• Title/Summary/Keyword: PKS

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Safety Evaluation of Filamentous Fungi Isolated from Industrial Doenjang Koji

  • Lee, Jin Hee;Jo, Eun Hye;Hong, Eun Jin;Kim, Kyung Min;Lee, Inhyung
    • Journal of Microbiology and Biotechnology
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    • v.24 no.10
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    • pp.1397-1404
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    • 2014
  • A few starters have been developed and used for doenjang fermentation but often without safety evaluation. Filamentous fungi were isolated from industrial doenjang koji, and their potential for mycotoxin production was evaluated. Two fungi were isolated; one was more dominantly present (90%). Both greenish (SNU-G) and whitish (SNU-W) fungi showed 97% and 95% internal transcribed spacer sequence identities to Aspergillus oryzae/flavus, respectively. However, the SmaI digestion pattern of their genomic DNA suggested that both belong to A. oryzae. Moreover, both fungi had morphological characteristics similar to that of A. oryzae. SNU-G and SNU-W did not form sclerotia, which is a typical characteristic of A. oryzae. Therefore, both fungi were identified to be A. oryzae. In aflatoxin gene cluster analysis, both fungi had norB-cypA genes similar to that of A. oryzae. Consistent with this, aflatoxins were not detected in SNU-G and SNU-W using ammonia vapor, TLC, and HPLC analyses. Both fungi seemed to have a whole cyclopiazonic acid (CPA) gene cluster based on PCR of the maoA, dmaT, and pks-nrps genes, which are key genes for CPA biosynthesis. However, CPA was not detected in TLC and HPLC analyses. Therefore, both fungi seem to be safe to use as doenjang koji starters and may be suitable fungal candidates for further development of starters for traditional doenjang fermentation.

Expression of protein kinase C in the spinal cords of rats with autoimmune encephalomyelitis (뇌염모델에서 Protein Kinase C의 발현에 관한 연구)

  • Shin, Tae-Kyun;Kim, Hyung-Min;Tanuma, Naoyuki;Matsumoto, Yoh
    • Korean Journal of Veterinary Pathology
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    • v.1 no.1
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    • pp.26-32
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    • 1997
  • Protein kinase C an enzyme of signal transduction has been known to regulate cell proliferation activation as well as apoptosis in some cancer cell lines. To explore the role of PKC in the course of cell mediated autoimmune disease such as experimental autoimmune encephalomyelitis (EAE) EAE was induced in Lewis rats(6-8 weeks old) with immunization of myelin basic protein supplemented with complete Freund's adjuvants and affected spinal cords were sampled at days 13 postimmunization(PI) as peak stage of EAE and at days 21 PI as recovery stage. The spinal cords with EAE were subjected to Northern blot analysis and insitu hybridization of PKC delta which is one of prominant isotypes of PKC in the haematopoietic cells. Northern blot analysis showed that levels of PKS delta mRNA in the spinal cords of rats withEAE was significantly increased at days 13 PI in which inflammatory cells including T cells and macrophages in the EAE lesions appeared. however the stage. By in situ hybridization signals of PKC delta in EAE lesions was intensely expressed on the delta is also expressed on some brain cells in normal rat central nervous system This finding suggests that PKC plays an important role on either activation of inflammatory cells including encephalitogenic T cells and macrophages or apoptotic elimination of some inflammatory cells depending on the stge of EAE.

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Multi-wavelength Study of Blazars Using Variability as a Tool

  • Baliyan, Kiran S.;Kaur, Navpreet;Chandra, Sunil;Sameer, Sameer;Ganesh, Shashikiran
    • Journal of Astronomy and Space Sciences
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    • v.33 no.3
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    • pp.177-183
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    • 2016
  • Active galactic nuclei (AGN) are too compact to be resolved by any existing optical telescope facility, making it difficult to understand their structure and the emission processes responsible for their huge energy output. However, variability, one of their characteristic properties, provides a tool to probe the inner regions of AGN. Blazars are the best candidates for such a study, and hence a considerable amount of effort is being made to investigate variability in these sources across the electromagnetic spectrum. Here, using the Mt. Abu infrared observatory (MIRO) blazar monitoring program, we present intra-night, inter-night, and long term aspects of the variability in S5 0716+71, 3C66A, and OJ 287. These stars show significant variability on short (a few tens of mins, to a few hours, to a few days) to long term (months to years) timescales. Based on the light travel time argument, the shortest variability timescales (micro-variability) provide upper limits to the size of the emission region. While S5 0716 shows a very high duty cycle of variability (> 80 %), 3C66A shows a much lower intra day variability (IDV) duty cycle (< 20 %). All three show rapid variations within 2.5 to 3.5 hr, which, perhaps, are generated near the vicinity of black holes. Assuming this, estimates of the masses of the black holes are made at ~109, 8×108, and 2.7×109 M for S5 0716+71, 3C66A, and OJ 287, respectively. Multi-wavelength light-curves for the blazar PKS 1510-089 are discussed to infer the emission processes responsible for the recent flaring episodes in this source.

Characterization of Aspergillus sojae Isolated from Meju, Korean Traditional Fermented Soybean Brick

  • Kim, Kyung Min;Lim, Jaeho;Lee, Jae Jung;Hurh, Byung-Serk;Lee, Inhyung
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.251-261
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    • 2017
  • Initially, we screened 18 Aspergillus sojae-like strains from Aspergillus spp. isolated from meju (Korean traditional fermented soybean brick) according to their morphological characteristics. Because members of Aspergillus section Flavi are often incorrectly identified because of their phylogenetic similarity, we re-identified these strains at the morphological and molecular genetic levels. Fourteen strains were finally identified as A. sojae. The isolates produced protease and ${\alpha}-amylase$ with ranges of 2.66-10.64 and 21.53-106.73 unit/g-initial dry substrate (U/g-IDS), respectively, which were equivalent to those of the koji (starter mold) strains employed to produce Japanese soy sauce. Among the isolates and Japanese koji strains, strains SMF 127 and SMF 131 had the highest leucine aminopeptidase (LAP) activities at 6.00 and 6.06 U/g-IDS, respectively. LAP plays an important role in flavor development because of the production of low-molecular-weight peptides that affect the taste and decrease bitterness. SMF 127 and SMF 131 appeared to be non-aflatoxigenic because of a termination point mutation in aflR and the lack of the polyketide synthase gene found in other A. sojae strains. In addition, SMF 127 and SMF 131 were not cyclopiazonic acid (CPA) producers because of the deletion of maoA, dmaT, and pks/nrps, which are involved in CPA biosynthesis. Therefore, A. sojae strains such as SMF 127 and SMF 131, which have high protease and LAP activities and are free of safety issues, can be considered good starters for soybean fermentations, such as in the production of the Korean fermented soybean products meju, doenjang, and ganjang.

Evaluation of Plant Performance during Biomass Co-firing in Pulverized Coal Power Plant (미분탄화력발전에서의 바이오매스 혼소 시 플랜트 성능특성 평가)

  • Mun, Tae-Young;Tefera, Zelalem Tumsa;Lee, Uendo;Lee, Jeung Woo;Yang, Won
    • Journal of the Korean Society of Combustion
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    • v.19 no.3
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    • pp.8-17
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    • 2014
  • The aims of this research were to evaluate effects of biomass co-firing to pulverized coal power plants and the variation of co-firing ratios on the plant efficiency related to power consumption of auxiliary system and flue gas characteristics such as production and component by process simulation based on the existing pulverized coal power plant. In this study, four kinds of biomass are selected as renewable fuel candidates for co-firing: wood pellet(WP), palm kernel shell(PKS), empty fruit bunch(EFB) and walnut shell(WS). Process simulation for various biomass fuels and co-firing ratios was performed using a commercial software. Gas side including combustion system and flue gas treatment system was considering with combination of water and steam side which contains turbines, condenser, feed water heaters and pumps. As a result, walnut shell might be the most suitable as co-firing fuel among four biomass since when 10% of walnut shell was co-fired with 90% of coal on thermal basis, flue gas production and power consumption of auxiliary systems were the smallest than those of other biomass co-firing while net plant efficiency was relatively higher than those of other biomass co-firing. However, with increasing walnut shell co-firing ratios, boiler efficiency and net plant efficiency were expected to decrease rather than coal combustion without biomass co-firing.

A Gene Cluster for the Biosynthesis of Dibenzodioxocinons in the Endophyte Pestalotiopsis microspora, a Taxol Producer

  • Liu, Yanjie;Chen, Longfei;Xie, Qiaohong;Yu, Xi;Duan, Anqing;Lin, Yamin;Xiang, Biyun;Hao, Xiaoran;Chen, Wanwan;Zhu, Xudong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.10
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    • pp.1570-1579
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    • 2019
  • The fungal products dibenzodioxocinones promise a novel class of inhibitors against cholesterol ester transfer protein (CEPT). Knowledge as to their biosynthesis is scarce. In this report, we characterized four more dibenzodioxocinones, which along with a previously described member pestalotiollide B, delimit the dominant spectrum of secondary metabolites in P. microspora. Through mRNA-seq profiling in $g{\alpha}1{\Delta}$, a process that halts the production of the dibenzodioxocinones, a gene cluster harboring 21 genes including a polyketide synthase, designated as pks8, was defined. Disruption of genes in the cluster led to loss of the compounds, concluding the anticipated role in the biosynthesis of the chemicals. The biosynthetic route to dibenzodioxocinones was temporarily speculated. This study reveals the genetic basis underlying the biosynthesis of dibenzodioxocinone in fungi, and may facilitate the practice for yield improvement in the drug development arena.

Assessment of The Biomass Potential Recovered from Oil Palm Plantation and Crude Palm Oil Production in Indonesia (인도네시아 오일 팜 바이오매스 잠재량 평가)

  • Ahn, Byoung-Jun;Han, Gyu-Seoung;Choi, Don-Ha;Cho, Sung-Taig;Lee, Soo-Min
    • Journal of the Korean Wood Science and Technology
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    • v.42 no.3
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    • pp.231-243
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    • 2014
  • In this study, the potential of biomass, which is generated from oil palm cultivation and crude palm oil (CPO) production of Indonesia was assessed in the aspect of energy content. The types of oil palm biomass were classified on the basis of the cultivation stage and the CPO production stage. In the cultivation stage, biomass is considered to be produced from its' root, trunk and frond. Other possible biomass resources such as empty fruit bunch (EFB), palm kernel shell (PKS) and fiber were included in the CPO production stage. As results, total biomass from damaged plantation area of Indonesia was estimated to be annually from 3 million to 16 million tons in 2011. From CPO mills, approximately 49 million tons/yr of biomass residues were estimated to be annually occurred. Their total energy content from each biomass source in cultivation stage was analyzed to be from 593,000 to 3,197,000 TOEs in terms of gross calorific value. In the case of CPO mills, around 22.7 million TOEs was estimated to be potential energy producible by biomass based on gross calorific value of dry basis. If moisture content considered, net calorific value was analyzed to be decreased to 16.3 million TOEs. Based on the results, the total energy contents of all oil palm biomass were estimated to be up to 25,919,000 TOE in terms of gross calorific value. CPO : Crude Palm Oil, EFB : Empty Fruit Bunch, FFB: Fresh Fruit Bunch, PKS : Palm Kernel Shell, OPF : Oil Palm Frond, PKOC : Palm Kernel Oil Cake, ISPO : Indonesia Sustainable Palm Oil Commission, TOE : Tone of Oil Equivalent.

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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Applicability of Various Biomasses to Pulverized Coal Power Plants in Terms of their Grindability (다양한 바이오매스의 분쇄도 실험을 통한 미분탄 화력발전 적용가능성 연구)

  • Kang, Byeol;Lee, Yongwoon;Ryu, Changkook;Yang, Won
    • Clean Technology
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    • v.23 no.1
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    • pp.73-79
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    • 2017
  • Recently usage of biomass is increased in pulverized coal power plants for reduction of $CO_2$ emission. Many problems arise when thermal share of the biomass is increased, and milling of the biomasses is one of the most important problems due to their low grindability when existing coal pulverizer is used. Grindability of coal can be measured through the HGI (Hardgrove grindability index) equipment as a standard, but method of measuring biomass grindability has not been established yet. In this study, grinding experiment of coal and biomass was performed using a lab-scale ball mill. One type of coal (Adaro coal) and six biomasses (wood pellet (WP), empty fruit bunch (EFB), palm kernel shell (PKS), walnut shell (WS), torrefied wood chip (TBC) and torrefied wood pellet (TWP)) were used in the experiment. Particle size distributions of the fuels were measured after being milled in various pulverization times. Pulverization characteristics were evaluated by portion of particles under the diameter of $75{\mu}m$. As a result, about 70% of the TBC and TWP were observed to be pulverized to sizes of under $75{\mu}m$, which implies that they can be used as alternative biomass fuels without modification of the existing mill. Other biomass was observed to have low grindability compared with torrefied biomass. Power consumption of the mill for various fuels was measured as well, and the results show that lower power was consumed for torrefied biomasses. This result can be used for characterization of biomass as an alternative fuel for pulverized coal power plants.

Evolutionary Explanation for Beauveria bassiana Being a Potent Biological Control Agent Against Agricultural Pests

  • Han, Jae-Gu
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.27-28
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    • 2014
  • Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

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