• Title/Summary/Keyword: protein heat treatment

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Simple Purification of shiga Toxin B Chain from Recombinant Escherichia coli

  • Oh, Young-Phil;Jeong, Seong-Tae;Kim, Dae-Weon;Kim, El-Chae;Yoon, Ki-Hong
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.986-988
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    • 2002
  • A plasmid expression vector of pEStxl encoding a mature form of the B chain of the Shiga toxin was constructed without a signal peptide under the control of an inducible n promoter. The encoded protein was purified to 90% by simple heat treatment, and then further purified to 95% by Phenyl-Sepharose and DEAE-Sepharose chromatographies, all in a single day. Accordingly, this expression system and heat treatment could facilitate the rapid purification of gram-scale amounts of the Shiga toxin B subunit from recombinant Escherichia coli cells.

Role of Bypass Protein in Feeding Ruminants on Crop Residue Based Diet - Review -

  • Garg, Manget Ram
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.2
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    • pp.107-116
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    • 1998
  • Measurement of DCP is considered inadequate and unsatisfactory means of assessing the protein value of the diet as no distinction is made between the digestion in ferestomach and in the small intestine. Protein meals should be classified on the basis of rumen degradable protein (RDP) and rumen undegradable protein (UDP). Usually, protein meals naturally available with high level of UDP or bypass protein value should be preferred for incorporation in the diet of lactating and growing animals. However, if such resources are non-available or are expensive, protein meals having high degradability can be carefully subjected to heat or formaldehyde treatment to achieve desired level of rumen bypassability. Various studies conducted the world over have revealed that bypass protein feeding to ruminants, especially when animals are fed on crop residue based basal diet, help increasing feed conversion efficiency in growing and lactating ruminants.

Rapid and Simple Method to Prepare Functional Pfu DNA Polymerase Expressed in Escherichia coli Periplasm

  • Chae, Young-Kee;Jeon, Woo-Chun;Cho, Kyoung-Suk
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.841-843
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    • 2002
  • Pfu DNA polymerase from Pyrococcus furiosus was expressed in the E. coli periplasm, and the fully active polymerase was partially purified by applying osmotic shock, ammonium sulfate precipitation, and heat treatment. This method represents a new way of expressing and purifying functional Pfu DNA polymerase without the use of chromatography.

Effect of Environmental Stress on Morphological Change of an Extremely Cadmium-Tolerant Yeast, Hansenula anomala B-7

  • Huh, Nam-Eung;Choi, Nack-Shick
    • Journal of Microbiology and Biotechnology
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    • v.9 no.1
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    • pp.70-77
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    • 1999
  • An extremely cadmium-tolerant budding yeast, Hansenula anomala B-7 underwent a morphological switch in response to either heat shock treatment or cadmium stress, respectively. It exhibited a morphological transition from a unicellular yeast form to a pseudohyphae-like coagulation when subjected to prolonged heat shock treatment. In contrast, the yeast cells showed an irregularity in surface morphology when given thermal stress for a short time. Patterns of proteins expressed in the pseudohyphae-like cells demonstrated that several proteins were overexpressed while others were underexpressed in comparison with those prepared from the cells in the yeast form. It was a striking feature, however, that nearly 40% of the proteins extracted from the cells in the pseudohyphae form appeared to be composed of a single polypeptide. This polypeptide was apparently overexpressed during the pseudohyphae phase and its molecular weight was estimated to be 58 kDa according to SDS-PAGE analysis. However, a significant level of the protein was not observed in the cells before transition to pseudohyphae. The architecture of the cell shape was also damaged when incubated in a medium containing more than 1,000 ppm (8.9mM) of cadmium ions, although able to proliferate at a slow rate. However, the irregularity in the cell morphology exerted either by the brief heat shock treatment or by the cadmium stress with the high concentrations of the metal ions was not repaired, even though the damaged cells were allowed to grow for sufficient time in fresh, cadmium-free medium.

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Ethanol Extract of Ulmus pumila Ameliorates Heat Stress through the Induction of Heat Shock Proteins Expression in RAW264.7 Macrophage Cells

  • dela Cruz, Joseph;Byambaragchaa, Munkhzaya;Choi, Seok-Geun;Hwang, Seong-Gu
    • Journal of Animal Environmental Science
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    • v.20 no.4
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    • pp.147-154
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    • 2014
  • Heat stress is a significant burden to animal production in most areas of the world. Improving our knowledge of physiological and metabolic mechanisms of acclimation may contribute to the development of procedures that may help to maintain health and production efficiency under hot temperature. The effect of Ulmus pumila (UP) extract in inducing Heat Shock Proteins (HSPs) expression in heat-stressed RAW264.7 macrophage cells was investigated. Cell viability assay showed a dose dependent increase in cells after treatment with UP for 24 hours. RT-PCR and western blot analysis showed that increasing concentrations of UP induce the expression of Heat Shock Factor 1 (HSF1) and dose dependently upregulated the expression of Heat shock protein 70 (Hsp70) and Hsp90. LPS-induced nitric oxide was dose-dependently reduced while phagocytic activity greatly recovered with UP treatment. These data demonstrated that UP can be a potential candidate in the development of cytoprotective agent against heat stress.

Artificial Induction of Environmental Mammary Stress by Temperature and Micro-organism Causing Mastitis and Modulation of Mammary Growth by Adenosine, IGF-I and Prolatin In Vitro (In Vitro내 유선조직에의 인위적인 온도 및 유방염 발생 미생물에 의한 환경스트레스 유기와 Adenosine, IGF-I 및 Prolactin에 의한 성장조절작용)

  • 정석근;장병배;이창수;박춘근;홍병주;여인서
    • Korean Journal of Animal Reproduction
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    • v.21 no.4
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    • pp.325-333
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    • 1997
  • Recent evidence indicates that growth factors modulate response of mammary epithelial cells to environmental stress. The objective of this study was to examine the cellular and biochemical responses of mammary tissue to environmental stress caused by artificial mastitis. For experimental a, pp.oach, toxins of most mastitis causing organisms(Staph. aureus or Strep. agalactiae) and heat stress(42$^{\circ}C$) were artificially exposed to mammary tissue. Effects of these environmental stresses on cell growth, cell death and heat shock protein synthesis were examined. Lactating mammary tissure were cultured under basal medium(DMEM) su, pp.emented with insulin(10$\mu\textrm{g}$/ml) and aldosterone(1$\mu\textrm{g}$/ml). All treatment groups in heat stress at 42$^{\circ}C$ incubation significantly decreased DNA synthesis rates in comparison with those at 39$^{\circ}C$(P<0.05), however, these decreased DNAa synthesis rates were recovered by addition of adenosine(10$\mu$M) and IGFI(10ng/ml). Similar results were obtained when tissue growth rates were measured by DNA content/tissue. Strep. agalactiae toxin did not significantly decreased DNA content/tissue in comparison with no treatment of bacterial toxin with or without heat stress, however, tended to decrease DNA contents/tissue without heat stress. In the fluorography analysis, heat stress(42$^{\circ}C$ incubation) slightly increased 35S-methoionine labelled 70kd protein synthesis. These results indicate that environmental stress caused by artificial mastitis slightly decreased mammary growth or mammary size, however, these results could be recovered by addition of adenosine and IGF-I.

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Effects of Specific Interaction Altering Reagents on Hardnesses of Succinylated Soy Protein Gel

  • Bae, Dongho;Jung, Hosun;Choi, Yong-Hee
    • Journal of Applied Biological Chemistry
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    • v.42 no.3
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    • pp.125-129
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    • 1999
  • The changes in gel characteristics of soy protein and succinylated soy protein due to various specific interaction-altering reagents which affect the formation and textural properties of gels, were studied. The reagents were added to 15% soy protein solutions prior to heat treatment. Succinylated soy protein formed harder gel without the addition of reagents. Hardly no gels were formed with urea, indicating that hydrogen bonds significantly contributed to the formation and hardness of the gel and the effects of urea on the hardness of succinylated soy protein gel were more significant. Disulfide bonds were important in the formation of hard gels whether they were succinylated or not, but the contributions of hydrophobic interactions to gel hardness were relatively insignificant. The hardness reducing effects of NaCl and NaSCN were more significant in succinylated soy protein gel. As such, electrostatic interactions were important for succinylated soy protein to form hard gel but not for unmodified soy protein.

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Effects of Heat Pre-Treatment and Reactor Configurations on the Anaerobic Treatment of Volatile Solids (열전처리와 반응조 형태가 고형 유기물의 혐기성 처리에 미치는 영향)

  • Hong, Young-Soek;Bae, Jae-Ho
    • Journal of Korean Society of Water and Wastewater
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    • v.10 no.2
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    • pp.104-116
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    • 1996
  • Anaerobic digestion is generally used for the treatment of volatile organic solids such as manure and sludge from waste water treatment plants. However, the reaction rate of anaerobic process is slow, and thus it requires a large reactor volume. To minimize such a disadvantage, physical and chemical pre-treatment is generally considered. Another method to reduce the reactor size is to adopt different reactor system other than CSTR. In this paper, the effects of heat pre-treatment and reactor configurations on the anaerobic treatability of volatile solids was studied. Carrot, kale, primary sludge, and waste activated sludge was chosen as the test materials, and the BMP method was used to evaluate the maximum methane production and first order rate constants from each sample. After the heat treatment at $130^{\circ}C$ for 30min., the measured increase in SCOD per gram VS was up to 394 mg/L for the waste activated sludge. However, the methane production potential per gram VS was increased for only primary and waste activated sludge by 17-23%, remaining the same for carrot and kale. The overall methane production process for the tested solids can be described by first order reactions. The increased in reaction constant after heat pre-treatment was also more significant for the primary and waste activated sludge than that for carrot and kale. therefore, the heat pre-treatment appeared to be effective for the solids with high protein contents rather than for the solids with high carbohydrate contents. Among the four reactor systems studied, CSTR, PFR, CSTR followed by PFR, and PFR with recycle, CSTR followed by PFR appeared to be the best choice considering methane conversion rate and the operational stability.

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An evaluation of heat on protein oxidation of soy protein isolate or soy protein isolate mixed with soybean oil in vitro and its consequences on redox status of broilers at early age

  • Zhang, Xianglun;Lu, Peng;Xue, Wenyue;Wu, Dawei;Wen, Chao;Zhou, Yanmin
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.8
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    • pp.1135-1142
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    • 2017
  • Objective: The objective of this study was to evaluate effects of heat treatment and soybean oil inclusion on protein oxidation of soy protein isolate (SPI) and of oxidized protein on redox status of broilers at an early age. Methods: SPI mixed with soybean oil (SPIO) heated at $100^{\circ}C$ for 8 h was used to evaluate protein oxidation of SPI. A total of two hundred and sixteen 1-day-old Arbor Acres chicks were divided into 3 groups with 6 replicates of 12 birds, receiving basal diet (CON), heat-oxidized SPI diet (HSPI) or mixture of SPI and 2% soybean oil diet (HSPIO) for 21 d, respectively. Results: Increased protein carbonyl, decreased protein sulfhydryl of SPI were observed as heating time increased in all treatments (p<0.05). Addition of 2% soybean oil increased protein carbonyl of SPI at 8 h heating (p<0.05). Dietary HSPI and HSPIO decreased the average daily gain of broilers as compared with the CON (p<0.05). Broilers fed HSPI and HSPIO exhibited decreased glutathione (GSH) in serum, catalase activity and total sulfhydryl in liver and increased malondialdehyde (MDA) and protein carbonyl in serum, advanced oxidation protein products (AOPPs) in liver and protein carbonyl in jejunal mucosa as compared with that of the CON (p<0.05). Additionally, broilers receiving HSPIO showed decreased glutathione peroxidase activity (GSH-Px) in serum, GSH and hydroxyl radical scavenging capacity in liver, GSH-Px activity in duodenal mucosa, GSH-Px activity and superoxide anion radical scavenging capacity in jejunal mucosa and increased AOPPs in serum, MDA and protein carbonyl in liver, MDA and AOPPs in jejunal mucosa (p<0.05). Conclusion: Protein oxidation of SPI can be induced by heat and soybean oil and oxidized protein resulted in redox imbalance in broilers at an early age.

Difference of Gene Expression between Hypertrophic Scar Keratinocytes and Normal Keratinocytes (비후성 반흔 각질세포와 정상 각질세포의 유전자 비교분석)

  • Choi, Sung-Won;Chung, Ho-Yun;Lim, Young-Kook;Kim, Hoon-Nam;Oh, Ji-Won;Kim, Moon-Kyu;Jeon, Sae-Hwa;Hong, Yong-Taek
    • Archives of Plastic Surgery
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    • v.37 no.4
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    • pp.317-322
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    • 2010
  • Purpose: There is no clear evidence of the original cause of hypertrophic scar, and the effective method of treatment is not yet established. Recently the steps of searching in gene and molecular level are proceeding. we are trying to recognize the difference between keratinocytes of hypertrophic scar and normal skin. Then we do support the comprehension of the scar formation mechanism and scar management. Methods: Total RNAs were extracted from cultured keratinocytes from 4 hypertrophic scars and normal skins. The cDNA chips were prepared. A total of 3063 cDNAs from human cDNA library were arrayed. And the scanning data were analyzed. Results: On microarray, heat shock protein, pyruvate kinase, tumor rejection antigen were more than 2 fold intensity genes. Among them, heat shock 70 kd protein showed the strongest intensity difference. Conclusion: In this study, it can be concluded that heat shock proteins play an important role in the process of wound healing and scar formation. This study provides basic biologic information for scar research. The new way of the prevention and treatment of scar formation would be introduced with further investigations.