• Title/Summary/Keyword: Enzyme I

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High Production of Thermostable Beta-galactosidase of Bacillus stearothemophilus in mesophiles

  • Okada, Hirosuke;Hirata, Haruhisa;Negoro, Seiji
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.509.1-509
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    • 1986
  • Recent advances in recombinant DNA techniques have provided a tool for breeding of microorganisms of hyper production. Enzyme production by cloned microorganism has some advantages. They are ⅰ) Enzymes can be produced by a microorganism easily cultured ⅱ) Hyper production. ⅲ) In some cases, such as thermophilic enzyme gene is cloned in a mesophilic bacteria, the enzyme purification procedure can be simplified. One example, production of thermophilic ${\beta}$-galactosidase in B. subtilis will be presented. Bacillus stearothermophilus IAM 11001 produced three ${\beta}$-galactosidases, ${\beta}$-galactosidase I, II and III (${\beta}$-gal-I, II and III). By connecting restriction fragments of the chromosomal DNA to plasmid vector, followed by transformation of Escherichia coli, two ${\beta}$-galactosidase genes (bgaA and bgaB) located close to each other on the chromosome were cloned.

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Angiotensin-Converting Enzyme Gene Polymorphism is not Associated with Myocardial Infarction in Koreans

  • Chai, Seok;Sohn, Dong-Ryul
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.5
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    • pp.645-650
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    • 1998
  • To assess the relationship between angiotensin-converting enzyme (ACE) gene polymorphism and myocardial infarction in Koreans, we recruited 112 healthy, unrelated subjects (mean age 53.4 years) and 104 myocardial infarction survivors (mean age 54.2 years) of both sexes. An insertion/deletion (I/D) polymorphism of the ACE gene was typed by polymerase chain reaction. The I allelic frequency of ACE gene in Korean subjects was irrelavant to myocardial infarction (patients, 65 control subjects 66%), as was true with the D allele. When compared with other populations, the frequency of D allele in Koreans (0.34) was lower than that in Caucasians, and was close to that of other Oriental populations. The data suggest that the ACE gene polymorphism is not an independent genetic risk factor for myocardial infarction in Koreans.

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Effects of Organophosphorus Fungicides on Cutinase Activity and Infection of Apple by Glomerella cingulata (사과 탄저병균 Glomerella cingulata에 의한 감염과 Cutinase의 활성에 미치는 유기인계 살균제의 효과)

  • 김기홍;이창은
    • Korean Journal Plant Pathology
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    • v.10 no.2
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    • pp.119-122
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    • 1994
  • Effects of organophosphorus fungicides on cutinase activity from Glomerella cingulata causing apple anthracnose and infection of apple were investigated. Diisoprophylfluorophosphate (DFP) inhibited the enzyme activity indicating that catalysis involves an active serine. In inhibition of the enzyme activity by organophosphorus fungicides, I50 (molar concentration of fungicide at which the enzyme activity is inhibited 50%) of Hinosan and Kitazin P were 26.3 $\mu$M and 427.7 $\mu$M, respectively. At concentration of 10-3 M DFP and organophosphorus fungicides, the infection of G. cingulata was inhibited to 5% in comparison with 15% infection at the unwounded healthy control, but increased to 30% when added with 1 mg/ml of cutinase. Mycelial growth was 36 mm in colony diameter on the medium added with 10-4M of hinosan in comparison with 90 mm of the untreated control, but was 90 mm on the medium added with 10-4M of kitazin P showing lower inhibition than hinosan. The spore germination was more than 60% at all the concentrations of both fungicides.

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A study on Alkalophilic Bacteria Producing $\beta$-Galactosidase. -Isolations and Cultural Characteristics (I)- ($\beta$-Galactosidase를 생산하는 호알카리성 세균에 관한 연구 -균주의 분리 및 배양조건 (I)-)

  • Yun, Seong-Sik;Min, Do-Sik;Yu, Ju-Hyeon
    • The Korean Journal of Food And Nutrition
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    • v.1 no.2
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    • pp.68-75
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    • 1988
  • A strain of alkalophilic Bacillus sp. YS-309 has been isolated from domestic soil. It belongs to genus Bacillus from its morphological and biochemical characteristics. The strain grows better in the alkaline media rather than in the neutral media. The optimum pH and temperature for growth were observed at 8.5 and 4$0^{\circ}C$, respectively. Glucose, lactose and maltose were appeared as good carbon source but soluble starch and fructose were utilized uneffectively for growth. Concentrations of lactose had affected both the cellular growth and the enzyme productions. The maximum growth and the highest enzyme productions were obtained at 0.5%(w/e) of lactose added in the media. B-Galactosidase from Bacillus sp. YS-309 was produced inducibly into the cell and total enzyme activities per ml were gradually decreased when the concentration of glucose increased.

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The Effect of Scoparone on the Hepatic Bromobenzene Metabolizing Enzyme System in Rats (간의 Bromobenzene 대사계에 미치는 Scoparone의 효과(I))

  • Kim, Eun-Ju;Lee, Chung-Kyu;Choi, Jong-Won
    • Korean Journal of Pharmacognosy
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    • v.23 no.2
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    • pp.81-88
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    • 1992
  • The effects of scoparone, one of coumarin derivative on the hepatic bromobenzene metabolizing enzyme system was estimated in rats. Scoparone pretreatment revealed dose-dependently the recovery of decrease in epoxide hydrolase activity due to the bromobenzene(310 mg/kg, i.p.) treatment. And also scoparone and scopoletin (each 5mg/kg, p.o.) pretreatments showed two times increase in the $V_{max}$ values compared to those of bromobenzene-treated group which were calculated from tripartite reciprocal plots. The mode of protective effect of scoparone against bromobenzene induced toxicity is considered to be due to the induction of microsomal enzyme activity by scopoletin, the intermediate metabolite of scoparone. The changes in cytochrome P-450 activity, aminopyrine N-demethylation, aniline hydroxylation and glutathione S-transferation in scoparone-treated group were not significantly different from those of the control group.

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Antioxidative Action of Enzymatic Hydrolysates of Mackerel Muscle Protein (고등어 근육단백질 효소 가수분해물의 항산화 작용)

  • 염동민;김영숙
    • The Korean Journal of Food And Nutrition
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    • v.7 no.2
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    • pp.128-136
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    • 1994
  • Mackerel muscle protein hydrolysates, which were prepared from defatted mackerel meal by proteases such as complex enzyme, alcalase, bromelain, pancrease, pepsin, w-chymotrypsin, trypsin and papain, were tested for the antioxidative action against linoleic acid. Among proteases tested, the hydrolysates obtained from the treatment of complex enzyme, bromelain and alcalase showed higher antioxidative effects. Also, the hydrolysates showed the synergistic effects with o-tocopherol and the inhibitory effects for peroxidation of metal ions(Fe3+, Cua+) From the profiles of fractionation of the hydrolysates with Bio-gel P-2 column, the most active fractions, part I(complex enzyme-derived) and part e(bromelain-derived), had below MW 1,400 and the antioxidative effects were closely related to the binding capacity with metal ion(Cua+). Amno acid composition of the part I was abundant in histidine, arginine, phenylalanine and lysine, and the part e was abundant in lysine, glutamic acid and leucine.

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Characterization of Antihypertensive Angiotensin I-Converting Enzyme Inhibitor from Recombinant E. coli (재조합 대장균으로부터 항고혈압 Angiotensin I-Converting Enzyme 저해제의 특성연구)

  • Kim, Jae-Ho;Jeong, Seung-Chan;Lee, Dae-Hyong;Lee, Jong-Soo
    • The Journal of Natural Sciences
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    • v.16 no.1
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    • pp.1-13
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    • 2005
  • The angiotensin I-converting enzyme (ACE) inhibitor has anti-hypertensive effects and has long been used as prevention or remedy of hypertension. This study were carried out to produce and purify a new ACE inhibitor from recombinant E. coli and further elucidate its structure-function relationship. Recombinant pGEX-4T-3 containing ACE inhibitory peptide gene of Saccharomyces cerevisiae was transformed into E. coli BL21(DE3). Glutathione-S transferase (GST) fusion protein from E. Coli BL21(DE3) harboring the recombination pGEX-4T-3 was obtained and the ACE inhibitory peptide was purified with Sephadex G-25 column chromatography. The purified ACE inhibitory peptide was a novel decapeptide with sequence Tyr-Asp-Gly-Gly-Val-Phe -Arg-Val-Tyr-Thr which shows very low similarity to the other ACE inhibitory peptide sequence. The purified ACE inhibitor competitively inhibited ACE.

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Enzymatic Production of Amylopectin Cluster Using Cyclodextrin Glucanotransferase (Cyclodextrin Glucanotransferase를 이용한 아밀로펙틴 클러스터의 생산)

  • Lee, Hye-Won;Jeon, Hye-Yeon;Choi, Hyejeong;Shim, Jae-Hoon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.9
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    • pp.1388-1393
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    • 2014
  • To enzymatically prepare amylopectin cluster (APC), cyclodextrin glucanotransferase (CGTase I-5) and its mutant enzyme from alkalophilic Bacillus sp. I-5 were employed, after which the hydrolysis patterns of CGTase wild-type and its mutant enzyme toward amylopectin were investigated using multi-angle laser light scattering. CGTase wild-type dramatically reduced the molecular weight of waxy rice starch at the initial reaction, whereas the mutant enzyme degraded waxy rice starch relatively slowly. Based on the results, the molecular weight of one cluster of amylopectin could be about $10^4{\sim}10^5g/mol$. To determine production of cyclic glucans from amylopectin, matrix-assisted laser desorption ionization time-of-flight mass spectrometry was performed. CGTase I-5 produced various types of cyclic maltooligosaccharides from amylopectin, whereas the mutant enzyme hardly produced any.

Expression Profiling of WSSV ORF 199 and Shrimp Ubiquitin Conjugating Enzyme in WSSV Infected Penaeus monodon

  • Jeena, K.;Prasad, K. Pani;Pathan, Mujahid Khan;Babu, P. Gireesh
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.8
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    • pp.1184-1189
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    • 2012
  • White spot syndrome virus (WSSV) is one of the major viral pathogens affecting shrimp aquaculture. Four proteins, WSSV199, WSSV 222, WSSV 249 and WSSV 403, from WSSV are predicted to encode a RING-H2 domain, which in presence of ubiquitin conjugating enzyme (E2) in shrimp can function as viral E3 ligase and modulate the host ubiquitin proteasome pathway. Modulation of host ubiquitin proteasome pathway by viral proteins is implicated in viral pathogenesis. In the present study, a time course expression profile analysis of WSSV Open Reading Frame (ORF) 199 and Penaeus monodon ubiquitin conjugating enzyme (PmUbc) was carried out at 0, 3, 6, 12, 24, 48 and 72 h post WSSV challenge by semi-quantitative RT-PCR as well as Real Time PCR. EF1${\alpha}$ was used as reference control to normalize the expression levels. A significant increase in PmUbc expression at 24 h post infection (h.p.i) was observed followed by a decline till 72 h.p.i. Expression of WSSV199 was observed at 24 h.p.i in WSSV infected P. monodon. Since the up-regulation of PmUbc was observed at 24 h.p.i where WSSV199 expression was detected, it can be speculated that these proteins might interact with host ubiquitination pathway for viral pathogenesis. However, further studies need to be carried out to unfold the molecular mechanism of interaction between host and virus to devise efficient control strategies for this chaos in the shrimp culture industry.

Peptide Inhibitors for Angiotensin I Converting Enzyme from Corn Gluten Digests. (옥수수 글루텐 효소 가수분해물의 Angiotensin I Converting Enzyme 활성 저해 펩타이드의 정제)

  • 오광석;이동건;홍정운;성하진
    • Microbiology and Biotechnology Letters
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    • v.31 no.1
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    • pp.51-56
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    • 2003
  • The angiotensin I converting enzyme (ACE) has an important role in the maintenance of blood pressure. The ACE inhibitory activities of foods have recently been studied. We tried to isolate ACE inhibitory peptides from the Flavourzyme (FZ), Pescalase (PE), and Thermolysine (TH) protease digests of corn gluten, which was restricted to the use the source of food for digestion problem. The FZ, PE, TH/PE protease hydrolyzed corn gluten and the inhibitory activities of the hydrolyzates for ACE were measured. Major fractions were isolated from the digests using ODS chromatography after treating with ethanol in step gradient. The ACE inhibitors were further purified by Bio-Gel P-2 column and reverse phase HPLC. Five inhibitory peptides were isolated. Their amino acids were sequenced as LPF ($IC_{50}$ = 40$\mu$M), GPP ($IC_{50}$ = 17.6$\mu$M), PNPY ($IC_{50}$ = 30.7$\mu$M), SPPPFYL ($IC_{50}$ = 63 $\mu$M), and SQPP ($IC_{50}$ = 17.2$\mu$M).