• Title/Summary/Keyword: Molecular weight degradation

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Purification and Characterization of Extracellular Aspartic Proteinase of Candida albicans

  • Na, Byoung-Kuk;Lee, Seong-Il;Kim, Sin-Ok;Park, Young-Kil;Bai, Gill-Han;Kim, Sang-Jae;Song, Chul-Yong
    • Journal of Microbiology
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    • v.35 no.2
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    • pp.109-116
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    • 1997
  • An extracellular proteinase of Candida albicans was purified by a combination of 0~75% ammonium sulfate precipitation, DEAE Sepharose Fast Flow ion exchange chromatography, and Sephacryl S-200 HR molecular sieve chromatography. Its mlecular weight was approximately 41 kDa on SDS-PAGE and isoelectric point was 4.4. The enzyme was inhibited by pepstain A. Optimum enzyme activity ranged from pH 2.0 to 3.5 with its maximum at pH 2.5 and a temperature of 45$^{\circ}C$. The addition of divalent cations, $Ca^{2+}$, Zn$^{2+}$ and $Mg^{2+}$, resulted in no significant inhibition of enzymatic activity. However, some inhibitory effects were observed by Fe$^{2+}$, Ag$^{2+}$ and Cu$^{2+}$. With BSA as substrate, an apparent $K_m$ was determined to be 7$\times$10$^{-7}$ M and $K_i$, using pepstatin A as an inhibitor, was 8.05$\times$10$^{-8}$ M. N-terminal amino acid sequence was QAVPVTLXNEQ. Degradation of BSA and fibronectin was shown but not collagen, hemoglobin, immunoglobulin G, or lysozyme. The enzyme preferred peptides with Glu and Leu at the P$_1$ position, but the enzyme activity was highly reduced when the P$_2$ position was phe or pro. This enzyme showed antigenicity against sera of patients with candidiasis.

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Isoaltion and characterization of petroleum degrading bacteria (원유분해세균의 분리 및 특성)

  • Song, Young-Hwan
    • Journal of fish pathology
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    • v.5 no.2
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    • pp.153-158
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    • 1992
  • From several sites of petroleum storage basement in South Coasts in Korea, various petroleum degrading bacteria have been isolated and characterized as Pseudomonas fluorescens, Acinetobacter baumanii, Pseudomonas maltophila and Pseudomonas aeruginosa, respectively. They show the ability of petroleum degradation on minimal media which contains petroleum as sole carbon source and loose the ability at high concentration of NaCl as increasing the concentration of NaCl from 0.5% to 6%. It has been confirmed that such bacteria have utilized the simple saturate hydrocarbon; n-decane, n-hexane, n-octane and n-decane because petroleum consists of various kinds of organic compounds. It has been also identified that petroleum degrading bacteria habor the plasmid and show the antibiotic resistance against ampicillin, tetracycline and chloramphenicol. These results strongly suggest that the petroleum degrading gene and antibiotic resistance gene might be located on the high molecular weight plasmid.

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Preparation and Characterization of PEG-impregnated Aloe Gel through DIS Processing of Aloe vera Leaf Slice (DIS 공정에 의한 Polyethylene Glycol 함침 알로에 베라 겔의 제조 및 특성화)

  • Kwon, Hye Mi;Hur, Won;Lee, Shin Young
    • KSBB Journal
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    • v.28 no.6
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    • pp.356-365
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    • 2013
  • The novel Aloe gels were prepared with dewatering and impregnation by soaking (DIS) processing of Aloe vera leaf slice at four different temperatures (25, 35, 45 and $55^{\circ}C$), using dehydration solution of 40% (w/v) polyethylene glycol (PEG4000). The PEG-impregnation to Aloe vera leaf slice during DIS was observed depending on immersion temperature, and the PEG-impregnated Aloe vera gel (PEG-i-AVG) obtained was characterized using $^1H$ NMR, FT-IR, GPC, XRD and TGA. The PEG-i-AVG had the higher levels of Aloe bioactives (glucomannan and O-acetyl contents) and better quality indices by $^1H$ NMR and FT-IR spectroscopy than those of native Aloe gel. Also, the obtained Aloe gel maintained the bimodal patterns in higher molecular weight region by GPC indicating no degradation of polysaccharide from native Aloe gel. The result observed by SEM confirmed a surface modification by forming the porous structure, and TGA result exhibited better thermal stability than that of native Aloe gel. XRD result revealed that the crystalline structure in Aloe gel was led by incorporation of PEG. Significant decrease of %insolubility and high enhancement of water solubility index were observed, respectively, and highly ordered conformation such as a helix structure was also indicated by Congo red reaction. We concluded that the modification effect for enhancing function of native Aloe gel was successfully obtained by DIS process using PEG as a dehydrating agent. These results suggested that this DIS process had a high potential for developing a new minimally processed product from Aloe vera leaf.

Purification and Characterization of Cyclodextrinase from Bacillus stearothermophilus KJ 16 (Bacillus stearothermophilus KJ16이 생산하는 Cyclodextrinase의 정제와 효소특성)

  • 권현주;유동주;김병우
    • Journal of Life Science
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    • v.8 no.5
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    • pp.497-503
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    • 1998
  • Cyclodextrinase from B. stearothemophilus KJ16 that can produce both cyclodextrin(CD) glucanotransferase and cyclodextrinase was purified 87.6-fold with 7% yield by ammonium sulfate precipitation, DEAE-cellulose chromatog-raphy, Sephadex G-100 chromatography, and FPLC. The molecular weight of the purified enzyme was about 68,000 dalton by SDS-PAGE. The optimal pH and temperature were 6.0 and 55$^{\circ}C$, respectively. The enzyme was stable at 5$0^{\circ}C$ for 2 hr in the pH range of 5.5 and 8.5. The enzyme activity was inhibited strongly by mercaptoethanol, di-thiothreitol, p-chloromercuribenzoate, N-bromosuccinimide, $Cu^{+2}$and $Hg^{+2}$. The purified enzyme hydrolyzed CDs with$\gamma$-CD>$\beta$-CD>$\alpha$-CD. The enzyme also hydrolyzed linear maltodextrins and polysaccharides, but the rates of hyd-rolysis for such substrates were slow as compared to that for $\gamma$-CD. The final degradation products with all substrates were maltose and glucose. Maltose was not further hydrolyzed.

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The Effect of Different Particle Size from PAHs Contaminated Sediment by Ultrasonic Irradiation (PAHs로 오염된 침전물의 초음파 처리시 입자크기가 미치는 영향)

  • Na, Seung-Min;Khim, Jee-Hyeong;Cui, Ming-Can;Ahn, Yun-Gyong;Weavers, Linda K.
    • Journal of Environmental Science International
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    • v.19 no.3
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    • pp.379-387
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    • 2010
  • Sediments of Little Scioto (LS) River in Ohio was contaminated by poor disposal of creosote from Baker Wood Creosoting Facility. Among the primary compounds of creosote, Polycyclic Aromatic Hydrocarbons (PAHs) are the most common ingredient PAHs are known for toxic, carcinogenic and mutagenic compounds. There are many difficulties to remove the PAHs in nature environment because their characteristics are having a less water-solubility, volatile and low mobility properties as increasing the molecular weight. The generation of hydroxyl radicals (${\cdot}OH$) and hydrogen peroxide ($H_2O_2$) forms as well as high temperature (5000 K) and pressure (1000 atm) by a physico-chemical effects of ultrasound during a cavitation collapse can promote the degradation and desorption of PAHs in sediment And it can also produces shock wave and microjets which are able to change the size and surface of particle in solid-liquid system as one of physical effects. Therefore, we explored to understand the role of particle size, the effect of elimination for PAHs concentration by ultrasound and optimize the conditions for ultrasonic treatment. The condition of various size of particles (> $150{\mu}m$, < $150{\mu}m$) and solid-liquid ratio (12.5g/L, 25g/L) for the treatment was considered and ultrasonic power (430 W/L) with liquid - hexane extraction and microwave extraction method were applied after ultrasound treatment.

Characterization of Two Algal Lytic Bacteria Associated with Management of the Cyanobacterium Anabaena flos-aquae

  • Kim, Jeong-Dong;Lee, Choul-Gyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.382-390
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    • 2006
  • Various microorganisms were isolated from the surface waters and sediments of eutrophic lakes and reservoirs in Korea to enable an investigation of bacteria having algal lytic activities against Anabaena flos-aquae when water blooming occurs and to study enzyme profiles of algal lytic bacteria. Two bacterial strains, AFK-07 and AFK-13, were cultured, characterized and identified as Acinetobacter johnsonii and Sinorhizobium sp., respectively. The A. johnsonii AFK-07 exhibited a high level of degradatory activities against A. flos-aquae, and produced alginase, caseinase, lipase, fucodian hydrolase, and laminarinase. Moreover, many kinds of glycosidase, such as ${\beta}-galactosidase,\;{\beta}-glucosidase,\;{\beta}-glucosaminidase,\;and\; {\beta}-xylosidase$, which hydrolyzed ${\beta}-O-glycosidic$ bonds, were found in cell-free extracts of A. johnsonii AFK-07. Other glycosidases such as ${\alpha}-galactosidase,\;{\alpha}-N-Ac-galactosidase,\;{\alpha}-mannosidase,\; and\;{\alpha}-L-fucosidase$, which cleave ${\alpha}-O-glycosidic$ bonds, were not identified in AFK-07. In the Sinorhizobium sp. AFK-13, the enzymes alginase, amylase, proteinase (caseinase and gelatinase), carboxymethyl-cellulase (CMCase), laminarinase, and lipase were notable. No glycosidase was produced in the AFK-13 strain. Therefore, the enzyme system of A. johnsonii AFK-07 had a more complex mechanism in place to degrade the cyanobacteria cell walls than did the enzyme system of Sinorhizobium sp. AFK-13. The polysaccharides or the peptidoglycans of A. flos-aquae may be hydrolyzed and metabolized to a range of easily utilized monosaccharides or other low molecular weight organic substances by strain AFK-07 of. A. johnsonii, while the products of polysaccharide degradation or peptidoglycans were more likely to be utilized by Sinorhizobium sp. AFK-13. These bacterial interactions may offer an alternative effective approach to controlling the water choking effects of summer blooms affecting our lakes and reservoirs.

Effects of ${\rho}-Chlorophenylalanine$ on the Synthesis of Pancreatic Amylase in Rats

  • Kwon, Hyeok-Yil;Eum, Won-Sik;Jang, Hyun-Woo;Lee, Yun-Lyul;Park, Hyoung-Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.4 no.2
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    • pp.129-135
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    • 2000
  • Previously, we have reported that ${\rho}-chlorophenylalanine$ (PCPA), a serotonin depletor, profoundly increased pancreatic fluid and bicarbonate secretion but remarkably inhibited pancreatic amylase secretion in anesthetized rats. The present study was performed to verify the detailed effects of PCPA on pancreatic amylase synthesis that is directly related to amylase exocrine secretion. PCPA significantly decreased pancreatic RNA and protein contents as well as the amylase activity. However, pancreatic DNA content, trypsin and chymotrypsin activities were not influenced by the treatment of PCPA. The rate of pancreatic amylase synthesis, which was assessed by the amount of incorporated $[^{35}S]-methionine$ into amylase for 1 h, was also significantly decreased by 44% in PCPA-treated rats. In order to determine whether the PCPA-induced decrease of amylase synthesis resulted from change in the level of amylase mRNA, Northern blot analysis was performed. The mRNA expression level of amylase was also decreased by 48% in the PCPA-treated rats, indicating that the inhibitory effect of PCPA on the synthesis of pancreatic amylase was mainly regulated at a step prior to translation. It was also revealed in SDS-polyacrylamide gel electrophoresis that the qualitative change of amylase was induced by PCPA. The 54 KDa amylase band seems to be degraded into small molecular weight protein bands in PCPA-treated rats, suggesting that the PCPA- induced decrease of amylase may be partly attributed to the degradation of synthesized amylase.

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Catalytic Recycling of Waste Polymer II. A Study of the Mechanism on the Catalytic Glycolysis of Flexible Polyurethane Foam (촉매를 이용한 폐고분자 물질의 자원화 II. 연질 폴리우레탄 폼의 글리콜분해반응 메카니즘)

  • Park, Chong-Rae;Kim, Sung-Ick;Kim, Young-Chul;Park, Nam-Cook;Seo, Gon
    • Applied Chemistry for Engineering
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    • v.10 no.3
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    • pp.388-393
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    • 1999
  • Polyurethane foams are polymeric material with repeating groups of urethane and urea. When these are heated with ethylene glycol and K acetate catalyst at $200^{\circ}C$, the transesterification of them leads to soluble products. The mechanisms of the reaction were investigated from the molecular weight and the component distributions of the products by GPC and IR analysis. The degradation of the urethane groups was faster than that of urea groups in transesterification reaction. K acetate catalyst accelerated the rate of the transesterification because it had a high ionization tendency. Each reaction, using K or Sr acetate as a catalyst, progressed in the same reaction path but yielded different compositions in products because of the difference of the reaction rate.

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Nutritional Quality and Physicochemical Characteristics of Defatted Bovine Liver Treated by Supercritical Carbon Dioxide and Organic Solvent

  • Kang, Sung-Won;Kim, Hye-Min;Rahman, M. Shafiur;Kim, Ah-Na;Yang, Han-Sul;Choi, Sung-Gil
    • Food Science of Animal Resources
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    • v.37 no.1
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    • pp.29-37
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    • 2017
  • Defatted bovine liver (DBL) is a potential source of protein and minerals. Supercritical carbon dioxide ($SC-CO_2$) and a traditional organic solvent method were used to remove lipid from bovine liver, and the quality characteristics of a control bovine liver (CBL), bovine liver defatted by $SC-CO_2$ ($DBLSC-CO_2$) at different pressures, and bovine liver defatted by organic solvent (DBL-OS) were compared. The $DBLSC-CO_2$ samples had significantly higher (p<0.05) protein, amino acid, carbohydrate, and fiber contents than CBL and DBL-OS. There was a higher yield of lipid from CBL when using $SC-CO_2$ than the organic solvent method. SDS-PAGE analysis demonstrated that the CBL and $DBLSC-CO_2$ had protein bands of a similar intensity and area, whereas DBL-OS appeared extremely poor bands or no bands due to the degradation of proteins, particularly in the 50 to 75 kDa and 20 to 25 kDa molecular weight ranges. In addition, $DBLSC-CO_2$ was shown to have superior functional properties in terms of total soluble content, water and oil absorption, and foaming and emulsification properties. Therefore, $SC-CO_2$ treatment offers a nutritionally and environmentally friendly approach for the removal of lipid from high protein food sources. In addition, $SC-CO_2$ may be a better substitute of traditional organic solvent extraction for producing more stable and high quality foods with high-protein, fat-free, and low calorie contents.

Preparation of BCNU-loaded PLGA Wafers and In Vitro Release Behavior (BCNU 함유 PLGA 웨이퍼의 제조와 생체외 방출거동)

  • 성하수;문대식;강길선;이정식;이해방
    • Polymer(Korea)
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    • v.26 no.1
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    • pp.128-138
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    • 2002
  • 1,3-Bis(2-chloroethyl)-1-nitrosourea (BCNU, Carmustine)-loaded poly(D, L-lactide-co-glycolide) (PLGA, lactide/glycolide mole ratio 75 : 25) microparticles were prepared and fabricated into wafers in an attempt to study the possibility for the treatment of malignant glioma by direct inserting the wafers to the tumor or the cavity remained after surgical resection of the tumor. SEM observation of the microparticles prepared by spray drying method revealed that the microparticles were spherical, i. e. microspheres. Significant reduction of the crystallinity of BCNU encapsulated in PLGA was confirmed by X-ray diffraction and differential scanning calorimetry analyses of the BCNU-loaded PLGA microparticles. Release pattern of BCNU was dependent on several preparation parameters, such as the molecular weight and concentration of PLGA, and initial BCNU loading amount, etc. In vitro release of BCNU was prolonged over 8 weeks with close to zero-order release pattern after initial burst effect. Observations of morphological change of wafers and pH change of release media during release test period confirmed that hydration and degradation of PLGA would be facilitated with an increase of BCNU-loading amount.