• Title/Summary/Keyword: thermostable

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Enzymatic Characterization of a Thermostable 4-α-Glucanotransferase from Thermotoga neapolitana (Thermotoga neapolitana 유래 내열성 4-알파-글루칸전이효소의 효소적 특성)

  • Choi, Kyoung-Hwa;Seo, Ja-Yeong;Kim, Ji-Eun;Cha, Jae-Ho
    • Journal of Life Science
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    • v.21 no.2
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    • pp.221-226
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    • 2011
  • The gene encoding 4-$\alpha$-glucanotransferase (mgtA) from Thermotoga neapolitana was cloned and expressed in Escherichia coli in order to investigate whether this enzyme was capable of producing cycloamylose for industrial applications. MgtA was purified to homogeneity by HiTrap Q HP and Sephacryl S-200 HR column chromatographies. The size of the enzyme as determined by SDS-PAGE was about 52 kDa, which was in good agreement with its deduced molecular mass of 51.9 kDa. The optimal temperature and pH for the activity of the 4-$\alpha$-glucanotransferase was found to be $85^{\circ}C$ and 6.5, respectively. The enzyme hydrolyzed the 1,4-$\alpha$-glucosidic bonds in oligomeric 1,4-$\alpha$-glucans and transferred oligosaccharides (maltotriose being the shortest one) to acceptor maltodextrins. However, the enzymes had no activity against pullulan, glycogen, and other di- or trioligosaccharides with rare types of $\alpha$-bond. MgtA is distinguished from 4-$\alpha$-glucanotransferase from Thermotoga maritima in that it can convert maltotriose into maltooligosaccharides. The treatment of glucoamylase after the reaction of MgtA with maltotriose, maltotetraose, maltopentaose, or maltohexaose as sole substrate revealed that MgtA yielded linear maltooligosaccharides instead of cycloamylose.

Removal of mycotoxin ochratoxin A by isolated bacteria (분리세균에 의한 진균독소 ochratoxin A의 제거)

  • Choi, Ho-Yeong;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.55 no.1
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    • pp.33-38
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    • 2019
  • Ochratoxin A (OTA), one of mycotoxins produced mainly by Aspergillus is a common contaminant of stored grains, posing health hazards to human and livestock. The aim of this study is to explore ability of isolated bacteria Bacillus subtilis AF13 and Streptomyces shenzhenensis YR226 to remove OTA. AF13 and YR226 could remove 94.23 and 97.73% of OTA ($100{\mu}g/L$), respectively during 24 h incubation in NB medium. When cultures of two strains were separated into washed cells and cell-free supernatant, the supernatant of both strains removed more than 90% of $100{\mu}g/L$ OTA, and 98.88% of OTA could be also removed by the washed cells of YR226. OTA removal occurred in a few second by the supernatant of both strains, and treatments of autoclaving, proteinase K and chymotrypsin did not affect the OTA removal by the culture supernatants, which indicate that some thermostable and non-proteinaceous substances secreted by these bacteria may be involved in OTA removal in these two bacteria. These results suggest that AF13 and YR226 can be used to remove OTA from OTA-contaminated grains and feeds, and therefore decrease economic damage in agriculture and feed industry.

Changes of Chlorophyll and SOD-like Activities of Chinese Chives Dehydrated at Different Heat Treatments (부추의 건조 온도 조건별 클로로필, Superoxide Dismutase 유사활성의 변화)

  • Kwak, Yeon-Ju;Kim, Jong-Sang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.7
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    • pp.879-884
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    • 2009
  • Allium tuberosum Rotter (Liliaceae, Chinese chives) is a perennial herb of which leaves are used for food. This study investigated the effect of pretreatment on quality of dehydrated Chinese chives. Chinese chives was blanched at $80^{\circ}C$ for 20 sec, followed by drying at $70^{\circ}C$, $80^{\circ}C$, $100^{\circ}C$, or drying at $100^{\circ}C$ for 30 min and subsequent drying at $70^{\circ}C$, or $100^{\circ}C$ for 60 min and subsequent drying at $70^{\circ}C$. Optimum drying temperature and time was $100^{\circ}C$ for 30 min and subsequent drying at $70^{\circ}C$, or $100^{\circ}C$ for 60 min and subsequent drying at $70^{\circ}C$. These conditions were shortened time for dehydration and showed smaller decrease than others in Hunter color L, a, b. Dehydrated Chinese chives showed a constant decrease in greenness with storage, probably due to destruction of chlorophyll by light. In the measurement of Hunter color L, a, b, these conditions showed smaller decrease than others in Hunter color for 15 week storage. Chlorophyll content and SOD (superoxide dismutase)-like activity in that condition was higher than others. It was assumed that a phenolic compound that forms its thermostable activity. The fitness of drying models was conducted in order to explain reducing chlorophyll loss and SOD (superoxide dismutase)-like activity loss. Based upon the chlorophyll content, SOD-like activity, and retention of green color of the vegetable, optimum drying conditions was $100^{\circ}C$ for 30 min followed by $100^{\circ}C$ for 30 min and subsequent drying at $70^{\circ}C$, or $100^{\circ}C$ for 60 min and subsequent drying at $70^{\circ}C$.

Studies on the Cold Attenuation and Protective Effects of a Thermostable Newscastle Disease Virus Isolated from Korean Pheasants (한국산 꿩으로부터 분리한 열안정성 뉴캣슬병 바이러스의 저온순화와 방어효과)

  • K. H. Kwak;S. C. Han;T. J. Kim;K. S. Chang;M. H. Jun;H. J. Song
    • Korean Journal of Poultry Science
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    • v.28 no.2
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    • pp.83-89
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    • 2001
  • Newcastle disease virus, CBP-1 strain isolated from Korean pheasants was passaged for 173 times by 9-day-old specific pathogenic free (SPF) embryonated eggs at $37^{\circ}C$ (parent strain) and subsequently passaged for 15 (cold attenuation (CA) -15) and 30(cold attenuation (CA) -30) times by 10-day-old of commercial broiler chicks embryonated eggs at $29^{\circ}C$, respectively, The Physical and chemical properties (sensitivity to lipid solvents, low pH and thermostability), pathogenicity (mean death time, intracerebral pathogenic index and intravenous patho-genic index), safety, booster or protective effect and characterization of temperature sensitivity were measured in cold attenuated CA-15 or 30 strain and compared to those of parent CBP-1 strain. NDV, CBP-1 CA-30 strain acquired cold attenuation and decreased infectivity at $41^{\circ}C$ compared to those of parent strain grown at $37^{\circ}C$. It lost hemagglutination activity (HA) and cell infectivity at $56^{\circ}C$ for 30, 60, and 120 Min. CA-30 strain treated with ethyl ether also lost its HA and cell infectivity. Both CA-30 and parent strains exhibited a little resistant to HA at pH 3.0 glycine HCI buffer. Intracerebral pathogenic index (ICPI) and intravenous pathogenic index (IVPI) of parent strain were 1.12 and 1.45, but decreased to 0.75 and 0.00 in CA-30 strain, respectively. The safety was evaluated by mortality in chicks inoculated with 10$^{4.0}$ EID$_{50}$ /0.1 ml. The mortalities of parent, CA-30 and commercial Bl strains were 17.5, 12.0 and 0.0%, respectively. The safety of CA-30 strain was higher than that of parent strain. The booster effects of CA-30 strain and parent strain performed in 4-week-old chicks after being vaccinated with primary commercial Bl strain.

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Characterization of Cellulases from Schizophyllum commune for Hydrolysis of Cellulosic Biomass (Schizophyllum commune에 의한 Cellulase 생산 및 섬유소계 바이오매스의 당화를 위한 효소적 특성)

  • Kim, Hyun-Jung;Kim, Yoon-Hee;Cho, Moon-Jung;Shin, Keum;Lee, Dong-Heub;Kim, Tae-Jong;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.38 no.6
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    • pp.547-560
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    • 2010
  • The optimum culture condition of Schizophyllum commune for the cellulase production and its enzymatic characteristics for saccharification of cellulosic biomass were analyzed. S. commune secrets ${\beta}$-1,4-xylosidase (BXL) and cellulases, including endo-${\beta}$-1,4-glucanase (EG), cellobiohydrolase (CBH), and ${\beta}$-glucosidase (BGL). The optimum reaction temperature for all cellulases was $50^{\circ}C$ and the thermostable range was $30{\sim}40^{\circ}C$C. The optimum reaction pH for all cellulases was 5.5 in a range of temperature from $0^{\circ}C$ to $55^{\circ}C$. The best nutritions for the cellulase production of S. commune among tested nutrients were 2% cellulose for the carbon source and corn steep liquor or peptone/yeast extract for the nitrogen source without vitamins. The environmental culture condition for the cellulase production was 5.5~6.0 for pH at $25{\sim}30^{\circ}C$. The enzyme activities of EG, BGL, CBH, and BXL were 3670.5, 631.9, 398.5, and 15.2 U/$m{\ell}$, respectively, after concentration forty times from the culture broth of S. commune which was grown at the optimized culture condition. Alternative filter paper unit assay showed 11 FPU/$m{\ell}$ enzyme activity. The saccharification tests using cellulase of S. commune showed the low saccharification rate on tested hardwoods but a high value of 50.5% on cellulose, respectively. The saccharification rate (50.5%) of cellulose by cellulase produced in this work is higher than 45.7% in the commercial enzyme (Celluclast 1.5L, 30 FPU/g, glucan).

Molecular Epidemiological Characteristics of Vibrio Parahaemolyticus as Recently wilde-spreaded in Korea (최근 한국에서 유행하는 장염비브리오균의 분자 역학적 특성)

  • 김상숙;이희무;이중복
    • KSBB Journal
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    • v.18 no.6
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    • pp.522-528
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    • 2003
  • The purpose of this study is to inquire into molecular epidemiological characteristics of Vibrio parahaemolyticus. For this study, 120 strains(120 strains of Vibrio parahaemolyticus sampled from diarrhea patients) were examined and analyzed for biochemical characteristics, TDH (thermostable direct hemolysin) antibiotics sensitivity and detection of toxR, gyrE, tdh, and tds gents. G-S PCR (Group Specific Polymerase), PFGE (Pulsed-field Gel Electrophoriesis) methods were performed on the materials from patients were results. 1 Vibrio parahaemolyticus didn't grow in 0% density of NaCl, but the fact was found that those grew in 8% density of NaCl. 2. O:K serotypes of Vibrio parahaemolyticus was turned up in domestic patients was 17 types. Among those O3:K6 was the most, it was 68.3%. 3. In 18 kinds of antibiotic tests resistant against Ampicillin, Ticacillin was comparatively high. the case of resistant against Ampicillin, Ticacillin, Vancomycin at the multiple resistant was 52.5%. 4. Toxin gene tdh had only 109 strains among 120 ones isolated from patients held the genes of 199bp size, and 11 strains was negativity 5. In the test of Kanagawa toxic productivity, 107 strains among strains isolated from patients appeared to be positivity reaction 6. The strain that held trh toxin was only 3, and those among test strains had the genes of 250bp size and that had tdh, trh genes at a time were 3 strains, and TDH toxic productivity of those were 16 times, and it was weak. 7. Group Specific-PCR appeared to be useful in the confirmation of O3:K6 serotype interrelations. 8. Three strains which showed difference of 7 DNA sequence even in the same serotype were detected by the result of analyzing the regular gene, toxRS DNA sequence. These strains are differ from general strains which carry infection easily. 9. These mutual dose epidemiological relations were classified into smaller-parts through PFGE method. As a result of such classify, 3 findings were found. V. parahaemolyticus sampled from diarrhea patients were classified into 3 types. And third, the result obtained through PFGE method can be used as a useful tool in a point of molecular-epidemiological view.

Genetic Characteristics and Virulence Factors of Pandemic Vibrio parahaemolyticus Isolated in South Korea and Japan (한국과 일본에서 유행하는 장염비브리오의 병원성 인자와 유전자의 특성)

  • Hong, Seuk-Won;Moon, Ji-Young;Lee, Bok-Kwon;Kim, Yung-Bu
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.386-395
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    • 2007
  • A total of 27 strains of Vibrio parahaemolyticus (18 strains isolated from Korea and 9 strains from Japan) were serotyped and examined for biochemical characteristics, antimicrobial susceptibility patterns, cytotoxicity assay, thermostable direct hemolysin (TDH) production and molecular epidemiology. Using polymerase chain reaction (PCR) method and DNA probe hybridization method, the strains were tested for toxR, tdh, trh and ORF 8 genes. The V. parahaemolyticus isolated from patients were belonged to 8 different serotypes : O3:K6, O1:K38, O3:K57, O4:K9, O4:Kl2, O4:K68, O5:Kl5 and O6:K46. Urease-positive strain possessed the trh gene, and conversely, urease-negative strains lacked the gene, indicating that urease production by V. parahemolyticus strains strongly correlates with the possession of the trh gene. Most strains showed multiple resistant to more than three antibiotics and the antibiogram could be classified into 6 group (I to VI). All of the O3:K6 strains isolated in South Korea and Japan producted TDH at high levels. The TDH titers ranged between 256 and 2.048, and the average titer was 1009. To distinguish the new and increasingly common V. parahaemolyticus strains from clinical isolates, ORF 8 is a useful genetic marker. After Southern hybridization, the HindIII restriction fragment patterns of the tdh gene were grouped one type, respectively. One type showed two bands one of which was 4.3kb and the other was 11.5kb in size. Variation between the O3:K6 serotype are minor when compared to the differences seen with the non O3:K6 strains. The migration patterns of Not I -digested of the total DNA of the O3:K6 strains were similar, and only slight variations were observed between the serotypes. By contrast, the O3:K6 strains and non O3:K6 had markedly different profiles. In conclusion, Random amplified polymorphic DNA (RAPD) profile using appropriate primers was an effective epidemiological marker.