• Title/Summary/Keyword: streptomyces

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Physiological Characteristics of Actinomycetes Isolated from Turfgrass Rhizosphere (잔디 근권에서 분리된 Actinomycetes균주의 생리학적 특성)

  • Lee, Jung Han;Min, Gyu Young;Shim, Gyu Yul;Jeon, Chang Wook;Kwak, Youn-Sig
    • Weed & Turfgrass Science
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    • v.4 no.4
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    • pp.348-359
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    • 2015
  • Total 443 isolates of actinomycetes were isolated from turfgrass rhizosphere as potential biological control agents. The two isolates (S11 and S4) showed highest cellulase activity with compared to the other isolates that exhibited a clear zone of 1.2 mm around the colony on cellulose agar medium. S12 strain appeared the most active chitin degrading, which exhibited a 1.2 mm of clear zone. The highest proteolytic activity on skim milk agar was which exhibited a 7.5 mm of clear zone by S2 strain. S1 strain from the soli showed siderophore production ability, which exhibited a 0.6 mm of large clear zone on chrome azurol S agar. The antifungal activity of the volatile compound producing by 4 selected actinomycetes was investigated that inhibition rate against Rhizoctonia solani AG2-2 and Sclerotinia homoeocarpa. Growth inhibition effect of S8 isolate against S. homoeocarpa was appeared to 94.8%, S2 to 76.9%, S5 to 46.1% and S12 to 43.5%. The significant inhibition effects on mycelial growth of S. homoeocarpa were shown on media with four strains. The inhibition effect was the highest with S8 strain treatment at 94.8%.

Functional Expression of Soluble Streptavidin in Escherichia coli (수용성 streptavidin의 Escherichia coli 에서 기능적 발현)

  • Han, Seung Hee;Kim, Hyeong Min;Lim, Myeong Woon;Kim, Jin-Kyoo
    • Journal of Life Science
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    • v.25 no.6
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    • pp.631-637
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    • 2015
  • Streptavidin, a protein produced by Streptomyces avidinii, strongly binds up to four molecules of vitamin H, d-biotin exhibiting the dissociation constant of about 10−15 M. This strong binding affinity has been applied for detection and characterization of numerous biological molecules suggesting expression and purification of functional streptavidin should be very useful for the application of this streptavidin-biotin interaction. To express a soluble streptavidin in Escherichia coli, We synthesized streptavidin genes and cloned into pET-22b plasmid, which uses T7 RNA polymerase/T7 promoter expression systems containing pelB leader for secretion into periplasmic space and six polyhistidine tags at C-terminus for purification of expressed proteins. Although streptavidin is toxic to Escherichia coli due to strong biotin binding property, streptavidin was expressed very sufficiently in a range of 10-20 mg/ml. In SDS-PAGE, the size of purified protein was shown as 17 kDa in denatured condition (boiling) and 68 kDa in native condition (without boiling) suggesting tetramerization of monomeric subunit by non-covalent association. Further analysis by size-exclusion chromatography supported streptavidin’s tetrameric structure as well. In addition, soluble streptavidin detected biotinylated proteins in westernblot indicating its functional activity to biotin. Taken these results together, it concluded that our simple expression system was able to show high yield, homotetrameric formation and biotin binding activity analogous to natural streptavidin.

Metabolic Engineering for Resveratrol Derivative Biosynthesis in Escherichia coli

  • Jeong, Yu Jeong;Woo, Su Gyeong;An, Chul Han;Jeong, Hyung Jae;Hong, Young-Soo;Kim, Young-Min;Ryu, Young Bae;Rho, Mun-Chual;Lee, Woo Song;Kim, Cha Young
    • Molecules and Cells
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    • v.38 no.4
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    • pp.318-326
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    • 2015
  • We previously reported that the SbROMT3syn recombinant protein catalyzes the production of the methylated resveratrol derivatives pinostilbene and pterostilbene by methylating substrate resveratrol in recombinant E. coli. To further study the production of stilbene compounds in E. coli by the expression of enzymes involved in stilbene biosynthesis, we isolated three stilbene synthase (STS) genes from rhubarb, peanut, and grape as well as two resveratrol O-methyltransferase (ROMT) genes from grape and sorghum. The ability of RpSTS to produce resveratrol in recombinant E. coli was compared with other AhSTS and VrSTS genes. Out of three STS, only AhSTS was able to produce resveratrol from p-coumaric acid. Thus, to improve the solubility of RpSTS, VrROMT, and SbROMT3 in E. coli, we synthesized the RpSTS, VrROMT and SbROMT3 genes following codon-optimization and expressed one or both genes together with the cinnamate/4-coumarate:coenzyme A ligase (CCL) gene from Streptomyces coelicolor. Our HPLC and LC-MS analyses showed that recombinant E. coli expressing both ScCCL and RpSTSsyn led to the production of resveratrol when p-coumaric acid was used as the precursor. In addition, incorporation of SbROMT3syn in recombinant E. coli cells produced resveratrol and its mono-methylated derivative, pinostilbene, as the major products from p-coumaric acid. However, very small amounts of pterostilbene were only detectable in the recombinant E. coli cells expressing the ScCCL, RpSTSsyn and SbROMT3syn genes. These results suggest that RpSTSsyn exhibits an enhanced enzyme activity to produce resveratrol and SbROMT3syn catalyzes the methylation of resveratrol to produce pinostilbene in E. coli cells.

Antifungal and Proteolytic Activity and Auxin Formation of Bacterial Strains Isolated from Highland Forest Soils of Halla Mountain (한라산 고지대 토양에서 분리한 미생물의 항균 및 단백질분해 활성, 오옥신 생산 특성)

  • Kim, Tack-Soo;Ko, Min-Jung;Lee, Se-Weon;Han, Ji-Hee;Park, Kyung-Seok;Park, Jin-Woo
    • The Korean Journal of Pesticide Science
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    • v.15 no.4
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    • pp.495-501
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    • 2011
  • Bacterial strains were isolated from forest soils of Halla mountain, Jeju island in Korea. The soil samples were collected at each altitude of 100m from 1,000 m above sea level. Total 398 strains were isolated and tested for their physiological characteristics of antagonistic and proteolytic activities, and auxin production. Among the isolates, 172 strains were selected as antifungal strains showing antagonistic activity against at least one of 8 plant fungal pathogens (Alternaria alternata, Botrytis cinerea, Collectotrichum acutatum, Fusarium oxysporum, Phytophthora capsici, Pythium ultimum and Sclerotinia sclerotiorum). In addition 203 strains for proteolytic activity and 26 strains for auxin production were characterized for further study. Je28-4 (Rhodococcus sp.) were showed 80% of control value against tomato gray mold in vivo. Thus, it is suggested that soil bacteria isolated from forest soils of Halla mountain can be important sources of bioactive compounds for improving plant growth or promising biocontrol agents.

A New Potato Cultivar "Jayoung", with High Concentration of Anthocyanin (Anthocyanin 함량이 높은 감자 신품종 "자영")

  • Park, Young Eun;Cho, Ji Hong;Cho, Hyun Mook;Yi, Jung Yun;Seo, Hyo Won;Chung, Myoung Gun
    • Korean Journal of Breeding Science
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    • v.41 no.1
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    • pp.51-55
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    • 2009
  • Jayoung was bred within the potato breeding program of Highland Agriculture Research Center (HARC), National Institute of Crop Science. It was originated from a cross made between Atlantic as a female and AG34314 as a male parent in 2003. Jayoung was evaluated as a clone number Daegwan 1-104 for the growth and tuber characteristics, yielding ability, and resistance to major potato diseases in the regional yield trials from 2006 to 2007. Finally, this clone was renamed as Jayoung and was presented to Korean Seed & Variety Service in 2007 for registration as a new potato cultivar. Jayoung has a oval shape, shallow eye depth, dark purple skinned and fleshed tuber characteristics. Its mean tuber yields from the regional yield trials was 37.3 MT/ha, and it showed the highest yield in spring cropping. Jayoung showed high resistance to both common scab (Streptomyces scabies) and potato leaf roll virus (PLRV), however it was susceptible to late blight (Phytophthora infestans). The dry matter content of Jayoung was fairy high 20.3%, therefore it is suitable for both chip processing and table usage. Its anthocyanin content was the highest in autumn cropping as 43.7 mg/100 g fresh weight and its mean anthocyanin content was 4.7 times higher than that of Jasim.

A New Potato Cultivar "Hongyoung", with Red Skin and Flesh Color, and High Concentrations of Anthocyanins (껍질과 속색이 붉고 Anthocyanin 함량이 높은 감자 "홍영")

  • Park, Young Eun;Cho, Ji Hong;Cho, Hyun Mook;Yi, Jung Yun;Seo, Hyo Won;Choung, Myoung Gun
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.502-506
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    • 2009
  • "Hongyoung" was bred within the potato breeding program of National Institute of Highland Agriculture (NIHA). It was originated from a cross made between Atlantic as a female and AG34314 as a male parent in 2003. "Hongyoung" was evaluated as a clone number Daegwan 1-102 for the growth and tuber characteristics, yielding ability, and resistance to major diseases in the regional yield trials from 2006 to 2007. Finally, this clone was renamed as "Hongyoung" and was presented to Korean Seed & Variety Service in 2007 for registration as a new potato cultivar. "Hongyoung" has a round shape, shallow eye depth, red skinned and fleshed tuber characteristics. Its mean tuber yields from the regional yield trials was 37.4 MT/ha, and it showed the highest yield in spring cropping. "Hongyoung" showed high resistance to common scab (Streptomyces scabies) and potato leaf roll virus (PLRV), however it was susceptible to both potato virus Y (PVY) and late blight (Phytophthora infestans). The dry matter content of "Hongyoung" was low to medium 17.7%, therefore it is suitable for table use. Its anthocyanin content was the highest in autumn cropping with 31.8 mg/100g fresh weight and it contained 3.6 times higher anthocyanin than the control, Jasim.

Investigation of Microbial Communities in Sulculus diversicolor supertexta Through 16S rRNA Sequencing and Antibacterial Monitoring of Harmful Strains (16S rRNA 염기서열 분석을 통한 오분자기(Sulculus diversicolor supertexta)내 미생물 군집 조사 및 인체유해 질병세균에 대한 항균활성 모니터링)

  • Kim, Min-Seon;Lee, Seung-Jong;Heo, Moon-Soo
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1477-1488
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    • 2018
  • This study investigated the muscles, intestines, and gonads of Sulculus diversicolor supertexta to examine the diversity of microbial communities within examples collected from the Jeju Coast. Using different media, initial pure isolation in MA, 1% BHIA, and 1% TSA indicated that the muscles, intestines, and gonads supported more communities, respectively. In analysis of relative similarity with 16s rRNA sequencing, 190 pure colonies were isolated, and further analysis with NBLAST identified 71 species, 39 genera, 25 families, and five phyla. Homogeny with the reference strain was 91-100%. Microbial communities in S. supertexta consisted of gamma and alpha Proteobacteria (48%), Actinobacteria (32.5%), Firmicutes (16.9%), Deinococcus-Thermus (1.3%), and Bacteroides (1.3%). In all tissue, Psychrobacter cibarius in Moraxellaceae was dominant. Alteromonadaceae, Enterobacteriaceae, Pasturellaceae, Moraxellaceae, Rhodobacteraceae, Geminicoccaceae, Dietziaceae, Intrasporangiaceae, Microbacteriaceae, Micrococcaceae, Micromonosporaceae, Streptomycetaceae, Aerococcaceae, Bacillaceae, Paenibacillaceae, Planococcaceae, and Staphylcoccaceae were commonly isolated across all tissues, and Flavobacteriaceae, Corynebacteriaceae, Yesiniaceae, Vibrionaceae, Hahellaceae, Pseudomonadaceae were also identified from the intestines. In microbial monitoring of four harmful bacteria, Streptomyces albus (96%) showed antibacterial activity against all four strains, and Agrococcus baldri (99%) and Psychrobacter nivimaris (99%) presented against E. Coli and E. aerogens. In addition, some strains with low homogeny were isolated and further experiments are therefore required, for example to refine the antimicrobial substances including new strain investigations. These additional experiments would aim to establish generic resources for the microbial communities in S. Supertexta and provide basic data for applied microbiological research.

Plant Growth Promotion and Biocontrol Potential of Various Phytopathogenic Fungi Using Gut Microbes of Allomyrina dichotoma Larva (장수풍뎅이 유충의 장내 미생물을 이용한 다양한 식물 균류병의 생물적 방제 및 생장촉진)

  • Kim, Joon-Young;Kim, Byung-Sup
    • Research in Plant Disease
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    • v.26 no.4
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    • pp.210-221
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    • 2020
  • This research was executed to select beneficial antagonists from digestive organ of Allomyrina dichotoma larva that can be put on environment friendly control against phytopathogenic fungi. We screened 38 bacterial strains inhibiting mycelial growth against eight plant pathogens through dual culture assay. The 10 strains among 38 bacterial strains were selected as beneficial microbes showing antifungal activity against Botrytis cinerea, Plasmodiophora brassicae, Colletotrichum acutatum and Phytophthora capsici through under greenhouse pot trials. The 10 bacterial strains that shown strongest antifungal activity were classified into 3 genera and 10 species, and identified as the genus Bacillus (DM146, DM152, DH2, and DH16), Paenibacillus (DF30, DH14, and DM142) and Streptomyces (DF137, DM48, and DH92) by morphological characteristics and 16s rRNA gene sequence. The 10 bacterial strains had solubilizing activity of insoluble phosphates, production of IAA (indole-3-acetic acid), β-1,3-glucanase and protease. Among the 10 bacterial strains, DM152 strain was produced significant enhancement of all growth parameters of chili pepper and tomato seedlings under greenhouse condition. Thus, this study demonstrated that gut microbes of Allomyrina dichotoma larva will be useful as a potential biocontrol agent against plant pathogens and biofertilizer.

Supplementation of protease and different nutrient density diets in growing-finishing pigs

  • Sehyun Park;Jihwan Lee;Won Yun;Seokman Hong;Hanjin Oh;Dongcheol Song;Seyeon Chang;Jaewoo An;Hyunah Cho;Kyeongho Jeon;Jinho Cho
    • Journal of Animal Science and Technology
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    • v.66 no.2
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    • pp.326-339
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    • 2024
  • This study was conducted to investigate the effects of protease supplementation and different nutrient density of diets in growing-finishing pigs. A total of one hundred-eight crossbred growing pigs ([Landrace × Yorkshire] × Duroc) with an initial body weight (BW; 18.74 ± 3.46 kg) were used for 15 weeks. Pigs were randomly assigned to six dietary treatments with 6 replicates of 3 pigs per pen in a 3 × 2 factorial through the following arrangement: Three groups of protease (1, Basal diets; 2, Protease A: 125 mg/kg protease derived from Streptomyces sps; 3, Protease B: 100 mg/kg protease derived from Bacillus licheniformis) at two different nutrient density diets (1, Basal requirement; 2, 0.94%-0.98% higher than requirement in dietary protein and 50 kcal/kg in energy). High nutrient (HN) diets showed higher average daily gain (ADG) (p < 0.05) and apparent total tract digestibility (ATTD) of crude protein (CP) (p < .0001) compared to basal nutrient (BN) diets during growing periods. Supplementation of protease showed higher BW (p < 0.05) and ADG (p < 0.05) compared to non-supplementation of protease during growing periods. Also, supplementation of protease showed higher ATTD of CP (p < 0.01), ATTD of gross energy (p < 0.05) and decreased blood urea nitrogen (BUN) level (p = 0.001) compared to non-supplementation of protease during finishing periods. Pigs which fed the protease showed decreased ammonia (NH3) emissions (p < 0.05) during experiment periods and decreased hydrogen sulfide (H2S) emissions (p < 0.01) during finishing periods. Interactions between nutrient density and protease were observed, which decreased the feed conversion ratio (p < 0.05) in HN diets without protease compared to BN diets without protease during weeks 4 to 6. Also, interaction between nutrient density and protease was observed, which resulted in improved ATTD of CP (p < 0.01) in response to PTA supplementation with HN diets during the finishing period. In conclusion, supplementation of protease reduces NH3 in feces and BUN in whole blood by increasing the digestibility of CP and improves growth performance. Also, diets with high nutrient density improved growth performance and nutrient digestibility in growing periods.

Effect of FS11052, an Inhibitor of Exocytosis, on Neurite Extension in Rat Hippocampal Neurons and PC12 Cells (신경전달물질 방출 저해제 FS11052가 신경세포와 PC12 세포의 돌기신장에 미치는 영향)

  • Lee Yun-Sik;Kim Dong-Seob
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.315-322
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    • 2006
  • FS11052, a novel microbial metabolite from Streptomyces spp. was identified as a small molecular substance and shown inhibition activities for the release of neurotransmitter from rat hippocampal neuron and PC12 cells. FS11052 is an inhibitor of tritiated norepinephrine ($[^3H]-NE$) release in high $K^+$ buffer solution containing ionomycin, indicating that FS11052 inhibits neurotransmitter release after the influx of $Ca^{2+}$ ions. When examined the effect of FS11052 on glucuronidase release from guinea pig neutrophils, FS11052 inhibited glucuronidase release: when treated with $5{\mu}g/ml$ of FS11052, which was not induced cellular cytotoxicity. The fact that the glucuronidase release in neutrophil and norepinephrine release in neuron was inhibited suggests the similarity in the locations and the mechanisms of FS11052 action targets. When treated with $5{\mu}g/ml$ of FS11052, $[^3H]-NE$ release and neurite extension for both rat hippocampal neurons and PC12 cells were prevented. These observations of FS11052 functioning as an inhibitor of neurotransmitter release suggest that FS11052 has an important role in synaptic transmission in neuron.