• Title/Summary/Keyword: growth modification

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Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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Software Reliability Growth Modeling in the Testing Phase with an Outlier Stage (하나의 이상구간을 가지는 테스팅 단계에서의 소프트웨어 신뢰도 성장 모형화)

  • Park, Man-Gon;Jung, Eun-Yi
    • The Transactions of the Korea Information Processing Society
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    • v.5 no.10
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    • pp.2575-2583
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    • 1998
  • The productionof the highly relible softwae systems and theirs performance evaluation hae become important interests in the software industry. The software evaluation has been mainly carried out in ternns of both reliability and performance of software system. Software reliability is the probability that no software error occurs for a fixed time interval during software testing phase. These theoretical software reliability models are sometimes unsuitable for the practical testing phase in which a software error at a certain testing stage occurs by causes of the imperfect debugging, abnornal software correction, and so on. Such a certatin software testing stage needs to be considered as an outlying stage. And we can assume that the software reliability does not improve by means of muisance factor in this outlying testing stage. In this paper, we discuss Bavesian software reliability growth modeling and estimation procedure in the presence of an imidentitied outlying software testing stage by the modification of Jehnski Moranda. Also we derive the Bayes estimaters of the software reliability panmeters by the assumption of prior information under the squared error los function. In addition, we evaluate the proposed software reliability growth model with an unidentified outlying stage in an exchangeable model according to the values of nuisance paramether using the accuracy, bias, trend, noise metries as the quantilative evaluation criteria through the compater simulation.

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Studies on the Estimation of K2O Requirement for rice through the Chemical Test Data of Paddy Top Soil (화학분석(化學分析)을 통(通)한 수도(水稻)의 가리적량(加里適量) 추정(推定)에 관한 연구(硏究))

  • Kim, Moon Kyu
    • Korean Journal of Agricultural Science
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    • v.2 no.1
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    • pp.61-100
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    • 1975
  • This study has been made to find out the possibilty of successfully using the following $K_2O$ recommended equation $K_2O\;kg/10a=(Ko/\sqrt{Ca+Mg}-Ks/\sqrt{Ca+Mg})sqrt{Ca+Mg}.\;47.\;B\;D$. where $Ko/sqrt{Ca+Mg}=0.03518+0.0007658\;Sio_2/O.M$. $K_Ssqrt{Ca+Mg}$=Exchangeable K me/100g/$\sqrt{Total\;soluble(Ca+Mg)me/100g\;in\;Soil}$ B. D. =Bulk density of top soil, when the dose of Nitrogen for rice is estimated from the following equation: $N\;kg/10a=(4.2+0.096\;SiO_2/O.M).F$ where $F=0.907+0.263x-0.013x^2$ $SiO_2/O.M=(available\;SiO_2=ppm)/(organic\;matter\;%)$in soil For this. two field experiments. one in sandy and the other in clay paddy soil. have been conducted using 3 levels of wollastonite (0, 500, 100kg/10a) as main treatments; 3 levels of $K_2O$ application were used as sub-plots. These were as follows : (1) 8kg of $K_2O$/10a regardless of the K activity-$K/\sqrt{Ca+Mg}$; (2) kg of $K_2O$/10a estimated from the above equation. and (3) same as (2) above plus additional 30% of $K_2O$. The dose of N kg/ 10a was determined from the above equation based on the value of $SiO_2$/O.M. ratio in each treatment. There were three replications. The leading variety of rice in Chung Chong Nam Do area. Akibare (introduced from Japan) was used. The data obtained. through soil and plant analysis and growth and yield observations. have been throughly examined to attain the following summarized conclusions. 1. The nitrogen dose. estimated from the above equation. was in excess for optimum growth of the rice variety Akibare; indicating the necessity of modification onthe value of "F" or the constants in the equation. The concept of using $SiO_2$/O.M. in the equation was shown to be applicable. 2. The dose of potash. estimated from the respective equation given above. also was in excess of the rice requirements indicating the necessity of minor change in the estimation of $Ko/\sqrt{Ca+Mg}$ value and some great modification in the calculation of $Ks/\sqrt{Ca+Mg}$ value for the equation; however the concept of using $K/\sqrt{Ca+Mg}$ as a basis of $K_2O$ recommendation was shown to be quite reasonable. 3. It was found. from the correlation study using the data of paddy yield and amount of $K_2O$ absorbed by rice plants that the substitution of the value of $Ks/\sqrt{Ca+Mg}$ in the equation for the vaule $Ks/\sqrt{Ca+Mg}=0.037+0.78K\;me/100g$ soil was much more applicable than using the value calculated from the data of soil and wollastonite analysis.

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Ligand Based Pharmacophore Identification and Molecular Docking Studies for Grb2 Inhibitors

  • Arulalapperumal, Venkatesh;Sakkiah, Sugunadevi;Thangapandian, Sundarapandian;Lee, Yun-O;Meganathan, Chandrasekaran;Hwang, Swan;Lee, Keun-Woo
    • Bulletin of the Korean Chemical Society
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    • v.33 no.5
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    • pp.1707-1714
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    • 2012
  • Grb2 is an adapter protein involved in the signal transduction and cell communication. The Grb2 is responsible for initiation of kinase signaling by Ras activation which leads to the modification in transcription. Ligand based pharmacophore approach was applied to built the suitable pharmacophore model for Grb2. The best pharmacophore model was selected based on the statistical values and then validated by Fischer's randomization method and test set. Hypo1 was selected as a best pharmacophore model based on its statistical values like high cost difference (182.22), lowest RMSD (1.273), and total cost (80.68). It contains four chemical features, one hydrogen bond acceptor (HBA), two hydrophobic (HY), and one ring aromatic (RA). Fischer's randomization results also shows that Hypo1 have a 95% significant level. The correlation coefficient of test set was 0.97 which was close to the training set value (0.94). Thus Hypo1 was used for virtual screening to find the potent inhibitors from various chemical databases. The screened compounds were filtered by Lipinski's rule of five, ADMET and subjected to molecular docking studies. Totally, 11 compounds were selected as a best potent leads from docking studies based on the consensus scoring function and critical interactions with the amino acids in Grb2 active site.

Health-related Dietary Behaviors and Lifestyle Factors associated with Sodium Hyperingestion in Korean Adults (한국 성인 나트륨 과다섭취군의 남녀별 건강관련 식이행태와 생활양식 특성요인)

  • Kim, Do-Woo;Lee, Moo-Sik;Na, Baeg-Ju;Hong, Jee-Young
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.7
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    • pp.3326-3337
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    • 2013
  • This study was performed to determine health-related dietary and lifestyle of characteristic factors associated with sodium hyperingestion of korean adults by sexual difference. Study subjects were 30~69years (n=1,622) adults who participated in the 2010 Korean National Health and Nutrition Examination Survey (KNHANES V-1). Subjects were divided into the hyperingestion group (HG, n=1,494) and normal ingestion group (NG, n=128) by 'WHO growth standard sodium intakes(2,000mg/day)' and the relationships between Comparison of demographics and physical measurement, eating behaviors and health-related factors were analyzed. By logistic regression method, risk factors for sodium hyperingestion group revealed in this study were gender(female), number of family', 'breakfast', 'lunch', 'dinner', 'nutrition labeling use'. and In case of male, risk factors for sodium hyperingestion group were 'lunch', 'dinner', 'self-rated body image' of eating behaviors. In case of female, Which were number of family' of demographics, 'breakfast', 'lunch', 'dinner' of eating behaviors. In these results, according to different sexual aspect of sodium hyperingestion, we suggest that nutrition label use and proper nutrition intake criteria among sexual, recognition promotion campaign along with education program for repressed sodium hyperingestion related to hypertension prevention of korean adults should include eating habits improvement and demographics lifestyle modification.

Larval Development of Mulberry Longicorn Beetle, Apriona germari Hope, on the Artificial Diet (실내 인공사료육에 의한 뽕나무하늘소(Apriona germari Hope) 유충의 발육)

  • 윤형주;박인균;마영일;설광열
    • Korean journal of applied entomology
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    • v.36 no.4
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    • pp.317-322
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    • 1997
  • Hatched-larvae of the mulberry longicorn beetle, Apriona germari Hope, collected from mulberry fields were reared on artificial diet at 25$^{\circ}$C with 14 h light and 10 h dark to study the larval developmental characteristics. Artificial diet developed for rearing silkworm was used with minor modification adding mulberry branch powder. In case of artificial diet rearing, the head width of larval instar from the I st to the 12th instars was ranged from 0.12 to 0.69cm, and growth rate of each instar was significantly high between the I st and the 2nd instars. In addition, the weight of the 8th instar larvae was increased approximately 176-fold in comparision with that of the 1st instar larvae. Larval duration of each instar took long with larval developmental stages, and that of the 1st to the 9th or the 12th instars was 186.03 or 304.58 days, respectively. The survival rate of larvae was 40.8% by the 8th instar. The pupation rate was approximately 32.4%. Furthermore, although pupation stage was broadly appeared from the 7th to the I lth instars, pupation was majorly observed at the 8th and the 9th instars.

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BIOLOGICAL RESPONSES OF OSTEOBLAST-LIKE CELLS TO DIFFERENT TITANIUM SURFACE BY ANODIZING MODIFICATION

  • Kim Myung-Joo;Kim Chang-Whe;Lim Young-Jun;Park Hyun-Joo
    • The Journal of Korean Academy of Prosthodontics
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    • v.43 no.6
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    • pp.751-763
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    • 2005
  • Statement of problem. To improve a direct implant fixation to the bone, various strategies have been developed focusing on the surface of materials. The surface quality of the implant depends on the chemical, physical, mechanical and topographical properties of the surface. The different properties will interact with each other and a change in thickness of the oxide layer may also result in a change in surface energy, the surface topography and surface, chemical composition. However, there is limited the comprehensive study with regard to changed surface and biologic behavior of osteoblast by anodization. Purpose of study. The aim of this study was to analyze the characteristics of an oxide layer formed and to evaluate the cellular biologic behaviors on titanium by anodic oxidation (anodization) by cellular proliferation, differentiation, ECM formation and gene expression. And the phospholipase activity was measured on the anodized surface as preliminary study to understand how surface properties of Ti implant are transduced into downstream cellular events. Methods and Materials. The surface of a commercially pure titanium(Grade 2) was modified by anodic oxidation. The group 1 samples had a machined surface and other three experimental specimens were anodized under a constant voltage of 270 V(Group 2), 350 V(Group 3), and 450 V(Group 4). The specimen characteristics were inspected using the following five categories; the surface morphology, the surface roughness, the thickness of oxide layer, the crystallinity, and the chemical composition of the oxide layer. Cell numbers were taken as a marker for cell proliferation. While the expression of alkaline phosphatase and Runx2 (Cbfa1) was used as early differentiation marker for osteoblast. The type I collagen production was determined, which constitutes the main structural protein of the extracellular matrix. Phospholipase $A_2$ and D activity were detected. Results. (1) The anodized titanium had a porous oxide layer, and there was increase in both the size and number of pores with increasing anodizing voltage. (2) With increasing voltage, the surface roughness and thickness of the oxide film increased significantly (p<0.01), the $TiO_2$phase changed from anatase to rutile. During the anodic oxidization, Ca and P ions were more incorporated into the oxide layer. (3) The in vitro cell responses of the specimen were also dependant on the oxidation conditions. With increasing voltage, the ALP activity, type I collagen production, and Cbfa 1 gene expression increased significantly (p<0.01), while the cell proliferation decreased. (4) In preliminary study on the relation of surface property and phospholipase, PLD activity was increased but $PLA_2$ activity did not changed according to applied voltage. Conclusion. The anodized titanium shows improved surface characteristics than the machined titanium. The surface properties acquired by anodization appear to give rise more mature osteoblast characteristics and might result in increased bone growth, and contribute to the achievement of a tight fixation. The precise mechanism of surface property signaling is not known, may be related to phospholipase D.

A Study on the Reducing Pollutants in Non-Ruminant Manure by Increasing Feed Utilization (사료이용율 증가에 따른 비반추가축의 분뇨에 의한 공해발생 감소에 관한 연구)

  • Nahm, K.H.
    • Korean Journal of Poultry Science
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    • v.28 no.3
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    • pp.245-257
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    • 2001
  • Localization of livestock facilities leads to concentration of livestock wastes and subsequent leakage of pollutants into the environment, resulting in public concern about their effects. Nitrogen (N) and phosphorus (P) are the most harmful components of animal manure, but odor from the manure itself and the livestock facilities is also a problem. Improving the nutrient efficiency of the livestock helps to decrease excretion of these environmental contaminants. Pigs and chickens are the main experimental models used in studies to improve nutrient efficiency. Addition of feed supplements and modifying feeding systems to improve nutrient efficiency can result in significant decrease in the N, P, odor and dry matter (DM) weight of manure. Examples of these methods include the following. 1) Addition of synthetic amino acids and reducing protein contents resulted N reductions of 10∼27% in broilers, 18∼35% in chicks and layers, 19∼62% in pigs, and a 9∼43% reduction in odor in pigs. 2) Enzyme supplementation resulted in a 12∼15% reduction in DM weight in broiler manure. 3) Phvtase supplementation resulted in P reductions of 25∼35% in chickens and 20∼60% in pigs. 4) Use of growth promoting substances resulted in a 5∼30% reduction in N and a 53∼56% reduction in odor of pigs. 5) Formulating diets closer to requirements (diet modification) reduced N and P by 10∼15% each in chickens and pigs, and odor by 28∼ 79% in pigs. 6) Phase feeding reduced N and P excretion by chicken and pigs from 10∼33% and 10∼13% each, as well as odor in growing and finishing pigs by 49∼79%. 7) Use of highly digestible raw materials in feed reduced N and P excretion by 5% in chickens and pigs.

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A Study on Improvement of Specialized High School of Electrical Field-to-Work Transition by SWOT Analysis (전기·전자·통신 분야 특성화고등학교의 SWOT 분석을 이용한 학교에서 일터로의 전이 개선 방안)

  • Kim, Jinkwon;Woo, Sangho;Lee, Yongjin;Kim, Jinsoo
    • 대한공업교육학회지
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    • v.32 no.1
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    • pp.55-72
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    • 2007
  • The purpose of this study is to improve specialized high school of electrical field-to-work transition by a SWOT Analysis. The subject analyzed the curricula of 12 special highschool and surveyed 192 vocational teachers of them. In this study, a SWOT matrix was filled to improve transition from electric special high school to work, and the results was gotten as follows: First, the government should offer the military service special case for students of electric special high school, strengthen the course of CEO for the school master, and make the system to expand job training. Second, the provincial office of education should aid a functional training with a administrative ability and financial support, offer a list of industry visiting teachers to specialized high school, and promote a modification of the department according to a growth industry of the area. Third, each school should present a school education goal agreed with the purpose of specialized department and intensify the counselling of job selection and the application of the approved textbook.

Conversion of Ginsenoside $Rb_1$ by Ginseng Soil Bacterium Cellulosimicrobium sp. Gsoil 235 According to Various Culture Broths (인삼 토양 미생물 Cellulosimicrobium sp. Gsoil 235의 배지조성에 따른 Ginsenoside $Rb_1$ 전환)

  • Na, Ju-Ryun;Kim, Yu-Jin;Kim, Se-Hwa;Kim, Ho-Bin;Shim, Ju-Sun;Kim, Se-Young;Yang, Deok-Chun
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.55-61
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    • 2009
  • Ginseng saponins (a secondary metabolite, termed ginsenosides) are the principal bioactive ingredients of ginseng, and modification of the sugar chains may markedly change the its biological activity. One of soil bacteria having $\beta$-glucosidase (to transform ginsenoside $Rb_1$) activity was isolated from soil of a ginseng field in Daejeon. 16S rRNA gene sequence analysis revealed that the isolate belonged to the genus Cellulosimicrobium, with highest sequence similarity (99.7%) to Cellulosimicrobium funkei ATCC BAA-$886^T$. The strain, Gsoil 235, could transform ginsenoside $Rb_1$ into Rd, $Rg_3$ and 3 of un-known ginsenosides by the analyses of TLC, HPLC. By investigating its deglycosylation progress, the optimal broth for, $\beta$-glucosidase was nutrient broth (In 48 hours, almost ginsenoside $Rb_1$ could be transformed into minor ginsenosides). On the contrary, the optimal broth for growth was determined as trypic soy broth (TSB).