell survival pathways. In Saccharomyces cerevisiae, Svf1 is required for the diauxic growth shift and survival under stress conditions. In this study, we characterized the role of FgSvf1, the Svf1 homolog in the homothallic ascomycete fungus Fusarium graminearum. In the FgSvf1 deletion mutant, conidial germination was delayed, vegetative growth was reduced, and pathogenicity was completely abolished. Although the FgSvf1 deletion mutant produced perithecia, the normal maturation of ascospore was dismissed in deletion mutant. The FgSvf1 deletion mutant also showed reduced resistance to osmotic, fungicide, and cold stress and reduced sensitivity to oxidative stress when compared to the wild-type strain. In addition, we showed that FgSvf1 affects glycolysis, which results in the abnormal vegetative growth in the FgSvf1 deletion mutant. Further, intracellular reactive oxygen species (ROS) accumulated in the FgSvf1 deletion mutant, and this accumulated ROS might be related to the reduced sensitivity to oxidative stress and the reduced resistance to cold stress and fungicide stress. Overall, understanding the role of FgSvf1 in F. graminearum provides a new target to control F. graminearum infections in fields.

Studies on the Induction of Available Mutant of Takju Yeast by UV light Irradiation Part III -On the Acid Productivity of the Mutant and Takju Brewing Utilized the Mutant- (자외선(紫外線) 조사(照射)에 의(依)한 탁주효모(濁酒酵母)의 변이주(變異株) 육성(育成)에 관(關)한 연구(硏究) (제3보)(第三報) -변이주(變異株)의 생산능(生酸能) 및 변이주(變異株)를 이용(利用)한 탁주양조(濁酒釀造)에 대(對)하여-)

  • Kim, C.J.;Oh, M.J.;Kim, S.Y.
    • Applied Biological Chemistry
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    • v.18 no.1
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    • pp.23-29
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    • 1975
  • This experiment was conducted to study the effects of temperature and pH upon the acid productivity of the acid producing mutant induced by the treatment of ultraviolet light, and to identify the producing acid by PPC and p-oxydiphenyl method. Chemical composition of Takju mash brewed with selected yeast and producing acid were observed and the results were as follows. 1) There was no apprecible difference in acid producing activity of mutant at $25^{\circ}C\;to\;30^{\circ}C$. 2) The acid producing activity of mutant was little below pH 4 and was gradually increased according to approach nenutral, and the accumulation of acid was amounted to 0.5-0.7% as a lactic acid at pH 5 to 7 within 48 hrs of fermentation. 3) The acid produced by mutant was detected to the lactic acid. 4) In the cases of the Takju was brewed with the starter from the acid producing mutant the requirement of Ipkuk was 5% for all the raw materials, on the contrary, using orginal strain the requirement of Ipkuk was 20%. 5) In the case of both starters from the acid producing mutant and orginal strain were added at different brewing times, and only Bunkuk was used as a saccharifying agent (without Ipkuk), Takju was able to brewed more repidly and successfully than the case of general process.

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In Vivo Analysis of fadB Homologous Enzymes Involved in Biosynthesis of Polyhydroxyalkanoates in Recombinant Escherichia coli (재조합 대장균에서 fadB 유사효소의 Polyhydroxyalkanoates 합성에 미치는 역할의 규명)

  • 최종일;박시재;이상엽
    • KSBB Journal
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    • v.19 no.4
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    • pp.331-334
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    • 2004
  • In vivo characterization of FadB homologous enzymes including PaaG, YdbU and YgfG for medium-chain-length (MCL) polyhydroxyalkanoate (PHA) biosynthesis was carried out in fadB mutant Escherichia coli. Previously, it was reported that amplification of FadB homologous enzymes such as PaaG and YdbU in fadB mutant E. coli resulted in enhanced biosynthesis of MCL-PHA by greater than two fold compared with control strain. In this study, we constructed paaG fadB double mutant E. coli WB114 and ydbU fadB double mutant E. coli WB115 to investigate the roles of PaaG and YdbU in biosynthesis of MCL-PHA. Inactivation of paaG and ydbU genes in fadB mutant E. coli harboring Pseudomonas sp. 61-3 phaC2 gene reduced the MCL-PHA production to 0.16 and 0.16 PHA g/L, respectively from 2 g/L of sodium decanoate, which are much lower than 0.43 PHA g/L obtained with fadB mutant E. coli WB101 harboring the phaC2 gene. Also, we identified new FadB homologous enzyme YgfG, and examined its roles by overexpression of ygfG and construction of ygfG fadB double mutant E. coli WB113.

Increased Carotenoid Production in Xanthophyllomyces dendrorhous G276 Using Plant Extracts

  • Kim, Soo-Ki;Lee, Jun-Hyeong;Lee, Chi-Ho;Yoon, Yoh-Chang
    • Journal of Microbiology
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    • v.45 no.2
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    • pp.128-132
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    • 2007
  • The red yeast Xanthophyllomyces dendrorhous (previously named Phaffia rhodozyma) produces astaxanthin pigment among many carotenoids. The mutant X. dendrorhous G276 was isolated by chemical mutagenesis. The mutant produced about 2.0 mg of carotenoid per g of yeast cell dry weight and 8.0 mg/L of carotenoid after 5 days batch culture with YM media; in comparison, the parent strain produced 0.66 mg/g of yeast cell dry weight and a carotenoid concentration of 4.5 mg/L. We characterized the utilization of carbon sources by the mutant strain and screened various edible plant extracts to enhance the carotenoid production. The addition of Perilla frutescens (final concentration, 5%) or Allium fistulosum extracts (final concentration, 1%) enhanced the pigment production to about 32 mg/L. In a batch fermentor, addition of Perilla frutescens extract reduced the cultivation time by two days compared to control (no extract), which usually required five-day incubation to fully produce astaxanthin. The results suggest that plant extracts such as Perilla frutescens can effectively enhance astaxanthin production.

Genetic Breeding of Korean Soy Sauce-Fermenting Bacillus by UV Mutation (돌연변이에 의한 한국간장균의 유전적 육종)

  • Kim, Jong-Kyu;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.31 no.4
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    • pp.346-350
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    • 1988
  • A mutant for Korean soy sauce which wilt improve the productivity of amylase and protease was obtained through the second mutation of the original strain using UV radiation. The original strain was the NTG treated mutant of the Bacillus sp. producing peculiar flavour which had been isolated from the korean soy sauce. The mutant could improve the productivity of amylase by 58% and that of protease by 41%. The enzyme produced in this way were similar in enzymatic properties such as optimal reaction pH and temperature. The reaction was not deterred by highly densed salt solution of 5 M and the enzyme productivity was not influenced in the concentration of up to 2 M.

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DNA Microarray and Gene Ontology Enrichment Analysis Reveals That a Mutation in opsX Affects Virulence and Chemotaxis in Xanthomonas oryzae pv. oryzae

  • Kim, Hong-Il;Park, Young-Jin
    • The Plant Pathology Journal
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    • v.32 no.3
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    • pp.190-200
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    • 2016
  • Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial leaf blight (BLB) in rice (Oryza sativa L.). In this study, we investigated the effect of a mutation in opsX (XOO1056), which encodes a saccharide biosynthesis regulatory protein, on the virulence and bacterial chemotaxis of Xoo. We performed DNA micro-array analysis, which showed that 63 of 2,678 genes, including genes related to bacterial motility (flagellar and chemotaxis proteins) were significantly downregulated ($<\;-2\;log_2$ fold changes) by the mutation in opsX. Indeed, motility assays showed that the mutant strain was nonmotile on semisolid agar swarm plates. In addition, a mutant strain (opsX::Tn5) showed decreased virulence against the susceptible rice cultivar, IR24. Quantitative real-time RT-PCR reaction was performed to confirm the expression levels of these genes, including those related to flagella and chemotaxis, in the opsX mutant. Our findings revealed that mutation of opsX affects both virulence and bacterial motility. These results will help to improve our understanding of Xoo and provide insight into Xoo-rice interactions.

Purification and Characterization of Phenoxazinone Synthase from Streptomyces sp. V-8 Mutant Producing Adenoside Deaminase Inhibitor (아데노신 탈아미노화 효소 억제제를 생산하는 Streptomyces sp. V-8의 변이종으로부터 페녹사지논 합성효소의 분리 및 특성)

  • 김경자;조성진
    • YAKHAK HOEJI
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    • v.43 no.1
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    • pp.68-76
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    • 1999
  • Phenoxazinone synthase catalyzes the oxidative condensation of two molecules of substituted o-aminophenol to the phenoxazinone chromophore of actinomycin. Mutant strain, Streptomyces sp. V-8-M-1 producing higher phenoxazinone synthase, was obtained from Streptomyces sp. V-8 by treatment of N-methyl-N'-nitro-N-nitrosoguanidine. The phenoxazinone synthase was purified from extract of mutant strain of Streptomyces sp. V-8-M-l by successive steps of streptomycin sulfate, ammonium sulfate precipitation. DEAE-cellulose and Sephadex G-200 column chromatography. Molecular weight of the enzyme was 360,000 daltons. The enzyme was composed of octamer of a single subunit of 45,000 daltons. The Km value and Vmax value for 3-HAA were $14.9{\;}{\mu}M$ and 9.5 mg/U, respectively. The optimal pH and temperature for the enzyme activity were 9.0 and $25~30^{\circ}C$, respectively. Treatment of the enzyme with group specific reagents, phenylglyoxal, p-hydroxymercury-benzoate, Nbromosuccinimide, 5.5'-dithiobis-nitrobenzoic acid and ethylmaleimide resulted in loss of enzyme activity, which shows arginine and cysteine residues are at or near the active site.

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Antifungal Activity of Root Colonizing Pseudomonas fluorescens MC07 is Responsible for Its Disease Suppression Ability (근권 정창 세균 Pseudomonas fluorescens MC07의 항진균 활성과 병 억제 능력)

  • 김진우;박병근;황인규;박창석
    • Korean Journal Plant Pathology
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    • v.14 no.6
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    • pp.606-611
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    • 1998
  • An antagonistic bacterium, Pseudomonas flurorescens MC07 inhibited the mycelial growth of Rhizoctonia solani, Pythium ultimum, Fusarium oxysporum, and Phytophthora capsici in on potato dextrose agan (PDA) and other media. The strain MC07 conlonizes various plant roots and possesses antifungal activity. To determine the role of antifungal activity of the bacterium in disease suppression, a mutant Okm3-4 which lost its activity was isolated after screening 2,500 colonies generated by Omegon-Km insertions. The mutant Okm3-4 showed diminished growth inhibition of R. solani, P. ultimum, F. oxysporum, and Ph. capsici in vitro and had reduced suppressive effects on sesame damping.-off compared to the parental strain. In soils, accumulation of the pathogens by continuous cropping, 90% of sesame plants were killed by natural infection of damping-off whereas, only 29% of plants grown from seeds treated with MC07 were killed. On the other hand, 85% of plants died when sesame seeds were treated with the Okm3-4 cells. This indicated that antifungal activity of MC07 in vitro is directly responsible for the suppression of damping-off disease. Emergence rates of sesame seeds in pots containing diseased soil were 33%. However, MC07 treatments on seeds significantly improved emergence rates, which has similar effects of Benomyl treatment. The mutant Okm3-4 exhibited 53% of emergence rate. This indicated that antifungal activity of MC07 also affects the emergence rate of sesame seeds.

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Production of 1,4-Androstadiene-3,17-dione by a Mutant Strain of Brevibacterium lipolyticum (Brevibacterium lipolyticum 변이주에 의한 1,4-Androstadiene-3, 17-Dione의 생성)

  • Choi, In-Wha;Lee, Kang-Man
    • YAKHAK HOEJI
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    • v.33 no.6
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    • pp.365-371
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    • 1989
  • Microbiological conversion of sterols to 17-ketosteroids has been recognized as a source for commercial preparation of steroidal drugs. In order to develop bacterial strains and process with Brevibacterium lipolyticum IAM 1398 capable of converting cholesterol to 1,4-Androstadiene-3,17-dione (ADD) at about 27% yield, we studied on strain improvement, fermentation condition and whole cell immobilization. By using UV and/or NTG as mutagens, a mutant to convert cholesterol to ADD with higher yield than 60% was selected. Better production of ADD was manifested in the case of maltose used as a supplemental carbon source, and yeast extract or soytone as a nitrogen source. Addition of tween 80 (0.05%) as a surfactant beneficial for increasing the productivity. The optimal initial pH of the medium was 6.5 and optimal culture temperature was $30^{\circ}C$. Whole cell immobilization by using carrageenan, agar, alginate and acrylamide was carried out and the activity of conversion was tested. In the case of carrageenan and agar, immobilized cells were active for at least two cycles of fermentation.

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A Phosphate Starvation-Inducible Ribonuclease of Bacillus licheniformis

  • Nguyen, Thanh Trung;Nguyen, Minh Hung;Nguyen, Huy Thuan;Nguyen, Hoang Anh;Le, Thi Hoi;Schweder, Thomas;Jurgen, Britta
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1464-1472
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    • 2016
  • The BLi03719 protein of Bacillus licheniformis DSM13 belongs to the most abundant extracellular proteins under phosphate starvation conditions. In this study, the function of this phosphate starvation inducible protein was determined. An amino-acid sequence analysis of the BLi03719-encoding gene showed a high similarity with genes encoding the barnase of Bacillus amyloliquefaciens FZB42 and binase-like RNase of Bacillus pumilus SARF-032. The comparison of the control strain and a BLi03719-deficient strain revealed a strongly reduced extracellular ribonuclease activity of the mutant. Furthermore, this knockout mutant exhibited delayed growth with yeast RNA as an alternative phosphate and carbon source. These results suggest that BLi03719 is an extracellular ribonuclease expressed in B. licheniformis under phosphate starvation conditions. Finally, a BLi03719 mutant showed an advantageous effect on the overexpression of the heterologous amyE gene under phosphate-limited growth conditions.