• Title/Summary/Keyword: Enzyme extract

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Optimization of Refolding Conditions for the Aklavinone 11-Hydroxylase of Streptomyces peucetius Overexpressed in Escherichia coli. (대장균에서 대량 발현된 Streptomyces peucetius유래 Aklavinone 11-Hydroxylase효소의 최적 가용화 조건)

  • 민우근;홍영수;최용경;이정준;홍순광
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.365-368
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    • 1998
  • The aklavinone 11-hydroxylase which was overexpressed using T7 promoter in E. coli could be detected in SDS-PAGE only in insoluble precipitate without any detectable enzyme activity. The insoluble enzyme was solubilized in 6M guanidine$.$HCl solution and their refolding ability was tested under various conditions. When the enzymatic activity was checked by the bioconversion experiment, stepwise dialysis against 6M, 3M, 1M guanidine$.$HCl and finally 100 mM potassium phosphate buffer of the solubilized protein gave the best bioconversion efficiency. The aklavinone 11-hydroxylase showed its enzymatic activity in the reaction buffer containing NADPH with vigorous shaking. The enzymatic activity was lost during partial purification and regained by the addition of crude extract of S. lividans in the reaction mixture. This effect was confirmed to due to some low-molecular weight component(s) in the crude extract, because the addition of dialyzed crude extract could not recover the enzymatic activity.

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The Effects of Irritating Spices on Drug Metabolizing Enzyme Activity -Effects on Hexobarbital Hypnosis in Mice- (자극성(刺戟性) 향신제(香辛劑)의 약물대사효소활성(藥物代謝酵素活性)에 미치는 영향(影響) -마우스의 Hexobarbital 수면시간(睡眠時間)에 미치는 영향(影響)-)

  • Woo, Won-Sick;Shin, Kuk-Hyun;Kim, In-Chull
    • Korean Journal of Pharmacognosy
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    • v.8 no.3
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    • pp.115-119
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    • 1977
  • Black pepper (Piper nigrum L.) among several irritating spices tested was highly effective on the duration of hexobarbital hypnosis in mice. Pretreatment of mice with the methanolic extract of black pepper (60mg/kg i.p.) prolonged markedly the duration of hexobarbital sleeping time. Three consecutive daily administrations of the same dose of black pepper extract, however, shortened (37%) the duration of hexobarbital sleeping time. The ether soluble fraction of black pepper extract caused most potent effects on the duration of hexobarbital hypnosis. From the above results, it was postulated that the lipid soluble components of black pepper might considerably change the drug action and metabolism by altering drug metabolizing enzyme systems.

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Enhancing Extraction Yield of Chlorella Extract by Enzyme Treatment

  • In, Man-Jin;Jang, Jae-Eun;Kim, Dong-Ho
    • Journal of Applied Biological Chemistry
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    • v.50 no.3
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    • pp.132-135
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    • 2007
  • An efficient production method of chlorella extract was developed by enzymatic treatment using cell lytic and proteolytic enzymes. The suitable dosage of Tunicase, a cell lytic enzyme, was found to be 1.0% (w/w). Proteolytic enzymes were screened to obtain high chlorella growth factor (CGF) index, which indicates crude CGF content and solid recovery. Among the seven tested proteases, Esperase, whose optimal dosage was 1.0% (w/w), was selected. By co-treatment using optimal dosages of Tunicase and Esperase, the highest CGF index and solid recovery were obtained. The CGF index and solid recovery of co-treatment were remarkably enhanced by 250 ($4.36{\rightarrow}15.21$) and 220% ($12.65%{\rightarrow}40.15%$), respectively, than those of the non-treated extracts.

Screening of Inhibitor of Thyroid Peroxidase, an Oxidative Coupling Enzyme from Natural Products (생약으로 산화적 결합 효소인 갑상선 peroxidase의 저해제 검색)

  • 이현정;장미영;김미리;배기환;석대은
    • YAKHAK HOEJI
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    • v.43 no.3
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    • pp.334-341
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    • 1999
  • Thyroid peroxidase is a biochemical target protein for the antithyroid drugs. Ethanol extracts from one hundred and thirty seven natural products were screened for the inhibition of thyroid peroxidase activity. Thyroid peroxidase was purified from porcine thyroids, and the inhibition of peroxidase activity was evaluated using guaiacol oxidation (C-C coupling) assay. Twenty one natural products expressed a remarkable inhibition (>50%) of peroxidase activity at $330{\mu\textrm{g}}$ solid weight/m. The 50% inhibitory concentration ($IC_{50}$) of 70% ethanol extract from six potent natural products ranged from 3.1 to $31.2{\;}{\mu\textrm{g}}$ solid weight/m, in contrast to the range ($0.33~0.54{\;}{\mu\textrm{g}}/ml$) of $IC_{50}$ values fro catechin and epigallocatechin gallate as positive controls. Noteworthy, the extract of Camellia taliensis showed irreversible inhibition of the enzyme. It is suggested that extract from some natural products such as Camellia taliensis, Rheum undulatum or Euphorbia perinensis, exhibiting a potent inhibition of peroxidase activity, may be developed as sources of potent antithyroid agents.

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Changes in Enzyme Activities and Population of Lactic Acid Bacteria during the Kimchi Fermentation Supplemented with Water Extract of Pine Needle (솔잎(Pinus densiflora Sieb. et Zucc.) 물추출물 첨가김치의 숙성 중 젖산균수와 효소활성의 변화)

  • 오영애;최경호;김순동
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.2
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    • pp.244-251
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    • 1998
  • To understand the effect of supplement of water extract of pine needle(WEPN) on shelf-life enhancement of the kimchi, activities of four enzymes and number of lactic acid bacteria, during fermentation of the kimchi, were assayed. Enzyme activities of kimchi fermented for 7 days with supplement by 2% water extract of pine needle showed amylase of 86.4%, protease of 85.8%, polygalacturonase of 61.5% and $\beta$-galactosidase of 58.8% against the control kimchi. WEPN showed weak inhibitory effect when it was applied to the isolated enzymes in vitro then those menifested by the kimchi in vivo. Number of total bacterial cell of WEPN supplemented kimchi increased by 10 folds than control between 7 to 14 days of fermentation. On contrast, number of lactic acid bacteria decreased maximaly to 21% of control by fermentation. The clear zone formed on paper disk by WEPN against L. plantarum was larger than that of Leu. mesenteroides.

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Effect of Cnidi rhizoma Boiling Extract Solution on Enzyme and Hormone of Plasma, and Liver in the Fatted Rats Induced by High Fat Dietary (천궁의 열수추출액이 고지방식이에 의한 흰쥐의 혈장 중 효소활성과 호르몬 및 간장의 지방축적에 미치는 영향)

  • 성태수;손규목
    • The Korean Journal of Food And Nutrition
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    • v.7 no.2
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    • pp.108-113
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    • 1994
  • This study was carried out to evaluate the effect of Cnidi rhizoma (CR) water extract on fat accumulation In fatted rats induced by the oral high fat administration for six weeks. To accomplish this evaluation, the serum and liver tissue have been examined for enzyme activity, cortisol and insulin level. The change of liver or tissue have been observed by the light microscope. GOT GPT and LDH activities were lower than the control group. Insulin and cortisol were higher than the control group, due to the fat accumulation. The liver of the control group observed by the tight microscope appeared to the fatty liver, but CR group showed some improvement of the fatty liver Based on the above results, it was shown that it is possible to improve fat accumulation induced by high fat dietary through using the oral administration of Cnidi rhizoma water extract.

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Biological activity of St. John's wort (Hypericum perforatum L.) (St. John's wort(Hypericum perforatum L.)의 생리활성 효과)

  • Cho, Young-Je;Chun, Sung-Sook;Yoon, So-Jung;Kim, Jeung-Hoan
    • Applied Biological Chemistry
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    • v.48 no.1
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    • pp.65-69
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    • 2005
  • The physiological activity of St. John's wort extracts were examined. Total phenol contents in the ethanol extracts $(246.0{\pm}10.5\;{\mu}g/ml)$ with St. John's wort leaf was higher than that in water extract $(237.4{\pm}13.2\;{\mu}g/ml)$. The electron donating ability in the water extracts and in the ethanol extracts were 95.0% and 95.2% respectively. Antioxidant protection factor of the ethanol extract was higher than that of the water extract. The water extract from St. John's wort leaves did not show an antimicrobial activity against Helicobacter pylori, but the ethanol extract revealed high antimicrobial activities such as 11 mm of clear zone in $100\;{\mu}g/ml$ of phenol content and 13 mm of clear zone in $150\;{\mu}g/ml$ of phenol content. The hot water extract showed an angiotensin converting enzyme inhibitory activity of 19.2%. The xanthin oxidase inhibitory activity of hot water and ethanol extract were very high, amounting to 84.8% and 100% respectively. The results suggested a possibility for developing the phenol compounds in St. John's wort as anti Helicobacter pylori, anti-oxidant and anti-gout agents.

Effect of Chongmyung-Tang Prescription Combination on the Production of Amyloid β protein and β-site amyloid precursor protein-cleaving enzyme Activity in vitro (In vitro에서 β-site amyloid precursor protein-cleaving enzyme 활성과 amyloid β protein 생산에 대한 총명탕가미방(聰明湯加味方)의 효과)

  • Lim, Jung-Hwa;Jung, In-Chul;Lim, Jong-Soon;Kim, Seung-Hyung;Lee, Sang-Ryong
    • Journal of Oriental Neuropsychiatry
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    • v.21 no.2
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    • pp.191-200
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    • 2010
  • Objectives : This experiment was designed to investigate the effect of Chongmyung-Tang Prescription Combination(CmTP-$C_{1-10}$) extract on the production of amyloid $\beta$ protein and $\beta$-site amyloid precursor protein-cleaving enzyme(BACE) activity. Methods : The effect of CmTP-$C_{1-10}$ extract on expression of APP mRNA, BACE2 mRNA in BV2 microglia cell line treated by lipopolysacchride(LPS) and amyloid $\beta$ protein fragment(A$\beta$ fragment) were investigated. The effect of CmTP-$C_{1-10}$ extract on production of amyloid $\beta$ protein(A$\beta$) in BV2 microglia cell line treated by LPS and A$\beta$ fragment were investigated. The effect of CmTP-$C_{1-10}$ extract on BACE activity were investigated. Results : 1. CmTP-$C_9$ extract the most significantly suppressed the expression of APP mRNA, BACE2 mRNA in BV2 microglia cell line treated by LPS and A$\beta$ fragment. 2. CmTP-$C_9$ extract significantly suppressed the production of A$\beta$ in BV2 microglia cell line treated by LPS and A$\beta$ fragment. 3. CmTP-$C_9$ extract the most significantly inhibited BACE activity. Conclusions : These results suggest that CmTP-$C_9$ may be effective for the prevention and treatment of Alzheimer's Disease. Investigation into clinical use of CmTP-$C_9$ for Alzheimer's Disease is suggested for future research.

EFFECT OF ZIZYPHI FRUCTUS EXTRACT ON THE BIOLOGICAL ACTIVITY OF GINGIVAL FIBROBLAST (대조 추출물분획이 치은 섬유아세포의 생물학적 활성화에 미치는 영향)

  • Yang, Chang-Ho;Lee, Yong-Moo;Cho, Ki-Yeong;Bae, Ki-Hwan;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.24 no.1
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    • pp.144-154
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    • 1994
  • Final goal of periodontal treatment is to reconstruct the destructed periodontal tissue as well as to remove the necrotic pathologic elements. The purpose of this study is to investigate on the effect of Zizyphi extract to the inhibitory ability on collagenolytic activity of P gingivalis, biologic activity of gingival fibroblasts, and on the collagen and protein synthesis of gingival fibroblasts. Gingival fibroblast from giniva of first bicuspids from patient for orthodontic treatment were used and cultured. For the measurement of inhibitory ability of collagenolytic activity, crude enzyme was extracted and used on the basis of modified Ono's method. On the inhibition of collagenolytic enzyme from herbal extracts, collagenokit CLN-100 were used. The cellular activity of gingival fibroblast, were studied using MTT solution and measured optical density on 570mm by ELISA reader. To measure the effects on the ability of whole protein and collagen synthesis, cell membrane was destructed with ultrasonic grinder after culturing, centrifuged and counted by liquid scintilation counter. The inhibitory effects on producing of $IL-l{\beta}$ by monocyte, after promotion of producing $IL-l{\beta}$ by LPS, were compared with the mixture of herbal extracts and other drugs using thymocyte stimulation assay. About inhibitory effects of $PGF_2$. by gingival fibroblasts, herbal extract was compared with the addition of the other control groups using enzyme imunoassay. On the inhibition of collagenolytic activity by P. gingivalis, benzene extracts showed the most efficient inhibitory effects among the $19{\mu}g/ml$ of the compared extracts and 40.5% by Tetracycline. On the cellular activity promoting effects, compared extracts showed a bit of more effects than PDGF of $100{\mu}g/ml$ concentration and IGF of $20{\mu}g/ml$ concentration. All of the PDGF, IGF, Zizyphi Fructus extract should increase in collagen synthesis, but especially 70% ethylalcohol extracts of Zizyphi Fructus showed comparably high effects among the compared extracts. Effects on whole protein synthesis were slightly increased on every extract but especially 70% ethylalcohol extract showed significantly effective than any other estract. On the inhibitory effects of Zizyphi Fructus $IL-l{\beta}$ production by monocyte, compared extracts showed 70% of highly inhibitory effect than that of 60% inhibition effects on controlled group and each extracts showed no significant difference. In $PGF_2$ production inhibitroy effect of Zizyphi Fructus gingival fibroblasts, Herbal extracts showed 70% of inhibition comparing with tat of 90.2% of controlled group, but each extracts showed similar effects excluding the $H_2O$ extracts. These results suggested that Zizyphi Fructus might be useful medicine for inhibition of inflammatory mediator including $IL-l{\beta}$ and $PGF_2$.

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Production and Characterization of Fibrinolytic Enzyme: Optimal Condition for Production of the Enzyme from Bacillus sp. KP-6408 Isolated from Chungkook-jang (새로운 혈전용해 효소의 생성 및 특성: 청국장에서 분리한 Bacillus sp. KP-6408로부터 효소 생성의 최적조건)

  • 박인식;길지은;김기남
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.1
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    • pp.51-56
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    • 1998
  • A bacteium, KP-6408, capable of hydrolyzing fibrin was isolated from Chungkook-jang, which was possibly identified as a strain of Bacillus sp. The effects of culture condition and medium composition on the enzyme production were investigated. Among nitrogen sources tested, yeast extract was the most effective for the enzyme production, and the level of the concentration for the optimal enzyme production was 0.2%(w/v). For carbon sources, glucose was the best for the enzyme production with the level of 2.0%(w/v). The enzyme was maximally produced by cultivating the enzyme production with the level of 2.0%(w/v). The enzyme was maximally produced by cultivating the organism at the liquid medium of the initial pH 8.0 and temperature of 4$0^{\circ}C$. In Chungkook-jang fermentation, the enzyme was maximally produced when incubated at 35$^{\circ}C$ for 24 hrs using soybean as a solid medium. The addition of various rice starch to the soybean in Chungkook-jang fermentation lowered the enzyme production.

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