• Title/Summary/Keyword: isolation of enzymes

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The Characteristics of Bacteria Isolated from Ordinary Korean Soy Sauce and Soybean Paste (한국(韓國) 재래식(在來式) 간장 및 된장에서 분리(分離)한 세균(細菌)의 특성(特性))

  • Kwon, Oh-Jin;Kim, Jong-Kyu;Chung, Yung-Gun
    • Applied Biological Chemistry
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    • v.29 no.4
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    • pp.422-428
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    • 1986
  • The bacteria were isolated from ordinary Korean soy sauce and soybean Paste. After isolation, we investigated the bacteria which produces the characteristic flavor of ordinary Korean soy sauce and soybean paste, nothing the aroma, presence of amino acids, and free sugars. The results were obtained as follows. The bacteria isolated from ordinary Korean soy sauce and soybean paste were various Bacillus species. The isolated bacteria produced characteristic orders: 'Meju' order, the characteristic saline odor of ordinary Korean soy sauce or ordinary Korean soybean paste order, and enzymes: protease and amylase. The good characteristic saline odor of ordinary Korean soy sauce was produced by Bacillus licheniformis$(SSB_3)$. The good odor of ordinary Korean soybean paste was produced by Bacillus polymyxa$(SSB_4)$, Bacillus species$(SPB_1)$, Bacillus brevis$(SPC_2)$, and Bacillus lichniformis$(SPC_{2-1})$.

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Isolation of Angiotensin I-Converting Enzyme Inhibitory Peptide from Chungkookjang (청국장으로부터 Angiotensin I 전환효소 저해 Peptide의 분리)

  • Matsui Toshiro;Yoo Hyung Jae;Hwang Jae Sung;Lee Dong Seok;Kim Han Bok
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.355-358
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    • 2004
  • Chunkookjang, Korean traditional fermented soybean food emerges as a functional food to improve intestinal function and blood circulation. During Chungkookjang fermentation, microorganisms, enzymes, and diverse bioactive compounds increase sharply. Chungkookjang contains diverse oligo-peptides. Formation of peptides was confirmed by SDS-PAGE. Solube fermented soybean in our sample contained Tyr, Gln-Lys, Trp, Gln, and Lys-Pro as major components. Lys-Pro (0.083 mg/100 g sample) was purified by HPLC analysis. Angiotensin I­converting enzyme (ACE) causes hypertension by converting angiotensin I to angiotension II. ACE inhibitory activity of Lys-Pro was determined to be $IC_{50}=32.1\;{\mu}M.$ Whether or not eating Chungkookjang can lower blood pressure was also determined. Sistolic blood pressure dropped by 15 mmHg, and diastolic blood pressure by 8 mmHg 2 hr after a single administration of 20 g of fermented soybean. Chungkookjang might be helpful in improving blood circulation since it has ACE inhibitor and antihypertenisve effect.

Isolation of a Thermophilic Mutant, Talaromyces luteus 2004 in relation to the Regulation of Carboxymethylcellulase Production and Enzymatic characteristics (고온성 변이균주 Talaromyces luteus 2004의 분리와 Carboxymethylcellulase의 생성 조절 및 효소의 특성)

  • Hong, Mi-Kyung;Han, Hyo-Young;Jung, Young-Hee;Min, Kyung-Hee
    • The Korean Journal of Mycology
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    • v.24 no.3 s.78
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    • pp.206-213
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    • 1996
  • Talaromyces luteus 2004, a thermophilic mutant of T. luteus 6112 was obtained by mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine. T. luteus 2004 produced thermophilic carboxymethylcellulase (CMCase), and other polysaccharide enzymes: avicellase, xylanase, and ${\beta}-glucosidase$. Induction of CMCase production was shown at the highest level in 3% carboxymethylcellulose (CMC) minimal broth, indicating that CMC could work as an inducer. However, glucose and D-cellobiose showed catabolite repression for CMCase production which was under the control of CMC utilization. Optimal conditions for CMCase activity were at $70^{\circ}C$ and pH 4.0, suggesting that CMCase of T. luteus 2004 was a thermophilic enzyme.

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A Simple Method for Extraction of High Molecular Weight DNA fromPorphyra Tenera (Rhodophyta) Using Diatomaceous Earth

  • Kim, Tae-Hoon;Hwang, Mi-Sook;Song, Ju-Dong;Oh, Min-Hyuk;Moon, Yong-Hwan;Chung, Ik-Kyo;Rhew, Tae-Hyoung;Lee, Choon-Hwan
    • ALGAE
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    • v.21 no.2
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    • pp.261-266
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    • 2006
  • The innate soluble polysaccharides and phenolic compounds of marine macroalgae are serious contaminants which interfere with experimental procedures such as restriction enzyme digestion, polymerase chain reaction (PCR) and other enzymatic reactions using extracted DNA samples. The viscous polysaccharides are co-precipitated with DNA samples by isopropanol or ethanol precipitation in conventional experiment. To overcome the problem, a method for the isolation of high molecular weight DNA from Porphyra tenera is developed with the application of diatomaceous earth column. The isolated DNAs by this method were about 50-100 kb in size and could be digested well with restriction enzymes. The nuclease activity seemed to be minimal, and high reproducibility in the arbitrary primed PCR for RAPD analyses was a distinctive feature. These results suggest that this method is very efficient in isolating nucleic acid from macroalgae including Porphyra.

Isolation, Identification, and Probiotic Properties of Lactobacillus reuteri HY701 from Human Feces

  • Kim, Jun-Tae;Jung, Hwang-Young;Lee, Na-Kyoung;Rhim, Seong-Lyul;Paik, Hyun-Dong
    • Food Science and Biotechnology
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    • v.15 no.5
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    • pp.677-682
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    • 2006
  • Strain HY701 was isolated from human feces for probiotic use by selecting highly resistant isolates to artificial gastric acid and bile acid. Strain HY701 was identified as Lactobacillus reuteri using 16S rDNA sequencing, and tentatively named L. reuteri HY701. The resistance of L. reuteri HY701 to artificial gastric acid (PH 2.5) was high with a survival rate of over 90%. L. reuteri HY701 also showed high tolerance to artificial bile acid after incubation in artificial gastric acid. Using the API ZYM test kit, the carcinogenic enzymes (${\beta}$-glucuronidase and (${\beta}$-glucosidase were not detected with L. reuteri HY70l, while the beneficial enzyme (${\beta}$-galactosidase was weakly detected. L. reuteri HY701 was sensitive to $100\;{\mu}g/mL$ nisin, $20\;{\mu}g/mL$ roxithromycin, $15\;{\mu}g/mL$ erythromycin, but resistant to $20\;{\mu}g/mL$ streptomycin, $10\;{\mu}g/mL$ tetracycline, $20\;{\mu}g/mL$ ciprofloxacin, $20\;{\mu}g/mL$ nystatin, $20\;{\mu}g/mL$ gentamycin, $10\;{\mu}g/mL$ doxycycline, $10\;{\mu}g/mL$ chloramphenicol, and $20\;{\mu}g/mL$ ampicillin. L. reuteri HY701 was shown to possess bactericidal activity as it inhibited the growth of Listeria monocytogenes ATCC 19111 and Escherichia coli JM109 completely within 24 hr of incubation. These results indicate that L. reuteri HY701 could be used as a probiotic strain.

Isolation and Characterization of Naturally Occuring Bacteria Carried TOL Plasmid (TOL 플라스미드 세균의 분리와 특성)

  • Nam, Cho-Byung;Cho, In-Sun;Rhee, Young-Ha;Ryu, Jae-Keun;Min, Kyung-Hee
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.321-326
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    • 1989
  • Eighty two bacterial strains have been isolated from five different soil and sewage samples by selective enrichment culture on m-toluate minimal medium. Two of these were identified as Pseudomonas capacia, one as P. putida, one as Yersinia intermedia, and one as Flavobaeterium odoratum. P. cepacia SUB37 appeared to carry plasmid superficially similar to TOL plasmid previously described in p. putida mt-2 and other two plasmids from Flavobacterium odorutum and Y. intermedia larger than that of p. putida mt-2. p. cepacia SUB37 was sensitive to streptomycin but resistant to rifampicin. P. cepacia SUB37 carrying plasmid metabolizes the hydrocarbons to benzoate and toluates via the corresponding alcohols and aldehydes. By the curing experiment, it appears that P. cepacia SUB37 carries TOL plasmid encoding for the enzymes responsible for the catabolism of toluene and xylene via benzoate and the toluates and then by meta pathway in the process of degradation of aromatic hydrocarbons. p. cepacia SUB37 degraded m-toluate rapidly to be very low level when it was fully grown.

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Studies on The Molds Affecting To The Cotton Textiles (면사제품(綿絲製品)에 번식(繁殖)하는 미생물(微生物)에 관(關)한 연구(硏究))

  • Han, Young-Ku
    • Applied Biological Chemistry
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    • v.13 no.3
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    • pp.243-256
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    • 1970
  • Nowadays, the damages caused by molds in cotton textile goods becomes influential. In our country, however, the relations between cotton goods and molds are not investigated and studied in detail. Two hundred and fifty seven kind of mold's samples were collected in ninety places through the whole country. The molds samples are mainly gathered according to each regions and seasons from molded cotton textiles. Out of this samples, we isolated six hundred and seventy two strains of molds and the results of isolation are following. 1. The distributed molds were Aspergillus sp., Rhizopus sp., Penicillium sp., etc. among them Aspergillus sp. were most widely distributed, and next were Rhizopus sp., Penicillium sp. etc. 2. The distribution of Aspergillus sp. abounded peculiarly in the dry season, while Rhizopus sp. in the rainy season. 3. The C.M.C, descomposing enzymes forming activity on molds were greatly concerned with intensity damage of cotton textile goods. 4. The formation of C.M.C. decomposing enzyme was only influenced by physiology of each strains. 5. Regarding to the growth. a. The molds which were saprophyting on the cotton textile goods were indicated vigorous growing. b. Among isolated six hundred and seventy two strains, there were above a hundred strains which produced pigment and nearly half of them fifty nine strains were Aspergillus sp. 6. Twenty one strains in isolated six hundred and seventy two strains were indentified which can heavily damage upon cotton textile. As a results of indentification of the selected strains, the following species was abtained, Aspergillus sydowi, wentii, niger, luchuensis, flavus, fumigatus, nidulans, Penicillium frequentants, roqueforti, chrysogenum, albicans, Rhizopus oligosporus, delemar, Mucor rouxii, mucedo, Neurospora sitophila, Monilia variabilis, fructigena, Cladsporium hurbarum and Aspergillus spp. Mucor spp.

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Halotolerant Spore-Forming Gram-Positive Bacterial Diversity Associated with Blutaparon portulacoides (St. Hill.) Mears, a Pioneer Species in Brazilian Coastal Dunes

  • Barbosa Deyvison Clacino;Irene Von Der Weid;Vaisman Natalie;Seldin Lucy
    • Journal of Microbiology and Biotechnology
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    • v.16 no.2
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    • pp.193-199
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    • 2006
  • Halotolerant spore-forming Gram-positive bacteria were isolated from the root, rhizosphere, and non-rhizosphere soil of Blutaparon portulacoides. The different isolates were characterized genetically using an amplified ribosomal DNA restriction analysis (ARDRA), and phenotypically based on their colonial morphology, physiology, and nutritional requirements. Three different 16S rRNA gene-based genotypes were observed at a 100% similarity using the enzymes HinfI, MspI, and RsaI, and the phenotypic results also followed the ARDRA groupings. Selected strains, representing the different ARDRA groups, were analyzed by 16S rDNA sequencing, and members of the genera Halobaeillus, Virgibacillus, and Oceanobacillus were found. Two isolates showed low 16S rDNA sequence similarities with the closest related species of Halobacillus, indicating the presence of new species among the isolates. The majority of the strains isolated in this study seemed to belong to the species O. iheyensis and were compared using an AP-PCR to determine whether they had a clonal origin or not. Different patterns allowed the grouping of the strains according to Pearson's coefficient, and the resulting dendrogram revealed the formation of two main clusters, denoted as A and B. All the strains isolated from the soil were grouped into cluster A, whereas cluster B was exclusively composed of the strains associated with the B. portulacoides roots. This is the first report on the isolation and characterization of halotolerant spore-forming Gram-positive bacteria that coexist with B. portulacoides. As such, these new strains may be a potential source for the discovery of bioactive compounds with industrial value.

Isolation and Characterization of Engineered Nucleoside Deoxyribosyltransferase with Enhanced Activity Toward 2'-Fluoro-2'-Deoxynucleoside

  • Yoo, Yeon-Jin;Choi, Kang-Hyun;Kim, Byoung-Kyun;Choi, Si-Sun;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.8
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    • pp.1041-1046
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    • 2022
  • Nucleoside deoxyribosyltransferase (NDT) is an enzyme that replaces the purine or pyrimidine base of 2'-deoxyribonucleoside. This enzyme is generally used in the nucleotide salvage pathway in vivo and synthesizes many nucleoside analogs in vitro for various biotechnological purposes. Since NDT is known to exhibit relatively low reactivity toward nucleoside analogs such as 2'-fluoro-2'-deoxynucleoside, it is necessary to develop an enhanced NDT mutant enzyme suitable for nucleoside analogs. In this study, molecular evolution strategy via error-prone PCR was performed with ndt gene derived from Lactobacillus leichmannii as a template to obtain an engineered NDT with higher substrate specificity to 2FDU (2'-fluoro-2'-deoxyuridine). A mutant library of 214 ndt genes with different sequences was obtained and performed for the conversion of 2FDU to 2FDA (2'-fluoro-2'-deoxyadenosine). The E. coli containing a mutant NDT, named NDTL59Q, showed 1.7-fold (at 40℃) and 4.4-fold (at 50℃) higher 2FDU-to-2FDA conversions compared to the NDTWT, respectively. Subsequently, both NDTWT and NDTL59Q enzymes were over-expressed and purified using a His-tag system in E. coli. Characterization and enzyme kinetics revealed that the NDTL59Q mutant enzyme containing a single point mutation of leucine to glutamine at the 59th position exhibited superior thermal stability with enhanced substrate specificity to 2FDU.

Studies on the Isolation, Purification and Characterization of a Cx Enzyme Produced by Pyricularia oryzae, $C-7^{+t}$ (도열병균에서 추출한 Cx효소의 순화 및 특성에 관한 연구)

  • Kim, Sang-Ho;Kim, W.S.
    • The Korean Journal of Mycology
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    • v.10 no.2
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    • pp.67-73
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    • 1982
  • The $(NH_4)_2\;SO_4$ (70%) treated crude enzymes from the culture filtrates of the$C-7^{+t}$ strain of Pyricularia oryzae which was grown on 2% CMC (carboxymethyl cellulose) for 8 days at $28^{\circ}C$ were chromatographied on Sephadex G-150 and DEAE-Sephadex A-25 columns. From the chromatography, three fractions of CMCase$(C_x)$ was examined using Na-CMC as substrate. The $C_x$ enzyme activity was optimal at pH 6.0 and $40^{\circ}C$, stable up to $40^{\circ}C$. The values of Km and Vmax of the enzyme were $2.8{\times}10\;mM$ and 5.9m moles/hour, respectively. The molecular weight determined by Sephadex G-150 column chromatography was around 80,000. Approximately sevenfold purified $C_x$ enzyme gave a single protein band on the polyacrylamide gel electrophoresis.

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