• Title/Summary/Keyword: protein heat treatment

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Gene Cloning, Nucleotide Sequence and Efficent Expression of Peptidyl proryl cis-trans Isomerase from Bacillus stearothermophilus (Bacillus stearothermophilus의 Peptidyl Prolyl cis-trans Isomerase 유전자 분리 염기배열 및 발현)

  • 김동주
    • The Korean Journal of Food And Nutrition
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    • v.9 no.4
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    • pp.452-458
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    • 1996
  • A PPIase gene of Bacillus stearothermophilus was screened from a genomic library by plaque hybridization using the A-1 primer as a probe. A PPIase positive plaque contained a 3.0kb insert of the chromosomal DNA. A 3.0kb fragment was subcloned into pUC18, resulting pPI1-40. A DNA fragment encoding the N-terminal portion of the PPIase in pPi-40 was amplified by polymerase chain reaction(PCR) method using the A-1 and B-2 primers. The amplified fragment was cloned into the Sma I site of pUC18 and recombinant plasmid was designated as pSN-18. The nucleotide sequence of 167bp fragment was determined. The deduced amino acid sequence of PPIase was completely matched with the determined N-terminal amino acid sequence of PPIase B. stearothermophilus. The translated protein sequence of PPIase B. stearothermophilus was compared with sequence from periplasmic PPIase from Escherichina coil ; homogies of 16 and 58%, respectively, were found. The clond PPIase gene was over-expressed in E. coil cell using pUC19 as an expression vector. The enzyme was partially purified by heat treatment and colum chromatochraphy on DEAE-Sepharose CL-6B. The molecular weight of the enzyme was dermined to be about 18.0 kDal by SDS-PAGE.

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Microbial Inhibition of Lactic Strains isolated from Kimchi (김치에서 분리한 젖산균의 미생물 생육 저해)

  • Park, Yun-Hee;Kwon, Jung-Joo;Jo, Do-Hyun;Kim, Su-Il
    • Applied Biological Chemistry
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    • v.26 no.1
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    • pp.35-40
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    • 1983
  • The inhibitory activity of 20 Lactic strains from Kimchi was tested against Escherichia coli and other microorganisms. Of the lactic strains investigated, A7 (Pediococcus cerevisiae) and C4(Leuconostoc spp.) were the most effective in restricting the growth of test organisms. The mixed culture inoculation of each selected lactic strain and Escherichia coli resulted in a drastic reduction in the plate count of Escherichia coli after 24 hours. Similar results were obtained when Staphylococcus aureus and Bacillus cereus were used as test organisms. For all test organisms, the presence of A7 caused a higher death rate constant than that of C4. Addition of catalase in the mixed culture did not prevent inhibition, suggesting that hydrogen peroxide did not cause the inhibition. The filtrate of A7 culture added to Escherichia coli showed identical inhibitory action, however heat treatment of filtrate at $80^{\circ}C$ 30min. destroyed the inhibitory activity. A7 filtrate treated with trypsin substantially lost the inhibitory effect, but not by pepsin. The results imply that the protein-like compound(s) is the principal inhibitor produced by this lactic strain.

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Inhibitory effect of Fagopyrum esculentum on degranulation and production of cytokine in RBL-2H3 cells (교맥의 RBL-2H3 비만세포 탈과립과 cytokine 생산 억제 효과)

  • Kang, Kyung-Hwa;Lee, Seung-Yeon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.25 no.3
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    • pp.1-12
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    • 2012
  • Objectives : Fagopyrum esculentum(FE) has been used for removal of inflammation of internal organs and treatment of sore and ulcer by heat toxin in Korean herbal medicines. In this study, To investigated the protective effect of FE on allergic response, we determined whether FE inhibits allergic response. Methods : The effect of FE was analyzed by ELISA, RT-PCR and Western blot in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase, as a marker of degranulation, cytokne, and intracellular ROS and MAPK and NF-${\kappa}B$ signaling. Results : We found that FE suppressed ${\beta}$-hexosaminidase release, the production of IL-4 and TNF-${\alpha}$ and intracellular ROS level in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. FE also significantly inhibited cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF in RBL-2H3. In addition, PF suppressed the phospholyation of ERK1/2, JNK1/2, p38 and $I{\kappa}B{\alpha}$ and NF-${\kappa}B$ signal transduction pathway. Conclusions : Our results indicate that FE protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and ROS via the suppression MAPK and NF-${\kappa}B$ of signal transduction. Abbrevations : FE, Fagopyrum esculentum; RBL-2H3, rat basophilic leukemia cell line; ROS, reactive oxygen species; MAPK, Mitogen-activated protein kinase; $NF{\kappa}B$, nuclear factor ${\kappa}B$; $TNF{\alpha}$, Tumor necrosis factor alpha; GM-CSF, Granulocyte macrophage colony-stimulating factor; ERK, extracellular-signal-regulated kinase; JNK, c-Jun NH2-terminal kinase; p38, p38 MAP kinase; $I{\kappa}B{\alpha}$, inhibitory-kappa B alpha.

Correlation analysis of muscle amino acid deposition and gut microbiota profile of broilers reared at different ambient temperatures

  • Yang, Yuting;Gao, Huan;Li, Xing;Cao, Zhenhui;Li, Meiquan;Liu, Jianping;Qiao, Yingying;Ma, Li;Zhao, Zhiyong;Pan, Hongbin
    • Animal Bioscience
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    • v.34 no.1
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    • pp.93-101
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    • 2021
  • Objective: Temperature could influence protein and amino acid deposition as well as gut microbiota profile and composition. However, the specific effects of ambient temperature on amino acids deposition and gut microbiota composition remain insufficiently understood. Methods: A total of 300 one-day-old Avian broilers were randomly divided into three groups and reared at high, medium, and low temperature (HT, MT, and LT), respectively. Breast muscle and fecal samples were collected for amino acid composition analysis and 16S rRNA gene sequence analysis. Results: Our data showed that compared to the MT group, there was a decrease of muscle leucine and tyrosine (p<0.05), as well as an increase of methionine in the HT group (p<0.05) and a decrease of serine in the LT group. Examination of microbiota shift revealed that at genus level, the relative abundance of Turicibacter and Parabacteroides was increased in the HT group (p<0.05) and that the relative abundances of Pandoraea, Achromobacter, Prevotella, Brevundimonas, and Stenotrophomonas in the LT group were higher than those in the MT group (p<0.05). In addition, there were substantial correlations between microbes and amino acids. In the HT group. Turicibacter was negatively correlated with aspartic acid and tyrosine, whereas Parabacteroides was positively correlated with methionine (p<0.05). In the LT group, there were multiple positive correlations between Achromobacter and arginine, isoleucine or tyrosine; between Prevotella and cysteine or phenylalanine; between Brevundimonas and cysteine; and between Stenotrophomonas and cysteine as well as a negative correlation between Stenotrophomonas and serine. Conclusion: Our findings demonstrated that amino acid content of breast muscle and intestinal microbiota profile was affected by different ambient temperatures. Under heat exposure, augmented abundance of Parabacteroides was correlated with elevated methionine. Low temperature treatment may affect muscle tyrosine content through the regulation of Achromobacter.

Efficacious Pleurodesis with OK-432 Plus Autoblood or OK-432 Against the Pneumothorax with Persistent Air Leak (지속성 기흉에서 OK-432와 자가혈액을 이용한 흉막 유착술의 효과)

  • Kim, Hyoung Soo;Choi, Goang Min
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.1
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    • pp.72-75
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    • 2006
  • Background : This report reviews our experience with persistent air leaks in the peumothorax that were not considered candidates for surgical treatment in order to evaluate the efficacy and risks of the OK-432 plus autoblood or OK-432 pleurodesis. Material & Methods : From March 2004 to July 2005, 8 consecutive patients who had an air leak in the pneumothorax over 5 days and had been treated with OK-432 plus autoblood or OK-432 pleurodesis. The patients were not considered candidates for surgical treatments because the chest CT findings revealed severe chronic lung disease with multiple bullae and/or bullous changes. A prolonged air leak with/without dead space was treated with either OK-432 plus autoblood or OK-432 pleurodesis. The efficacy and side effects of OK-432 pleurodesis were assessed by determining the duration of the air leak, the number of pleurodesis, the patients' symptoms, measurements of the white blood cell count and the c-reactive protein level. Results : All of eight patients were male and the mean age was $72.4{\pm}8.5$. The mean number of pleurodesis was $1.9{\pm}1.1$ and the mean duration of the air leak was $4.6{\pm}4.6days$ after pleurodesis. Side effects after pleurodesis were encountered in 7 patients, which included a chilling sensation in 7 cases, chest pain in 5 cases, headache in 3 cases, local heat sensation in 2 cases, and fever in 1 case. Leukocytosis was observed in 6 patients, and the mean of WBC count and CRP were $14500{\pm}2100$ and $21.9{\pm}11.4mg/dL$, respectively. Conclusion : Either OK-432 plus autoblood or OK-432 pleurodesis has acceptable side effects, and can be considered a treatment option for persistent air leaks in the pneumothorax that are not candidates for surgical treatment.

Effect of Keratinocyte Derived Exosome on Proliferation and Migration on Human Skin Keratinocyte (각질형성세포 유래 엑소좀이 피부각질형성세포의 증식과 이주에 미치는 영향)

  • Kim, Do Yoon;Yu, Ho Jin;Hwang, Dae Il;Jang, Sang Hee;Lee, Hwan Myung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.4
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    • pp.359-366
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    • 2016
  • Exosome, a small vesicle secreted from cells, has diverse functions depending on cell origins and tissue types and plays a important role in cell viability and intercellular communication. Recently, many researchers have demonstrated the use of exosomes for the treatment of cancers and immune diseases, and the development of diagnostic biomarker. However, the secretion mechanism of exosome from skin cell and its physiological functions in skin remain unclear. Thus, this study aimed to explore whether keratinocyte-derived exosome affects proliferation and migration in HaCaTs. Exosomes were isolated from HaCaTs by ExoQuick-TC and then boiled or unbolied. Boiled and unboiled exosome induced proliferation in HaCaTs in a dose-dependant manner ($0.1{\sim}20{\mu}g/mL$), respectively. Boiled and unboiled exosome at concentration of $20{\mu}g/mL$ increased proliferation level in HaCaTs by $186.96{\pm}3.87%$ and $193.48{\pm}10.48%$ compared with control group. Unboiled exosome stimulated migration in HaCaTs in a dose-dependent manner ($0.1{\sim}20{\mu}g/mL$), which reached a maxium at concentration of $20{\mu}g/mL$ ($179.39{\pm}4.89%$ of control), but boiled exosome did not affect HaCaT migration. In addition, unboiled exosome ($0.1{\sim}20{\mu}g/mL$) dose-dependently stimulated sprout outgrowth in HaCats. These results demonstrate that in exosome from HaCaTs, heat-stable components such as lipid may induce HaCaT proliferation and heat-unstable components such as protein may stimulate migration and sprout outgrowth in HaCaTs, thereby leading to reepithelialization and skin-wound healing activities. It is concluded that exosomes from HaCaTs may be used as cosmetic materials.

Properties of Dietary Fiber Extract from Rice Bran and Application in Bread-making (미강에서 추출한 식이섬유추출물의 특성 및 제빵에의 응용)

  • Kim, Young-Soo;Ha, Tae-Youl;Lee, Sang-Hyo;Lee, Hyun-Yu
    • Korean Journal of Food Science and Technology
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    • v.29 no.3
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    • pp.502-508
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    • 1997
  • Rice bran dietary fiber extract, which was obtained after termamyl treatment of defatted rice bran contained $27.3{\sim}30.5%$ protein, $49.7{\sim}54.1%$ insoluble dietary fiber, and $1.9{\sim}2.7%$ soluble dietary fiber. Extrusion decreased the insoluble dietary fiber content but increased the soluble dietary fiber content, while roasting did not. Influence those content. Each mineral element content was depended upon heat processing method. Extrusion increased the water binding capacity and L value, while roasting reduced the water binding capacity and L value. Scanning electron microscopy showed damaged cell walls for extruded sample compared to roasted one which had fully collapsed cell walls. The increase of water absorption, developing time, and stability and the of MTI of wheat flour-dietary fiber extract composites with addition of dietary fiber extract were observed by Farinograph. Rice bran dietary fiber extract had an effect on the bread making resulting in increase of bread weight and color of crumb and crust, and decrease of bread volume and texture. As a result of sensory evaluation, appearance, texture, overall acceptability were significantly different from control but flavor and taste were not different significantly up to 6% level. Heat treated samples had differences in mean values, but not significant differences statistically.

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Allergenicity Change of Soybean Proteins by Thermal Treatment Methods (열처리 방법에 따른 품종별 콩 단백질의 항원성 변화)

  • Seol, Hui-Gyeong;Ko, Yu-Jin;Kim, Eun-Jung;Lee, Gyeong-Lan;Kim, Do-Gyeong;Lee, Jeong-Ok;Ahn, Kang-Mo;Ryu, Chung-Ho
    • Journal of Life Science
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    • v.22 no.4
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    • pp.524-531
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    • 2012
  • Soybean is one of the most common food materials causing food hypersensitivity reactions in Korea. In this study, we have investigated the effect of roasting and fermentation on the allergenicity of soybean. Three kinds of soybean ($Daepung$, $Daewon$, and $Taegwang$) were prepared as raw, roasted, and fermented by $Bacillus$ $subtilis$ GSK 3580, and then their proteins were extracted. The proteins were separated using SDS-PAGE, and the detection of IgE specific to soybean proteins was performed by immunoblotting using 7 sera of soybean allergy patients and non-allergic control individuals. Serum specific IgE to soybean was measured by ELISA. The SDS-PAGE of raw soybean proteins showed various-sized bands ranging from 9 to 76 kDa, which are known as major allergens. In particular, 9, 21, 34, 52, 72, and 76 kDa proteins are known as LTP, Kunits trypsin inhibitor, $Gly$ m Bd 30K, ${\beta}$-subunit, ${\alpha}$-subunit, and ${\alpha}$'-subunit of ${\beta}$-conglycinin, respectively; these are major allergens in soybean. In contrast, only peptides of less than 35 kDa were found in roasted and fermented soybeans. IgE immunoblot analysis of three roasted species of soybeans commonly detected at 38-40 kDa and 10-15 kDa. The protein bands in fermented soybean showed very weak signals or were not detected. In addition, the reactivity of most patients' sera to soybean was decreased after roasting and fermentation. With these results, it may be concluded that the allergenicity of soybeans is reduced by the roasting and fermentation processes. It is supposed that allergenic proteins in soybean were degraded by heat treatment methods and proteolytic enzymes were secreted from fermenting microorganisms.

New Technologies for the Removal of Bacteriophages Contaminating Whey and Whey Products as Cheese by-Products: A Review (치즈 부산물인 유청과 유청 제품에 감염된 박테리오파지 제거를 위해 새롭게 개발된 기술: 총설)

  • Kim, Dong-Hyeon;Chon, Jung-Whan;Kim, Hyun-Sook;Kim, Hong-Seok;Song, Kwang-Young;Hwang, Dae-Geun;Yim, Jin-Hyuk;Kang, Il-Byung;Lee, Soo-Kyung;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.32 no.2
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    • pp.93-100
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    • 2014
  • In general, whey obtained from various cheese batches is being reused, so as to improve the texture and to increase the yield and the nutrient value of the various final milk-based products. In fact, re-usage of whey proteins, including whey cream, is a common and routine procedure. Unfortunately, most bacteriophages can survive heat treatments such as pasteurization. Hence, there is a high risk of an increase in the bacteriophage population during the cheese-making process. Whey samples contaminated with bacteriophages can cause serious problems in the cheese industry. In particular, the process of whey separation frequently leads to aerosol-borne bacteriophages and thus to a contaminated environment in the dairy production plant. In addition, whey proteins and whey cream reused in a cheese matrix can be infected by bacteriophages with thermal resistance. Therefore, to completely abolish the various risks of fermentation failure during re-usage of whey, a whey treatment that effectively decreases the bacteriophage population is urgently needed and indispensable. Hence, the purpose of this review is to introduce various newly developed methods and state-of-the-art technologies for removing bacteriophages from contaminated whey and whey products.

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Isolation of Marine Bacteria Killing Red Tide Microalgae -IV. Characteristics of Algicidal Substances, Produced from Micrococcus sp. LG-5 and the Effects on Marine Organisms- (적조생물 살조세균 탐색 -IV. 살조세균 Micrococcus sp. LG-5가 생산하는 살조물질의 특성과 해양생물에 미치는 영향-)

  • JEONG Seong-Youn;PARK Young-Tae;KIM Mu-Chan;CHOI Seok-Cheol;SEONG Hee-Kyung;KIM Jai-Young;KIM Tae-Un;LEE Won-Jae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.4
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    • pp.339-347
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    • 2000
  • An algicidal bacterium, Micrococcus sp. LG-5 against the harmful dinoflagellate, Cochlodinium polykrikoides was isolated. The optimal conditions for the highest algicidal activity of bacterial culture filtrate showed in the range of $20{\~}30^{\circ}C$, at pH 7.0 and $3.0{\%}$ of NaCl concentration. In addition, $IC_(50)(mean of 50{\%} inhibitory concentration)$ of the culture filtrate against C. polykrikoides after incubation of 5 days was $0.482{\%}$. To investigate heat and pH stability of the culture filtrate of Micrococcus sp. LG-5, the culture filtrate ($pore size, 0.1 {\mu}m$) was heated to $121^{\circ}C for 15 min$ and adjusted pH from 2.0 to 10.0. There were no significant changes in algicidal activity by heat treatment and the pH change between pH from 5.0 to 10.0. The algicidal substances produced from Micrococcus sp. LG-5 were mainly detected in the fraction of $10,000{\~}1,000$ MWCO (molecular weight cut-off). The culture filtrate of Micrococous sp. LG-5 showed antimicrobial activity against Enterococcus faecalis, Escheiichia coli, Uebsiella pneunioniae and Vibrio altinolyticus, but did not show against Pseudomonas aeminosa, P. Buorescens, Salmonella typhi, Staphylococcus aureus, V. cholerae and V parahaemolyicus. The culture filtrate of Micrococcus sp. LG-5 was examined against 16 phytoplankton species and showed the algicidal activity against Ajexandzium tuarense, Eutreptiella Drnnastin, Gymnodinium catenatum, G. mikimotoi, G. sanguineum, eyodinium impuaicum, Heterocapsa triquetra, Heterosipa akashiwo, Prorocentrum micans and Pyraminonas sp.. However no algicidal effects of Micrococcus sp. LG-5 were observed against Chlamydomonas sp., Cylindrotheoa closterium, P. mininum, P. triestimum, Pseudonieschia sp. and Sczipuiella trochoidea. On the other hand, algicidal activity on the tested marinelivefood was not detected except for Isochrysis galbana. In addition, physiological responses of cultured olive flounder (Paralichthys oliraceus) exposed to $1 and 10{\%}$ of the culture filtrate of Micrococcus sp. LG-5 were measured. There were no clear changes in AST, GGT, creatinine, urea, total cholesterol, total protein, albumine, $Mg^(+2), Ca^(+2), Na^+, K^+, and Cl^-$. These results indicate that olive flounders were not affected when they were exposed to the culture filtrate of Micrococcus sp. LG-5.

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