Since ${\beta}-aminoethylphosphonic$ acid was discovered in the living organism, the biosynthesis and biological function of aminophosphonic acids have been extensively studied. The purpose of this project consists in the two parts: 1)the preparation of DL-1-amino-2-phenylethylphosphonic acid (Phenylalanine aminophosphonic acid) and DL-1-amino-3-methylbutyl-phosphonic acid (Isoleucine aminophosphonic acid) by the method of Chamber and Isbell. 2) the study of metabolism and biological functions of those synthetic materials by the animal experiment (white rats) The importance of this project proved to be the first experience fed by animals for the elucidation of biochemical and metabolic functions in the animal body. The following organic synthesis of DL-1-amino-3-methylbutylphosphonic acid and DL-1-amino-2-phenylethylphosphonic acid are studied. 1)Synthesis of DL-1-amino-3-methylbutylphosphonic acid a) Synthesis of Iso-butylbromide b) Synthesis of Ethyl iso-butylmalonate c) Synthesis of Iso-caproic acid d) Synthesis of $Ethyl-{\alpha}-bromo$ iso-caproate e) Synthesis of $Triethyl-{\alpha}-phosphono$ iso-caproate f) Synthesis of DL-1-amino-3-methylbutylphosphonic acid 2)Synthesis of DL-1-amino-2-phenylethylphosphonic acid a) Synthesis of Diethyl phosphite b) Synthesis of Ethylchloro acetate c) Synthesis of Triethyl phospho acetate d) Synthesis of Triethyl benzyl phospho acetate e) Synthesis of DL-1-amino-2-phenylethylphosphonic acid The synthetic compounds; DL-1-amino-3-methylbutylphosphonic acid and DL-1-amino-2-phenyl ethylphosphonic acid which are essential amino acid (isoleucine, phenylalanine)analogue are supplemented to the animal diet at the level of 0.2% and 0.4% for isoleucine analogue and 0.35% and 0.7% for phenylalanine analogue. The plain isoleucine and phenylalanine at the same level in the diet are fercilitated as comparable groups in this study. Two sets of experience including 100 male rats were carried out for seven weeks each total 14 weeks. During this period, urine samples, and each big organs were collected for the analysis of total nitrogen, phosphorus, and glycogen contents in the individual samples by Micro Kjeldahl Fisk & Subbarow and Nelson Somogye, method. 1) The result of the project a) The yield of DL-1-amino-3-methylbutylphosphonic acid and DL-1-amino-2-phenylethylphosphonic acid showed low tendency at the level of 12.5% and 20% Melting point of those two compounds were very high and the ${\alpha}-amino$ group in the synthetic compounds showed positive reaction with ninhydrin in the violet color. b) Ail the experimental groups included in this study revealed statistically no significant difference in the organ weight, total body nitrogen retention and urinary phosphorus excretion This means isoleucine aminophosphonic acid and Phenylalanine aminophosphonic acid were utilized in the body as much as the plain amino acids, isoleucine and phenylalanine did. c) The glycogen contents in the liver of the phenylalaine aminophosphonic acid gruop showed higher statistically significant(p<0.05) in the comparision with the group of the Phenylalanine and the Standard-2. It was noteworthy that the higher glycogen content in the liver might indicate the significance in the incorporation of phenylalanine aminophosphonic acid into the intermediate of tricarboxylic acid cycle as activated state.
Advances in bacterial and fungal genome mining uncover a plethora of cryptic secondary metabolite biosynthetic gene clusters. Guided by the genome information, targeted transcriptional derepression could be employed to determine the product of a cryptic gene cluster and to explore its biological role. Monascus spp. are food grade filamentous fungi popular in eastern Asia and several genome data belong to them are now available. We achieved transcription activation of a cryptic fungal polyketide synthase-nonribosomal peptide synthase gene Mpfus1 in Monascus purpureus ${\Delta}MpPKS5$ by inserting Aspergillus gpdA promoter at the upstream of Mpfus1 through double crossover gene replacement. The gene cluster with Mpfus1 show a high similarity to those for the biosynthesis of conjugated polyene derivatives with 2-pyrrolidone ring and the mycotoxin fusarin is the representative member of this group. The ${\Delta}MpPKS5$ is incapable of producing azaphilone pigment, providing an excellent background to identify chromogenic and UV-absorbing compounds. Activation of Mpfus1 resulted in a yellow hue on mycelia and its methanol extract exhibit a maximum absorption at 365 nm. HPLC analysis of the organic extracts indicated the presence of a variety of yellow compounds in the extract. This implies that the product of MpFus1 is metabolically or chemically unstable. LC-MS analysis guided us to predict the MpFus1 product and to propose that the Mpfus1-containing gene cluster encode the biosynthesis of a desmethyl analogue of fusarin. This study showcases the genome mining in Monascus and the possibility to unveil new biological activities embedded in it.
Journal of Korean Society of Environmental Engineers
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v.33
no.7
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pp.523-529
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2011
This study has been performed to examine the influence of the size of particles on the stabilization in the aerobic stabilization equipment connected with MBT system. The biodegradable waste inside the reactor (60% of food waste, 25% of paper waste, 2% of wood waste and 5% of compost) has been charged in same composition. The degree of stabilization was compared and analyzed after charging with adjustment of particle size in 5 mm, 10 mm, 20 mm, 50 mm, 100 mm and state of no separation. The experiment revealed that highest temperature beyond $65^{\circ}C$ was shown in the particle size of less than 50 mm in change of temperature and the highest temperature was about $50^{\circ}C$ in reactor of 100 mm and no separation. The proportionality between generated quantity of $CO_2$ and particle size was not observed, even the highest in generated quantity was shown in over 100 mm. The weight changes based on wet and dry conditions in the reaction process showed the 30% and 46% of reduction in the smallest particle size of 5 mm and it showed the trend of the lower reduction rate at the bigger particle size. The water soluble $COD_{Cr}$ and TOC showed the reduction rate of 60% in reactor of particle size in 100 mm and no separation while the reduction rate comparing to the initial stage of reaction in the reactor of less than 50 mm was 80%. Such result derived the conclusion of acceleration in the decomposing stabilization of biodegradable material due to the decomposing rate of organic substance as the particle size of biodegradable waste gets smaller. It is concluded as necessary to react in adjustment under 50 mm of particle size as much as possible.
Kim, You-Ah;Lee, Jung-Im;Kim, Hae-Jin;Kong, Chang-Suk;Nam, Taek-Jeong;Seo, Young-Wan
Journal of Applied Biological Chemistry
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v.52
no.4
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pp.180-186
/
2009
Whole plants of Vitex rotundifolia were extracted for 2 days with methylene chloride ($CH_2Cl_2$) followed by extraction of the residue for an additional 2 days. The same procedure was also applied using methanol (MeOH). The two crude extracts were combined and partitioned between $CH_2Cl_2$ and $H_2O$. The organic layer was further partitioned between n-hexane and 85% aq. MeOH, and the aqueous layer was also further fractionated with n-BuOH and $H_2O$, successively. From the 85% aq. MeOH fraction, one compound was isolated through the repeated HPLC. According to the results of physicochemical data including NMR and MS, the chemical structure of the compound was determined as artemetin (1). The antiproliferative effects of the crude extracts, fractions, and compound against HT1080, AGS, MCF-7 and HT-29 human cancer cells were compared with the control by using MTT assay. In the comparative analysis, the 85% aq. MeOH fraction exhibited the strongest antiproliferative effects on human cancer cell lines in a dose-dependent manner (p<0.05). In addition, exposure of compound 1 isolated from 85% aq. MeOH fraction led to strong antiproliferative effect in HT1080 cancer cell lines. These results suggest that the extracts and compound isolated from V. rotundifolia may be used as potential chemopreventive and chemotherapeutic agents.
Purpose: Skin pH, an indicator of skin health, is maintained by various organic factors, which include lactate, free amino acid (FAA), and free fatty acid (FFA). As skin ages or with illness, skin pH becomes less acidic, and functional food has been developed to maintain the acidic pH of skin. In this study, we determined the dietary effect of green tea extract (GTE) on skin pH of photo-aged mice, as measured by epidermal levels of lactate, FAA, and FFA. The protein expression and activity of lactate dehydrogenase (LDH), an enzyme of pyruvate reduction for lactate generation, was further determined. Methods: Albino hairless mice were fed a control diet (group UV+) or a diet with 1% GTE (group GTE) in parallel with UV irradiation for 10 weeks. A normal control group was fed a control diet without UV irradiation for 10 weeks (group UV-). Results: Skin pH was higher (less acidic) in group UV+ than in group UV-. In parallel, epidermal levels of lactate and FFA, as well as of LDH protein expression and activity, were reduced in group UV+. Dietary supplementation of GTE (group GTE) reduced skin pH to similar to the level of group UV-, and inversely increased epidermal levels of lactate, LDH protein expression and activity, but not of FFA. Although epidermal levels of FAA were similar in groups UV- and UV+, it was increased in group GTE to a level higher than in group UV-. In further analysis of major FFA, epidermal levels of palmitic acid [16:0], oleic acid [18:1(n-9)], and linoleic acid [18:2(n-6), but not of stearic acid [18:0] in group GTE were similar to or lower than those in group UV+. Conclusion: Dietary GTE normalized skin pH with increased levels of lactate and FAA, as well as with increased protein expression and activity of LDH in the epidermis of UVB irradiated hairless mice.
Kim, Joo-Seok;Lee, Yea-Ji;Yang, Jinfeng;Sa, Yeo-Jin;Kim, Myeong-Ok;Park, Jong-Hyuk;Park, Dong-Sik;Yu, Chang-Yeon;Kim, Myong-Jo
Korean Journal of Plant Resources
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v.26
no.1
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pp.111-118
/
2013
The objective of this study was to determine the biological activities of Sorglum bicolor extracts. Organic fractions, including n-Hexane, EtOAc, and n-BuOH fractions were obtained from the methanol extract of Sorglum bicolor M.. In DPPH radical scavenging activity, $SC_{50}$ values of methanol extract and EtOAc fraction were exhibited $0.66{\pm}0.26{\mu}g/mL$ and $1.03{\pm}0.02{\mu}g/mL$, respectively. Contents of total polyphenol and flavonoids in EtOAc fraction, which were much higher than those of other fractions, were 58.12 mg/g and 4.79 mg/g respectively. Also, effects of reducing power was strongly showed in EtOAc fraction. ${\alpha}$-Glucosidase and ${\alpha}$-amlyase inhibition activities were showed the higher effect in D.W. fraction ($2.83{\mu}g/mL$, $36.64{\mu}g/mL$). In MTT assay in the AGS, HT29 and HCT116 cell lines were significantly higher in the n-BuOH fraction than in the other fractions at $50{\mu}g/mL$ concentration of extracts.
Industrial wastes from pulp and food plants were treated with microorganisms to clarify organic waste-water and to produce cells as animal feed, and results were summarized as follows. (1) Waste-water from pulp, beer, bread yeast, and ethanol distillation plants contained $1.4{\sim}1.5%$ of total sugar, $0.25{\sim}0.35%$ nitrogen, and biological oxygen demand (BOD) was $400{\sim}25,000$, chemical oxygen demand (COD), $500{\sim}28,000$, and pH, $3.8{\sim}7.0$. The BOD and COD were highest in waste-water from ethanol distillation plants among others. (2) Bacterial and yeast counts were $4{\times}10^4-1{\times}10^9,\;2{\times}10^2-7{\times}10^4/ml$ in waste-water. (3) Bacteria grew better in pulp waste and yeasts in beer, bread yeast, and ethanol distillation waste. (4) Saccharomyces cerevisiae SAFM 1008 and Candida curvata SAFM 70 were the most suitable microorganisms for clarification of ethanol distillation waste. (5) When liquid and solid waste from ethanol distillation were treated with microbial cellulase, xylanase, and pectinase, solid waste was reduced by 36%, soluble waste was increased, and recuding sugar content was increased by 1.3 times which provided better medium than untreated waste for cultivation of yeasts. (6) Optimum growth conditions of the two species of yeast in ethanol distillation waste were pH 5.0, $30^{\circ}C$, and addition of 0.2% of urea, 0.1% of $KH_2PO_4$ and 0.02% of $MgSO_4$. (7) Minimum number of yeast for proper propagation was $1.8{\times}10^5/ml$. (8) C. curvata70 was better than cerevisae for the production of yeast cells from ethanol distillation waste treated with microbial enzymes. (9) S. cerevisiae produced 16 g of dried cell per 1,000ml of ethanol distillation waste and reduced BOD by 46%. C. curvata produced 17.6g of dried cell and reduced BOD by 52% at the same condition. (10) Yeast cells produced from the ethanol distillation waste contained 46-52% protein indicating suitability as a protein source for animal feed.
To investigate the characteristics of anodic film formed on metals and the reactivities of organic inhibitor Di-iso-butylnitrosoamine (DBNA) in sea water. the cathodic reactions of anodic film formed on metals were carried out by using the potential drop method and galvanostatic method at $25^{\circ}C$. The investigated results are as follows: The anodic films formed on aluminum and zinc in 60mM $DBNA+9\%_{\circ}$ sea water did not show-changes of potential drop. However, those formed on lead and copper were reasonable electrodes. It was concluded that cathodic reactivities of proton through the anodic film in aqueous solution were constant without regard to the kinds of metals used with anodic film electrode at operated current density ranges, because the values of transition time obtained in 0.5M sodium chloride solution and $9\%_{\circ}$ sea water nearly coincided. The values of transition time of the first step by the galvanostatic method were obtained from 0.22 to 1.40 sec ranges far less than one minute. Therefore, it suggested that cathodic reactions of proton through the anodic film were mainly controlled by diffusion/adsorption process. The differences of between $\tau_{1}/4$ in $9\%_{\circ}$ sea water and $\tau_{1}/4$ in 60mM $DBNA+9\%_{\circ}$ sea water, and between $E_{1}/4$ in sea water and $E_{1}/4$ in 60mM $DBNA+9\%_{\circ}$ sea water at the constant current density with $1.9\times10^{-4}\sim5.0{\times}10^{-6}\;amp/cm^2$ were 0.06 sec and 0.53 v. respectively and cathodic reactions of DBNA on the anodic film electrodes were chiefly controlled by adsorption/diffusion process. The reason that adsorption quantities of proton on anodic film formed on aluminum and zinc in aqueous solutions were much more than those on lead and copper, seems to lie due mostly to the number of porosity produced on anodic film used.
Ko, Dae-Geun;Choi, Ji-Woong;Lim, Byung Jin;Park, Jong Hwan;An, Kwang-Guk
Korean Journal of Environment and Ecology
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v.26
no.6
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pp.892-901
/
2012
The purpose of this research was to analyze fish fauna, distribution patterns, and fish community structures in the Juksan-Weir regions of upper weir(R-UW) and lower weir(R-LW), which was constructed in 2011 on Yeongsan River watershed. Our outcomes will provide valuable information in the impact analysis of weir constructions next decades, even if the current research is limited to distributions and fauna studies of fishes. The fishes observed were 8 family and 30 species in all the sites, and the number of the species in the regions of R-UW and R-LW was 23(938 individuals) and 27(1,680 individuals), respectively, indicating greater abundance in the lower region of Juksan-Weir. Most dominant species was Opsarichthys uncirostris amurensis with relative abundance of 42%, and this species had greater abundance in the R-LW(49%) than in the R-UW(30%). Coilia nasus, was the migratory fish sampled in this region and had no differences of abundance between the two regions. The extic species observed were Carassius cuvieri, Lepomis macrochirus, and Micropterus salmoides. The two formers of Carassius cuvieri(1.2%) and Lepomis macrochirus(3.7%) dominated in the R-UW, whereas Micropterus salmoides had greater dominance in the region of R-LW. According to tolerance guild analysis, the proportion of sensitive species ranged from 0.4 to 0.5%, and that of tolerant species ranged from 65% to 70%, implying a severe pollution of nutrients and organic matters in the water body. In the mean time, according to trophic guild analysis, the proportion of carnivores(57 - 77%) was evidently higher than that of insectivores(10 - 20%), indicating a dominance of the carnivores. Especially, high abundance of the bass, exotic species, may disturb the fish overall food chain, so the efficient ecosystem management may be required for the conservation of endemic species. When we examined fish data files of last 60 years, the species of Culter brevicauda, designated as endangered species, was observed firstly in our research area and the total number was 5 individuals(0.2% of the total). So, this region observed should be conserved as a protection zone and further monitoring studies should be done in the future.
Oh, Hyun Hee;Huh, Chang Ki;Choi, Ha Nuel;Yang, Hee Sun;Bae, In Hyu;Lee, Jai Sung;Jeong, Yong Seob;Lee, Nam Keun;Jung, Hoo Kil
Journal of Dairy Science and Biotechnology
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v.31
no.2
/
pp.133-141
/
2013
This study was performed to identify the cheese starter potential of antifungal lactic acid bacteria isolated from Kimchi. Eight fungi were isolated from cheese or the cheese ripening room, and identified as Penicillium and Cladosporium by ITS-5.8S rDNA analysis. Twenty-two lactic acid bacteria species with antifungal activity were isolated from Kimchi, and identified as Lactobacillus and Pediococcus by 16S rRNA sequence analysis. Six lactic acid bacteria species were selected (L. sakei subsp. ALJ011, L. sakei subsp. ALI033, L. sakei subsp. ALGy039, P. pentosaceus ALJ015, P. pentosaceus ALJ024, and P. pentosaceus ALJ026) based on higher antifungal activity from the initial 22 species. Out of the six identified species, L. sakei subsp. ALI033 had the highest antifungal activity. For growth of the six lactic acid bacteria, optimal temperature and pH were $30{\sim}37^{\circ}C$ and 7.0, respectively. Proteolytic activities of the six lactic acid bacteria were almost as strong as the commercial strain Str. thermophilus Body-1. Coagulative activities of L. sakei subsp. ALI033, P. pentosaceus ALJ015, and P. pentosaceus ALJ024 were higher than those of L. sakei subsp. ALJ011, L. sakei subsp. ALGy039, and P. pentosaceus ALJ026. The acid resistance of L. sakei subsp. was higher than that of P. pentosaceus. The major organic acid component of the lactic acid bacteria culture medium was lactic acid.
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