• Title/Summary/Keyword: isolation of enzymes

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Isolation and identification of tumor-inducing (Ti) Plasmids (식물종양유도 플라스미드의 분리 및 동정)

  • 음진성;김계현;심웅섭
    • Korean Journal of Microbiology
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    • v.24 no.1
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    • pp.1-6
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    • 1986
  • Tumor-inducing (Ti) plasmid which may be used in genetic engineering of higher plants, was isolated from Agrobacterium tumefaciens in Korea. Among the 7 strains of A. tumefaciens isolated from various regions in Korea, KU12, KU13, KU14, and KU49 strains were confirmed to contain plasmid. The isolated Ti plasmids were digested with 4 kinds of restriction enzymes, respectively. According to the result of the cleavage patterns, we confirmed that these plasmids in the KU12, KU13, KU14 and KU49 strains are different.

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Isolation and Analysis of the Enzymatic Properties of Thermophilic Fungi from Compost

  • Lee, Hanbyul;Lee, Young Min;Jang, Yeongseon;Lee, Sangjoon;Lee, Hwanhwi;Ahn, Byoung Jun;Kim, Gyu-Hyeok;Kim, Jae-Jin
    • Mycobiology
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    • v.42 no.2
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    • pp.181-184
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    • 2014
  • To the best of our knowledge, this is the first report on thermophilic fungi isolated in Korea. Three species of thermophiles were isolated from compost and were identified as Myriococcum thermophilum, Thermoascus aurantiacus, and Thermomyces lanuginosus. They can grow at temperatures above $50^{\circ}C$ and produce high levels of cellulolytic and xylanolytic enzymes at high temperatures. Notably, the considerable thermostability of the endo-glucanase produced by T. aurantiacus has made the fungus an attractive source of industrial enzymes.

Uniqueness of Microbial Cutinases in Hydrolysis of p-Nitrophenyl Esters

  • KIM, YANG-HOON;JEEWON LEE;SEUNG-HYEON MOON
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.57-63
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    • 2003
  • Using fungal (Fusarium solani f. pisi) and bacterial (Pseudomonas mendocina) cutinases, the initial hydrolysis rate of p-nitrophenyl esters was systematically estimated for a wide range of enzyme and substrate concentrations using a 96-well microplate reader. Both cutinases exhibited a high substrate specificity; i.e. a high hydrolytic activity on p-nitrophenyl butyrate (PNB), yet extremely low activity on p-nitrophenyl palmitate (PNP). When compared to the hydrolysis of PNB and PNP by other hydrolases [lipases and esterases derived from different microbial sources, such as bacteria (Pseudomonas cepacia, Psedomonas furescens, Baciilus stearothermophilus), molds (Aspeillus niger, mucor miehei), and yeasts (Candida rugosa, Candida cylindracea)], the above substrate specificity would seem to be a unique characteristic of cutinases. Secondly, the hydrolytic activity of the cutinases on PNB appeared much faster than that of the other hydrolytic enzymes mentioned above. Furthermore, the current study proved that even when the cutinases were mixed with large amounts of other hydrolases (lipases or esterases), the Initial hydrolysis rate of PNB was determined only by the cutinase concentration for each PNB concentration. This property of cutinase activity would seem to result from a higher accessibility to the substrate PNB, compared with the other hydrolytic enzymes. Accordingly, these distinct properties of cutinases may be very useful in the rapid and easy isolation of various natural cutinases with different microbial sources, each of which may provide a novel industrial application with a specific enzymatic function.

Isolation and Characterization of White Rot Fungi for Decolorization of Several Synthetic Dyes (염료의 색도 제거능력이 우수한 백색부후균 분리 및 특성연구)

  • 오광근;김현수;조무환;채영규;전영중
    • Microbiology and Biotechnology Letters
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    • v.27 no.6
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    • pp.500-508
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    • 1999
  • Several white-rot fungi collected from the mountains of Korea were evaluated for their ability to decolorize azo, polymeric, and reactive dyes. Strains CJ-105, CJ-212 and CJ-315, identified as Trametes sp., Pleurotus sp. and Fomes sp., respectively, showed higher potential for decolorization of those dyes in either solid or liquid media. For Trametes sp. CJ-105, 100ppm of Remazol Brilliant blue R and 500ppm of Acid Red 264 were completely decolorized after 2 days under liquid culture. The dominating ligninolytic enzyme existing in the culture broth was laccase (E.C. 1.10.3.2). Also, Pleurotus sp. CJ-212 and Fomes sp. CJ-315 showed similar patterns in decolorization of Remazol Brilliant Blue R and Acid Red 264. The extent of decolorization of the dyes in liquid culture was found to be proportional to the activities of the ligninolytic of decolorization of the dyes in liquid culture was found to be proportional to the activities of the ligninolytic enzymes produced by each strain. In addition to that Trametes sp. CJ-105 was highly effective in degradation of polycyclic aromatic hydrocarbons and pentachlorophenol by the activity of the ligninolytic enzymes produced. In this study, we found that white-rot fungi, Trametes sp. CJ-105(KFCC 10941), Pleurotus sp. CJ-212(KFCC 10943) and Fomes sp. CJ-315(KFCC 10942), were effective in decolorizing a wide range of structurally different synthetic dyes, as well as some chemical compounds which are known to be hardly degradable.

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Restriction endonuclease maps of three plasmids from bacillus thuringiensis serovar israelensis 4Q1 (Bacillus thuringiensis serovar israelensis 4Q1로부터 분리된 plasmid 제한효소지도 작성)

  • Faust, R.M.;;;C.L.Meyers-Dowling;P.E.McCawley
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.122-128
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    • 1985
  • Bacillus thuringiensis serovar israelensis 4Q1 contains 8 different covalently closed-circular (CCC) plasmids of molecular weight 204, 267, 109, 103, 16, 7.6, 6.4, and 5.0kb. The three smallest plasmids, designated pBti6, and pBti8 may prove to be useful as cloning vectors because of thier size and ease of isolation. The three plasmids were incubated separately with 9 different restriction enzymes and 7 of the enzymes tested cleaved one or more of the plasmids. Plasmid pBti6 has a single site for Bg1 II, Pst I and Pvu II, two sites for Bc1 I and Eco RI, and five sites for Hind III. Plasmid pBti7 has a single site for Bam HI and Pst I, two sites for Hind III, and three sites for PvuII. Plasmic pBti8 has a single site for Bam HI, BelI and Hind III, two sites for Eco RI, and three sites for Bgl II and Pvu II. Composite restriction enzyme maps for pBti6, pBti7 and pBti8 were constructed. The sites of restriction enzyme cleavage were determined by single, double and partial digests of the plasmid DNA. All the restriction sites were aligned relative to the single Bgl II(pBti6), Pst I(pBti7) or Hind III(pBti8) site, respectively.

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Isolation and Characterization of Microbial Strains with Hydrolytic Enzyme Profile from Clay Minerals

  • Lee, Sulhee;Cho, Eui-Sang;Nam, Young-Do;Park, So-Lim;Lim, Seong-Il;Seo, Dong-Ho;Kim, Jae-Hwan;Seo, Myung-Ji
    • Microbiology and Biotechnology Letters
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    • v.48 no.1
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    • pp.64-71
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    • 2020
  • A total of 262 bacterial strains were isolated from clay minerals, bentonite and zeolite, in Gyeongsangbukdo, Republic of Korea, and their hydrolytic enzyme activities were analyzed. Most of the isolated strains belonged to Micrococcales and Bacillales order. Of strains, 96 strains produced α-amylase activity, 42 strains showed cellulase activity, 111 strains had pectinase activity, and 70 strains showed protease activity. Among them, 177 isolates exhibited one or more of the hydrolytic enzyme activities and in particular Bacillus cereus MBLB1321, B. albus MBLB1326 and KIGAM017, B. mobilis MBLB1328, MBLB1329 and MBLB1330 showed all of the enzyme activities. These results demonstrate the diversity of functional Bacillus species in clay minerals as vital sources for the discovery of industrially valuable hydrolytic enzymes, which have a great commercial prospect in various bio-industrial applications.

Rapid and Efficient Isolation of Genes for Biosynthesis of Peptide Antibiotics from Gram-positive Bacterial Strains

  • Lee, Soon-Youl;Rhee, Sang-Ki;Kim, Chul-Ho;Suh, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • v.8 no.4
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    • pp.310-317
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    • 1998
  • Peptide synthetases are large multifunctional enzyme complexes that catalyze the nonribosomal synthesis of a structurally diverse family of peptide antibiotics. These enzymes are composed of functionally independent domains with independent enzymatic activities. Their specific linkage order of domains forms the protein template that defines the sequence of the incorporated amino acids. Within each domain, several motifs of highly conserved sequences have been identified from the sequence alignment of the various peptide synthetases [30]. Taking advantage of the conserved nucleotide sequence of Core 1 and Core 2, we designed PCR primers to amplify the peptide synthetase genes from three different gram-positive bacterial strains. Nucleotide sequence analysis of the amplified PCR products from those three strains showed significant homology to various peptide synthetase genes, suggesting that the PCR products are parts of peptide synthetase genes. Therefore, this rapid and efficient PCR technique can be used for the isolation of peptide synthetase genes from various strains.

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In Vitro Culture of Endothelial Cell and Smooth Muscle Cell for Studying Vascular Diseases

  • Kim, Joo-Young
    • Journal of Yeungnam Medical Science
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    • v.27 no.2
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    • pp.91-97
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    • 2010
  • Endothelial cells play a key role in pathological processes such as cancer cell metastasis, atherosclerosis, and diabetic retinopathy. Vascular smooth muscle cells directly involve in the formation of atheroma in atherosclerosis. Some kinds of the endothelial cells are simply harvested from the umbilical veins, the tunica intima of aortic walls, the retina using various enzymes solutions. Those purely isolated cells provide a powerful tool in vitro studies of the endothelial cell related diseases. In this context, the cultured smooth muscle cells after the isolation from the tunica media of aortic walls are also used for elucidating the pathogenesis of atherosclerosis. Here, I briefly introduce articles that include the isolation of human umbilical vein endothelial cells(HUVEC), aortic endothelial and smooth muscle cells, retinal microvascular endothelial cells(RMEC), as well as the diseases' applications of these cells.

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Isolation and Characterization of Chlorella Virus from Fresh Water in Korea and Application in Chlorella Transformation System

  • Park, Hye-Jin;Yoon, Hong-Mook;Jung, Heoy-Kyung;Choi, Tae-Jin
    • The Plant Pathology Journal
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    • v.21 no.1
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    • pp.13-20
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    • 2005
  • Chlorella viruses are large icosahedral, plaque-forming, dsDNA viruses that infect certain unicellular, chlorellalike green algae. The genomic DNA of over 300 kb contains many useful genes and promoters. Over 40 chlorella viruses have been isolated from fresh water in Korea since 1998. The viruses were amplified initially in chlorella strain NC64A, and pure isolates were obtained by repeated plaque isolation. SDS-PAGE analysis revealed similar but distinct protein patterns, both among the group of purified viruses and in comparison with the prototype chlorella virus PBCV-1. Digestions of the 330- to 350-kb genomic DNAs with 10 restriction enzymes revealed different restriction fragment patterns among the isolates. The tRNA-coding regions of 8 chlorella viruses were cloned and sequenced. These viruses contain 14-16 tRNA genes within a 1.2- to 2-kb region, except for the SS-1 isolate, which has a 1039-bp spacer in a cluster of 11 tRNA genes. Promoter regions of several early genes were isolated and their activities were analyzed in transformed chlorella. Some promoters showed stronger activity than commonly used CaMV 35S promoter and chlorella transformation vectors for heterologous protein are beings constructed using these promoters.