• Title/Summary/Keyword: Acid.Alkaline treatment

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Purification and Characterization of Fibrinolytic Enzyme Produced by Bacillus subtilis K7 Isolated from Korean Traditional Soy Sauce (한국재래간장 발효균 Bacillus subtilis K7 유래의 혈전용해 Protease의 정제 및 특성)

  • Kim, Doo-Young;Lee, Eun-Tag;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.46 no.3
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    • pp.176-182
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    • 2003
  • An alkaline fibrinolytic protease-producing bacteria was isolated front Korean traditional soy sauce and identified as Bacillus subtilis K7 from the results of analyses of its morphological and physiological properties, $API^{\circledR}$, and Biolog system. The enzyme was purified by 75% ammonium sulfate fractionation, QAE-Sephadex anion and SP-Sephadex cation exchange column chromatography and Sephadex G-100 gel filtration. The specific activity of the purified enByme was 233.9 unit/mg protein and the yield of enzyme was 3.8%. The homogeneity of the purified enzyme was confirmed by polyacrylamide gel electrophoresis. Molecular mass of the enzyme was estimated about 21,500 Da by SDS-polyacrylamide get electrophoresis and gel chromatography. The optimum temperature and pH for the enzyme activity were $40^{\circ}C$ and 9.0, respectively. The enzyme was stable in a pH range of 5.0 to 12.0, and 60% of its activity was lost on heat treatment at $50^{\circ}C$ for 20 min. The activity of the purified enzyme was inhibited by the presence of $Fe^{2+},\;Ag^{2+},\;Cu6{2+}$, iodoacetate, ethylene diamine tetraacetic acid (EDTA), and trans-1,2-diaminocycloheane-N,N,N',N'-tetraacetic acid (CDTA). The results indicates that the enzyme requires a metal ion for its enzymatic activity.

Protein Carboxyl Ο-Methylation in Porcine Liver and Testis (돼지 간 및 정소에서 단백질 카르복실메칠화 현상)

  • 조재열;김성수;이향우;홍성렬
    • YAKHAK HOEJI
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    • v.45 no.1
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    • pp.46-54
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    • 2001
  • Protein carboxyl Ο-methylation is a kind of enzymatic reaction producing carboxyl methylester catalyzed by protein carboxyl Ο-methyltransferases at the carboxyl group of amino acid residues in polypeptide. Since the finding of carboxyl methylesterl many studies have been focused on the under-standing of biological functions in eukaryotes but still not clear except for roles in Ras attachment to membrane and protein repair. In this study, we investigated the protein carboxyl methylation in porcine liver and testis in respect of identification and characterization of carboxyl methylesters and natural proteinous substrates using pH stability of the esters and electrophoresis under acidic and basic conditions. We detected several kinds of methyl esters, 3 kinds each in cytosolic fractions from liver and testis. Under the treatment of strong acid and base, the ratio between base-stable substrates and unstable ones in liver (4 : 6) was different from the ratio obtained in testis (6 : 4). The methyl accepting capacities were affected by enzymatic proteolysis between the range of 55 to 65% in liver and of 35 to 45% in testis. Separation of the methylated proteins by acidic electrophoresis in the presence of urea and SDS revealed distinctively natural substrates of 26, 33 and 80 kD in the cytosol from liver and of 14, 25, 32 and 86 kD from testis. Most of the labelling, however were lost following electrophoresis under moderate alkaline condition, except for molecules of newly detected 7 and 17 kD in livers and 15, 29, 40 and 80 kD in testis. From these results, it was proposed that protein carboxyl Ο-methylation in each organs may be catalyzed by different classes of protein carboxyl Ο-methyltransferases. In addition, it is suggested that the protein carboxyl methylation in liver and testis may have different patterns in respect of natural substrates.

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Effect of Ssangwha-tang Fermented by Lactobacillus fermentum on Osteoclast Differentiation and Osteoporosis of Ovariectomized Rats (Lactobacillus fermentum으로 발효한 쌍화탕의 파골 세포 분화와 난소 적출한 랫트의 골다공증에 미치는 영향)

  • Shim, Ki-Shuk;Lee, Ji-Hye;Lee, Jae-Hoon;Ma, Jin-Yeul
    • Korean Journal of Oriental Medicine
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    • v.16 no.1
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    • pp.149-155
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    • 2010
  • Objective : Ssangwha-tang is a traditional medicine formula widely prescribed for a decrease of fatigue after an illness in Korea. The aim of this study is to investigate the effect of Ssangwha-tang fermented by Lactobacillus fermentum (SF) on osteoclast differentiation in vitro and on bone metabolism of an ovariectomized rat in vivo. Methods : Tartrate-resistant acid phosphatase activity and staining were applied to evaluate the formation of osteoclasts. Ovariectomized rats were orally administrated with SF (30 ml/kg/day) for 12 weeks. Serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, total cholesterol, high-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol, triglyceride, phosphate, calcium levels were determined. Effect of SF on bone loss were studied by histological analysis and the measurement of bone mineral density. Results : SF significantly inhibited tartrate-resistant acid phosphatase activity and formation of osteoclasts in RAW264.7 cells stimulated by receptor activator for nuclear factor ${\kappa}B$ (NF-${\kappa}B$) ligand (RANKL). In addition, SF significantly decreased the level of triglyceride and increased the level of low-density lipoprotein. Moreover, the decrease of trabeculae of the femur was partially prevented in ovariectomized rats administrated with SF. Conclusions : SF treatment could prevent ovariectomy induced bone loss and its effects could be medicated by the inhibition of osteoclastogenesis.

EFFECTS OF POLYPHENOLS OF Cocos Nucifera HUSK FIBRE ON SELECTED KIDNEY FUNCTION INDICES IN MICE

  • Adebayo, Joseph Oluwatope;Owolabi, O.O.;Adewumi, O.S.;Balogun, E.A.;Malomo, S.O.
    • CELLMED
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    • v.9 no.1
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    • pp.2.1-2.6
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    • 2019
  • Decoction of Cocos nucifera husk fibre is used indigenously in Nigeria for malaria treatment. Polyphenols have been identified as the phytochemicals responsible for the antimalarial activity of Cocos nucifera husk fibre, though their toxicity has not been evaluated. The polyphenols of Cocos nucifera husk fibre were therefore evaluated for their effects on selected kidney function indices in mice. Fifty mice were randomly divided into five groups (A-E) of ten mice each. Mice in group A were orally administered 5% DMSO solution while those in groups B, C, D and E were orally administered 31.25, 62.5, 125 and 250 mg/Kg body weight of the polyphenols respectively for seven days. Serum urea, creatinine and uric acid concentrations were determined. Serum levels of sodium, potassium, chloride and calcium ions and kidney alkaline phosphatase (ALP), glutamate dehydrogenase (GDH) and gamma-glutamyltransferase (GGT) activities were also determined. The results showed that the polyphenols significantly reduced (p<0.05) urea concentration at 250 mg/Kg body weight and creatinine concentration at all doses compared to controls. The polyphenols caused no significant alteration (p>0.05) in serum uric acid concentration and kidney ALP, GGT and GDH activities compared to controls. There was significant increase (p<0.05) in serum sodium ion concentration at 31.25, 125 and 250 mg/Kg body weight of polyphenols whereas significant increase (p<0.05) in serum potassium and chloride ions was observed at 62.5 and 250 mg/Kg body weight compared to controls. Thus, polyphenols of Cocos nucifera husk fibre may adversely affect some osmoregulatory functions of the kidney, especially at higher concentrations.

Protective Effect of Jaboyangyeong-hwan Water Extracts on CCl4-Induced Liver Damage (자보양영환의 물추출물이 사염화탄소로 유발된 간 손상에 미치는 영향)

  • 전병훈;이형철;황상구;남은영;김대근;박정원;이영찬;박승택
    • Journal of Life Science
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    • v.12 no.2
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    • pp.136-143
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    • 2002
  • Jaboyangyeong-hwan (IAE) has been known as a traditional medicine for the treatment of debility, fatigue, and liver diseases. The hepatoprotective effect of the water extract of Jaboyangyeong-hwan was investigated against carbon tetrachloride ($CCl_4$)-induced hepatic damage. A single intraperitoneal injection of $CCl_4$produced liver damage in rats as manifested by the significant rise of aspartate aminotransferase (AST, alanine aminotransferase (ALT), and alkaline phosphatase (ALP) in serum as compared to those of untreated normal group. Pretreatments of rats with the JAE extract (300, 600, and 1200 mg/kg for 7 days) were significantly reduced AST, ALT, and ALP levels compared with $CCl_4$-treated control group. Treatment of rats with $CCl_4$led to significantly increase in lipid peroxidation and significantly decrease in cytochrome P450 and P450 reductase. The oral administration of the JAE extract significantly inhibited the accumulation of microsomal thiobarbituric acid reactive substance (TBARS) and increased the cytochrome P450 and P450 reductase activity. All these biochemical alterations resulting from $CCl_4$administration were inhibited by the pretreatment with JAE extract. These results suggest that JAE water extract can be useful as a hepatoprotective agent.

The Effects of Hydrogen Peroxide Pretreatment on Rice Straw Fermentation for Feed (볏짚 발효사료 제조시 과산화수소 전처리 효과)

  • Choi, Yoon-Hee;Lee, Sang-Bok;Kim, Myeong-Sook;Hong, Jai-Sik
    • Applied Biological Chemistry
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    • v.37 no.5
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    • pp.326-333
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    • 1994
  • Pleurotus florida with high cellulase activity as well as lignin degradability was selected out among strains for fermentation of the rice straw to improve the nutritive value. When the rice straw was fermented by P. florida, the contents of hemicellulose, cellulose and lignin were decreased to 22.5%, 11.4% and 28.1%, respectively, whereas the contents of rice straw fermented after pretreatment with $H_2O_2$ or alkaline hydrogen peroxide were decreased much in the lower concentration. The content of T-N (total-nitrogen) and crude fat was increased with the longer fermentation period. The amino acid content of rice straw fermented by P. florida in 30 days was increased to 28.9% and 35.1% as the rice straw was fermented after pretreatment without and with 4% $H_2O_2$, respectively. The crystalline intensity of rice straw was decreased by pretreatment with 4% $H_2O_2$ and fermentation by P. florida. However, the crystall intensity was increased by treatment with alkaline hydrogen peroxide and the more when the straw was washed after the treatment. When the rice straw was fermented by P. florida for 30 days, the in vitro organic matter digestibility was increased up to 6% of $H_2O_2$ pretreatment.

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Quality Changes in Peeled Lotus Roots Immersed in Electrolyzed Water Prior to Wrap- and Vacuum-Packaging (전기분해수 침지처리 박피연근의 랩 및 진공포장 저장 중의 품질변화)

  • Park, Kee-Jai;Jeong, Jin-Woong;Lim, Jeong-Ho;Kim, Bum-Kun;Jeong, Seong-Won
    • Food Science and Preservation
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    • v.15 no.5
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    • pp.622-629
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    • 2008
  • This study investigated changes in quality characteristics of wrap- and vacuum-packaged peeled lotus roots treated with strong acidic electrolyzed acid water (SAEW pH 2.58, ORP 1,128 mV, HClO 105.0 ppm) or low alkaline electrolyzed water (LAEW pH 8.56, ORP 660 mV, HClO 73.8 ppm) as immersion liquids prior to packaging and storage at 5C. Immersion of peeled lotus roots in SAEW and LAEW reduced initial microbial load by about 1 log compared to treatment with tap water (TW). Hardness differences on storage were observed. However, reduction in PPO activity by electrolyzed water was not reproducible. Changes in Hunter's color value and the color difference value ($\Delta$) of peeled lotus roots immersed in 0.5% (w/v) sodium metabisulfite (SMS) and electrolyzed water were smaller than those of roots treated with TW prior to storage. Sensory characteristics measured during storage were best-preserved in lotus roots previously immersed in 0.5% (w/v) SMS or electrolyzed water, compared to TW. Immersionin electrolyzed water and vacuum packaging preserves the quality of peeled lotus roots in terms of microbial, visual, and sensory aspects, at levels comparable to those offered by storage after treatment with 0.5% (w/v) SMS.

Proteolytic Enzymes Distributed in the Tissues of Dark Fleshed Fish 1. Comparison of the Proteolytic Activity of the Tissue Extracts from the Meat of Mackerel and Sardine (혈합육어의 조직중에 분포하는 단백질분해효소 1. 고등어와 정어리 육조직중의 단백질분해효소의 활성비교)

  • PYEUN Jae-Hyeung;KIM Hyeung-Rak;CHO Jin-Guen
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.5
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    • pp.469-476
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    • 1986
  • Proteolytic activity of the tissue extracts from the muscle of mackerel, Scomber japonicus, and sardine, Sardinops melanostcta, was comparence with referenced to the optimum reaction condition. Thermal stability and change of proteolytic activity of the tissue extracts during storage were investigated. The existence of acid, weak acid and alkaline proteinase was identified in the ordinary and dark muscle of the mackerel and sardine. Specific activity of acid proteinase was stronger than weak acid or alkaline proteinase in the both fish. The proteolytic activity of the tissue extracts on the optimum reaction condition was: ordinary muscle of mackerel, 0.12 nM-Tyr. eq./mg-prot. /min. at pH 3.0 and $50^{\circ}C$; dark muscle of mackerel, 0.36 nM-Tyr. eq./mg-prot. /min. at pH 3.0 and $45^{\circ}C$; ordinary muscle of sardine, 0.45 nM-Tyr. eq./mg-prot. /min. at pH 2.4 and $45^{\circ}C$; dark muscle of sardine, 0.24 nM-Tyr. eq./mg-port. /min. at pH 2.4 and $45^{\circ}C$. The proteinases distributed in the muscle of mackerel and sardine were stable with the heat treatment at $45^{\circ}C$ for 5 minutes, but those in the dark muscle of mackerel was stable with the treatment at $5^{\circ}C$ for 5 minutes. The proteinases from the muscle were slowly inactivated with the whole storage days at $5^{\circ}C\;and\;-15^{\circ}C$, those were more stable at $-15^{\circ}C\;than\;5^{\circ}C$ storage.

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Study on the Pretreatment of Rice Hull to Enhance Enzymatic Saccharification Efficiency (효소 당화효율 증진을 위한 왕겨의 전처리 방법 연구)

  • Bark, Surn-Teh;Koo, Bon-Cheol;Moon, Youn-Ho;Cha, Young-Lok;Yoon, Young-Mi;Kim, Jung Kon;An, Gi Hong;Park, Kwang-Geun;Park, Don-Hee
    • Applied Chemistry for Engineering
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    • v.23 no.4
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    • pp.399-404
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    • 2012
  • The objective of this study was to investigate the efficient pretreatment method for bioethanol production from rice hull. Ammonia and sodium hydroxide as an alkaline solution and dilute sulfuric acid as an acidic solution were used in a batch reactor under high-temperature and high-pressure conditions. The highest enzymatic saccharification efficiency of 82.8% and ash removal rate of 94.7% were obtained in the dilute sulfuric acid treated sample after the sodium hydroxide solution treatment. The enzymatic saccharification efficiencies and ash removals of pretreated rice hull samples have very similar variation tendency. This means that the maximum obstructive factor for the enzymatic saccharification of rice hull is the ash (silicate) content in biomass. The findings suggest that the combined sodium hydroxide-dilute sulfuric acid treatment system under high-temperature and high-pressure conditions is a promising pretreatment method to enhance the enzymatic saccharification of the silica-rich biomass.

Accumulation of the Conjugated Linoleic Aacid (CLA) in Tilapia ( Tilapia nilotica) Fed Diets on Various Levels of CLA (CLA (Conjugated linoleic acid) 급이수준에 따른 역돔의 CLA 축적량)

  • CHOI Byeong-Dae;KANG Seok-Joong;HA Young-Lae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.5
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    • pp.509-514
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    • 2001
  • Effects of conjugated linoleic acid (CLA), known as an effective anticarcinogen in several animal models, on the tilapia were investigated. The CLA was made from safflower oil by alkaline isomerization method. Isomers in CLA such as cis-9, trans-11 and trans-10, cis-12 occupied over $80\%$, and other isomers was below $20\%$. In experiment, 250 fishes (average weight is 32 g) were divided into 15 fishes per five treatment and triplicate group for 8 weeks: control, $1\%$ CLA, $2.5\%$ CLA, $5.0\%$ CLA, and $10\%$ CLA diets. Daily growth rate and feed coefficiency were measured every week. The most effective diet for the growth rate and feed coefficiency of tilapia was $1.0\%$ CLA diet group. Every two weeks, sampled and determined the contents of CLA in the muscle and liver, After 8 weeks, $1.0\%$ and $10.0\%$ of CLA fed group accumulated the CLA as 41.3 and 180.9 mg/g of fat in their muscle respectively, Also, n-9 and n-3 fatty acid (FA) compositions were almost not changed in the muscle and liver. But n-6 fatty acid was changed according to the contents of fed CLA. The $1.0\%$ CLA fed group was shown the highest contents of n-6 FA and the $10.0\%$ CLA group was shown the lowest contents of n-6 FA.

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