• Title/Summary/Keyword: Total Effective Strain

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Antagonistic Activities of Bacillus spp. Strains Isolated from Tidal Flat Sediment Towards Anthracnose Pathogens Colletotrichum acutatum and C. gloeosporioides in South Korea

  • Han, Joon-Hee;Shim, Hongsik;Shin, Jong-Hwan;Kim, Kyoung Su
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.165-175
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    • 2015
  • Anthracnose is a fungal disease caused by Colletotrichum species that is detrimental to numerous plant species. Anthracnose control with fungicides has both human health and environmental safety implications. Despite increasing public concerns, fungicide use will continue in the absence of viable alternatives. There have been relatively less efforts to search antagonistic bacteria from mudflats harboring microbial diversity. A total of 420 bacterial strains were isolated from mudflats near the western sea of South Korea. Five bacterial strains, LB01, LB14, HM03, HM17, and LB15, were characterized as having antifungal properties in the presence of C. acutatum and C. gloeosporioides. The three Bacillus atrophaeus strains, LB14, HM03, and HM17, produced large quantities of chitinase and protease enzymes, whereas the B. amyloliquefaciens strain LB01 produced protease and cellulase enzymes. Two important antagonistic traits, siderophore production and solubilization of insoluble phosphate, were observed in the three B. atrophaeus strains. Analyses of disease suppression revealed that LB14 was most effective for suppressing the incidence of anthracnose symptoms on pepper fruits. LB14 produced antagonistic compounds and suppressed conidial germination of C. acutatum and C. gloeosporioides. The results from the present study will provide a basis for developing a reliable alternative to fungicides for anthracnose control.

Construction of a Shuttle Vector for Protein Secretory Expression in Bacillus subtilis and the Application of the Mannanase Functional Heterologous Expression

  • Guo, Su;Tang, Jia-Jie;Wei, Dong-Zhi;Wei, Wei
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.431-439
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    • 2014
  • We report the construction of two Bacillus subtilis expression vectors, pBNS1/pBNS2. Both vectors are based on the strong promoter P43 and the ampicillin resistance gene expression cassette. Additionally, a fragment with the Shine-Dalgarno sequence and a multiple cloning site (BamHI, SalI, SacI, XhoI, PstI, SphI) were inserted. The coding region for the amyQ (encoding an amylase) signal peptide was fused to the promoter P43 of pBNS1 to construct the secreted expression vector pBNS2. The applicability of vectors was tested by first generating the expression vectors pBNS1-GFP/pBNS2-GFP and then detecting for green fluorescent protein gene expression. Next, the mannanase gene from B. pumilus Nsic-2 was fused to vector pBNS2 and we measured the mannanase activity in the supernatant. The mannanase total enzyme activity was 8.65 U/ml, which was 6 times higher than that of the parent strain. Our work provides a feasible way to achieve an effective transformation system for gene expression in B. subtilis and is the first report to achieve B. pumilus mannanase secretory expression in B. subtilis.

Experimental validation of a multi-level damage localization technique with distributed computation

  • Yan, Guirong;Guo, Weijun;Dyke, Shirley J.;Hackmann, Gregory;Lu, Chenyang
    • Smart Structures and Systems
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    • v.6 no.5_6
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    • pp.561-578
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    • 2010
  • This study proposes a multi-level damage localization strategy to achieve an effective damage detection system for civil infrastructure systems based on wireless sensors. The proposed system is designed for use of distributed computation in a wireless sensor network (WSN). Modal identification is achieved using the frequency-domain decomposition (FDD) method and the peak-picking technique. The ASH (angle-between-string-and-horizon) and AS (axial strain) flexibility-based methods are employed for identifying and localizing damage. Fundamentally, the multi-level damage localization strategy does not activate all of the sensor nodes in the network at once. Instead, relatively few sensors are used to perform coarse-grained damage localization; if damage is detected, only those sensors in the potentially damaged regions are incrementally added to the network to perform finer-grained damage localization. In this way, many nodes are able to remain asleep for part or all of the multi-level interrogations, and thus the total energy cost is reduced considerably. In addition, a novel distributed computing strategy is also proposed to reduce the energy consumed in a sensor node, which distributes modal identification and damage detection tasks across a WSN and only allows small amount of useful intermediate results to be transmitted wirelessly. Computations are first performed on each leaf node independently, and the aggregated information is transmitted to one cluster head in each cluster. A second stage of computations are performed on each cluster head, and the identified operational deflection shapes and natural frequencies are transmitted to the base station of the WSN. The damage indicators are extracted at the base station. The proposed strategy yields a WSN-based SHM system which can effectively and automatically identify and localize damage, and is efficient in energy usage. The proposed strategy is validated using two illustrative numerical simulations and experimental validation is performed using a cantilevered beam.

A Near-tip Grid Refinement for the Effective and Reliable Crack Analysis by Natural Element Method (효율적이고 신뢰성있는 자연요소 균열해석을 위한 균열선단 그리드 세분화기법)

  • Cho, Jin-Rae
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.32 no.3
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    • pp.183-190
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    • 2019
  • This paper introduces a near-tip grid refinement and explores its usefulness in the crack analysis by the natural element method(NEM). As a sort of local h-refinement in finite element method(FEM), a NEM grid is locally refined around the crack tip showing high stress singularity. This local grid refinement is completed in two steps in which grid points are added and Delaunay triangles sharing the crack tip node are divided. A plane strain rectangular plate with symmetric edge cracks is simulated to validate the proposed local grid refinement and to examine its usefulness in the crack analysis. The crack analysis is also simulated using a uniform NEM grid for comparison. Unlike the uniform grid, the refined grid provides near-tip stress distributions similar to the analytic solutions and the fine grid. In addition, the refined grid shows higher convergence than the uniform grid, the global relative error to the total number of grid points.

Korean Red Ginseng enhances pneumococcal △pep27 vaccine efficacy by inhibiting reactive oxygen species production

  • Lee, Si-On;Lee, Seungyeop;Kim, Se-Jin;Rhee, Dong-Kwon
    • Journal of Ginseng Research
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    • v.43 no.2
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    • pp.218-225
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    • 2019
  • Background: Streptococcus pneumoniae, more than 90 serotypes of which exist, is recognized as an etiologic agent of pneumonia, meningitis, and sepsis associated with significant morbidity and mortality worldwide. Immunization with a pneumococcal pep27 mutant (${{\Delta}}pep27$) has been shown to confer comprehensive, long-term protection against even nontypeable strains. However, ${{\Delta}}pep27$ is effective as a vaccine only after at least three rounds of immunization. Therefore, treatments capable of enhancing the efficiency of ${{\Delta}}pep27$ immunization should be identified without delay. Panax ginseng Mayer has already been shown to have pharmacological and antioxidant effects. Here, the ability of Korean Red Ginseng (KRG) to enhance the efficacy of ${{\Delta}}pep27$ immunization was investigated. Methods: Mice were treated with KRG and immunized with ${{\Delta}}pep27$ before infection with the pathogenic S. pneumoniae strain D39. Total reactive oxygen species production was measured using lung homogenates, and inducible nitric oxide (NO) synthase and antiapoptotic protein expression was determined by immunoblotting. The phagocytic activity of peritoneal macrophages was also tested after KRG treatment. Results: Compared with the other treatments, KRG significantly increased survival rate after lethal challenge and resulted in faster bacterial clearance via increased phagocytosis. Moreover, KRG enhanced ${{\Delta}}pep27$ vaccine efficacy by inhibiting reactive oxygen species production, reducing extracellular signal-regulated kinase apoptosis signaling and inflammation. Conclusion: Taken together, our results suggest that KRG reduces the time required for immunization with the ${{\Delta}}pep27$ vaccine by enhancing its efficacy.

Failure and Flexural Behavior of Reinforced Concrete Beams Strengthened with CFRP Strips (탄소섬유판(CFRP Strip)으로 보강된 철근콘크리트 부재의 파괴거동 및 휨 거동 특성)

  • Lim, Dong Hwan;Park, Sung Hwan
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.28 no.2A
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    • pp.289-295
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    • 2008
  • The purpose of this study was to examine the flexural behavior of reinforced concrete beams strengthened with CFRP strips. A total of 12 rectangular beams were tested. Test variables in this study were the shapes, bonded length and the number of longitudinal layers of CFRP strips. From the experimental study, flexural capacity of the beams strengthened with CFRP strips significantly increased compared to the reinforced concrete beam without a CFRP strip. Maximum increase of ultimate strength was found about 120% more than the control beam. In this test, most of the strengthened beams failed suddenly due to the debonding of CFRP strips. It is also observed that the debonding of the strip was initiated in the flexural zone of the beam and propagated rapidly to the end of the beam. The ultimate tensile strains of CFRP strips in this test were occurred at the level of 36% of rupture tensile strength of the CFRP strip, and an analytical approach to compute the flexural strength of reinforced beams strengthened with CFRP strips based on the effective stresses was conducted.

Selection for Duration of Fertility and Mule Duck White Plumage Colour in a Synthetic Strain of Ducks (Anas platyrhynchos)

  • Liu, H.C.;Huang, J.F.;Lee, S.R.;Liu, H.L.;Hsieh, C.H.;Huang, C.W.;Huang, M.C.;Tai, C.;Poivey, J.P.;Rouvier, R.;Cheng, Y.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.5
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    • pp.605-611
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    • 2015
  • A synthetic strain of ducks (Anas platyrhynchos) was developed by introducing genes for long duration of fertility to be used as mother of mule ducklings and a seven-generation selection experiment was conducted to increase the number of fertile eggs after a single artificial insemination (AI) with pooled Muscovy semen. Reciprocal crossbreeding between Brown Tsaiya LRI-2 (with long duration of fertility) and Pekin L-201 (with white plumage mule ducklings) ducks produced the G0. Then G1 were intercrossed to produce G2 and so on for the following generations. Each female duck was inseminated 3 times, at 26, 29, and 32 weeks of age. The eggs were collected for 14 days from day 2 after AI. Individual data regarding the number of incubated eggs (Ie), the number of fertile eggs at candling at day 7 of incubation (F), the total number of dead embryos (M), the maximum duration of fertility (Dm) and the number of hatched mule ducklings (H) with plumage colour were recorded. The selection criterion was the breeding values of the best linear unbiased prediction animal model for F. The results show high percentage of exhibited heterosis in G2 for traits to improve (19.1% for F and 12.9% for H); F with a value of 5.92 (vs 3.74 in the Pekin L-201) was improved in the G2. Heritabilities were found to be low for Ie ($h^2=0.07{\pm}0.03$) and M ($h^2=0.07{\pm}0.01$), moderately low for Dm ($h^2=0.13{\pm}0.02$), of medium values for H ($h^2=0.20{\pm}0.03$) and F ($h^2=0.23{\pm}0.03$). High and favourable genetic correlations existed between F and Dm ($r_g=0.93$), between F and H ($r_g=0.97$) and between Dm and H ($r_g=0.90$). The selection experiment showed a positive trend for phenotypic values of F (6.38 fertile eggs in G10 of synthetic strain vs 5.59 eggs in G4, and 3.74 eggs in Pekin L-201), with correlated response for increasing H (5.73 ducklings in G10 vs 4.86 in G4, and 3.09 ducklings in Pekin L-201) and maximum duration of the fertile period without increasing the embryo mortality rate. The average predicted genetic response for F was 40% of genetic standard deviation per generation of selection. The mule ducklings' feather colour also was improved. It was concluded that this study provided results for a better understanding of the genetics of the duration of fertility traits in the common female duck bred for mule and that the selection of a synthetic strain was effective method of improvement.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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Effects of Gentiana scabra Bunge var.buergeri Max. Extract on Lipid Metabolism of Serum in Dietary Hyperlipidemic Rats (용담(Gentiana scabra Bunge var.buergeri Max.) 추출액이 식이성 고지혈증 흰쥐의 혈청 지질대사에 미치는 영향)

  • 김한수;김용균;김석환;권오창;손병구;최영환
    • Journal of Life Science
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    • v.8 no.5
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    • pp.532-536
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    • 1998
  • This study was designed to observe the effects of the feeding Gentiana scabra Bunge var.buergeri Max. extract on the improvement of the blood glucose, lipids in the serum of dietary hyperlipidemic rats (S.D. strain, ♂) fed the experimental diets for 4 weeks. Concentration of blood glucose was significantly higher in the cholesterol administration groups (group 2(cholesterol+water), 3(cholesterol+Gentiana scabra Bunge extract) than those in the control group(group 1 (water)). Blood glucose concentration was rather lower in the group 3(cholesterol+Gentiana scabra Bunge extract) than in the group 2(cholesterol+water). There was almost little significance among the groups. Concentration of total cholesterol in serum was significantly lower in the group 3 than those in the group 2. Concentration of LDL-cholesterol in serum was significantly lower in the group 3 than that in the group 2. In the ratio of HDL-cholesterol concentration to total cholesterol concentration, Gentiana scabra Bunge extract administration group was higher percentage than in the group 2. Concentration of free cholesterol and cholesteryl ester in serum was significantly lower in the group 3 than in the group 2. Concentrations of triglyceride and phospholipid in serum were decreased in the group 3 than in the high fat diet group. The activities of aspartate aminotransferase and alanine aminotransferase in serum were increased in the all experimental groups than in the control group. From the above research, the Gentiana scabra Bunge var.buergeri Max. exacts were effective on the improvement of the lipid composition in serum of dietary hyperlipidemic rats.

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Invitro and Virtual Screening of Bioactive Molecule from Mycelium of Trichoderma atroviride Inhibit the UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine Deacetylases (LpxC) for Treatment of Bacterial Infection

  • Saravanakumar, Kandasamy;Park, Cheol-Ho;Wang, Myeong-Hyeon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.67-67
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    • 2018
  • Trichoderma species are a rich source of metabolites, but less known for biomedical potential. This work deals with antibacterial and antioxidant potentials of intracellular non-cytotoxic metabolites, extracted from Trichoderma atroviride (KNUP001). A total of 53 fractions was collected by column chromatography and tested for cytotoxicity by MTT assay. Only one fraction (F41) was found to be non-toxic to Vero cells with $95.4{\pm}0.61%$ of survival. The F41 was then subjected to chemical analysis, antibacterial and antioxidant assays. The F41 at $500{\mu}g.ml^{-1}$ showed the total antioxidant of $48.70{\pm}2.90%$, DPPH radical scavenging activity of $37.25{\pm}2.25$, nitric oxide (NO) radical scavenging activity of $54.55{\pm}1.95$ and $H_2O_2$ radical scavenging activity of $43.75{\pm}3.21$. The F41 at $25{\mu}g.ml^{-1}$ displayed antibacterial activity against E. coli ($14.25{\pm}0.2mm$), P. mirabilis ($10.4{\pm}0.6mm$), S. dysenteriae ($18.6{\pm}03mm$), S. paratyphi A ($14.1{\pm}1.1mm$), E. aerogenes ($5.6{\pm}0.4mm$) and S. marcescens ($14.25{\pm}0.2mm$). GC-MS analysis revealed the dominant presence of oleic acid C 18.1 (63.18%), n-hexadecanoic acid (6.17%), and ethyl oleate (4.93%) and potent molecules such as 8-[(2E)-2-(3-hydroxybenzylidene)hydrazinyl]-1,3,7-trimethyl-3,7-dihydro-1H-purine-2,6-dione, 2-(Dimethylamino)ethyl (1Z)-N-hydroxy-2-(4-morpholinyl)-2-oxoethanimidothioate, Fluorene in the F41, and virtual study revealed that these molecules are likely responsible for the antibacterial activities of F41. Hence, further investigation deserves on purification and characterization of the active metabolites from T. atroviride strain KNUP001 towards developing molecular leads to effective antibacterial drugs, and non-toxic to host cells.

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