• Title/Summary/Keyword: protein heat treatment

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Morphological, Physiological and Biochemical Responses of Gerbera Cultivars to Heat Stress

  • Chen, Wen;Zhu, Xiaoyun;Han, Weiqing;Wu, Zheng;Lai, Qixian
    • Horticultural Science & Technology
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    • v.34 no.1
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    • pp.1-14
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    • 2016
  • Heat stress is an agricultural problem for Gerbera jamesonii, and it often causes poor seedling growth, reduced flower yield and undesirable ornamental characteristics of flowers. However, little is known about the effect of heat stress on the morphological, physiological and biochemical characteristics of gerbera plants. Here, the responses of six cultivars of Gerbera jamesonii to heat stress were investigated. Under a 1-d heat treatment at $45^{\circ}C$, the leaves of gerbera cultivars showed yellowing, wilting, drying and death to varying degrees. The heat treatment also resulted in increased electrical conductivity, decreased soluble protein and chlorophyll contents, and the accumulation of malondialdehyde (MDA) and proline in leaves. Moreover, heat tolerance differed among the six tested gerbera cultivars. Our results demonstrated that among the six gerbera cultivars, 'Meihongheixin' is a heat-resistant cultivar, whereas 'Beijixing' is a heat-sensitive one. 'Shijihong' and 'Linglong' are relatively heat-resistant cultivars, and 'Dadifen' and 'Taiyangfengbao' are relatively heat sensitive.

Purification and Characterization of Peptidyl Prolyl cis-trans Isomerase (PPlase) from Bacillus stearothermophilus SIC1

  • KIM Dong-Ju
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.6
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    • pp.728-735
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    • 1995
  • The peptidyl prolyl cis-trans isomerase(PPlase, EC 5.2.2.8) from Bacillus stearothermophilus SIC1 was extracted from the cells treated with by lysozyme. PPlase was purified from the cell extracts by heat treatment, ammonium sulfate precipitation, ion exchange chromatography and finally gel filtration (FPLC). The purity of purified the enzyme after Superose 12 column chromatography was examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE). The molecular weight of the purified PPlase was estimated as 18,000 by SDS-PAGE. The 39 amino acid residues from the N-terminus were determined by the protein sequencer. The enzyme showed the optimum pH at 8.0 and was stable at the range of pH 7.0 to 8.0. The enzyme was considerably stable after heat treatment at $60^{\circ}C$ for 30 minutes, and the enzyme was quite stable up to $65^{\circ}C$. The presence of the PPlase in the refolding solution accelerated the isomerization rate of the assay peptide.

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Characterization of an antimicrobial Chitinase Purified from the Grapefruit Extract (자몽 추출물로부터 분리된 항균성 Chitinase의 특성)

  • 김외연;정나은;제대엽;이동철;김재원;조성환;이상열
    • Korean Journal Plant Pathology
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    • v.10 no.4
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    • pp.277-283
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    • 1994
  • An antimicrobial chitinase was purified from grapefruit extract and its properties were characterized. The chitinase was purified with a single step chromatography on regenerated chitin affinity gel column. The molecular weight of the purified chitinase was 29 kDa. The grapefruit extract contained the chitinase protein more than 50% of its total soluble proteins measured by coomassie stained protein bands. When the purified chitinase was incubated with polymers of N-acetylglucosamine (NAG), such as mycelia of Fusarium oxysproum and swollen chitin, they were degraded to oligosaccharides, and the oligosaccharides were then further hydrolyzed by the same enzyme to monomer and dimer of NAG. This result suggests that the chitinase contained both endo- and exo- chitinase activities. The chitinase was stable to heat and pH treatment; its activity was not diminished by the heat treatment upto 7$0^{\circ}C$ for 1 hr, and it showed a pH stability in the range of pH 4.0 to 12.0.

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Effects of Heating Time and Temperature on Functional Properties of Proteins of Yellow Mealworm Larvae (Tenebrio molitor L.)

  • Lee, Ha-Jung;Kim, Ji-Han;Ji, Da-Som;Lee, Chi-Ho
    • Food Science of Animal Resources
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    • v.39 no.2
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    • pp.296-308
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    • 2019
  • Although the yellow mealworm (Tenebrio molitor L.) is a promising alternative protein source, the effects of processing conditions on functional properties are unclear. In this study, a protein extract of yellow mealworm larvae (PEYM) was subjected to different heat temperature ($55^{\circ}C$, $75^{\circ}C$, and $95^{\circ}C$) with different time (20, 40, and 60 min) to evaluate the functional properties and protein oxidation. Different heat temperature treatment significantly affected the exposure of surface hydrophobicity of the proteins and protein molecule aggregation, which reached maximum levels at $95^{\circ}C$ for 60 min. Protein oxidation was inversely proportional to the temperature. Both the highest carbonyl value (1.49 nmol/mg protein) and lowest thiol value (22.94 nmol/mg protein) were observed at $95^{\circ}C$ for 60 min. The heating time-temperature interaction affected several functional properties, including solubility, emulsifying potential, and gel strength (GS). Solubility decreased near the isoelectric point (pH 5 to 6). As the temperature and heating time increased, emulsifying properties decreased and GS increased. The oil absorption capacity and foaming properties decreased and the water absorption capacity increased. These results confirmed that PEYM is a suitable source of proteins for processing and applications in the food industry.

A study of the Emulsifying Properties of Kidney Bean Protein Isolate (분리 강남콩 단백질의 유화특성에 관한 연구)

  • 최희령;손경희;민성희
    • Korean journal of food and cookery science
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    • v.5 no.2
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    • pp.9-17
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    • 1989
  • This study was carried out in order to study the emulsifying properties of kidney bean protein isolate. Kidney bean protein isolate was tested for the purpose of finding out the effect of pH, addition of NaCl, and heat treatment on the solbulity and emulsion capacity, emulsion stability, surface hydropobicity and emulsion viscosity. The results were summarized as follows. 1 The solubility of kidney bean protein isolate was affected by pH and showed the lowest value at pll 4.5 which is isoelectric point of kidney bean isolate. When the kidney bean protein isolate was heated, the highest value observed at pH 2 and pH 7 was 96.11%, 97.41% respectively. 2. The emulsion capacity of kidney bean protein isolate was not significantly different with each pH. With addition of NaCl, emulsion capacity decreased steadily. When heated thr highest value observed at pH 2 and pH 7 was 82.91 ml oil/100 mg protein ($60^{\circ}C$), 82.08 m1 oil/100 mg protein ($80^{\circ}C$) respectively. 3. The emulsion stability was significantly higher at pH 4.5 than that of pH 2 and pH 7 (p 0.05) When NaCl was added, emulsion stability was generally increased after 2hrs. When heated, the highest value observed at pH 2 and pH 7 was 21.25% ($80^{\circ}C$),23.7%($100^{\circ}C$) respectively after 2hrs. 4. Surface hydrophobicity increased sharply as 0.2 M NaCl was added to pH 4.5. When heated, the surface hydrophobicity increased as the temperature increased. 5. The highest value of emulsion viscosity was observed at pH 4.5 and pH 7 when 0.2 M NaCl was added. Under heat treatment, the highest value was 48,000 cps at pH 4.5 ($40^{\circ}C$). In the case of pH 7, the highest value was 105,000 cpa at $100^{\circ}C$.

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The Effects of Injinchunggan-tang(Yinchenqinggan-tang) on DMN-induced Liver Damage by Applying Proteomics (인진청간탕(茵蔯淸肝湯)이 DMN 유발 간섬유화와 단백질 발현에 미치는 영향)

  • Park, Sang-Baek;Kim, Young-Chul;Lee, Jang-Hoon;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.29 no.1
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    • pp.200-218
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    • 2008
  • Objectives : The purpose of this study was to investigate the effects of Injinchunggan-tang (Yinchenchinggan-tang) on DMN-induced liver damage by applying proteomics. Materials and Methods : Sprague-Dawley rats were used in this experiment and were divided into the normal group (normal saline), the control group (DMN) and the sample group (DMN+IJCGT). DMN was injected i.p. once a day three times a week for 3 weeks in the control group. Normal saline instead of DMN was administered to the normal group. In the sample group, Injinchunggan-tang (Yinchenchinggan-tang) extract was orally administered once a day for 10 days after DMN was induced. The livers of each group were processed and analyzed by histology, Western blot, $Oxyblot^{TM}$, CBB and 2-dimensional electrophoresis. Results : In the histological findings of the liver, IJCGT reduced collagen deposition and liver damage in DMN-induced hepatic fibrosis. IJCGT increased MMP-13 protein production assessed by western blot. Protein oxidation induced by DMN treatment was decreased by IJCGT. In the 2-dimensional electrophoresis finding, the level of the increased proteins induced by DMN treatment such as GRP 75, 58kDa glucose regulated protein and heat shock 70kDa protein 5 were decreased by IJCGT. IJCGT was considered to have the protective effects on hepatotoxicity induced by DMN. In the 2-dimensional electrophoresis finding, the level of increased oxidized proteins such as heat shock 70 protein, mitochondrial malonyltransferase, calreticulin precursor, actin, NADP-isocitrate dehydrogenase, ankyrin repeat and SOCS box protein 11 were decreased by IJCGT. IJCGT was considered to have protective effect on the protein production induced by DMN treatment. Conclusion : Injinchunggan-tang (Yinchenchinggan-tang) exerts an inhibitory effect against the fibrosis and protein oxidation induced by DMN treatment in the rat liver. IJCGT was considered to have protective effects on the hepatotoxicity and protein production induced by DMN treatment.

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Protective Effect of Saururus chinensis Ethanol Extract against Styrene in Mouse Spermatocyte Cell Line (마우스 정모세포주에서 스티렌에 대한 삼백초 에탄올 추출물의 보호 효과)

  • Yoon, Ji Hye;Sohn, Sang Hyun;Lee, Eun Young;Kim, Geum Soog;Lee, Seung Eun;Lee, Dae Young;Seo, Kyung Hye;Lee, Sang Won;Kim, Hyung Don
    • Korean Journal of Medicinal Crop Science
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    • v.25 no.1
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    • pp.45-51
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    • 2017
  • Background: This study was performed to evaluate the protective effect of Saururus chinensis ethanol extract (SCE) against styrene toxicity in mouse spermatocyte cells [GC-2spd (ts) cell line]. Methods and Results: Cytotoxicity in mouse spermatocyte cells was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Generation of reactive oxygen species (ROS) was determined using 2',7'-dichlorodihydrofluorescein diacetate (DCF-DA) assay. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and western blotting were performed to quantify the mRNA and protein expression levels, resepectiviely, of stress or apoptosis-related genes including p21, p53, heat shock protein 70 (Hsp70), heat shock protein 90 (Hsp90), Bax, Bcl-2, and caspase-3. The results of the MTT assay showed that $50 {\mu}g/m{\ell}$ SCE did not affect cell viability. ROS generation in mouse spermatocyte cells increased by treatment with $100{\mu}M$ styrene, and decreased by co-treatment with SCE. SCE repressed the mRNA expression of stress-related genes, which increased by styrene treatment. In addition, SCE inhibited the apoptosis of mouse spermatocyte cells by ameliorating mRNA and protein levels of apoptotic genes that were altered by styrene treatment. Conclusions: These results suggest that SCE may alleviate styrene toxicity in mouse spermatocyte cells by reducing ROS stress and regulating genes related to styrene toxicity.

Inhibitory Effects of Tannic Acid on the Skin Toxicity and Heat Shock Protein Induction by UVB Irradiation in Hairless Mouse (자외선 B 파로 유도된 Hairless Mouse에서 타닌의 피부 독성 억제효과 및 Heat Shock Protein 70의 생성억제 효과)

  • 이세윤;이민경;장동덕;안령미;안형수
    • Toxicological Research
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    • v.13 no.1_2
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    • pp.79-86
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    • 1997
  • Inhibitory effects of tannic acid on skin toxicity and heat shock protein induced by UVB were investigated. Tannic acid was administered either topically or orally for 3 days to hairless mice, which were previously irradiated with UVB. UVB was found to cause skin erythema . However, the skin erythema was decreased when tannic acid was administered either topically or orally. The heat shock proteins, Hsp-78 kDa and 70 kDa, were induced by UVB irradiation, but the induction was decreased by treatment of tannic acid in both topically and orally administered groups. The hsp induction was more prominent in orally administered groups than in topically administerd groups. However, the difference between two groups was not statistically significant. The route of administrations, topical and oral, does not affect the activity of tannic acid. In the skin tissue observation, tannic acid regenerated the epithelial cells with 7-9 cell layers which were injured by UVB. In conclusion, tannic acid has an ability to protect against UVB irradiation and regenerate the skin.

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Purification of Hydrogenases from Purple Sulfur Bacterium Thiocapsa roseopersicina Using Various Applications of Chromatography (홍색 유황세근 Thiocapsa roseopersicina로 부터 생산되는 Hydrogenase의 각종 크로마토그래피에 의한 정제)

  • Choi, Eun-Hye;Oh, You-Kwan;Kim, Mi-Sun
    • Transactions of the Korean hydrogen and new energy society
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    • v.19 no.2
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    • pp.124-131
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    • 2008
  • Crude cytoplasmic fraction of phototrophic purple sulfur bacterium, Thiocapsa roseopersicina NCIB 8347, were initially prepared and purified by sonication, ultracentrifugation, ammonium sulfate fractionation and heat-treatment and it has been previously reported. Using various applications of chromatography far the purification of membrane-bound and soluble hydrogenases from heat-treated enzyme fraction were studied at present report. When the heat-treated enzyme preparation was applied to the anion column chromatography using Q-sepharose, Fraction I and II, which were extracted with the KCl 0-0.5 M gradient, showed the specific evolution hydrogenase activity 3.86 and 2.27 U/mg-protein respectively. Specific hydrogenase activitys of Fraction I and II were further increased to 4.35 and 7.46 U/mg-protein for Fraction I and to 2.49 and 4.41 U/mg-protein fur Fraction II respectively, when hydrophobic interaction column, Phenyl superose, and anion exchange column, Mono-Q, were applied. Size exclusion chromatography using superdex 200 concentrated the hydrogenase Fraction I and II to 9.19 and 7.84 U/mg-protein respectively at the final step of purification.

Coaggregation between Porphyromonas gingivalis and Tannerella forsythia (Porphyromonas gingivalis와 Tannerella forsythia의 응집반응)

  • Um, Heung-Sik;Lee, Seok-Woo;Park, Jae-Hong;Nauman, R.K.
    • Journal of Periodontal and Implant Science
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    • v.36 no.1
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    • pp.265-272
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    • 2006
  • Dental plaque, a biofilm consisting of more than 500 different bacterial species, is an etiological agent of human periodontal disease, It is therefore important to characterize interactions among periodontopathic microorganisms in order to understand the microbial pathogenesis of periodontal disease. Previous data have suggested a synergistic effect of tow major periodontal pathogens Porphyromonas gingivalis and Tannerella forsythia in the periodontal lesion. In the present study, to better understand interaction between P. gingivalis and T. forsythia, the coaggregation activity between these bacteria was characterized. The coaggregation activity was observed by a direct visual assay by mixing equal amount (1 ${\times}$ $10^9$)of T. forsythia and P. gingivaJis cells. It was found that the first aggregates began to appear after 5-10 min, and that the large aggregates completely settled within 1 h. Electron and epifluorescence microscopic studies confirmed cell-cell contact between two bacteria. The heat treatment of P. gingivalis completely blocked the activity, suggesting an involvement of a heat-labile component of P. gingivalis in the interaction. On the other hand, heat treatment of T. forsythia significantly increased the coaggregation activity; the aggregates began to appear immediately. The coaggregation activity was inhibited by addition of protease, however carbohydrates did not inhibit the activity, suggesting that coaggregation is a protein-protein interaction. The results of this study suggest that coaggregation between P. gingivalis and T. forsythia is a result of cell-cell physical contact, and that coaggregation is mediated by a heat-labile component of P. gingivalis and T. forsythia component that can be activated on heat treatment.