• Title/Summary/Keyword: enzyme secretion

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Construction of Secretion Vectors Using the $\alpha$-amylase Signal Sequence of Bacillus subtilis NA64

  • Kim, Sung-Il;Lee, Se-Yong
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.74-81
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    • 1996
  • Two secretion vectors, pUBA240 and pUB340 were constructed by using the promoter and secretory signal region of the .alpha.-amylase gene from an .alpha.-amylase hyperproducing strain, Bacillus subtilis NA64. In this secretion vector system, various restriction enzyme sites are located immediately after the proregion of the .alpha.-amylase gene for easy replacement of various foregn structural genes. To evaluate this secretion vectors, the .betha.-lactamase gene of pBR322 was used as a reporter gene. The expressed and biologically active .betha.-lactamase was secreted into the culture broth from B. subtilis LKS86 transformants harboring each .betha.-lactamase secreting plasmid, pUBAbla and pUBSble. In both cases, more than 92% of expressed .betha.- lactamases were located idn the culture medium. The amount of the secreted .betha.-lactamase was about 80% of the total secreted proteins in the culture medium.

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Preliminary Characterization of Keratinolytic Enzyme of Aspergillus flavus K-03 and Its Potential in Biodegradation of Keratin Wastes

  • Kim, Jeong-Dong
    • Mycobiology
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    • v.31 no.4
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    • pp.209-213
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    • 2003
  • Aspergillus flavus K-03 isolated from poultry forming soil in Korea was studied for its ability to produce extracellular proteases on basal medium containing 2%(w/v) chicken feathers. The fungus was observed to be a potent producer of such enzymes. Keratinolytic enzyme secretion was the best at 15 days of incubation period at pH 9 and temperature $40^{\circ}C$. No relationship existed between the enzyme yield and increase of biomass. Enzyme production was suppressed by exogenous sugars in descending order arabinose>maltose>mannose>fructose. But glucose did not influence the enzyme activity. The keratinolytic enzyme released by the fungus demonstrated the ability to decompose keratin substrates as chicken feather when exogenous glucose was present. The keratinolytic activity was inhibited by $HgCl_2$ and serine-protease inhibitors such as phenymethylsulfonyl fluoride(100%), chymostain(88%), crystalline soybean trypsin inhibtor(80%), antipain(45%) and aprotinin(40%), and was not by cystein-protease and aspartyl-protease inhibitors. The enzyme activity is only partially inhibited by metallo-protease inhibitor. Thus, the enzyme secreted by A. flavus K-03 belongs to the alkaline serine-type protease.

Effect of CCK and Carbachol on Enzyme Secretion From the Isolated Pancretic Acinar Cells of Rats fed Heated or Raw Soybean Diet (CCK와 Carbachol 이 익힌 대두와 생대두를 먹인 쥐에서 분리한 췌장세포의 외분비기능에 미치는 영향)

  • 이상선
    • Journal of Nutrition and Health
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    • v.19 no.6
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    • pp.374-381
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    • 1986
  • The effects of the hormonal factor (CCK) a and neural factor(carbachol) on the exocrine function of the pancreas were studied in th is experiment. A superfusion technique was used for in vitro study of stimulus-secreti- o on coupling in isolated pancreatic acinarce 11s frQm the rats fed heated or raw soybean diet. Chymotrypsin secretion was higher in cells from the raw soybean group than in those from the heated soybean group with both kinds of stimulants(CCK and carbachol), whereas, amylase secretion was higher inthe h heated soybean group than in the raw soy­b bean group. This indicated that chymotrvpsin a and amylase secretion from the acinar cells are not parallel with CCK and carbachol st­i imulation.

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Anti-inflammatory and Antioxidant Effects of Zizyphus jujuba Miller var. inermis extracts (대추 추출물의 항산화 및 아토피 피부염 관련 항염증 효과)

  • Hong, Chang-Eui;Chun, Young-Hee;Lyu, Su-Yun
    • Korean Journal of Pharmacognosy
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    • v.50 no.2
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    • pp.86-95
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    • 2019
  • In the present study, the effect of Z. jujuba Miller var. inermis Rehder extracts on the secretion of atopic dermatitis (AD)-related cytokines and hyaluronidase activity was investigated. We prepared four fractions, butylene glycol (JB), ethanol (JE), and water (JW), with Z. jujuba Miller var. inermis extracts. JW significantly reduced the secretion of interleukin-8 and JE reduced the secretion of tumor necrosis factor-alpha in human keratinocyte HaCaT cells. Also, hyaluronidase activity was measured by enzyme assay and the fractions inhibited the activity in a dose-dependent manner. In addition, the human dermal fibroblast, HDF-n cells were treated with the extracts and antioxidant activities were measured. The results showed that the extracts increased the free radical scavenging activity and the superoxide dismutase activity. Taken together, Z. jujuba Miller var. inermis extracts reduced the secretion of AD-related cytokines and inhibited the hyaluronidase. In addition, the extracts showed antioxidant activity.

Dietary Protein Restriction Alters Lipid Metabolism and Insulin Sensitivity in Rats

  • Kang, W.;Lee, M.S.;Baik, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.9
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    • pp.1274-1281
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    • 2011
  • Dietary protein restriction affects lipid metabolism in rats. This study was performed to determine the effect of a low protein diet on hepatic lipid metabolism and insulin sensitivity in growing male rats. Growing rats were fed either a control 20% protein diet or an 8% low protein diet. Feeding a low protein diet for four weeks from 8 weeks of age induced a fatty liver. Expression of acetyl-CoA carboxylase, a key lipogenic enzyme, was increased in rats fed a low protein diet. Feeding a low protein diet decreased very low density lipoprotein (VLDL) secretion without statistical significance. Feeding a low protein diet down-regulated protein expression of microsomal triglyceride transfer protein, an important enzyme of VLDL secretion. Feeding a low protein diet increased serum adiponectin levels. We performed glucose tolerance test (GTT) and insulin tolerance test (ITT). Both GTT and ITT were increased in protein-restricted growing rats. Our results demonstrate that dietary protein restriction increases insulin sensitivity and that this could be due to low-protein diet-mediated metabolic adaptation. In addition, increased adiponectin levels may influences insulin sensitivity. In conclusion, dietary protein restriction induces a fatty liver. Both increased lipogenesis and decreased VLDL secretion has contributed to this metabolic changes. In addition, insulin resistance was not associated with fatty liver induced by protein restriction.

Effect of Eriobotryae folium extract on glucokinase and hexokinase activities of diabetes mellitus mice induced by interleukin-$1{\beta}$ (IL-1${\beta}$ 처리(處理) 당뇨병(糖尿病) 마우스의 췌장(膵臟) glucokinase 및 hexokinase 활성(活性)에 대(對)한 비파엽(枇杷葉)의 영향(影響))

  • Yoon, Cheol-Ho;Shin, Hyeon-Chul;Jeong, Ji-Cheon
    • The Journal of Internal Korean Medicine
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    • v.19 no.1
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    • pp.466-476
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    • 1998
  • We investigated the in vivo effect of an aqueous extract (referred to as EF) from Eriobotryae folium on glucokinase and hexokinase activities of diabetes mellitus induced by $interleukin-1{\beta}\;(IL-1{\beta})$. After 1 week of $IL-1{\beta}$ injection, the levels of serum glucose concentration and insulin secretion were dramatically increased. However, the insulin secretion was decreased with administration of EF. The level of glucose concentration was decreased by EF administration. Furthermore, it was observed that EF was effective in recovering the levels of insulin secretion. Enzyme activities of the glucokinase and hexokinase, which are key enzymes of glucose phosphorylastion, were decreased by $IL-1{\beta}$. EF administration to the mice allowed proportional increasing by stimulation of induction of enzyme activities as high as normal group. These results suggested that EF is highly effective in treatment of diabetes mellitus induce by $IL-1{\beta}$.

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Heterologous Expression of ${\alpha}$-Amylase Gene of Bifidobacterium adolescentis Int57 in Bacillus polyfermenticus SCD

  • Paik, Hyun-Dong;Kim, Il-Gi;Lee, Jin-Hyoung;Lee, Jang-Hyun;Park, Kyu-Yong;Ji, Geun-Eog;Jin, Tae-Eun;Rhim, Seong-Lyul
    • Food Science and Biotechnology
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    • v.16 no.4
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    • pp.655-658
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    • 2007
  • Bacillus polyfermenticus SCD was transformed by the recombinant shuttle vector for Bacillus and Escherichia coli containing 3 antibiotic resistant genes and an ${\alpha}$-amylase gene from Bifidobacterium adolescentis Int57. The ${\alpha}$-amylase gene fused to a secretion sequences was expressed under the control of the promoter of amylase gene from B. subtilis var. natto. The recombinant plasmid was maintained stably in the transformants producing the ${\alpha}$-amylase. The enzyme was secreted to outside of the cell and showed the similar enzyme activity as that of Bacillus subtilis BD170 under the same conditions of pH and growth temperature. Because of the relatively easy transformation and the secretion of the enzyme, the transformants of B. polyfermenticus SCD may give a new strategy in the production of foreign genes.

Optimum culture conditions for production of extracellular cytosine deaminase by bacellus polymyxa YL 38-3 (Bacillus polymyxa YL38-3의 세포외 cytosine deaminase 생성의 최적 배양 조건)

  • 유대식;김대현;박정문;송형익;정기택
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.362-367
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    • 1988
  • The strain YL 38-3, which was capable of producing extracellular cytosine deaminase, was isolated and taxonomically examined. The isolated strain was identified to be Bacillus polymyxa YL 38-3. The optimal conditions for the enzyme production from Bacillus polymyxa YL 38-3 were investigated. The enzyme production was reached maximum level in the medium containing 0.5% glucose, 0.2% beef extract, 0.5% NaCl and 0.1% $KH_{2}PO_{4}$ (pH 6.0). And the enzyme showed the highest activity when the strain YL 38-3 was cultivated at $35^{\circ}C$ for 24 gours under the initial pH 6.0. By the additions of peptone the extracellular enzyme production was inhibited, meanwhile the intracellular enzyme production was highly stimulated. It was, therefore, deduced that peptone was related to the secretion mechanism of the enzyme from this bacterial cell.

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Effects of Haengso-tang and Chwiyeon-tang on Expression of Respiratory Mucin Gene and Secretion of Airway Mucus (행소탕(杏蘇湯) 및 취연탕(取淵湯)이 호흡기 뮤신 유전자의 발현과 점액분비에 미치는 영향)

  • Kang, Won-Je;Seo, Un-Kyo
    • The Journal of Korean Medicine
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    • v.29 no.3
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    • pp.76-87
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    • 2008
  • Objectives: In this study, the author investigated whether Haengso-tang (HST) and Chwiyeon-tang (CHT) affect both in vitro mucin secretion and MUC5AC gene expression in airway epithelial cells and in vivo mucin secretion from animal model for airway mucus hypersecretion. Materials and Methods: Confluent HTSE cells (non-labeled) were chased for 30 min in the presence of HST and CHT to assess the effects of the agents on mucin secretion by enzyme-linked immunosorbent assay (ELISA), with removal of oriental herbal medicine extract from each agent-treated sample by centrifuge microfilter. Also, the effects of the agents on TNF- or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. The author also induced hypersecretion of airway mucus by exposure of rats to SO2 for 3 weeks. Effects of orally-administered HST and CHT during 1 week on in vivo mucin secretion from tracheal goblet cells of rats were assessed using ELISA. Results: (1) HST significantly decreased in vitro mucin secretion from cultured HTSE cells. However, CHT did not affect in vitro mucin secretion from HTSE cells; (2) CHT significantly inhibited the expression levels of EGF- or TNF-alpha-induced MUC5AC gene in NCI-H292 cells. However, HST did not affect the expression levels of EGF- or TNF-alpha-induced MUC5AC gene in NCI-H292 cells; (3) CHT significantly inhibited hypersecretion of in vivo mucin. However, HST did not affect hypersecretion of in vivo mucin. Conclusion: These results suggest that CHT can not only affect the secretion of mucin but also the expression of the mucin gene and could be helpful for treating pulmonary disease caused by secretion of mucin.

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Comparative effects of angiotensin II and angiotensin-(4-8) on blood pressure and ANP secretion in rats

  • Phuong, Hoang Thi Ai;Yu, Lamei;Park, Byung Mun;Kim, Suhn Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.6
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    • pp.667-674
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    • 2017
  • Angiotensin II (Ang II) is metabolized from N-terminal by aminopeptidases and from C-terminal by Ang converting enzyme (ACE) to generate several truncated angiotensin peptides (Angs). The truncated Angs have different biological effects but it remains unknown whether Ang-(4-8) is an active peptide. The present study was to investigate the effects of Ang-(4-8) on hemodynamics and atrial natriuretic peptide (ANP) secretion using isolated beating rat atria. Atrial stretch caused increases in atrial contractility by 60% and in ANP secretion by 70%. Ang-(4-8) (0.01, 0.1, and $1{\mu}M$) suppressed high stretch-induced ANP secretion in a dose-dependent manner. Ang-(4-8) ($0.1{\mu}M$)-induced suppression of ANP secretion was attenuated by the pretreatment with an antagonist of Ang type 1 receptor ($AT_1R$) but not by an antagonist of $AT_2R$ or $AT_4R$. Ang-(4-8)-induced suppression of ANP secretion was attenuated by the pretreatment with inhibitor of phospholipase (PLC), inositol triphosphate ($IP_3$) receptor, or nonspecific protein kinase C (PKC). The potency of Ang-(4-8) to inhibit ANP secretion was similar to Ang II. However, Ang-(4-8) $10{\mu}M$ caused an increased mean arterial pressure which was similar to that by 1 nM Ang II. Therefore, we suggest that Ang-(4-8) suppresses high stretch-induced ANP secretion through the $AT_1R$ and $PLC/IP_3/PKC$ pathway. Ang-(4-8) is a biologically active peptide which functions as an inhibition mechanism of ANP secretion and an increment of blood pressure.