• Title/Summary/Keyword: Substrate culture

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Screening of New Antibiotics Inhibiting Bacterial Peptide Deformylase (PDF) (세균의 Peptide Deformylase(PDF)를 억제하는 새로운 항균물질의 스크리닝)

  • 곽진환;김현주;설민정;서병선;이종국;최수영
    • YAKHAK HOEJI
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    • v.47 no.3
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    • pp.184-189
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    • 2003
  • Peptide deformylase (PDF) is essential and unique to bacteria, thus making it an attractive target for the discovery of novel antibacterial drugs. PDF deformylates the N-formylmethionine of newly synthesized polypeptides in prokaryotes. In this study, a pdf gene from Staphylococcus aureus 6538p was cloned in pET-14b vector and PDF protein was over-produced in Escherichia coli BL21 (DE3). NH$_2$-terminal His-tagged PDF protein was purified by nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography. Enzymatic activity of purified 6xHis-tagged PDF was tested on the substrate (formyl-Methionine-Alanine-Serine) by formate dehydrogenase-coupled spectrometric assay of peptide deformylase. For the discovery of new PDF inhibitors from chemical libraries and culture broths of soil bacteria, a target-oriented screening system using a 96-well plate was developed. About 3,000 commercial chemical libraries were tested in this screening system, and 2 chemicals (0.07%) among them showed an inhibitory activity against PDF enzyme. This result showed that a new screening system can be used for the discovery of new PDF inhibitors.

Isolation of Antimicrobial Substances from Hericium erinaceum

  • Kim, Dong-Myong;Pyun, Chul-Woo;Ko, Han-Gyu;Park, Won-Mok
    • Mycobiology
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    • v.28 no.1
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    • pp.33-38
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    • 2000
  • Mycelium of Hericium erinaceum isolate KU-1 was cultured in liquid medium (HL medium) and solid medium (Ko medium) at pH 4.0 in $28^{\circ}C$. 1.0% glucose or fructose was the most favorable carbon source, and 0.2% amonium acetate or $NaNO_3$ was an exellent nitrogen source for mycelial growth as well as production of antimicrobial substances. The mixture of saw dust 70% with rice bran 30% (SR medium) was the substrate for formation of sporophores. The active substrates in extracts from mycelium, culture filtrate and fruiting body were separated by TLC. The solvent for TLC was EtOAc: Chloroform: MeOH (10 : 5 : 10). Phenol-like substances appeared at Rf $0.5{\sim}0.9$, and fatty acid-like substances appeared at Rf $0.1{\sim}0.2$. The purified materials from the extracts showed antimicrobial effects to Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Aspergillus niger, Candida albicans and Microsporum gypseum. The S. aureus was the most inhibited. Minimal inhibitory concentration (MIC) of purified white powder and the Hercenone derivatives against S. aureus were $5.65\;{\mu}g/ml$ and $1.85\;{\mu}g/ml$, respectively.

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Discovery of Two Chrysosporium Species with Keratinolytic Activity from Field Soil in Korea

  • Gurung, Sun Kumar;Adhikari, Mahesh;Kim, Sang Woo;Bazie, Setu;Kim, Hyun Seung;Lee, Hyun Goo;Kosol, San;Lee, Hyang Burm;Lee, Youn Su
    • Mycobiology
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    • v.46 no.3
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    • pp.260-268
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    • 2018
  • In an ongoing survey of Korean indigenous fungi, two fungal strains (KNU16-74 and KNU16-99) belonging to the genus Chrysosporium were isolated from field soil in Gyeongnam, Korea. Morphological characterization and phylogenetic analysis using sequence of the internal transcribed spacer regions were carried out to confirm its precise identification. These strains were identified as Chrysosporium indicum (KNU16-74) and Chrysosporium fluviale (KNU16-99). To examine the keratin degradation efficiency of these two fungal species, human hair strands were incubated with fungus culture. Results revealed that these two fungal species have the ability to degrade keratin substrate. This is the first report of these two species in Korea.

A Specific Short Dextrin-Hydrolyzing Extracellular Glucosidase from the Thermophilic Fungus Themoascus aurantiacus 179-5

  • Carvalho Ana Flavia Azevedo;Goncalves Aline Zorzetto;Silva Roberto da;Gomes Eleni
    • Journal of Microbiology
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    • v.44 no.3
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    • pp.276-283
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    • 2006
  • The thermophilic fungus Thermoascus aurantiacus 179-5 produced large quantities of a glucosidase which preferentially hydrolyzed maltose over starch. Enzyme production was high in submerged fermentation, with a maximal activity of 30 U/ml after 336 h of fermentation. In solid-state fermentation, the activity of the enzyme was 22 U/ml at 144 h in medium containing wheat bran and 5.8 U/ml at 48 h when cassava pulp was used as the culture medium. The enzyme was specific for maltose, very slowly hydrolyzed starch, dextrins (2-7G) and the synthetic substrate (${\alpha}$-PNPG), and did not hydrolyze sucrose. These properties suggest that the enzyme is a type II ${\alpha}$-glucosidase. The optimum temperature of the enzyme was $70^{\circ}C$. In addition, the enzyme was highly thermostable (100% stability for 10 h at $60^{\circ}C$ and a half-life of 15 min at $80^{\circ}C$), and stable within a wide pH range.

Degradation of Phenolic Resin, Resole by Microbial Consortia (미생물 컨소시엄에 의한 페놀수지 Resole의 분해)

  • 오계헌;최원식
    • KSBB Journal
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    • v.13 no.2
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    • pp.220-222
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    • 1998
  • Three microbial consortia were screened for their ability to degrade phenolic resin, resole as a sole carbon source. These microbial consortia were derived from soil samples collected from a phenolic resin manufacturing plant site. Among the consortia, the test consortium, designated as MS2, displayed approximately 70% degradation of the substrate, 100 mg of resole per liter, within the fist twelve days of incubation but the degradation was inhibited. During the incubation period, pH was decreased from 7.0 to 2.7, and the resole degradation became inhibited under the conditions. UV-scans of spent culture showed that the wavelength of maximum absorption was 261 nm for resole.

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Production of Chiral (S)-styrene Oxide by Rhodosporidium sp. SJ-4 which has an Epoxide Hydrolase Activity (에폭사이드 가수분해효소를 갖는 Rhodosporidium sp. SJ-4를 이용한 광학활성 (S)-styrene Oxide의 생산)

  • Yoo, Seung-Shick;Lee, Eun-Yeol;Kim, Hee-Sook;Kim, Jung-Sun;Oh, You-Kwan;Park, Sung-Hoon
    • Journal of Institute of Control, Robotics and Systems
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    • v.11 no.10
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    • pp.857-863
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    • 2005
  • A yeast strain utilizing styrene epoxide as a sole carbon and energy source was isolated from soil samples for the production of enantiopure of styrene epoxide by kinetic resolution. The strain, identified as a Rhodosporidium toruloides SJ-4, expressed an epoxide hydrolase which preferentially converted (R)-styrene epoxide into the corresponding diol. A maximum activity of 135 U/L was observed when biomass (dry cell mass) reached 6.7 g/L at 21 h of batch culture. Under the partially optimized reaction conditions ($35^{\circ}C$ and pH 8.0), the optically pure (S)-styrene epoxide was obtained with the yield of 21% when the initial substrate concentration was 100 mM. The reaction was completed at 9 h.

Fabrication of Hydrophilic Poly(dimethylsiloxane)with Periodic Wrinkling Surface and Its Application (일정한 주름을 갖는 친수성 PDMS 제작 및 응용)

  • Lee, Dong-Guk;Oh, Chang-Kyu;Yang, Sung-Ho;Han, Seung-Jin;Jeong, Ok-Chan
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.63 no.5
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    • pp.671-675
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    • 2014
  • This paper presents a fabrication of hydrophilic Poly(dimethylsiloxane) (PDMS) with periodic wrinkling surface. The proposed periodic wrinkling surface was fabricated using the sequential processes of typical curing process of PDMS, cutting process, platinum deposition process, and wrinkling transfer process. The surface morphology of the fabricated wrinkling surface was observed by using optical and dynamic atomic force microscopy and discussed. The measured period and amplitude of wrinkling was about $2.2{\mu}m$ and $0.31{\mu}m$, respectively. And, the contact angles of water droplets on the wrinkled surface were measured in order to understand effect of the wrinkling surface on surface modification of hydrophobic PDMS. Our new finding was that the proposed wrinkling surface was hydrophilic and the measured contact angle was about $62^{\circ}$. Moreover, it was found out from the simple cell culture test that the fabricated wrinkling surface was more effective for cell spreading and adhesion than the case of native PDMS substrate.

Purification and Characterization of a Major Fibrinolytic Enzyme from Bacillus amyloliquefaciens MJ5-41 Isolated from Meju

  • Jo, Hyeon-Deok;Lee, Hwang-A;Jeong, Seon-Ju;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.11
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    • pp.1166-1173
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    • 2011
  • Meju is a traditional Korean fermented soy product used as a key element for soy sauce and doenjang. Bacilli with antimicrobial activity were isolated from meju prepared by traditional methods at Sunchang county, Jeollabukdo, Korea. Six isolates were identified as Bacillus amyloliquefaciens by recA gene sequencing and RAPD-PCR. One isolate, B. amyloliquefaciens MJ5-41, showed the strongest fibrinolytic activity. A 27 kDa active fibrinolytic enzyme, AprE5-41, was purified from the culture supernatant of MJ5-41 grown on LB by chromatographic methods. The optimum pH and temperature for purified AprE5-41 were 7.0 and $45^{\circ}C$, respectively. AprE5-41 quickly degraded $A{\alpha}$ and $B{\beta}$ chains but not the ${\gamma}$-chain of fibrinogen. AprE5-41 exhibited the highest specificity for N-succinyl-Ala-Ala-Pro-Phe p-nitroanilide, a known substrate for ${\alpha}$-chymotrypsin, cathepsin G, and subtilisin BPN'. The structural gene, aprE5-41, was cloned by PCR and successfully expressed in B. subtilis.

Enhancing Factors of Electricity Generation in a Microbial Fuel Cell Using Geobacter sulfurreducens

  • Kim, Mi-Sun;Cha, Jaehwan;Kim, Dong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1395-1400
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    • 2012
  • In this study, we investigated various cultural and operational factors to enhance electricity generation in a microbial fuel cell (MFC) using Geobacter sulfurreducens. The pure culture of G. sulfurreducens was cultivated using various substrates including acetate, malate, succinate, and butyrate, with fumarate as an electron acceptor. Cell growth was observed only in acetate-fed medium, when the cell concentrations increased 4-fold for 3 days. A high acetate concentration suppressed electricity generation. As the acetate concentration was increased from 5 to 20 mM, the power density dropped from 16 to $13mW/m^2$, whereas the coulombic efficiency (CE) declined by about half. The immobilization of G. sulfurreducens on the anode considerably reduced the enrichment period from 15 to 7 days. Using argon gas to create an anaerobic condition in the anode chamber led to increased pH, and electricity generation subsequently dropped. When the plain carbon paper cathode was replaced by Pt-coated carbon paper (0.5 mg $Pt/cm^2$), the CE increased greatly from 39% to 83%.

Formation of Cheese Flavor and EMC Technology (치즈 풍미성분의 형성과 EMC 제조기술)

  • Han, Kyeong-Sik;Jeon, Woo-Min
    • Journal of Dairy Science and Biotechnology
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    • v.21 no.2
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    • pp.88-96
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    • 2003
  • Cheese flavor is derived from three main pathways, that are proteolysis, lipolysis and glycolysis, the extent of which varies according to the cheese variety. Proteolysis is the most complex of the three primary events during cheese ripening. The basis of EMC technology is the use of specific enzymes acting at optimum conditions to produce required cheese flavors from suitable substrates. These enzymes consist of proteinases, peptidases, lipases, esterases. The key factors in EMC production are the type of cheese flavor required, the type and specificity of enzyme or cultures used, their concentration and some processing parameters, such as pH, temperature, agitation, aeration, and incubation time. The emulsifiers, bacteriocins, flavor compounds, and precursors also effect to it importantly. The dosage of enzyme or starter culture used is dependent on the intensity of flavor required, processing time and temperature and the quality of the initial substrate. To produce a consistent EMC product it is necessary to have a highly controlled process, and a detailed knowledge of the enzymatic reactions under the conditions used must be fully understood.

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