• Title/Summary/Keyword: High temperature incubation

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Studies on the Production and Characteristics of Glucose Isomerase from Steptomyces sp. GI 32. (Streptomyces GI 32 방선균의 Glucose Isomerase 생산과 효소특성)

  • 서형주;김진만;이태경;양한철
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.198-201
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    • 1989
  • Steptomyces sp. GI 32 with high production of glucose isomerase was isolated from soil. The maximum enzyme production was observed in the culture medium containing 1% sorbitol, 0.6% tryptone, 0.4% yeast extract, 1mM Fe$_2$(SO$_4$)$_3$ with initial pH 7.0 when the cell was cultured at 35$^{\circ}C$ about 18 hours with shaking. The enzyme was partially purified by ammonium sulfate fractionation and DEAE cellulose chromatography. The enzyme was also appeared to be relatively thermostable, and no apopreciable inactivation was observed after incubation at 7$0^{\circ}C$ for 1 hour. The optimal pH and temperature of the enzyme were pH 8.0 and 7$0^{\circ}C$, respectively.

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Properties of β-Galactosidase from Lactobacillus salivarius subsp. salivarius Nam27

  • Bae, Hyoung-Churl;Renchinkhand, Gereltuya;Nam, Myoung-Soo
    • Food Science of Animal Resources
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    • v.27 no.1
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    • pp.110-116
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    • 2007
  • Lactobacillus salivarius subsp. salivarius Nam27 with a high ${\beta}$-galactosidase activity was selected for enzymatic characterization. For purification, cell pellet was disrupted by Bead Beater, by DEAE-Sepharose and Mono-Q chromatography. The specific activity of the purified enzyme was 5,312 units/mg. The molecular weight of native monomeric ${\beta}$-galactosidase was estimated to be 30,000 dalton (monomer) by the SDS-PAGE. The optimum temperature and optimum pH were $50^{\circ}C$ and 5.0, respectively. This enzyme was stable between 35 and $55^{\circ}C$. ${\beta}$-galactosidase activity was lost rapidly above pH 7.0. But ${\beta}$-galactosidase was more stable at pH 4.0 (acidic conditions). And ${\beta}$-galactosidase activity was lost rapidly above $65^{\circ}C$ after 10 min incubation. $Ca^{2+}$ and $Zn^{2+}$ metal ions enhanced ${\beta}$-galactosidase activity by 164.09% and 127.37% while $Cu^{2+}$, $Fe^{3+}$ and $Mn^{2+}$ lowered ${\beta}$-galactosidase activity by 58.29%,85.10% and 77.66% respectively. Other metal ions didn't affect ${\beta}$-galactosidase activity significantly.

Deamidation on Glutaminyl and Asparaginyl Residues of Protein by Neutrase (Neutrase에 의한 glutamine과 aspargine 잔기의 탈아미드화)

  • Kim, Hyo-Sun;Kang, Yeung-Joo
    • Korean Journal of Food Science and Technology
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    • v.27 no.5
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    • pp.794-798
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    • 1995
  • Deamidation by neutrase on glutaminyl and asparaginyl residues of protein was examined. The optimum pH and temperature for BSA(bovine serum albumin) deamidation by neutrase were 10 and $20^{\circ}C$, respectively. The incubation for 3 hrs under the optimum condition removed 24% of amide groups and hydrolyzed 2.9% of peptide bonds. Deamidation by neutrase was superior to that by pronase, bromelain, or ficin. Deamidation degrees of egg albumin, soy protein isolate and casein by neutrase under the optimum condition were about 20%, 14% and 14%, respectively. However, relatively high degree of peptide hydrolysis was accompanied with casein deamidation.

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Effect of Reaction Conditions on the Thiamine Decomposition by Bracken (고사리의 Thiamine 분해에 미치는 반응조건(反應條件)의 영향)

  • Yoon, Jae-Young;Lee, Su-Rae
    • Korean Journal of Food Science and Technology
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    • v.20 no.4
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    • pp.600-605
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    • 1988
  • Antithiamine activity of raw and cooked brackens(Pteridium aquilinum) was evaluated under various reaction conditions by means of the thiochrome fluorescence method. The effects of caffeic acid and cysteine on the thiamine decomposition were also determined by thiochrome fluorescence and Lactobacillus viridescens bioassay methods. A water extract of raw bracken exhibited a high antithiamine activity which was increased with higher pH, temperature, incubation time and concentration of bracken. The influence of reaction conditions was less apparent in cooked bracken than in raw bracken. Caffeic acid stimulated the thiamine decomposition whereas cysteine showed a suppressive effect. The effect of cysteine was lower in the decomposition of thiamine by bracken extract.

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Screening of Microorganisms Secreted High Efficient Enzymes and Properties of Enzymatic Deinking for Old Newsprint(IV) -Culture conditions of fungal extracellular enzyme production for biological deinking system- (고효율 효소를 분비하는 균주의 선발 및 신문고지의 효소탈묵 특성(제4보) -고지탈묵용 Fungal Cellulase와 Xylanase의 생산-)

  • Park Seong-Cheol;Kang Jin-Ha;Lee Yang-Soo
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.37 no.1 s.109
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    • pp.38-46
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    • 2005
  • This study was focused on the optimum culture condition in CMCase, FPase and xylanase activities of two fungal strains that secret extracellular enzymes for using enzymatic deinking agent to old newsprint. The results of this study were as follows. When Fusarium pallidoroseum was grown on the medium, containing of rice bran+xylan $2.0\%,\;peptone\;0.6\%,\;KH_2PO_4\;0.075\%\;and\;MnSO_4\;0.06\%\;with\;pH\;9.0,\;at\;29^{\circ}C$ for 6 days, the quantitative degree of extracellular enzyme production was the highest. Optimum culture condition for Aspergillus niger was pH 5.0, $27^{\circ}C$ incubating temperature and 7 days incubation period on liquid medium, containing of CMC+xylan $2.5\%,\;yeast\;extract\;0.4\%,\;K_3PO_4\;0.05\%\;and\;CaCl_2+FeSO_4\;0.08\%$. Aspergillus niger was fairly higher FPase and xylanase activities than Trichoderma reesei ATCC 28217.

Bioproduction and Anticancer Activity of Biosurfactant Produced by the Dematiaceous Fungus Exophiala dermatitidis SK80

  • Chiewpattanakul, Paramaporn;Phonnok, Sirinet;Durand, Alain;Marie, Emmanuelle;Thanomsub, Benjamas Wongsatayanon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1664-1671
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    • 2010
  • A new biosurfactant producer was isolated from palm-oil-contaminated soil and later identified through morphology and DNA sequencing as the yeast-like fungus Exophiala dermatitidis. Biosurfactant production was catalyzed by vegetable oil, supplemented with a basal medium. The culture conditions that provided the biosurfactant with the highest surface activity were found to be 5% palm oil with 0.08% $NH_4NO_3$, at a pH of 5.3, with shaking at 200 rpm, and a temperature of $30^{\circ}C$ for a 14-day period of incubation. The biosurfactant was purified, in accordance with surfactant properties, by solvent fractionation using silica gel column chromatography. The chemical structure of the strongest surface-active compound was elucidated through the use of NMR and mass spectroscopy, and noted to be monoolein, which then went on to demonstrate antiproliferative activity against cervical cancer (HeLa) and leukemia (U937) cell lines in a dose-dependent manner. Interestingly, no cytotoxicity was observed with normal cells even when high concentrations were used. Cell and DNA morphological changes, in both cancer cell lines, were observed to be cell shrinkage, membrane blebbling, and DNA fragmentation.

Simple Screening Method for Double-strand DNA Binders Using Hairpin DNA-modified Magnetic Beads

  • Jo, Hun-Ho;Min, Kyoung-In;Song, Kyung-Mi;Ku, Ja-Kang;Han, Min-Su;Ban, Chang-Ill
    • Bulletin of the Korean Chemical Society
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    • v.32 no.1
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    • pp.247-250
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    • 2011
  • We designed an effective screening method for double strand DNA (dsDNA) binders using DNA-modified magnetic particles. Hairpin DNA was immobilized on the surface of magnetic particle for a simple screening of dsDNA binding materials in a solution containing various compounds. Through several magnetic separation and incubation processes, four DNA-binding materials, DAPI, 9AA, AQ2A, and DNR, were successfully screened from among five candidates. Efficiency of screening was demonstrated by HPLC analysis using a C2/18 reverse-phase column. In addition, their relative binding strengths were verified by measuring the melting temperature ($T_m$). If hairpin DNA sequence is modified for other uses, this magnetic bead-based approach can be applied as a high-throughput screening method for various functional materials such as anti-cancer drugs.

Protoplast fusion between saccharomyces cerevisiae and candida cariosilignicola (Saccharomyces cerevisiae와 candida cariosilignicola사이의 세포융합에 관한 연구)

  • 이재동;임하선
    • Korean Journal of Microbiology
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    • v.26 no.1
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    • pp.37-43
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    • 1988
  • This research was focused on investigation of the condition for protoplast formation and regeneration of protoplast fusion between Saccharomyces cerevisiae which has fermentation ability and Candida cariosilignicola which can grow at high temperature and utilize methanol. The results obtained were as follows; The highest production was collected in exponential growth phase. Ninety-nine% protoplast formation of C. cariosilignicola was obtained in glycin-NaOH buffer (pH10.0) containing Zymolyase 0.5mg/ml at $35^{\circ}C$ for 1hr incubation. The highest regeneration was produced when protoplast wuwpension containing 0.5% soft agar in buffered 50mM $CaCl_{2}$ was poured as a soft overlay onto 2% agar plates. Equal amuont of protoplast suspension of two strains was mixed and centrifuged. The subsequent pellet was added to 2ml of 35% polyethylene glycol (MW 4,000) containing 50mM $CaCl_{2}$, and incubated at $30^{\circ}C$ for 10min. Then 0.1ml of the suspension of aggregated protoplast was immediately covered with minimal medium and incubated at $40^{\circ}C$ for 5-7 days. As results, $SC_{1}$, $SC_{2}$, and $SC_{3}$ fusants were obtained. The physiological characteristics of fusants produced by protoplast fusion were; $SC_{1}$, and $SC_{2}$ utilized maltose, galactose, methanol, potassium nitrate. $SC_{3}$ utilized all the above materials except galactose.

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Organic-Inorganic Hybrid Nanoflowers as Potent Materials for Biosensing and Biocatalytic Applications

  • Tran, Tai Duc;Kim, Moon Il
    • BioChip Journal
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    • v.12 no.4
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    • pp.268-279
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    • 2018
  • Flower-shaped organic-inorganic hybrid nanostructures, termed nanoflowers, have received considerable recent attention as they possess greatly enhanced activity, stability, durability, and even selectivity of entrapped organic biomolecules, which are much better than those from the conventional methods. They can be synthesized simply via co-incubation of organic and inorganic components in aqueous buffer at room temperature and yield hierarchical nanostructures with large surface-to-volume ratios, allowing for low-cost production by easy scale-up, as well as the high loading capacity of biomolecules without severe mass transfer limitations. Since a pioneering study reported on hybrid nanoflowers prepared with protein and copper sulfate, many other organic and inorganic components, which endow nanoflowers with diverse functionalities, have been employed. Thanks to these features, they have been applied in a diverse range of areas, including biosensors and biocatalysis. To highlight the progress of research on organic-inorganic hybrid nanoflowers, this review discusses their synthetic methods and mechanisms, structural and biological characteristics, as well as recent representative applications. Current challenges and future directions toward the design and development of multi-functional nanoflowers for their widespread utilization in biotechnology are also discussed.

A simple chemical method for conversion of Turritella terebra sea snail into nanobioceramics

  • Sahin, Yesim Muge;Orman, Zeynep;Yucel, Sevil
    • Journal of Ceramic Processing Research
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    • v.19 no.6
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    • pp.492-498
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    • 2018
  • In this study, a sea shell was converted into bioceramic phases at three different sintering temperatures ($450^{\circ}C$, $850^{\circ}C$, $1000^{\circ}C$). Among the obtained bioceramic phases, a valuable ${\beta}-TCP$ was produced via mechanochemical conversion method from sea snail Turritella terebra at $1000^{\circ}C$ sintering temperature. For this reason, only the bioceramic sintered at $1000^{\circ}C$ was concentrated on and FT-IR, SEM/EDX, BET, XRD, ICP-OES analyses were carried out for the complete characterization of ${\beta}-TCP$ phase. Biodegradation test in Tris-buffer solution, bioactivity tests in simulated body fluid (SBF) and cell studies were conducted. Bioactivity test results were promising and high rate of cell viability was observed in MTT assay after 24 hours and 7 days incubation. Results demonstrated that the produced ${\beta}-TCP$ bioceramic is qualified for further consideration and experimentation with its features of pore size and ability to support bone tissue growth and cell proliferation. This study suggests an easy, economic method of nanobioceramic production.