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Purification of an Antibacterial Peptide from the Gills of the Pufferfish Takifugu pardalis (졸복의 아가미로부터 항균성 펩타이드의 정제)

  • Kim, Tae Young;Go, Hye-Jin;Park, Nam Gyu
    • Journal of Life Science
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    • v.27 no.1
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    • pp.50-56
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    • 2017
  • An antibacterial peptide was purified from an acidified gill extract of the pufferfish Takifugu pardalis. The acidified gill extract was put through a Sep-Pak C18 solid phase extraction cartridge using a stepwise gradient and divided into a flow-through (F.T.) and 60% methanol fraction (RM 60). Among the eluents, RM 60 had potent antibacterial activity against Bacillus subtilis KCTC 1021. RM 60 was partially purified on a cationic-exchange column (SP-5PW) by a linear gradient, and the antibacterial peptide was then further purified, using a series of cationic-exchange and $C_{18}$ reversed-phase HPLC columns. For characterization of the purified peptide, its molecular weight and amino acid sequence were analyzed by MALDI-TOF MS and Edman degradation. The molecular weight of the peptide was about 1171.6 Da. The amino acid sequence of the peptide was partially determined as: STKEKAPRKQ. A comparison of the N-terminal amino acid sequence of the purified peptide with that of other known polypeptides revealed high homology with the N-terminus of the histone H3 protein, which belongs to the histone H3 family. Thus, this peptide was designated as a puffer fish gill (PFG)-related antimicrobial peptide. This is the report to describe an antimicrobial function for the N-terminus of histone H3 of an animal species. The findings suggest that this peptide plays a significant role in the innate defense system of the pufferfish.

Gardenia jasminoides Exerts Anti-inflammatory Activity via Akt and p38-dependent Heme Oxygenase-1 Upregulation in Microglial Cells (소교세포에서 heme oxygenase-1 발현 유도를 통한 치자(Gardenia jasminoides)의 항염증 효과)

  • Song, Ji Su;Shin, Ji Eun;Kim, Ji-Hee;Kim, YoungHee
    • Journal of Life Science
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    • v.27 no.1
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    • pp.8-14
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    • 2017
  • Died Gardenia jasminoides fruit is used as a dye in the food and clothes industries in Asia. The present study investigated the anti-inflammatory effects of aqueous extract of G. jasminoides fruits (GJ) in BV-2 microglial cells. GJ inhibited lipopolysaccharide-induced nitric oxide (NO) secretion, inducible nitric oxide synthase (iNOS) expression, and reactive oxygen species production, without affecting cell viability. Furthermore, GJ increased the expression of heme oxygenase-1 (HO-1) in a dose-dependent manner. Moreover, the inhibitory effect of GJ on iNOS expression was abrogated by small interfering RNA-mediated knock-down of HO-1. In addition, GJ induced nuclear translocation of nuclear factor E2-related factor 2 (Nrf2), a transcription factor that regulates HO-1 expression. GJ-mediated expression of HO-1 was suppressed by LY294002, a phosphoinositide 3-kinase (PI-3K) inhibitor, and SB203580, a p38 kinase inhibitor, but not by the extracellular signal-regulated kinase (ERK) inhibitor PD98059 or c-Jun N-terminal kinase (JNK) inhibitor SP600125. GJ also enhanced the phosphorylation of Akt and p38. These results suggest that GJ suppresses the production of NO, a pro-inflammatory mediator, by inducing HO-1 expression via PI-3K/Akt/p38 signaling. These findings illustrate a novel molecular mechanism by which extract from G. jasminoides fruits inhibits neuroinflammation.

Hepatoprotective Effect of Bacillus subtilis-fermented Silkworm (Bombyx mori L.) Extract on an Alcoholic Fatty Liver in Rats (고초균 발효누에 추출물이 알코올성 지방간 유발 흰쥐에 미치는 간 기능 개선 효과)

  • Kim, Tae-Hoon;Ahn, Hee-Young;Kim, Young-Wan;Sim, So-Yeon;Seo, Kwon-Il;Cho, Young-Su
    • Journal of Life Science
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    • v.28 no.6
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    • pp.697-707
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    • 2018
  • The aim of this study was to investigate the potential effects of extracts from silkworm (Bombyx mori L.) that was fermented with Bacillus subtilis KACC 91157 at the levels of 1%(v/w), 2.5%(v/w), and 5%(v/w) in alcohol-fed rats. Sprague-Dawley rats were divided into seven groups: normal group (N), alcohol treated group (C), silymarin treated group (PC, positive control), 5% silkworm powder treated group (SP5), 1% Bacillus subtilis-fermented silkworm powder treated group (BSP1), 2.5% BSP treated group (BSP2.5), and 5% BSP treated group (BSP5). The activities of AST, ALT, ALP, and LDH in the serum and the triglyceride concentrations in the liver and serum were increased by alcohol feeding but were reduced in the BSP5 group. In addition, the contents of total lipids, free fatty acids, and total cholesterol were increased in the alcohol-fed group but were reduced in the BSP5 group. The activities of ADH, ALDH and ADH, ALDH protein levels in the liver were increased in the BSP5 group. The TBARS contents in the liver, serum, liver mitochondria, and liver microsomes were slightly decreased in the BSP5 group compared to the C group. The hepatocytes in alcohol-fed rats contained numerous large droplets; however, BSP5 treatment prevented alcohol-induced lipid droplet accumulation in the hepatocytes. Based on these results, extracts from Bacillus subtilis-fermented silkworm (Bombyx mori L.) have significant potential for development into a functional health food that can improve alcoholic fatty liver conditions.

Rapid Detection of the pathogenic agent of Bacterial white enteritis of Larval and Juvenile Stages in Olive flounder (Paralichthys olivaceus) (넙치(Paralichthys olivaceus)자치어 장관백탁증(Bacterial white enteritis) 원인균의 신속 검출)

  • Mun, Yeong-Geon;Park, Geun-Tae;Son, Hong-Ju;Lee, Sang-Hyeon;Lee, Jeong-Min;Heo, Mun-Su
    • Journal of fish pathology
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    • v.17 no.3
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    • pp.159-169
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    • 2004
  • Bacterial wihte enteritis ocurred by infection of V. ichthyoenteri is a devastating disease in olive flounder (Paralichthys olivaceus) hatcheries in Korea. Since white enteritis has been a problem in aquqtic industries, necessity of a rapid detection method is increased. In an attempt to develop rapid PCR method the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. The intergenic spacers were amplified by primers complementary to conserved region of 16S and 23S rRNA genes. The intergenic spacer region between the 16S and 23S rRNA genes of V. ichthoenteri were investigated by PCR fragment length typing and DNA sequencing. Analysis of the ISR sequences showed that V. ichthyoenteri contains one types of polymorphic ISRs. The size of ISRs ranged 348bp length and not contains tRNA genes. Mutiple alignment of representative sequences from different Vibrio species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of Vibrio ichthyoenteri. PCR. The specific of the primer was examined using genomic DNA prepared from 19 different Vibrio species, isolated 18group Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

The Effect of Nutritional Balance between Carbon and other Nutrient Sources on the Growth of Sporobolomyces holsaticus (탄소원과 다른 영양원간의 영양균형이 Sporobolomyces holsaticus의 균체생육도에 미치는 영향)

  • Park, Wan-Soo;Koo, Young-Jo;Shin, Dong-Hwa;Min, Byung-Yong
    • Korean Journal of Food Science and Technology
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    • v.15 no.1
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    • pp.56-61
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    • 1983
  • Direct production of biomass from starch using amylolytic yeast, Sporobolomyces holsaticus FRI Y-5 was studied with varying the ratios between carbon and other nutrient sources in the medium. It was investigated under condition of constant C/P and C/S ratio to influence the initial concentration of starch $(S_o)$ and C/N ratio on its growth which is described as the specific growth rate $({\mu})$, cell yield (Y), the maximum concentration of cell $(X_m)$, and productivity (P). They were very dependent on both $S_o$ and C/N ratio. The form of the relationship between and ${\mu}$ and $S_o$ was observed to be similar to saturation kinetics at C/N = 100 but presented substrate inhibition at other C/N ratios. As $S_o$ was changed from 22.5 to 90 g/l, Y was observed to vary with C/N ratios but seemed to decrease as a wholes. $X_m$ was linearly related to $S_o$ at more than C/N = 50 but at less than C/N = 10 substrate inhibition was presented. P increased suddenly to $S_o$ = 45 g/l and then changed decreasingly at less than C/N = 50, but at more than C/N = 100 it changed increasingly. The effect of C/P ratio and C/S ratio on the yeast growth was also investigated at constant $S_o$ and C/N ratio. ${\mu}$ was dependent on C/P and C/S ratios, but Y, independent on them. But $X_m$ was reliant upon C/P ratio but not upon C/S ratio.

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Rhizoctonia Blight of Azolla japonica Caused by Rhizoctonia solani (Rhizoctonia solani에 의한 큰물개구리밥(Azolla japonica) 마름병)

  • Lee, Jung-Han;Cha, Jea-Yul;Noh, Gil-Han;Han, Ki-Soo;Bae, Dong-Won;Kwon, Young-Sang;Lim, Chae-Shin;Jeong, Sung-Woo;Kwon, Jin-Hyeuk;Park, Chung-Gyoo;Kwak, Youn-Sig
    • Research in Plant Disease
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    • v.17 no.3
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    • pp.405-409
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    • 2011
  • Azolla Lam. is a small aquatic fern with deeply bilobed leaves, which are consisted of a thick greenish, with chlorophyll, upper (dorsal) lobe and a thinner, translucent lower (ventral) lobe, without chlorophyll, submerged in the water. Azolla blight was observed at a lotus pond. Mycological characteristics of the fungus associated with Azolla blight was immediately determined as Rhizoctonia sp. by the thickness and branching of hypha at right angles at the point toward the distal end of septa, with branching hypha is constricted. The fungus produced brown mycelia and dark brown sclerotia on PDA. The optimum temperature for mycelial growth and sclerotia formation were $25^{\circ}C$ and $30^{\circ}C$, respectively. The optimum temperature for fungal infection was $30^{\circ}C$, when spray inoculated. Phylogenetic analysis of rDNA-ITS revealed that the fungus was identified as Rhizoctonia solani (AG-1 IA) closest to one causing rice sheath blight disease. This is the first report on the blight disease of Azolla caused by R. solani in Korea.

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.

Occurrence of Gray Mold on Yacon Caused by Botrytis cinerea (Botrytis cinerea에 의한 야콘 잿빛곰팡이병의 발생)

  • Kim, Jeom-Soon;Lee, Young-Gyu;Kim, Su-Jeong;Hong, Sung-Kee;Choi, Hyo-Won
    • Research in Plant Disease
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    • v.16 no.3
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    • pp.316-319
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    • 2010
  • The gray mold disease occurred on tuberous roots of yacon in storage facilities in Gangneung, Korea, in March 2010. Symptoms typically appeared as in the form of dark brown discoloration on the surface of tuberous roots and water-soaked brown lesions in cross sections of the affected portions. A total of five isolates of Botrytis sp. were obtained from the symptomatic portions. All isolates on potato-dextrose agar (PDA) produced abundant conidia which were pale brown, one-celled, mostly ellipsoid or ovoid in shape and $8.2{\sim}14.8{\times}6.5{\sim}9.9\;{\mu}m$ in size. Large numbers of round to irregular, smooth, black, hard sclerotia were produced on PDA over time. The optimal temperature for mycelial growth and sclerotia formation of the fungal isolates was $20^{\circ}C$. On the basis of morphological and cultural characteristics, all the fungal isolates were identified as Botrytis cinerea. Pathogenicity test on host plants showed that the fungus could infect not only tuberous roots but also leaves and petioles of yacon. This is the first report on gray mold of yacon (Smallanthus sonchifolius) caused by Botrytis cinerea in Korea.

Diffusible and Volatile Antifungal Compounds Produced by an Antagonistic Bacillus velezensis G341 against Various Phytopathogenic Fungi

  • Lim, Seong Mi;Yoon, Mi-Young;Choi, Gyung Ja;Choi, Yong Ho;Jang, Kyoung Soo;Shin, Teak Soo;Park, Hae Woong;Yu, Nan Hee;Kim, Young Ho;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • v.33 no.5
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    • pp.488-498
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    • 2017
  • The aim of this study was to identify volatile and agardiffusible antifungal metabolites produced by Bacillus sp. G341 with strong antifungal activity against various phytopathogenic fungi. Strain G341 isolated from four-year-old roots of Korean ginseng with rot symptoms was identified as Bacillus velezensis based on 16S rDNA and gyrA sequences. Strain G341 inhibited mycelial growth of all phytopathogenic fungi tested. In vivo experiment results revealed that n-butanol extract of fermentation broth effectively controlled the development of rice sheath blight, tomato gray mold, tomato late blight, wheat leaf rust, barley powdery mildew, and red pepper anthracnose. Two antifungal compounds were isolated from strain G341 and identified as bacillomycin L and fengycin A by MS/MS analysis. Moreover, volatile compounds emitted from strain G341 were found to be able to inhibit mycelial growth of various phytopathogenic fungi. Based on volatile compound profiles of strain G341 obtained through headspace collection and analysis on GC-MS, dimethylsulfoxide, 1-butanol, and 3-hydroxy-2-butanone (acetoin) were identified. Taken together, these results suggest that B. valezensis G341 can be used as a biocontrol agent for various plant diseases caused by phytopathogenic fungi.

Corn Cultivation to Reduce the Mycotoxin Contamination (곰팡이 독소 오염 경감을 위한 옥수수 재배법)

  • Kim, Yangseon;Kang, In Jeong;Shin, Dong Bum;Roh, Jae Hwan;Jung, Jingyo;Heu, Sunggi;Shim, Hyeong Kwon
    • Research in Plant Disease
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    • v.23 no.3
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    • pp.256-261
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    • 2017
  • The effects of insecticide and fungicide treatment were investigated to reduce mycotoxin contamination of corn (Zea mays L.) seeds. Deoxynivalenol and zearalenone contents were reduced in the treated seeds, but aflatoxin, ochratoxin A, fumonisin, and T-2 toxin were not effective by chemical treatments. The chemical treatment did not affect the growth of saprophyte, but inhibited the pathogenic fungi such as Fusarium verticillioides, F. graminearum and F. equiseti. Myotoxin contents at different harvesting time were compared. As the harvest time was delayed, both levels of deoxynivalenol and zearalenone and frequency of Fusarium spp. increased. However, the major nutrient contents of corn seeds were not affected by harvesting period. These results show that chemical treatments are necessary to reduce the fungal contamination of corn and harvest without delay is important as well.