• Title/Summary/Keyword: novel strains

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Chemical Constituents of the Culture Broth of Panus rudis

  • Song, Ja-Gyeong;Ha, Lee Su;Ki, Dae-Won;Choi, Dae-Cheol;Lee, In-Kyoung;Yun, Bong-Sik
    • Mycobiology
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    • v.49 no.6
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    • pp.604-606
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    • 2021
  • In our ongoing search for new secondary metabolites from fungal strains, one novel compound (1) and nine known compounds (2-10) were isolated from the EtOAc-soluble layer of the culture broth of Panus rudis. The culture broth of P. rudis was extracted in acetone and fractionated by solvent partition; column chromatography using silica gel, Sephadex LH-20, and Sephadex G-10; MPLC; and HPLC. The structures of isolated compounds were elucidated by one- and two-dimensional NMR and LC-ESI-mass measurements. One new compound, panepoxydiol (1), and nine known compounds, (E)-3-(3-hydroxy-3-methylbut-1-en-1-yl)-7-oxabicyclo[4.1.0]hept-3-ene-2,5-diol (2), isopanepoxydone (3), neopanepoxydone (4), panepoxydone (5), panepophenanthrin (6), 4-hydroxy-2,2-dimethyl-6-methoxychromane (7), 6-hydroxy-2,2-dimethyl-3-chromen (8), 2,2-dimethyl-6-methoxychroman-4-one (9), 3,4-dihydroxy-2,2-dimethyl-6-methoxychromane (10), were isolated from the culture broth of P. rudis. This is the first report of isolation of a new compound panepoxydiol (1) and nine other chemical constituents (2-5, 7-10) from the culture broth of P. rudis.

Inhibition of Verticillium Wilt in Cotton through the Application of Pseudomonas aeruginosa ZL6 Derived from Fermentation Residue of Kitchen Waste

  • Qiuhong Niu;Shengwei Lei;Guo Zhang;Guohan Wu;Zhuo Tian;Keyan Chen;Lin Zhang
    • Journal of Microbiology and Biotechnology
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    • v.34 no.5
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    • pp.1040-1050
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    • 2024
  • To isolate and analyze bacteria with Verticillium wilt-resistant properties from the fermentation residue of kitchen wastes, as well as explore their potential for new applications of the residue. A total of six bacterial strains exhibiting Verticillium wilt-resistant capabilities were isolated from the biogas residue of kitchen waste fermentation. Using a polyphasic approach, strain ZL6, which displayed the highest antagonistic activity against cotton Verticillium wilt, was identified as belonging to the Pseudomonas aeruginosa. Bioassay results demonstrated that this strain possessed robust antagonistic abilities, effectively inhibiting V. dahliae spore germination and mycelial growth. Furthermore, P. aeruginosa ZL6 exhibited high temperature resistance (42℃), nitrogen fixation, and phosphorus removal activities. Pot experiments revealed that P. aeruginosa ZL6 fermentation broth treatment achieved a 47.72% biological control effect compared to the control group. Through activity tracking and protein mass spectrometry identification, a neutral metalloproteinase (Nml) was hypothesized as the main virulence factor. The mutant strain ZL6ߡNml exhibited a significant reduction in its ability to inhibit cotton Verticillium wilt compared to the strain P. aeruginosa ZL6. While the inhibitory activities could be partially restored by a complementation of nml gene in the mutant strain ZL6CMߡNml. This research provides a theoretical foundation for the future development and application of biogas residue as biocontrol agents against Verticillium wilt and as biological preservatives for agricultural products. Additionally, this study presents a novel approach for mitigating the substantial amount of biogas residue generated from kitchen waste fermentation.

Construction of the Genomic Expression Library of Bacillus anthracis for the Immunomic Analysis (면역체 분석을 위한 탄저균 유전자 발현 라이브러리의 구축)

  • Park, Moon-Kyoo;Jung, Kyoung-Hwa;Kim, Yeon-Hee;Rhie, Gi-Eun;Chai, Young-Gyu;Yoon, Jang-W.
    • Korean Journal of Microbiology
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    • v.46 no.1
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    • pp.21-26
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    • 2010
  • As the causative agent of Anthrax, Bacillus anthracis causes an acute fatal disease in herbivores such as cattle, sheep, and horses as well as humans. The therapeutics and prevention of anthrax currently available are based on antibiotics and the live attenuated vaccine strains, which may be problematic due to the emergency of antibiotic resistant strains or residual virulence in those vaccine strains. Therefore, it has been required to develop novel therapeutics and vaccines which are safer and applicable to humans. Recently, the development of the multivalent vaccine targeting both spores and vegetative cells of B. anthracis along with anthrax toxin has been reported. In our attempts to screen potential candidates for those multivalent vaccines, the whole genomic expression library of B. anthracis was constructed in this study. To the end, the partial digests of the genomic DNA from B. anthracis (ATCC 14578) with Sau3AI were ligated with the inducible pET30abc expression vectors, resulting in approximately $1{\times}10^5$ clones in E. coli BL21(DE3). The redundancy test by DNA nucleotide sequencing was performed for the randomly selected 111 clones and found 56 (50.5%) B. anthracis genes, 17 (15.3%) vector sequences, and 38 (34.2%) unknown genes with no sequence homology by BLAST. An inducible expression of the recombinant proteins was confirmed by Western blot. Interestingly, some clones could react with the antiserum against B. anthracis. These results imply that the whole genomic library constructed in this study can be applied for analyzing the immunomes of B. anthracis.

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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Diversity and Antimicrobial Activity of Actinomycetes from Fecal Sample of Rhinoceros Beetle Larvae (장수풍뎅이 유충의 분변에 존재하는 방선균의 다양성 및 항균활성)

  • Lee, Hye-Won;Ahn, Jae-Hyung;Kim, Minwook;Weon, Hang-Yeon;Song, Jaekyeong;Lee, Sung-Jae;Kim, Byung-Yong
    • Korean Journal of Microbiology
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    • v.49 no.2
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    • pp.156-164
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    • 2013
  • Actinomycetes produce diverse secondary metabolites which have the primary importance in medicine, agriculture and food production, and key to this is their ability to interact with other organisms in natural habitats. In this study, we have investigated the taxonomical and functional diversity of actinomycetes in fecal sample of rhinoceros beetle larvae (Allomyrina dichotoma L.) by using culture-dependent and -independent approaches. For the culture-independent approach, the community DNA was extracted from the sample and 16S rRNA genes of actinomycetes were amplified using actinomycetes-specific PCR primers. Thirty-seven clones were classified into 15 genera and 24 species of actinomycetes. For the culture-dependent approach, 53 strains were isolated from larval feces, of which 27 isolates were selected based on morphological characteristics. The isolates were classified into 4 genera and 14 species, and 24 isolates (89%) were identified as the genus Streptomyces. Many of the representative isolates had antimicrobial activities against plant pathogenic fungi and Gram-positive bacteria. In addition, most of the isolates (78%) showed biochemical properties to hydrolyze cellulose and casein. The results demonstrated that diverse and valuable actinomycetes could be isolated from insect fecal samples, indicating that insect guts can be rich sources for novel bioactive compounds.

Diversity and Physiological Characteristics of Culturable Bacteria from Marine Sediments of Ross Sea, Antarctica (남극 로스해 퇴적물로부터 분리된 세균의 다양성 및 생리학적 특성)

  • Lee, Yung Mi;Jung, You-Jung;Hong, Soon Gyu;Kim, Ji Hee;Lee, Hong Kum
    • Korean Journal of Microbiology
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    • v.50 no.2
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    • pp.119-127
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    • 2014
  • The affiliations and physiological characteristics of culturable bacteria isolated from the sediments of Ross Sea, Antarctica were investigated. Sixty-three isolates obtained by cultivation were grouped into 21 phylotypes affiliated with the phyla Actinobacteria and Bacteroidetes and with the classes Alphaproteobacteria and Gammaproteobacteria by phylogenetic analysis of 16S rRNA gene sequences. Based on phylogenetic analysis (<98.65% sequence similarity), approximately 49% of total isolates represented potentially novel species or genus. Among them, extracellular protease, lipase, and exopolysaccharide activities at $10^{\circ}C$ or $20^{\circ}C$ were detected in approximately 46%, 25%, and 32% of the strains, respectively. Forty-three isolates produced at least one type of extracellular material and 21 of them produced at least two extracellular protease, lipase, and/or exopolysaccharides. Our findings indicate that culturable bacterial diversity present within the marine sediments of Ross Sea, Antarctica may contribute to the hydrolysis of the major organic constituents which is closely related with carbon and nitrogen cycling in this environment.

Characterization of Paenibacillus sp. MBT213 Isolated from Raw Milk and Its Ability to Convert Ginsenoside Rb1 into Ginsenoside Rd from Panax ginseng

  • Renchinkhand, Gereltuya;Cho, Soo Hyun;Urgamal, Magsar;Park, Young W;Nam, Joong Hyeon;Bae, Hyung Churl;Song, Gyu Yong;Nam, Myoung Soo
    • Food Science of Animal Resources
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    • v.37 no.5
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    • pp.735-742
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    • 2017
  • This study was conducted to isolate and characterize Paenibacillus sp. MBT213 possessing ${\beta}$-glucosidase activity from raw milk, and examine the enzymatic capacity on the hydrolysis of a major ginsenoside ($Rb_1$). Strain MBT213 was found to have a high hydrolytic ability on ginsenoside $Rb_1$ by Esculin Iron Agar test. 16S rDNA analysis revealed that MBT213 was Paenibacillu sp. Crude enzyme of MBT213 strain exhibited high conversion capacity on ginsenoside $Rb_1$ into ginsenoside Rd proven by TLC and HPLC analyses. The API ZYM kit confirmed that Paenibacillu sp. MBT213 exerted higher ${\beta}$-glucosidase and ${\beta}$-galactosidase activity than other strains. Optimum pH and temperature for crude enzyme were found at 7.0 and $35^{\circ}C$ in hydrolysis of ginsenoside $Rb_1$. After 10 d of optimal reaction conditions for the crude enzyme, ginsenoside $Rb_1$ fully converted to ginsenoside Rd. Ginseng roots (20%) were fermented for 14 d, and analyzed by HPLC showed that amount of ginsenoside $Rb_1$ significantly decreased, while that of ginsenoside Rd was significantly increased. The study confirmed that the ${\beta}$-glucosidase produced by Paenibacillus sp. MBT213 can hydrolyze the major ginsenoside $Rb_1$ and convert to Rd during fermentation of the ginseng. The ${\beta}$-glucosidase activity of this novel Paenibacillus sp. MBT213 strain may be utilized in development of variety of health foods, dairy foods and pharmaceutical products.

Insect Pest Resistance to Insecticides and Future Researches (해충의 살충제저항성과 금후대책)

  • Choi Seung Yoon
    • Korean journal of applied entomology
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    • v.22 no.2 s.55
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    • pp.98-105
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    • 1983
  • The rapid increase in cases of insect resistance to insecticides indicates that the contribution of present chemical control practices inevitably leads to exhaustion of available insecticide resources against key insect species. Now the problem of insecticide resistance exists worldwide among insects and mites affecting field crops and animals including human beings, ranging from minimal or absent in some developing countries, where use of insecticides has been low, to extremely severe in many developed countries. Since the occurrence of insect resistance to insecticides was firstly recognized in 1908, the increase in recent decades has been almost linear and now the number of species of insects and acarines in which resistant strains have evolved have been increased to a total of 432. Of these, $261(60\%)$ are agricultural importance and $171(40\%)$ of medical/veterinary importance. The phenomenon of insecticide resistance is asserting itself as the greatest challenge to effective chemical control of many important insect pests. Resistance of insects to insecticides has a history of nearly 80 years, but its greatest increase and its strongest impact have occurred during the last 40 years following the discovery and extensive use of synthetic organic insecticides and acaricides. The impact of resistance should be considered not only in terms of greater cost of pest control due to increased dosages and number of applications but also in terms of the ecological disruption of pest-beneficial species density relationships, the loss of investment in the development of the insecticides concerned, and socio-economic disruption in agricultural communities. Despite its grave economic consequences, the phenomenon of insecticide resistance has received surprisingly little attention in Korea. Since the study of insecticides started firstly in 1963, many entomologists have been concerned with this study. According to their results, some of the rice pests and some of the mites on orchard trees, for example, have developed worrisome level of resistance in several areas of this peninsula. With many arthropods, considerable advances in the developed countries have been made in the study of the biochemical and physiological mechanisms of resistance. Progress involves the biochemical characteristics of specific defense mechanisms, their genetics, interactions, and their quantitative and qualitative contribution to resistance. But their studies arc still inadequately known and relatively little have been contributed in terms of unique schemes of population management in achieving satisfactory pest control. It is apparent that there is no easy solution to resistance as a general phenomenon. For future challenging to effective control of insect pests which are resistant to the insecticides concerned, new insecticide groups with distinctly novel mode of action are urgently needed. It is clear, however, that a great understanding of the factors which govern the intensity of selection of field population for resistance could lead to far more permanently successive use of chemicals within the framework of integrated pest management than heretofore practiced.

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Isolation, Characterization, and Molecular Cloning of the cDNA Encoding a Novel Phytase from Aspergillus niger 113 and High Expression in Pichia pastoris

  • Xiong, Ai Sheng;Yao, Quan-Hong;Peng, Ri-He;Li, Xian;Fan, Hui-Qin;Guo, Mei-Jin;Zhang, Si-Liang
    • BMB Reports
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    • v.37 no.3
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    • pp.282-291
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    • 2004
  • Phytases catalyze the release of phosphate from phytic acid. Phytase-producing microorganisms were selected by culturing the soil extracts on agar plates containing phytic acid. Two hundred colonies that exhibited potential phytase activity were selected for further study. The colony showing the highest phytase activity was identified as Aspergillus niger and designated strain 113. The phytase gene from A. niger 113 (phyI1) was isolated, cloned, and characterized. The nucleotide and deduced amino acid sequence identity between phyI1 and phyA from NRRL3135 were 90% and 98%, respectively. The identity between phyI1 and phyA from SK-57 was 89% and 96%. A synthetic phytase gene, phyI1s, was synthesized by successive PCR and transformed into the yeast expression vector carrying a signal peptide that was designed and synthesized using P. pastoris biased codon. For the phytase expression and secretion, the construct was integrated into the genome of P. pastoris by homologous recombination. Over-expressing strains were selected and fermented. It was discovered that ~4.2 g phytase could be purified from one liter of culture fluid. The activity of the resulting phytase was 9.5 U/mg. Due to the heavy glycosylation, the expressed phytase varied in size (120, 95, 85, and 64 kDa), but could be deglycosylated to a homogeneous 64 kDa species. An enzymatic kinetics analysis showed that the phytase had two pH optima (pH 2.0 and pH 5.0) and an optimum temperature of $60^{\circ}C$.

Identification and Phylogenetic Analysis of Culturable Bacteria in the Bioareosol from Several Environments (환경 유형에 따른 바이오에어로졸 중 배양성 세균 동정 및 계통분석)

  • Lee, Siwon;Chung, Hyen-Mi;Park, Su Jeong;Choe, Byeol;Kim, Ji Hye;Lee, Bo-Ram;Joo, Youn-Lee;Kwon, Oh Sang;Jheong, Weon Hwa
    • Microbiology and Biotechnology Letters
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    • v.43 no.2
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    • pp.142-149
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    • 2015
  • Bioaerosols are comprised of particles 0.02-100 μm in size that originate in natural and artificial environments, and as a result of human activities. They consist of microorganisms including viruses, bacteria, fungi, and protozoa; fungal spores; microbial toxins; pollen; plant or animal material; expectorated liquid from humans; and glucans (peptidoglycan and β-glucan). Bioaerosols can cause respiratory and other diseases in humans and animals. In this study, bioaerosol samples acquired from agricultural sources, livestock, a sewage treatment plant, a beach, and a pristine area were analyzed to identify and phylogenetically characterize culturable microorganisms. The isolated bacteria exhibited regional differences, with different species dominating. However, Bacillus cereus was isolated in all samples, with a total of 31 strains isolated from all areas, and Acinetobacter baumannii was isolated from an indoor poultry farm. In addition, bacteria determined to be of novel genus or species of the genera Domibacillus, Chryceobacterium, Nocardioides and family Comamonadaceae were isolated from the agricultural, livestock and beach environments.