• Title/Summary/Keyword: High temperature incubation

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The Bactericidal Effect of High Temperature Is an Essential Resistance Mechanism of Chicken Macrophage against Brucella abortus Infection

  • Arayan, Lauren Togonon;Reyes, Alisha Wehdnesday Bernardo;Hop, Huynh Tan;Xuan, Huy Tran;Baek, Eun Jin;Min, Wongi;Kim, Suk
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1837-1843
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    • 2017
  • Knowledge of avian host responses to brucellosis is critical to understanding how birds resist this infection; however, this mechanism is not well established. On the other hand, temperature has a major involvement in the physiology of living organisms, and cell death induced by heat is attributed to protein denaturation. This study demonstrates the direct bactericidal effect of a high temperature ($41^{\circ}C$) on Brucella abortus that resulted in the gradual reduction of intracellular bacteria and inhibited bacterial growth within avian macrophage HD11 in an increasing period of time. On the other hand, this study also revealed that high temperature does not affect the rate of bacterial uptake, as confirmed by the bacterial adherence assay. No significant difference was observed in the expression of target genes between infected and uninfected cells for both temperatures. This study suggests the susceptibility of B. abortus to bacterial death under a high temperature with an increased period of incubation, leading to suppression of bacterial growth.

Production of Mn-Dependent Peroxidase from Bjerkandera fumosa and Its Enzyme Characterization

  • Jarosz-Wilkolazka, Anna;Luterek, Jolanta;Malarczyk, Elzbieta;Leonowicz, Andrzej;Cho, Hee-Yeon;Shin, Soo-Jeong;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.2
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    • pp.85-95
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    • 2007
  • Manganese dependent peroxidase (MnP) is the most ubiquitous enzyme produced by white-rot fungi, MnP is known to be involved in lignin degradation, biobleaching and oxidation of hazardous organopollutants. Bjerkandera fumosa is a nitrogen-unregulated white-rot fungus, which produces high amounts of MnP in the excess of N-nutrients due to increased biomass yield. The objective of this study was to optimize the MnP production in N-sufficient cultures by varying different physiological factors such as Mn concentration, culture pH, and incubation temperature. The growth of fungus was optimal in pH 4.5 at $30^{\circ}C$, $N_2$-unregulated white-rot fungus produces high amounts of MnP in the excess N-nutrients. The fungus produced the highest level of MnP (up to $1000U/{\ell}$) with $0.25g/{\ell}$ asparagine and $1g/{\ell}$ $NH_4Cl$ as N source at 1.5 mM $MnCl_2$ concentration, pH value of 4.5 at $30^{\circ}C$. Purification of MnP revealed the existence of two isoforms: MnPl and MnP2. The molecular masses of the purified MnPl and MnP2 were in the same range of 42~45 kDa. These isoforms of B. fumosa strictly require Mn to oxidize phenolic substrates. Concerned to kinetic constants of B. fumosa MnPs, B. fumosa has similar Km value and Vmax compared to the other white-rot fungi.

Optimization of Tannase Production by Aspergillus niger in Solid-State Packed-Bed Bioreactor

  • Rodriguez-Duran, Luis V.;Contreras-Esquivel, Juan C.;Rodriguez, Raul;Prado-Barragan, L. Arely;Aguilar, Cristobal N.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.9
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    • pp.960-967
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    • 2011
  • Tannin acyl hydrolase, also known as tannase, is an enzyme with important applications in the food, feed, pharmaceutical, and chemical industries. However, despite a growing interest in the catalytic properties of tannase, its practical use is very limited owing to high production costs. Several studies have already demonstrated the advantages of solid-state fermentation (SSF) for the production of fungal tannase, yet the optimal conditions for enzyme production strongly depend on the microbial strain utilized. Therefore, the aim of this study was to improve the tannase production by a locally isolated A. niger strain in an SSF system. The SSF was carried out in packed-bed bioreactors using polyurethane foam as an inert support impregnated with defined culture media. The process parameters influencing the enzyme production were identified using a Plackett-Burman design, where the substrate concentration, initial pH, and incubation temperature were determined as the most significant. These parameters were then further optimized using a Box-Behnken design. The maximum tannase production was obtained with a high tannic acid concentration (50 g/l), relatively low incubation temperature ($30^{\circ}C$), and unique low initial pH (4.0). The statistical strategy aided in increasing the enzyme activity nearly 1.97-fold, from 4,030 to 7,955 U/l. Consequently, these findings can lead to the development of a fermentation system that is able to produce large amounts of tannase in economical, compact, and scalable reactors.

Effects of Processing Methods and Variety of Rapeseed Meal on Ruminal and Post Ruminal Amino Acids Digestibility

  • Chen, Xibin;Qin, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.6
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    • pp.802-806
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    • 2005
  • The objective of this study was to determine the effects of processing method and rapeseed variety on ruminal and intestinal protein digestibility of rapeseed meal in steers. Intestinal amino acid digestibility was assessed with an in situ ruminal incubation and precision-fed rooster bioassay. In this experiment one traditional rapeseed meal sample (sample A, prepress extraction) and three double low rapeseed meal samples (sample B, prepress extraction, sample C, screw press and sample D, low temperature press) were placed in polyester bags(8 cm${\times}$12 cm) and suspended in the ventral rumen of steers for 16 h. The residues of in situ incubations were intubated to roosters. Total excreta were collected for 48 h after incubation and then desiccated and amino acid concentrations were determined. Results showed that in ruminal incubation the degradation rate of amino acid and crude protein was higher for traditional rapeseed meal sample A than for double low rapeseed meal sample B, but was much lower than for double low sample C and D. In the group of double low rapeseed meal samples, sample D processed by low temperature press had the highest degradation rate of amino acids in the rumen. For all amino acids, the digestibility of the residual protein as measured by the precision-fed rooster bioassay tended to be lower for sample B than for sample A, which had the same processing method with sample B, and in the group of double low rapeseed meals, sample B had similar digestibility of amino acid in residual protein to sample D and higher than that of sample C. However, although the total amino acid availability involving the digestibility of amino acids in the rumen and rooster bioassay of double low rapeseed meal sample D (low temperature press) was higher than those of the other three samples by 7 to 9 percent, there were no significant differences. Results indicated that processing method markedly affected ruminal and post ruminal amino acid digestibility of rapeseed meal when the temperature exceeded 110$^{\circ}C$. Rapeseed meal that had a high content of fiber was not suitable for dry heat treatment at higher temperatures or the amino acids digestibility in rumen and total availability of amino acids could be reduced. Results also suggested the variety of rapeseed meal had no significant effect on the digestibility and availability of amino acids.

Characterization of a Thermophilic Lignocellulose-Degrading Microbial Consortium with High Extracellular Xylanase Activity

  • Zhang, Dongdong;Wang, Yi;Zhang, Chunfang;Zheng, Dan;Guo, Peng;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.305-313
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    • 2018
  • A microbial consortium, TMC7, was enriched for the degradation of natural lignocellulosic materials under high temperature. TMC7 degraded 79.7% of rice straw during 15 days of incubation at $65^{\circ}C$. Extracellular xylanase was effectively secreted and hemicellulose was mainly degraded in the early stage (first 3 days), whereas primary decomposition of cellulose was observed as of day 3. The optimal temperature and initial pH for extracellular xylanase activity and lignocellulose degradation were $65^{\circ}C$ and between 7.0 and 9.0, respectively. Extracellular xylanase activity was maintained above 80% and 85% over a wide range of temperature ($50-75^{\circ}C$) and pH values (6.0-11.0), respectively. Clostridium likely had the largest contribution to lignocellulose conversion in TMC7 initially, and Geobacillus, Aeribacillus, and Thermoanaerobacterium might have also been involved in the later phase. These results demonstrate the potential practical application of TMC7 for lignocellulosic biomass utilization in the biotechnological industry under hot and alkaline conditions.

Synthesis of MnFeP1-xAsx Nanocrystalline Powders by High-Energy Ball Milling (고에너지볼밀링을 이용한 MnFeP1-xAsx 나노분말의 합성)

  • 조영환
    • Journal of Powder Materials
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    • v.10 no.2
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    • pp.129-135
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    • 2003
  • Nanocrystalline powders of $MnFeP_{1-x}As_x$(x=0.45-0.6) have been synthesized by mechanochemical reaction at room temperature using high-energy ball milling from mixtures of Mn, Fe, P, and As Powders. It has been found that a mechanically induced self-propagating reaction (MSR) occurs within 2 hours of milling and it produces very fine polycrystalline powder having a hexagonal $Fe_2P$ structure. Further milling up to 24 hours did not change the crystalline and average particle sizes or the phase composition of the milling product. When x is 0.65, no reaction among the reactants has been observed even after 24 hours of milling. As the P content decreases in $MnFeP_{1-x}As_x$, the incubation time for the MSR has increased and the lattice constants in both a and c axes have changed.

LOW DISSIPATION OF EXCITATION ENERGY IN THE PHOTOSYNTHETIC MACHINERY OF CHILLING-SENSITIVE PLANTS DURING LOWTEMPERATURE PHOTOINHIBITION

  • Moon, Byoung Yong;Lee, Shin Bum;Gong, Yong-Gun;Kang, In-Soon
    • Journal of Photoscience
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    • v.5 no.2
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    • pp.53-61
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    • 1998
  • Using a squash plant, a chilling-sensitive species, and a spinach plant, a chilling-resistant one, effects of chilling temperature on the photosynthetic machinery were studied in terms of chlorophyll fluorescence. When thylakoid membranes were isolated and subjected to incubation at different temperatures, spinach showed stable photosystem II activity at the low temperature side, in contrast to squash which showed quite severe inactivation at low temperature. When parameters of chlorophyll fluorescence were examined, chilling in darkness did not affect either Fv/Fm or photochemical and non-photochemical quenching, in both types of plants. However, chilling of squash plants under irradiance of medium intensity caused a specific decrease in Fv/Fm accompanied by a decline in energy-dependent quenching. Contrastingly, photosystem li of spinach plants were not much affected by light-chilling. When the pool size of zeaxanthin was examined after exposure to high light at different temperatures, squash plants was shown to have a much lower content of antheraxanthin + zeaxanthin, as compared to spinach plants, during low-temperature photoinhibition. These results suggest that chilling-sensitive plants have low capacity to dissipate excitation energy nonradiatively, when they are exposed to low-temperature photoinhibition, and, as a consequence, more vulnerable to photoinhibitory, damage to the photosynthetic apparatus.

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Thermostable Xylanase from Marasmius sp.: Purification and Characterization

  • Ratanachomsri, Ukrit;Sriprang, Rutchadaporn;Sornlek, Warasirin;Buaban, Benchaporn;Champreda, Verawat;Tanapongpipat, Sutipa;Eurwilaichitr, Lily
    • BMB Reports
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    • v.39 no.1
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    • pp.105-110
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    • 2006
  • We have screened 766 strains of fungi from the BIOTEC Culture Collection (BCC) for xylanases working in extreme pH and/or high temperature conditions, the so-called extreme xylanases. From a total number of 32 strains producing extreme xylanases, the strain BCC7928, identified by using the internal transcribed spacer (ITS) sequence of rRNA to be a Marasmius sp., was chosen for further characterization because of its high xylanolytic activity at temperature as high as $90^{\circ}C$. The crude enzyme possessed high thermostability and pH stability. Purification of this xylanase was carried out using an anion exchanger followed by hydrophobic interaction chromatography, yielding the enzyme with >90% homogeneity. The molecular mass of the enzyme was approximately 40 kDa. The purified enzyme retained broad working pH range of 4-8 and optimal temperature of $90^{\circ}C$. When using xylan from birchwood as substrate, it exhibits $K_m$ and $V_{max}$ values of $2.6{\pm}0.6\;mg/ml$ and $428{\pm}26\;U/mg$, respectively. The enzyme rapidly hydrolysed xylans from birchwood, beechwood, and exhibited lower activity on xylan from wheatbran, or celluloses from carboxymethylcellulose and Avicel. The purified enzyme was highly stable at temperature ranges from 50 to $70^{\circ}C$. It retained 84% of its maximal activity after incubation in standard buffer containing 1% xylan substrate at $70^{\circ}C$ for 3 h. This thermostable xylanase should therefore be useful for several industrial applications, such as agricultural, food and biofuel.

Inoculum Sources to Generate High Mechanical Transmission of Barley yellow mosaic virus

  • Jonson, Gilda;Kim, Yang-Kil;Kim, Mi-Jung;Park, Jong-Chul;Hyun, Jong-Nae;Kim, Jung-Gon
    • The Plant Pathology Journal
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    • v.23 no.2
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    • pp.103-105
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    • 2007
  • Mechanical transmission of barley seedlings with barley yellow mosaic virus (BaYMV) is generally inefficient and is the major constraint for testing cultivar resistance to the virus. To explore mechanical transmission, BaYMV-infected barley plants were grown at different conditions and used as inoculum sources to seedlings of susceptible barley cultivar Baegdong. Extracts prepared from BaYMV-infected Baegdong plants at 47, 53, 74, and 90 days after symptom appearance (DASA) and grown at 10 and $12^{\circ}C$ gave 10, 30, 68 and 76% infection, respectively on inoculated susceptible barley cv. Baegdong seedlings. While Jinyangbori, another susceptible cultivar obtained 95% infection rate inoculated with extracts from 90 DASA disease source and grown at $10/12^{\circ}C$. However, low infection rates were obtained when the virus sources were grown in a greenhouse at $15-18^{\circ}C$. Our results indicate that longer incubation period and lower temperature are required for virus accumulation and stability.

Production and Characterization of an Alkaline Protease from Bacillus licheniformis MH31

  • Yu, Jeong-Hyeon;Jin, Hyun-Seok;Choi, Woo-Young;Yoon, Min-Ho
    • Journal of Applied Biological Chemistry
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    • v.49 no.4
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    • pp.135-139
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    • 2006
  • A alkalophilic strain, Bacillus licheniformis MH31 producing an alkaline protease was isolated from mine soil of Boryeong in Korea. Production of a high level of alkaline protease was achieved 42 h after incubation when the bacterium was grown at pH 9.0 and $35^{\circ}C$ in Horikoshi medium supplemented with 0.5%(w/v) starch and 1%(w/v) skim milk as carbon and nitrogen source, respectively. The molecular weight of partially purified enzyme was estimated to be 30 kDa by SDS-PAGE and its optimum pH was pH 10. The enzyme showed optimum temperature at $50^{\circ}C$, and was stable up to $60^{\circ}C$ after 1 h incubation. The protease was strongly inhibited by 1 mM of PMSF which was known well as strong inhibitor of serine proteases, but almost not inhibited by 5 mM of EDTA and 1,10-phenanthroline. When the protein hydrolysis products of 1% skim milk by partially purified protease was compared with available commercial proteases using HPLC analysis, most of hydrolysis products were detected below molecular weight of 10,000 and the hydrolysis ratio of purified enzyme was 24.8% lower than those(above 32%) of commercial proteases.