• Title/Summary/Keyword: enzyme resource

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Anti-allergic Effects of Castanea crenata Leaf Tea (밤나무 잎차(茶)의 항알레르기 효과)

  • Choi, Ok-Beom;Kim, Kyung-Man;Yoo, Gyurng-Soo;Park, Keun-Hyung
    • Korean Journal of Food Science and Technology
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    • v.30 no.2
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    • pp.468-471
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    • 1998
  • Regarding the characterstics of allergic diseases, preventive and continuous treatment is desirable, and tea would be the one of the best functional food formula for it. Here we report the development of tea processing method for the leaves of Castnaea crenata. Two forms of Castnaea crenata leaf tea were prepared, non-fermented steaming tea and semi-fermented rolling tea. Anti-allergic actions of Castanea crenata leaf tea were asessed by testing their effects on the degranulation of mast cells. For this, hexosaminidase release (degranulation marker) from RBL-2H3 cells (mast cell line) was used. At the concentration of $300\;{\mu}g/mL$ of the water extract, the degranulation of RBL-2H3 cells were inhibited 50.4% and 35.4% by non-fermented steaming tea and semi-fermented rolling tea, respectively. These results suggest that the tea processing method we developed could provide a valuable resource for the treatment of allergic diseases.

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Production of Cellulolytic Enzymes by Trichoderma harzianum FJ1 in Solid State Fermentation. (Trichoderma harzianum FJ1의 고체상태배양에 의한 섬유소분해효소의 생산)

  • 유승수;김경철;김성준
    • Microbiology and Biotechnology Letters
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    • v.31 no.3
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    • pp.257-263
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    • 2003
  • The cellulases production in solid state fermentation (SSF) of Trichoderma harzianum FJ1 with high cellulases productivity using cellulosic wastes was investigated. Physical and chemical conditions of the fermentation, such as moisture content, initial pH, and composition of mixed substrate (wine waste, rice straw, and soybean flour) on FPase (Filter paper activity) production were examined. The enzyme production was optimized in the conditions of moisture content of 70%, pH 5.0, 3$0^{\circ}C$, and 1:1:1 composition of mixed substrate containing wine waste, rice straw, and soybean flour. The highest activities of FPA, CMCase, Xylanase, $\beta$-glucosidase, and Avicelase in the optimized culture conditions were 15.2, 69.1, 83.9, 29.2, and 4.2 unit/g-SDW in 5 day cultivation, respectively. Economical and efficient production of cellulolytic enzymes by T harzianum FJ1 using cellulosic wastes in solid state fermentation will contribute to the biological saccharification of cellulosic wastes with enormous potential resource value in future.

Itaconic and Fumaric Acid Production from Biomass Hydrolysates by Aspergillus Strains

  • Jimenez-Quero, A.;Pollet, E.;Zhao, M.;Marchioni, E.;Averous, L.;Phalip, V.
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1557-1565
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    • 2016
  • Itaconic acid (IA) is a dicarboxylic acid included in the US Department of Energy's (DOE) 2004 list of the most promising chemical platforms derived from sugars. IA is produced industrially using liquid-state fermentation (LSF) by Aspergillus terreus with glucose as the carbon source. To utilize IA production in renewable resource-based biorefinery, the present study investigated the use of lignocellulosic biomass as a carbon source for LSF. We also investigated the production of fumaric acid (FA), which is also on the DOE's list. FA is a primary metabolite, whereas IA is a secondary metabolite and requires the enzyme cis-aconitate decarboxylase for its production. Two lignocellulosic biomasses (wheat bran and corn cobs) were tested for fungal fermentation. Liquid hydrolysates obtained after acid or enzymatic treatment were used in LSF. We show that each treatment resulted in different concentrations of sugars, metals, or inhibitors. Furthermore, different acid yields (IA and FA) were obtained depending on which of the four Aspergillus strains tested were employed. The maximum FA yield was obtained when A. terreus was used for LSF of corn cob hydrolysate (1.9% total glucose); whereas an IA yield of 0.14% was obtained by LSF of corn cob hydrolysates by A. oryzae.

Selection of High Laccase-Producing Coriolopsis gallica Strain T906: Mutation Breeding, Strain Characterization, and Features of the Extracellular Laccases

  • Xu, Xiaoli;Feng, Lei;Han, Zhenya;Luo, Sishi;Wu, Ai'min;Xie, Jun
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1570-1578
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    • 2016
  • Commercial application of laccase is often hampered by insufficient enzyme stocks, with very low yields obtained from natural sources. This study aimed to improve laccase production by mutation of a Coriolopsis gallica strain and to determine the biological properties of the mutant. The high-yield laccase strain C. gallica TCK was treated with N-methyl-N-nitro-N-nitrosoguanidine and ultraviolet light. Among the mutants isolated, T906 was found to be a high-production strain of laccases. The mutant strain T906 was stabilized via dozens of passages, and the selected ones were further processed for optimization of metallic ion, inducers, and nutritional requirements, which resulted in the optimized liquid fermentation medium MF9. The incubation temperature and pH were optimized to be 30℃ and 4.5, respectively. The mutant strain T906 showed 3-times higher laccase activity than the original strain TCK under optimized conditions, and the maximum laccase production (303 U/ml) was accomplished after 13 days. The extracellular laccase isoenzyme 1 was purified and characterized from the two strains, respectively, and their cDNA sequence was determined. Of note, the laccase isoenzyme 1 transcription levels were overtly increased in T906 mycelia compared with values obtained for strain TCK. These findings provide a basis for C. gallica modification for the production of high laccase amounts.

Genetic Association of the Porcine C9 Complement Component with Hemolytic Complement Activity

  • Khoa, D.V.A.;Wimmers, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.9
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    • pp.1354-1361
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    • 2015
  • The complement system is a part of the natural immune regulation mechanism against invading pathogens. Complement activation from three different pathways (classical, lectin, and alternative) leads to the formation of C5-convertase, an enzyme for cleavage of C5 into C5a and C5b, followed by C6, C7, C8, and C9 in membrane attack complex. The C9 is the last complement component of the terminal lytic pathway, which plays an important role in lysis of the target cells depending on its self-polymerization to form transmembrane channels. To address the association of C9 with traits related to disease resistance, the complete porcine C9 cDNA was comparatively sequenced to detect single nucleotide polymorphisms (SNPs) in pigs of the breeds Hampshire (HS), Duroc (DU), Berlin miniature pig (BMP), German Landrace (LR), Pietrain (PIE), and Muong Khuong (Vietnamese potbelly pig). Genotyping was performed in 417 $F_2$ animals of a resource population (DUMI: $DU{\times}BMP$) that were vaccinated with Mycoplasma hyopneumoniae, Aujeszky diseases virus and porcine respiratory and reproductive syndrome virus at 6, 14 and 16 weeks of age, respectively. Two SNPs were detected within the third exon. One of them has an amino acid substitution. The European porcine breeds (LR and PIE) show higher allele frequency of these SNPs than Vietnamese porcine breed (MK). Association of the substitution SNP with hemolytic complement activity indicated statistically significant differences between genotypes in the classical pathway but not in the alternative pathway. The interactions between eight time points of measurement of complement activity before and after vaccinations and genotypes were significantly different. The difference in hemolytic complement activity in the both pathways depends on genotype, kind of vaccine, age and the interaction to the other complement components. These results promote the porcine C9 (pC9) as a candidate gene to improve general animal health in the future.

Changes in the levels of $Ca^{2+}$/calmodulin - binding proteins and glutamate decarboxylase during the growth of tobacco suspension cells (담배 배양 세포의 성장과정 중 칼슘/칼모듈린-결합단백질 및 glutamate decarboxylase의 생성변화)

  • Han, Kwang-Soo;Oh, Suk-Heung
    • Applied Biological Chemistry
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    • v.43 no.4
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    • pp.231-235
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    • 2000
  • The changes of calmodulin levels, calmodulin-binding proteins, and $Ca^{2+}$/calmodulin-dependent glutamate decarboxylase during the growth of tobacco suspension cells were investigated. Tobacco cells exhibited a typical growth curve, including an exponential growth phase between 3 and 5 days after inoculation, and an apparent stationary phase occurring after 5 day. Although slight changes were observed from sample to sample, calmodulin protein levels remained similar during the phases of culture growth. Several $Ca^{2+}-dependent$ calmodulin-binding proteins including 56, 46, 36, and 32-kDa proteins were detected in tobacco cell extracts. The 56-kDa protein was identified as glutamate decarboxylase by Western-blot analysis using an anti-GAD monoclonal antibody. The levels of GAD protein and the specific activity of GAD enzyme were highest during the middle exponential phase of the culture growth cycle. These data suggest that $Ca^{2+}$/calmodulin-dependent glutamate decarboxylase is modulated during the growth of tobacco suspension cells.

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Differentially Expressed Genes by Inhibition of C-terminal Src Kinase by siRNA in Human Vascular Smooth Muscle Cells and Their Association with Blood Pressure

  • Hong, Kyung-Won;Shin, Young-Bin;Kim, Koan-Hoi;Oh, Berm-Seok
    • Genomics & Informatics
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    • v.9 no.3
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    • pp.102-113
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    • 2011
  • C-terminal SRC kinase (CSK) is a ubiquitously expressed, cytosolic enzyme that phosphorylates and inactivates several SRC family protein tyrosine kinases. Recent genomewide association studies have implicated CSK in the regulation of blood pressure. The current study aim is to determine the blood pressure association of the genes regulated by CSK down-regulation. The CSK mRNA expression was downregulated in vascular smooth muscle cells using small interfering RNA (siRNA). CSK mRNA levels fell by 90% in cells that were treated with CSK siRNA; the RNA from these cells was examined by microarray using the Illumina HumanRef-8 v3 platform, which comprises 24,526 reference mRNA probes. On treatment with CSK siRNA, 19 genes were downregulated by more than 2-fold and 13 genes were upregulated by more than 2-fold. Three (CANX, SLC30A7, and HMOX1) of them revealed more than 3 fold differential expression. Interestingly, the HMOX1 SNPs were associated with diastolic blood pressure in the 7551 Koreans using Korea Association REsource data, and the result was supported by the other reports that HMOX1 linked to blood vessel maintenance. Among the remaining 29 differentially expressed genes, seven (SSBP1, CDH2, YWHAE, ME2, PFTK1, G3BP2, and TUFT1) revealed association with both systolic and diastolic blood pressures. The CDH2 gene was linked to blood pressures. Conclusively, we identified 32 differentially expressed genes which were regulated by CSK reduction, and two (HOMX1 and CDH2) of them might influence the blood pressure regulation through CSK pathway.

Nonspecific Mouse Hepatitis Virus Positivity of Genetically Engineered Mice Determined by ELISA

  • Han, Dae Jong;Kim, Hyuncheol;Yeom, Su-Cheong
    • Biomedical Science Letters
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    • v.21 no.1
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    • pp.9-14
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    • 2015
  • Mouse hepatitis virus (MHV) is a major pathogen in laboratory mice that usually leads to fatal diseases, such as hepatitis, multiple sclerosis, encephalitis, and respiratory disease. MHV has a high infection rate, and it needs to be detected as soon as possible to prevent its spread to other facilities. However, MHV detection by enzyme-linked immunosorbent assay (ELISA) often gives false positives; thus, it is very important that the results are confirmed as true positives in the early infection stage or distinguished as false positives with more accurate, reliable methods. Under microbiological screening, MHV ELISA-positive mice were found in four GFP-tagging transgenic mice. To verify the detection of the MHV antigen directly, reverse transcription polymerase chain reaction (RT-PCR) was performed, and the mice were determined to be MHV negative. Additional serum antibody-based screening was conducted with three different ELISA kits, and multiplexed fluorometric immunoassay (MFIA) was performed to confirm their accuracy/sensitivity. In brief, the ELISA kit for A59 nucleocapsid protein (MHV-A59N) revealed MHV ELISA positivity, while other ELISA kits (MHV-S lysate and MHV-JHM lysate) demonstrated MHV negativity. In MFIA, only the test for the recombinant A59 nucleocapsid antigen was MHV positive, which was consistent with the ELISA results. These results suggest that the ELISA kit with the recombinant A59 nucleocapsid antigen might induce non-specific MHV ELISA positivity and that confirmation is therefore essential.

Effect of Dioscorea Rhizoma on Gastritis by Acute Gastric Ulcer in Rats (산약이 흰쥐의 급성 위궤양에 의한 위장 장애에 미치는 효과)

  • Park, Young Mi;Lim, Jae Hwan;Seo, Eul Won
    • Korean Journal of Plant Resources
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    • v.28 no.1
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    • pp.1-8
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    • 2015
  • In order to verify the effects of Dioscorea Rhizoma on gastrointestinal damages, we investigated the protective role of the crude extracts on induced gastric ulceration in rats. Rats administered of Dioscorea Rhizoma extracts showed gradually decreased congestion and hemorrhage, but control group did not show any symptom in gastric tissue. Moreover, Dioscorea Rhizoma extract had a role in lowering gastrin and histamine levels in gastric ulcer rats, thereby inhibiting the gastric tissue damages. Excessive production of malondialdehyde shown in gastric ulcer rats was declined in all rats administered with Dioscorea Rhizoma extract as well as the levels of SOD and GPX surged by acute gastric ulcer. Also, the increased activity of CAT showed an effect in activation of antioxidant enzyme to normal state. All data suggest that Dioscorea Rhizoma extract was verified to be highly effective resource in improving the gastrointestinal function of rats by preventing from gastric tissue damage in acute gastric ulcer and restoring the activities of plasma substances and antioxidant enzymes.

Status of Research and Development of Foot and Mouth Disease Diagnosis (Review) (구제역 진단법 연구개발 현황 (총설))

  • Kwak, Kyeongrok;Choi, So-Young;Kim, Eunyoung;Hwang, Choon Hong;Lee, Sung-Jin
    • ANNALS OF ANIMAL RESOURCE SCIENCES
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    • v.28 no.2
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    • pp.78-96
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    • 2017
  • Foot-and-mouth disease (FMD) is a infection that can easily spread when it occurs and causes serious economic damage because of the existence of multiple serotypes of the virus and extreme contagiousness. The most effective method in preventing the transmission of FMD virus (FMDV) is the culling of livestock and additional vaccination in the other areas depending on the spreading rate and situation. Diagnostic methods are utilized not only for the definite diagnosis of FMD but also for identification of serotype, and confirmation of antibody production after vaccination. Although many methods have been developed to diagnose, they are not still enough to detect accurately the disease in a short time. Therefore, it has been needed new diagnostic methods improved from existing methods. Previous methods were based on the enzyme-linked immunosorbent assay (ELISA) as a serological diagnostic method, or polymerase chain reaction (PCR), which is a molecular genetic method. The recent technology has been performing about the combination of both methods and how to make it faster, less costly, more sensitive and accurate way.