• Title/Summary/Keyword: Organic Solvents

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Characterization and Purification of Subtilosin A Produced by Bacillus vallismortis MCBL 1012 Isolated from Seasoned Dried Radish

  • Se-Yeon Lee;Dae-Ook Kang
    • Journal of Life Science
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    • v.34 no.8
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    • pp.576-587
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    • 2024
  • In this study, diverse bacterial strains were isolated from fermented foods to screen those with antibacterial activity. Among them, one strain, identified as Bacillus vallismortis MCBL 1012 through 16S rRNA gene sequence analysis, was selected for its bacteriocin production. The culture supernatant of B. vallismortis MCBL 1012 showed antibacterial activity, mainly against Gram-positive bacteria. Scanning electron microscopy (SEM) revealed that bacteriocin treatment led to cellular content leakages in Listeria monocytogenes KCCM 40307, Enterococcus faecium KCCM 12118, and Streptococcus mutans KCTC 3065. PCR analysis confirmed B. vallismortis MCBL 1012 harbored subtilosin A gene (sbo A). Antibacterial activity was decreased by proteolytic enzymes like proteinase K, subtilisin A, and α-chymotrypsin. The bacteriocin demonstrated stability at 40℃ and 60℃ for 120 min, and up to 80℃ for 60 min, with rapid activity loss at 100℃. It retained full antibacterial activity within a pH range of 4.0 to 8.0 and was not affected by up to 100% organic solvents like ethanol, methanol, acetonitrile, and tetrahydrofuran. Nevertheless, activity decreased with more than 40% isopropanol and 80% acetone. Most tested inorganic salts and detergents had no effect on antibacterial activity except, CuSO4 and NiSO4 at specified concentrations. The bacteriocin exerted its antibacterial effect through bactericidal action against L. monocytogenes KCCM 40307. The bacteriocin was purified by ammonium sulfate precipitation, DEAE anion exchange chromatography, and RP-HPLC. The purification resulted in a final yield of 0.03% and a 283.7-fold increase in specific activity. MALDI-TOF MS analysis determined the exact molecular weight of purified bacteriocin to be 3,326.1 Da.

Characterization of a Psychrophilic Metagenome Esterase EM2L8 and Production of a Chiral Intermediate for Hyperlipemia Drug (메타게놈유래의 저온성 에스터라제 EM2L8의 효소적 특성과 이를 활용한 고지혈증 치료제 키랄소재의 생산)

  • Jung, Ji-Hye;Choi, Yun-Hee;Lee, Jung-Hyun;Kim, Hyung-Kwoun
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.118-124
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    • 2009
  • Esterase EM2L8 gene isolated from deep sea sediment was expressed in Escherichia coli BL21 (DE3) and the esterase activity of the cell-free extract was assayed using p-nitrophenyl butyrate-spectrophotometric method. Its optimum temperature was $40-45^{\circ}C$ and 45% activity of the maximum activity was retained at $15^{\circ}C$. The activation energy at $15-45^{\circ}C$ was calculated to be 4.9 kcal/mol showing that esterase EM2L8 was a typical cold-adapted enzyme. Enzyme activity was maintained for 6 h and 4 weeks at $30^{\circ}C$ and $4^{\circ}C$, respectively. When each ethanol, methanol, and acetone was added to the reaction mixture to 15% concentration, enzyme activity was maintained. In the case of DMSO, enzyme activity was kept up to 40% concentration. (S)-4-Chloro-3-hydroxy butyric acid is a chiral intermediate for the synthesis of Atorvastatin, a hyperlipemia drug. When esterase EM2L8 (40 U) was added to buffer solution (1.2 mL, pH 9.0) containing ethyl-(R,S)-4-chloro-3-hydroxybutyrate (38 mM), it was hydrolyzed into 4-chloro-3-hydroxy butyric acid with a rate of $6.8\;{\mu}mole/h$. The enzyme hydrolyzed (S)-substrate more rapidly than (R)-substrate. When conversion yield was 80%, e.e.s value was 40%. When DMSO was added, hydrolysis rate increased to $10.4\;{\mu}mole/h$. The plots of conversion yield vs e.e.s in the presence or absence of DMSO were almost same, implying that the reaction enantioselectivity was not changed by the addition of DMSO. Taken together, esterase EM2L8 had high activity and stability at low temperatures as well as in various organic solvents/aqueous solutions. These properties suggested that it could be used as a biocatalyst in the synthesis of useful pharmaceuticals.

A Study on the Establishment of Management Methods about Occupational Dermatoses (직업성 피부질환에 대한 현황 파악 및 관리 대책 수립을 위한 연구)

  • Lim, Hyun-Sul;Cheong, Hae-Kwan;Choi, Byung-Soon;Kim, Ji-Yong;Sung, Yeol-Oh;Kim, Yang-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.29 no.3 s.54
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    • pp.617-637
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    • 1996
  • Occupational dermatosis is one of the most prevalent occupational disorders. However, the extent of the occupational dermatoses including incidences and prevalencies of each disease entity, and etiologic materials are not yet welt stated in Korea. Authors reviewed the literatures on the statistic data and reports on the occupational dermatoses, and surveyed on the occupational dermatoses in two factories, and surveyed the physicians responsible to the occupational dermatoses with formed questionnaire. The results are as follows; 1. Among medical journals published since 1964, there were 31 articles on the occupational dermatoses. Of 31 articles, 18 were case reports and all others were review articles. Of 18 case reports, 9 were epidemiologic survey. The Workers' Periodic Health Examinations revealed that prevalence of the occupational dermatoses was highest(4.36 per 10,000 workers) in 1974, but number of the cases reported were decreased sharply since 1978 with some tendency to increase since 1981. There were 2,240 reported cases of occupational dermatoses between 1966 and 1992, which is 1.90% of all the reported occupational diseases. Skin infection and injuries due to chemicals were most frequent and there were 6 cases of skin cancer. 2. In an epidemiological survey on the dermatoses among 995 workers in a metal product manufacturing factory and 225 workers in a coal chemical factory, there were 794 with dermatomycosis, 296 workers with acne, 130 workers with scar, 123 workers with deformity of toe nails. Scars, photosensitivity dermatitis, deformity of finger and toe nails, and acne were more prevalent in the metal product manufacturing factory(p<0.05). In the metal prouct manufacturing factory, workers treating organic solvents and oils had more dermatoses than those without treating the materials(p<0.05). On the skin patch performed on 16 workers in the metal product manufacturing factory, there were 8 cases of irritation dermatitis and 5 cases of contact dermatitis. Prevalence of contact dermatitis in the metal product manufacturing factory was 1.3%. 3. On the questionnaire survey, 34 dermatologists, 29 doctors of preventive medicine, and 22 family physician replied. The proportion of occupational etiology among all dermatoses assumed by the physicians were below 9%, and the most important occupational dermatosis in Korea was contact dermatitis. Main etiologic materials related to the occupational dermatosis were organic solvent, acid and alkali, and metals. The reason for the scarcity of report of occupational dermatoses were difficulty in diagnosis and physician's ignorance of the occupational etiology. They replied that to prevent the occupational dermatoses in the workplace, the use of protective devices was most important, and development of diagnostic criteria on the occupational dermatoses is urgent. Above results shows us that there is many workers with occupational dermatoses, but they are mostly unreported. Measures to prevent and manage the occupational dermatoses are not satisfactory at present. Hence, authors suggest measures for the precise diagnosis, report and prevention of the occupational dermatoses. a. Dermatologist, preventive physician, and industrial hygienist should work as a team to examine the high risk group and establish the preventive measures. b. Disease entities, diagnostic criteria of occupational dermatoses should be listed, criteria for the compensation and job fitting at recruitment should be established, and manual for the proper treatment and effective prevention of each occupational dermatosis should be developed. c. Patch test antigens against each occupational category should be developed and it should be available to any physicians responsible. d. To facilitate the diagnosis of occupational dermatoses by the doctors responsible for the Workersr Periodic Health Examination, development of standardized questionnaire, education on the techniques of the patch test, and cooperation with the dermatologist in diagnosis of occupational dermatoses is essential.

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Free Radical Scavenging Effect and Extraction Condition of Ethanol Extracts of Epimedium Koreanum Nakai Containing Different Icariin Quantity (Icariin 함량에 따른 삼지구엽초 추출물의 추출조건 및 항산화 효과)

  • Kim, Seo-Jin;Oh, Deog-Hwan
    • Journal of Food Hygiene and Safety
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    • v.22 no.4
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    • pp.359-364
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    • 2007
  • This study was conducted to determine the optimum condition of extraction and antioxidant activity of ethanol extracts of Epimedium koreanum Nakai based on the icariin quantity. Also, further organic solvent fractions of hexane, chloroform, ethyl acetate and butanol were obtained from the ethanol extract of Epimedium koreanum Nakai at different temperatures. Total ethanol extraction yield of wild grape seed ranged from 11.8% to 39.3% depending on the concentration of icariin as well as different ethanol concentration, extraction temperature and time condition. Among different extraction temperatures and time, the highest extraction yield of 39.3% was obtained at 70% ethanol for 3 hour at $50^{\circ}C$ in the sample containing the 0.596% icariin (KE9412). In the meantime, the strongest free radical scavenging effect $(RC_{50})$ with $18.9{\mu}g/mL$ in the KE9412 sample was observed in 70% ethanol extract of Epimedium koreanum Nakai extracted for 7 hour at $70^{\circ}C$, while $RC_{50}$ with $35.2{\mu}g/mL$ was observed in the KE9405 (0.20% icariin content) sample at same condition. Also, antioxidant activity of ethanol extracts of Epimedium koreanum Nakai increased as icariin concentration increased. Among each fraction obtained from organic solvents, butanol fraction was found to have the strongest $RC_{50}\;(39.2{\mu}g/mL)$ and followed by ethylacetate $(49.0{\mu}g/mL)$, water $(118.8{\mu}g/mL)$, ethylacetate $(119.8{\mu}g/mL)$, and chloroform fraction $(138.5{\mu}g/mL)$ respectively.

Studies on the storage of fresh fruits and vegetables by plastic coating (1) on Rall's Janet Apple- (Plastic Coating에 의(依)한 청과물저장연구(靑果物貯藏硏究) (제1보(第1報)) -사과(국광(國光))에 대(對)하여-)

  • Park, Nou-Pung
    • Applied Biological Chemistry
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    • v.12
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    • pp.89-97
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    • 1969
  • These studies were made on coating of fresh fruits and vegetables with PVC under view on preservation of fresh fruits and vegetables due to restrain water evaporation of them and control gas metabolisms. The results obtained, as selection of PVC materials availble for preservation of them, procedure of the coating, useful organic solvents, concentration of PVC solution and the time of dipping, and effects of the preservation of them, were summarized as follows: 1) PVC powder 222 and PVC powder 443 were surveyed as suitable materials for coating apple, and later was excellent in lustrous aspect. 2) The coating procedure which dipt into PVC solution was suitable to treat much within a short space of time, and using of ventillator accelerate rather evaporation of the organic solvent. 3) Aceton and methyethylketon as solvent of the PVC were, in purpose of storage only, avaible, while aceton was, in view ef taste, better. 4) 10% of the PVC solutions were better to preserve the freshness of apple and 5, 4, 1 and 0.5% of them in order were gradually decreased in preservability, and 15% of them as high concentration were looked like fermentation in one week after the coating. 5) The dipping time was also better n minute than 10 seconds in a preservation but 1-2 minutes could be applied owing to be not affected of the taste due to coating. 6) The freshness of treatment groups were extend about 48 days after coating in room temperature but control lost market value with 12 days. 7) Weight of control was decreased to 10-13% but treatment which was dipped into 10% of coating solution for one minute was decreased to 3.5-4.6% and treatment for four minutes was did to 2.9-3.0%. 8) Change in respiration was less is treatment groups than control in exhaustion of $CO_2$, and water soluble sugar, reduced sugar and pH were not changed almost due to coating. 9) Pannel discussion of the taste was indicated that control was better than treatments soon after coating, but treatments were rather than control last period of storage and treatment with aceton solvent specially was better than others.

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THE EFFECT OF PRIMING ETCHED DENTIN WITH SOLVENT ON THE MICROTENSILE BOND STRENGTH OF HYDROPHOBIC DENTIN ADHESIVE (산 부식된 상아질에 대한 용매를 이용한 프라이밍이 소수성 상아질 접착제의 미세인장접착강도에 미치는 영향)

  • Park, Eun-Sook;Bae, Ji-Hyun;Kim, Jong-Soon;Kim, Jae-Hoon;Lee, In-Bog;Kim, Chang-Keun;Son, Ho-Hyun;Cho, Byeong-Hoon
    • Restorative Dentistry and Endodontics
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    • v.34 no.1
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    • pp.42-50
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    • 2009
  • Deterioration of long-term dentin adhesion durability is thought to occur by hydrolytic degradation within hydrophilic domains of the adhesive and hybrid layers. This study investigated the hypothesis that priming the collagen network with an organic solvent displace water without collapse and thereby obtain good bond strength with an adhesive made of hydrophobic monomers and organic solvents. Three experimental adhesives were prepared by dissolving two hydrophobic monomers, bisphenol-A-glycidylmethacrylate (Bis-GMA) and triethyleneglycol dimethacrylate (TEGDMA), into acetone, ethanol or methanol. After an etching and rinsing procedure, the adhesives were applied onto either wet dentin surfaces (wet bonding) or dentin surfaces primed with the same solvent (solvent-primed bonding). Microtensile bond strength (MTBS) was measured at 48 hrs, 1 month and after 10,000 times of thermocycles. The bonded interfaces were evaluated using a scanning electron microscope (SEM). Regardless of bonding protocols, well-developed hybrid layers were observed at the bonded interface in most specimens. The highest mean MTBS was observed in the adhesive containing ethanol at 48 hrs. With solvent-primed bonding, increased MTBS tendencies were seen with thermo cycling in the adhesives containing ethanol or methanol. However, in the case of wet bonding, no increase in MTBS was observed with aging.

Free Radical Scavenging Effect and Extraction Condition of Ethanol Extracts and Fractions of Wild Grape Seed (Vitis coignetiea) (머루종자의 적정 추출조건 및 추출 분획물의 Free Radical 소거능 효과)

  • Kim, Nan-Young;Kim, Young-Kuk;Bae, Ki-Ja;Choi, Jae-Ho;Moon, Jea-Hak;Park, Geun-Hyung;Oh, Deog-Hwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.755-758
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    • 2005
  • Wild grape is a traditional medicine plant in north-eastern part of Asia and has been known to have healing properties for various illnesses. This study was to determine the optimum extraction condition and antioxidant activity of ethanol extracts of wild grape (V. coignetiae) seed. Also, organic solvent fractions of hexane, chloroform, ethyl acetate and butanol were obtained from the ethanol extract of wild grape seed at different temperatures. Total ethanol extraction yield of wild grape seed ranged from $4\%\;to\;12\%$ depending on the ethanol concentration, extraction temperature and time condition. The highest extraction yield of $11.9\%$ was obtained at $90\%$ ethanol condition for 12 hour at $70^{\circ}C$. However, the strongest free radical scavenging effect $(RC_{50})$ with $20.93\mu g/mL$ was observed in $70\%$ ethanol extract of wild grape seed extracted for 6 hour at $70^{\circ}C,\;while\;RC_{50}\;with\;40.42$\mu g/mL$ was observed in $90\%$ ethanol extract for 12 hour at $70^{\circ}C$. Antioxidant activity of ethanol extracts of wild grape seed increased as total phenol contents increased. Among each fraction obtained from organic solvents, ethyl acetate fraction was found to have the strongest $RC_{50}\;(8.6\mu g/mL)$ and 636.77 mg GAE/g phenol contents .

A Study on the Extraction of Monasil PCA using Liquid CO2 (액체 이산화탄소 이용한 Monasil PCA 추출에 대한 연구)

  • Cho, Dong Woo;Oh, Kyoung Shil;Bae, Won;Kim, Hwayong;Lee, Kab-Soo
    • Korean Chemical Engineering Research
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    • v.50 no.4
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    • pp.684-689
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    • 2012
  • Poly(acrylic acid) (PAA) microspheres is one of the widely-used polymeric materials for the bio-field application and the electric materials. For the synthesis of PAA microspheres, the polymerization technique using surfactants is applied. After the synthesis, the purification and separation processes are required for the removal of surfactant. When general organic solvents were used, many problems, such as huge amount of waste solvent, additional separation processes, and the possibility of residual media, were occurred. Thus, High-pressure Soxhlet extraction using liquid $CO_2$ was developed to solve these problems. In this study, High-pressure Soxhlet extraction of the synthesized PAA microspheres using liquid $CO_2$ was conducted for the removal of Monasil PCA which is used for the dispersion polymerization of acrylic acid in compressed liquid Dimethyl ether (DME). The morphology of the extracted PAA particles was checked by field emission scanning electron microscopy (FE-SEM) and the residual concentration of Monasil PCA was analyzed by inductively coupled plasma - Optical Emission Spectrometer (ICP-OES). For studying the effect of the solvent effect, Soxhlet extraction was conducted using n-hexane, liquid DME, and liquid $CO_2$. In case of n-hexane, some extracted PAA microspheres were produced. However, deformation was also occurred due to the high thermal energy of n-hexane vapor. Liquid DME could not remove Monasil PCA. When using liquid $CO_2$, the extracted PAA microspheres which were free for the residual solvent were produced without deformation. For finding the optimum operating condition, high-pressure Soxhlet extraction was conducted for 8 hours with changing the temperature of reboiler and condenser. When the extractor temperature is $19.6{\pm}0.2^{\circ}C$ and the pressure is $51.5{\pm}0.5$ bar, the best removal efficiency was obtained.

Urinary Metabolites of Dimethylformamide, Methyl Ethyl Ketone, and Toluene exposed Workers in Synthetic Leather Factories (일부 합성피혁 근로자들의 Dimethylformamide, Methyl Ethyl Ketone, Toluene 노출에 따른 요중 대사물질)

  • Choi, Ho-Chun;Kim, Kang Yoon;An, Sun-Hee;Lee, Young-Ja;Chung, Kyou-Chull
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.11 no.2
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    • pp.135-144
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    • 2001
  • This study was performed to measure airborne dimethylformamide(DMF), methyl ethyl ketone(MEK) and toluene and their urinary metabolites concentrations and to determine the relationship between airborne and urinary concentration. Airborne samples and their urinary metabolites were measured 98 male workers who work for 8 synthetic leather factories in a portion of Kyoung-In area. Urine samples were collected at end-of-shift to estimate the exposure levels. 1. The concentration of airborne DMF by process was 8.81 ppm for wet-mixing, 15.05 ppm for wet-coating, 6.03 ppm for dry-mixing, 5.58 ppm for dry-coating, 5.37 ppm for printing, and 9.03 ppm for total. There was statistically significant difference by process. Urinary NMF concentrations of wet-mixing, wet-coating, dry-mixing, dry-coating and printing were $90.55mg/{\ell}$, $79.80mg/{\ell}$, $39.86mg/{\ell}$, $25.23mg/{\ell}$, and $38.15mg/{\ell}$, respectively, and total geometric mean was $56.24mg/{\ell}$. There was significant difference by process. 2. The concentration of airborne MEK by process was 1.89 ppm for wet-mixing, 1.96 ppm for wet-coating, 10.33 ppm for dry-mixing, 29.24 ppm for dry-coating, 14.98 ppm for printing, and 4.87 ppm for total. There was statistically significant difference by process. Urinary MEK concentrations of wetmixing, wet-coating, dry-mixing, dry-coating and printing were $0.93mg/{\ell}$, $0.70mg/{\ell}$, $3.29mg/{\ell}$, $3.29mg/{\ell}$, and $1.06mg/{\ell}$, respectively, and total geometric mean was $1.25mg/{\ell}$. There was statistically significant difference by process. Urinary MEK 3. The concentration of airborne toluene by process was 0.35ppm for wet-mixing, 0.42ppm for wet-coating, 2.95ppm for dry-mixing, 11.67ppm for dry-coating, 4.88ppm for printing, 1.24ppm for total. There was statistically significant difference by process. Urinary hippuric acid concentrations of wet-mixing, wet-coating, dry-mixing, dry-coating and printing were 0.24g/g creatinine, 0.21g/g creatinine, 0.34g/g creatinine, 0.52g/g creatinine, and 0.29g/g creatinine, respctively and total geometric mean was 0.28g/g creatinine. There was statistically significant difference by process. 4. No. of exceeded KPEL was 40 workers(40.8%) for DMF(10ppm), 1 worker(1.0%) for MEK(200ppm), and no worker for toluene(100ppm). No. of exceeded KBEI was 62 workers(63.3%) for urinary NMF($40mg/{\ell}$), 29 workers(29.6%) for urinary MEK, 1 worker(1.0%) for urinary hippuric acid. 5. The regression equations were Log(NMF)=0.4094*Log(DMF)+1.3587(r=0.4516) for DMF, Log(MEKU)=0.1859*Log(MEK)-0.0324(r=0.3303) for MEK, Log(HA)=0.2106*Log(Toluene)-0.5685(r=0.4497) for toluene. Synthetic leather factory workers expose to 3 kinds of organic solvents which are DMF, MEK and toluene. Their urinary NMF and MEK levels were higher than their concentration levels through respiratory. It seems that the urinary levels were affected skin absorption for working habit and alcohol intake.

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Anti-oxidative and Anti-cancer Activities of Extracting of Yacon (Yacon 추출물의 항산화 및 항암 활성)

  • Min, Kyung-Jin;Cheon, Jeong-Un;Cha, Chun-Geun
    • Journal of Food Hygiene and Safety
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    • v.23 no.2
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    • pp.163-168
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    • 2008
  • This study investigated the anti-oxidative and anti-cancer activities by extracting of Yacon by fraction with organic solvents. The content of total phenol was the highest by 45.53% in ethyl acetate fraction and the results of electron donating abilities were 65.20% ($100\;{\mu}g/ml$), 91.81% ($500\;{\mu}g/ml$) and 95.06% ($1000\;{\mu}g/ml$) and those of nitrite scavenging abilities were 11.71% ($100\;{\mu}g/ml$), 36.81% ($500\;{\mu}g/ml$), 59.70% ($1000\;{\mu}g/ml$) in ethyl acetate fraction, which were higher concentration than the control group. Xanthine oxidase inhibitory activities were 23.74% ($100\;{\mu}g/ml$) and 43.41% ($500\;{\mu}g/ml$) in low concentration, which showed higher activities than the control group. After measuring the growth inhibitory activities for cancer cells, it has been found that hexane fraction had the highest growth inhibitory activities by 23.75% ($10\;{\mu}g/ml$), 34.67% ($50\;{\mu}g/ml$), 54.21% ($100\;{\mu}g/ml$) for SNU-1. Meanwhile, hexane fraction demonstrated high growth inhibitory activities by 41.38% ($10\;{\mu}g/ml$), 50.53% ($50\;{\mu}g/ml$), 60.91% ($100\;{\mu}g/ml$) and butanol fraction had 17.05% ($10\;{\mu}g/ml$), 43.87% ($50\;{\mu}g/ml$), 62.99% ($100\;{\mu}g/ml$) for HeLa. Therefore, it has been believed that there were peculiar properties of anti-oxidative and anti-cancer activities depending on each fraction. Furthermore, it has been expected that further studies on the development of functional food and concrete components depending on the components of each fraction to be conducted.