• Title/Summary/Keyword: Enzyme Efficiency

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Effects of Dietary Fructooligosaccharide on Digestive Enzyme Activities, Intestinal Microflora and Morphology of Growing Pigs

  • Xu, Z.R.;Zou, X.T.;Hu, C.H.;Xia, M.S.;Zhan, X.A.;Wang, M.Q.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1784-1789
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    • 2002
  • One hundred and twenty-eight growing barrows (Jiaxing Black${\times}$Duroc${\times}$Landrace) at an average BW of 20.8 kg were allocated to four treatments for 42 days, each of which was replicated four times with eight pigs per replicate and used to investigate the effects of fructooligosaccharide (FOS) on digestive enzyme activities, intestinal microflora and morphology of growing pigs. The pigs received the same basal corn-soybean meal diet and FOS was added to the basal diet at 0, 2, 4, 6 g/kg diet at the expense of corn, respectively. As compared to control, supplementation with 4 and 6 g/kg FOS significantly improved average daily gain and feed efficiency. Addition of FOS enhanced the growth of Bifidobacterium and Lactobacillus, but inhibited Clostridium and Escherichia coli in the small intestinal and proximal colonic contents. Supplementation with 4 and 6 g/kg FOS significantly improved the activities of total protease, trypsin and amylase in the small intestinal contents. However, FOS had no significant effect on the activity of lipase in the small intestinal contents as well as the digestive enzymes in pancreas. Morphological measurement of jejunal mucosa did show response to consumption of FOS. Villus height and the villus height to crypt depth ratio at the jejunal mucosa were significantly higher with 4 and 6 g/kg FOS supplementation as compared to control.

Enzyme-assisted extraction of Ecklonia cava fermented with Lactobacillus brevis and isolation of an anti-inflammatory polysaccharide

  • Lee, Won-Woo;Ahn, Ginnae;Wijesinghe, W.A.J.P.;Yang, Xiudong;Ko, Chang-Ik;Kang, Min-Cheol;Lee, Bae-Jin;Jeon, You-Jin
    • ALGAE
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    • v.26 no.4
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    • pp.343-350
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    • 2011
  • Fermentation and enzyme-assisted extraction (EAE) improve nutritional and functional properties of foods by increasing the extraction of active compounds, ingestion rates, and body absorption. In this study, we investigated whether applying the EAE process improves the extraction and isolation efficiency of a polysaccharide from fermented Ecklonia cava (FE), which inhibited NO production in lipopolysaccharide (LPS)-activated RAW 264.7 cells. The results showed that the FE using the fungi Candida utilis and two different bacteria, namely Lactobacillus brevis and Saccharomyces cerevisiae increased protein and carbohydrate contents in comparison with those in non-fermented E. cava (NE). Aqueous extracts of fermented E. cava increased extraction yields and carbohydrate content, compared with the aqueous extract of NE. In addition, treating LPS-stimulated RAW 264.7 cells with aqueous extracts resulted in reduced NO production compared to that in LPS-treated cells. Ten EAEs of L. brevis-fermented E. cava (LFE) improved NO inhibitory effects in LPS-activated RAW 264.7 cells and the Viscozyme extract (VLFE) from the resulting extracts showed the highest NO inhibitory effect. We found that the >30 kDa fraction of VLFE led to markedly high inhibition of LPS-induced NO production as compared to that in the <30 kDa fraction. The crude polysaccharide isolated from >30 kDa fraction (VLFEP) consisted of fucose and markedly decreased NO production induced by LPS stimulation. VLFEP could be useful as an anti-inflammatory agent to suppress macrophage activation.

Effects of Soyosangamibang on antitumor chemotherapy and lysosomal enzymes of tumor cel (소요산가미방(逍遙散加味方)이 항암화학요법제(抗癌化學療法劑)의 항종양효과(抗腫瘍效果) 및 종양세포(腫瘍細胞)의 lysosomal enzymes에 미치는 영향(影響))

  • Cho, Hyun-Ju;Won, Bong-Hui;Mun, Gu;Mun, Seok-Jae;Jeon, Byeong-Hun
    • The Journal of Korean Medicine
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    • v.18 no.2
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    • pp.119-136
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    • 1997
  • In order to investigate the effects of Soyosangamibang Extract(逍遙散加味方抽出液) on antitumor effects after human cell lines(A549, hep3B, Caki-1, Ehrlich) transplantation into the peritoneal cavity or right groin in mice induced by RPMI1640 and GIBCO etc., the extracts of its herbal medicines were orally administered for 10 or 12 days. Experimental studies were performed for measurance of antitumor effect of MMC(Mitomycin C) and lysosomal enzyme's activities using colony forming efficency, SRB assay which were regarded as a valuable method for antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows: 1. The change of colony-forming efficiency and SRB assay of Caki-1 cells, hep3B and A549 cells after exposure to the extract of Soyosangamibang extract depressed the growth of tumor cells by concentration of Soyosangamibang, 2. Antitumor activity of the ethanol extract from Soyosangamibang extract and MMC on ascites form of Ehrlich carcinoma in mice is a little improved. Especially mean survival times of the group of 200mg/kg and MMC 0.1mg/kg is improved Over 50%. 3. WhenSoyosangamibang extract and MMC are administrated together, the weight of turnor is more decreased than MMC alone. 4. The lysosomal enzyme's activities of the Soyosangarmibang extract and MMC are more significantly improved than MMC alone. According to the above results, it could be suggested that Soyosangamibang extract has indirect antitumor effect by strengthen the effect of MMC.

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Differences of Soil Enzyme Activity after Incorporation with Chinese Milk Vetch Litter Cut at Different Growth Stages

  • Lee, Ji-Hyun;Shim, Sang-In
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.3
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    • pp.341-347
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    • 2007
  • Chinese milk vetch (CMV) is a winter legume that is commonly used as cover crop in Korea. Kill date of cover crop for addition into soil affects N content in cover crop and N availability in soil. This study was conducted to evaluate the effect of CMV as green manure cover crop according to kill dates before growing corn without artificial fertilizer. Top of CMV cut three times on 13 April, 27 April, and 11 May were added into soil at a rate of 600 kg per 10a. Sugar content in CMV litter was persistently decreased from mid-April to late-May. The decrease of sugar content might be due to the transformation into starch and/or other storage or structural constituents. The decreased amount of sugars was greater than 12% and the increased amount of starch was less than 0.2%. Concentration of $NH_4^+$ in soil treated by CMV litter cut on May 11 was slightly higher than that in the treatment with early-cut (April 13) CMV, the concentration at 28 and 49 DAT (days after treatment) was higher in the treatment with late-cut CMV litter. Regardless of cut (kill) date of CMV, the phosphatase activity in the treatment of CMV litter was higher compared to the untreated control. Soil dehydrogenase activity was increased steadily by addition of CMV litter implying total microbial activities in the soil were increased. Our results demonstrate that the status of cover crop species at kill date is an important factor influencing soil enzyme activities derived from microorganisms. Therefore, the optimal kill date of cover crop should be examined to improve the efficiency of cover crop as green manure crop regarding the practical sequence in cropping system.

Effects of Chunggukjang and Greentea-Chunggukjang on Lipid Profile and Antioxidative Enzyme Activity of Liver Tissue in Growing Rats Fed Cholesterol (청국장과 녹차청국장이 고콜레스테롤 식이를 섭취한 성장기 쥐의 Lipid Profile 및 항산화효소 활성에 미치는 영향)

  • Jung, Yun-Jung;Choi, Mi-Ja
    • Journal of the East Asian Society of Dietary Life
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    • v.25 no.2
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    • pp.278-286
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    • 2015
  • The overall purpose of this study was to investigate the effects of Chunggukjang and Greentea-Chunggukjang on the lipid profile, lipid peroxidation and antioxidative enzyme activities of liver tissue in growing male rats fed cholesterol. Twenty seven rats were divided into three treatment groups (Control, Chunggukjang and Greentea-Chunggukjang) and were given experimental diets with 1% cholesterol for 9 weeks. All rats in this study were fed a casein-based diet. Chunggukjang groups were fed diet containing 33.1% Chunggukjang powder. The Chunggukjang and Greentea-Chunggukjang groups showed significantly lower weight gain, food efficiency ratio than the control group regardless of Chunggukjang type. Serum total cholesterol was significantly lower in the Chunggukjang group than in the control group, whereas serum triglyceride and atherogenic index were significantly lower in the Greentea-Chunggukjang group than in the control group. Hepatic triglyceride contents was not significantly different among the diets. However, hepatic cholesterol content was significantly lower in the Greentea-Chunggukjang group than in the control group. Lipid peroxidation of malondialdehyde (MDA) contents was significantly lower in the Chunggukjang and Greentea-Chunggukjang groups than in the control group. Activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) in liver tissue of the Chunggukjang and Greentea-Chunggukjang groups were not significantly different. It can be concluded that Chunggukjang and Greentea-Chunggukjang influence lipid profile and hepatic malondialdehyde contents in growing male rats fed cholesterol.

Effects of Green and Black Korean Teas on Lipid Metabolism in Diet-Induced Hyperlipidemic Rats (한국산 녹차와 홍차가 고지혈증 유도 쥐에 있어서 혈청 지질 대사에 미치는 영향)

  • Jung, Young-Hee;Han, Sung-Hee;Shin, Mee-Kyung
    • Journal of the East Asian Society of Dietary Life
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    • v.16 no.5
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    • pp.550-558
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    • 2006
  • The study investigated the serum lipid metabolism and enzyme activities of Korean teas for their preventative activity against chronic disease and obesity. Male Sprague-Dawley rats were raised for 8 weeks on four experimental diets: normal diet, hyperlipidemic diet, and hyperlipidemic diet to which green and black teas (2% each) were added. Various biological actions, including lipid metabolism and enzyme activities of the serum, were investigated. Diet-induced, hyperlipidemic rats fed with green and black teas, showed significant decrease in food efficiency ratio, triglyceride, total lipid, and phospholipid compared to control, i.e. the normal and diet-induced, hyperlipidemic rats. Total cholesterol, LDL-cholesterol, Al(atherogenic index), LHR, VLDL-cholesterol, ester-cholesterol, and free-cholesterol also showed a significant decrease. However, there was no significant difference between the tea-fed, diet-induced, hyperlipidemic dieted groups. HDL-cholesterol concentration was increased significantly in the tea-dieted and normal groups compared to the control. There was a little difference in lipase activity between the normal and control groups, although green and black tea-dieted experimental groups were both increased compared to the control. The contents of total lipid, triglycerides, and total cholesterol were decreased in the normal and experimental groups compared to the control. The GOT, GPT, ALP and LDH serum enzyme activities of the experimental groups were significantly reduced compared to those of the control groups.

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Role of the Salt Bridge Between Arg176 and Glu126 in the Thermal Stability of the Bacillus amyloliquefaciens ${\alpha}$-Amylase (BAA)

  • Zonouzi, Roseata;Khajeh, Khosro;Monajjemi, Majid;Ghaemi, Naser
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.7-14
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    • 2013
  • In the Bacillus amyloliquefaciens ${\alpha}$-amylase (BAA), the loop (residues 176-185; region I) that is the part of the calcium-binding site (CaI, II) has two more amino acid residues than the ${\alpha}$-amylase from Bacillus licheniformis (BLA). Arg176 in this region makes an ionic interaction with Glu126 from region II (residues 118-130), but this interaction is lost in BLA owing to substitution of R176Q and E126V. The goal of the present work was to quantitatively estimate the effect of ionic interaction on the overall stability of the enzyme. To clarify the functional and structural significance of the corresponding salt bridge, Glu126 was deleted (${\Delta}$E126) and converted to Val (E126V), Asp (E126D), and Lys (E126K) by site-directed mutagenesis. Kinetic constants, thermodynamic parameters, and structural changes were examined for the wild-type and mutated forms using UV-visible, atomic absoption, and fluorescence emission spectroscopy. Wild-type exhibited higher $k_{cat}$ and $K_m$ but lower catalytic efficiency than the mutant enzymes. A decreased thermostability and an increased flexibility were also found in all of the mutant enzymes when compared with the wild-type. Additionally, the calcium content of the wild-type was more than ${\Delta}E126$. Thus, it may be suggested that ionic interaction could decrease the mobility of the discussed region, prevent the diffusion of cations, and improve the thermostability of the whole enzyme. Based on these observations, the contribution of loop destabilization may be compensated by the formation of a salt bridge that has been used as an evolutionary mechanism or structural adaptation by the mesophilic enzyme.

Enzymatic Properties of a Thermostable ${\alpha}$-Glucosidase from Acidothermophilic Crenarchaeon Sulfolobus tokodaii Strain 7

  • Park, Jung-Eun;Park, So Hae;Woo, Jung Yoon;Hwang, Hye Sun;Cha, Jaeho;Lee, Heeseob
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.56-63
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    • 2013
  • We have characterized the putative ${\alpha}$-glucosidase gene (st2525) selected by total genome analysis from the acidothermophilic crenarchaeon Sulfolobus tokodaii strain 7. The ORF was cloned and expressed as a fusion protein in Escherichia coli, and recombinant ST2525 was purified by Ni-NTA affinity chromatography. Maximum activity was observed at $95^{\circ}C$ and pH 4.0, and the enzyme exhibited stability with half-lives of 40.1 min and 7.75 min at extremely high temperatures of $100^{\circ}C$ and $105^{\circ}C$, respectively. The enzyme retained at least 85% of its maximal activity in the pH range of 4.0-11.0. ST2525 exclusively hydrolyzed ${\alpha}$-1,4-glycosidic linkages of oligosaccharides in an exo-type manner, with highest catalytic efficiency toward maltotriose. The enzyme also displayed transglycosylation activity, converting maltose to isomaltose, panose, maltotriose, isomaltotriose, etc. From these results, ST2525 could be potentially useful for starch hydrolysis as well as novel synthesis of oligosaccharides in industry.

Hydrolysis of Cellulose by Immobilized Cellulase in a Packed Bed Reactor (충진층 반응기에서 고정화 cellulase에 의한 셀룰로스 가수 분해)

  • Kang, Byung Chul;Lee, Jong Baek
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1365-1370
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    • 2013
  • Immobilized cellulase on weak ion exchange resin showed a typical Langmuir adsorption isotherm. Immobilized cellulase had better stability with respect to pH and temperature than free cellulase. Kinetics of thermal inactivation on free and immobilized cellulase followed first order rate, and immobilized cellulase had a longer half-life than free cellulase. The initial rate method was used to characterize the kinetic parameters of free and immobilized enzyme. The Michaelis-Menten constant $K_m$ was higher for the immobilized enzyme than it was for the free enzyme. The effect of the recirculation rate on cellulose degradation was studied in a recycling packed-bed reactor. In a continuous packed-bed reactor, the increasing flow rate of cellulose decreased the conversion efficiency of cellulose at different input lactose concentrations. Continuous operation for five days was conducted to investigate the stability of long term operation. The retained activity of the immobilized enzymes was 48% after seven days of operation.

Purification, Characterization and Immobilization of Lipase from Proteus vulgaris OR34 for Synthesis of Methyl Oleate

  • Misbah, Asmae;Koraichi, Saad Ibnsouda;Jouti, Mohamed Ali Tahri
    • Microbiology and Biotechnology Letters
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    • v.48 no.4
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    • pp.491-505
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    • 2020
  • A newly isolated strain, Proteus vulgaris OR34, from olive mill waste was found to secrete an alkaline extracellular lipase at 11 U·ml-1 when cultivated on an optimized liquid medium. This lipase was purified 94.64-fold with a total yield of 9.11% and its maximal specific activity was shown to be 3232.58 and 1777.92 U·mg-1 when evaluated using the pH-stat technique at 55℃ and pH 9 and Tributyrin TC4 or olive oil as the substrate. The molecular mass of the pure OR34 lipase was estimated to be around 31 kDa, as revealed by SDS-PAGE and its substrate specificity was investigated using a variety of triglycerides. This assay revealed that OR34 lipase preferred short and medium chain fatty acids. In addition, this lipase was stable in the presence of high concentrations of bile salt (NaDC) and calcium ions appear not to be necessary for its activity. This lipase was inhibited by THL (Orlistat) which confirmed its identity as a serine enzyme. In addition, the immobilization of OR34 lipase by adsorption onto calcium carbonate increased its stability at higher temperatures and within a larger pH range. The immobilized lipase exhibited a high tolerance to organic solvents and retained 60% of its activity after 10 months of storage at 4℃. Finally, the OR34 lipase was applied in biodiesel synthesis via oleic acid mediated esterification of methanol when using hexane as solvent. The best conversion yield (67%) was obtained at 12 h and 40℃ using the immobilized enzyme and this enzyme could be reused for six cycles with the same efficiency.