• Title/Summary/Keyword: Functional characterization

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Antioxidant Activity and Agronomic Characteristics of Colored Waxy Corns (유색 찰옥수수의 항산화력 및 작물학적 특성)

  • Lee, Hee-Bong;Park, Bo-Young;Ji, Hee-Chung;Cho, Jin-Woong;Kim, Seog-Hyung;Mo, Eun-Kyung;Lee, Mi-Ra
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.179-184
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    • 2006
  • This study was carried out to evaluate major agronomical characterization and phenol compound contents, Xanthine oxidase inhibitory activity (XO), Catalase activity, Superoxide dismutase activity (SOD) and 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) radical sacvenging activity were analyzed in colored waxy corns. The mean of stem height and ear length were 248.8 cm and 18.6 cm, respectively. The pericarp thickness in CNU108 $(30.3{\mu}m)$ was thinner than other hybrids. The period of tasseling days in CNU69 and CNU202 were very shorter than other hybrids (59 days). 100-kernel weight of CNU50 was 35.6 g and heavier than the others. The antioxidant activities such as xanthin oxidase (XO), catalase and superoxide dismutase (SOD) were higher in CNU70 and CNUI 38. Especially highest EDA (electron donating ability) in DPPH radical scavenging effect was 94.8% and 94.6% in CNU160 and CNU193, respectively. In the results, the antioxidant enzyme activity and antioxidant acticity were higher in CNU109 and CNU 34 hybrids. The hybrids, CNU34, CNU70, CNU108, CNU 138 and CNU193 may be considered higher functional color waxy corn.

Purification and Characterization of $\beta$-Xylosidase B of Bacillus stearothemophilus No.236 Produced by Recombinant Escherichia coli. (재조합 균주 Escherichia coli가 생산하는 Bacillus stearothermophilus No.236 $\beta$-Xylosidase B의 정제 및 특성)

  • 장욱진;조쌍구;최용진
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.297-302
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    • 1998
  • $\beta$-Xylosidase B was produced by Escherichia coli HB101/pKMG12 carrying the xylB gene of Bacillus stearothermophilus No.236 on its recombinant plasmid. The $\beta$-xylosidase B produced was purified by ammonium sulfate fractionation, DEAE-Sepharose CL-6B, Sephacryl S-200 and Superdex 200 HR gel filtration. The purified enzyme showed the highest activity at pH 6.5 and 5$0^{\circ}C$. But, the enzyme was observed to be very sensitive to the pH and temperature of the reaction mixture. The enzyme was activated about 35% of its original activity in the presence of 1 mM of $Mn^{2+}$ but it was completely inhibited by $Ag^{+}$, $Cu^{2+}$and $Hg^{2+}$ions. In contrast with the $\beta$-xylosidase A, the B enzyme was found to have $\alpha$-arabinofuranosidase activity though the activity was fairly low compared with the $\alpha$-arabinofuranosidase produced from the arfI gene of the same Bacillus stearothermophilus. Therefore, $\beta$-xylosidase B is considered to be more suitable than $\beta$-xylosidase A at least for the biodegradation of arabinoxylan. The $K_{m}$ and V$_{max}$ values of the $\beta$-xylosidase B for o-nitrophenyl-$\alpha$-D-xylopyranoside were 6.43 mM and 1.45 $\mu$mole/min, respectively. Molecular mass of the enzyme was determind to be about 54 kDa by SDS-PAGE and 160 kDa by Superdex 200HR gel filtration, indicating that the functional $\beta$-xylosidase B was composed of three identical subunits.s.

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A Commensal Thermophile, Symbiobacterium toebii: Distribution, Characterization, and Genome Analysis

  • Bae Jin-Woo;Kim Kwang;Song Jae Jun;Ha Jae Seok;Kim Joong-Jae;Kang Gwan-Tae;Kim Mi-Hwa;Hong Seung-Pyo;Sung Moon-Hee
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2001.11a
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    • pp.46-53
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    • 2001
  • A commensal thermophile, Symbiobacterium toebii, isolated from hay compost (toebii) in Korea commensally interacted with a thermophilic Geobacillus toebii sp. nov., which was a new species within the genus Geobacillus on the basis of the phenotypic traits and molecular systematic data. S. toebii required the crude extracts and/or culture supernatant of the Geobacillus toebii for axenic growth and could grow on the temperature between 45 and $70^{\circ}C$ (optimum: $60^{\circ}C$; 2.4 h doubling time) and pH 6.0 and 9.0 (optimum: pH 7.5). The G+C content of the genomic DNA was $65 mol\%$, and the major quinones were MK-6 and MK-7. A phylogenetic analysis of its 16S rDNA sequence indicated that Symbiobacterium toebii was closely related with solely reported Symbiobacterium thermophilum. The presence of the commensal thermophile 16S rDNA and accumulation of indole in all the enriched cultures indicate that Symbiobacterium toebii is widely distributed in the various soils. The genome of S. toebii constituted a circular chromosome of 3,280,275 base pairs and there was not an extra-chromosomal element (ECE). It contained about 4,107 predicted coding sequences. Of these protein coding genes, about $45.6\%$ was encoded well-known proteins and annotated the functional assignment of 1,874 open reading frames (ORFs), and the rest predicted to have unknown functions. The genes encoding thermostable tyrosine phenol-lyase and tryptophan indole-lyase were cloned from the genomic DNA of S. toebii and the enzymatic production of L-tyrosine and L-tryptophan was carried out with two thermostable enzymes overexpressed in recombinant E. coli.

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Functional Characterization of sll1556 of Synechocystis sp. PCC6803 as Type II Isopentenyl Diphosphate Isomerase (Type II Isopentenyl Diphosphate Isomerase로서 Synechocystis sp. PCC6803의 sll1556의 작용 특성)

  • Cho, Kab-Yeon
    • The Korean Journal of Food And Nutrition
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    • v.23 no.4
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    • pp.526-530
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    • 2010
  • Isopentenyl diphosphate(IPP) isomerization to dimethylallyl diphosphate(DMAPP) is an important step for the efficient production of isoprenoids such as lycopene, ${\beta}$-carotene, astaxanthin, etc. The type II isopentenyl diphosphate isomerase gene from Synechocystis sp. PCC6803(sll1556, Syidi2) was cloned and expressed in Escherichia coli $DH5{\alpha}$. When E. coli $DH5{\alpha}$ harboring lycopene synthesis genes, crtE, crtB, and crtI and mevalonate pathway genes, MvK1, MvK2, and Mvd, was cultured on LB medium containing mevalonate, the strain grew very slowly be due to the toxicity of isopentenyl diphosphate derived from mevalonate. When Syidi2 was introduced to E. coli $DH5{\alpha}$ harboring the lycopene synthesis genes and mevalonate pathway genes, growth on mevalonate medium was fully restored and the colony showed red color indicating lycopene formation. The growth rate of the mutant strain, E. coli $DH5{\alpha}$(idi::${\Delta}km$), was very slow because of IPP accumulation and DMAPP deprivation. Ultimately the idi mutant was complemented by introducing the Syidi2 gene.

Identification and Characterization of a KDR-positive Mesoderm Population Derived from Human Embryonic Stem Cells Post BMP4 Treatment (BMP4 처리에 의한 인간 배아줄기세포 유래 KDR 양성 중배엽성 세포군의 분화 양상 조사)

  • Kim, Jung-Mo;Son, On-Ju;Cho, Youn-Jeong;Lee, Jae-Ho;Chung, Hyung-Min
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.9-15
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    • 2011
  • The functional cardiovascular system is comprised of distinct mesoderm-derived lineages including endothelial cells, vascular smooth muscle cells and other mesenchymal cells. Recent studies in the human embryonic stem cell differentiation model have provided evidence indicating that these cell lineages are developed from the common progenitors such as hemangioblasts and cardiovascular progenitor cells. Also, the studies have suggested that these progenitors have a common primordial progenitor, which expresses KDR (human Flk-1, also known as VEGFR2, CD309). We demonstrate here that sustained activation of BMP4 (bone morphogenetic protein 4) in hESC line, CHA15 hESC results in $KDR^+$ mesoderm specific differentiation. To determine whether the $KDR^+$ population derived from hESCs enhances potential to differentiate along multipotential mesodermal lineages than undifferentiated hESCs, we analyzed the development of the mesodermal cell types in human embryonic stem cell differentiation cultures. In embryoid body (EB) differentiation culture conditions, we identified an increased expression of $KDR^+$ population from BMP4-stimulated hESC-derived EBs. After induction with additional growth factors, the $KDR^+$ population sorted from hESCs-derived EBs displays mesenchymal, endothelial and vascular smooth muscle potential in matrix-coated monolayer culture systems. The populations plated in monolayer cultures expressed increased levels of related markers and exhibit a stable/homologous phenotype in culture terms. In conclusion, we demonstrate that the $KDR^+$ population is stably isolated from CHA15 hESC-derived EBs using BMP4 and growth factors, and sorted $KDR^+$ population can be utilized to generate multipotential mesodermal progenitors in vitro, which can be further differentiated into cardiovascular specific cells.

Characterization of a Chitinase Gene Exhibiting Antifungal Activity from a Biocontrol Bacterium Bacillus licheniformis N1

  • Lee, Kwang-Youll;Heo, Kwang-Ryool;Choi, Ki-Hyuck;Kong, Hyun-Gi;Nam, Jae-Sung;Yi, Young-Byung;Park, Seung-Hwan;Lee, Seon-Woo;Moon, Byung-Ju
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.344-351
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    • 2009
  • A biocontrol bacterium Bacillus licheniformis N1 grown in nutrient broth showed no chitinolytic activity, while its genome contains a gene which encodes a chitinase. The gene for chitinase from B. licheniformis N1 was amplified by PCR and the deduced amino acid sequence analysis revealed that the chitinase exhibited over 95% identity with chitinases from other B. licheniformis strains. Escherichia coli cells carrying the recombinant plasmid displayed chitinase activity as revealed by the formation of a clear zone on chitin containing media, indicating that the gene could be expressed in E. coli cells. Chitinase gene expression in B. licheniformis N1 was not detected by RT-PCR analysis. The protein was over-expressed in E. coli BL21 (DE3) as a glutathione S-transferase fusion protein. The protein could also be produced in B. subtilis 168 strain carrying the chitinase gene of N1 strain. The crude protein extract from E. coli BL21 carrying GST fusion protein or culture supernatant of B. subtilis carrying the chitinase gene exhibited enzyme activity by hydrolyzing chitin analogs, 4-methylumbelliferyl-$\beta$-D-N,N'-diacetylchitobioside and 4-methylumbelliferyl-$\beta$-D-N,N',N"-triacetylchitotrioside. These results indicated that even though the chitinase gene is not expressed in the N1 strain, the coding region is functional and encodes an active chitinase enzyme. Furthermore, B. subtilis 168 transformants expressing the chitinase gene exhibited antifungal activity against Fulvia fulva by suppressing spore germination. Our results suggest that the proper engineering of the expression of the indigenous chitinase gene, which will lead to its expression in the biocontrol strain B. licheniformis N1, may further enhance its biocontrol activity.

Characterization of a Nitrous Oxide-reducing Bacterial Consortium (아산화질소 환원 세균 컨소시움의 특성)

  • Park, Hyung-Joo;Kwon, Ji-Hyeon;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.630-638
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    • 2019
  • Nitrous oxide (N2O) is a greenhouse gas with a global warming potential 310 times higher than that of carbon dioxide. In this study, an N2O-reducing consortium was obtained by enrichment culture using advanced treatment sludge as the inoculum. The dominant bacteria in the consortium were Sulfurovum (17.95%), Geobacter (14.63%), Rectinema (11.45%), and Chlorobium (8.24%). The consortium displayed optimal N2O reducing activity when acetate was supplied as the carbon source at a carbon/nitrogen ratio (mol·mol-1) of 6.3. The N2O reduction rate increased with increasing N2O concentration at less than 3,000 ppm. Kinetic analysis revealed that the maximum N2O reduction rate of the consortium was 163.9 ㎍-N·g-VSS-1·h-1. Genes present in the consortium included nosZ (reduction of nitrous oxide to N2), narG (reduction of nitrate to nitrite), nirK (reduction of nitrite to nitric oxide), and norB (reduction of nitric oxide to nitrous oxide). These results indicate that the N2O-reducing consortium is a promising bioresource that can be used in denitrification and N2O mitigation.

Characterization of Mouse Interferon-Induced Transmembrane Protein-1 Expression in Mouse Testis

  • Lee, Ran;Park, Hyun Jung;Lee, Won Young;Kim, Ji Hyuk;Kim, In Chul;Kim, Dong Woon;Lee, Sung Dae;Jung, Hyun Jung;Kim, Jong Moon;Yoon, Hyung Moon;Kwon, Hyuk Jung;Song, Hyuk
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.225-230
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    • 2012
  • Interferon induced transmembrane protein-1 (Ifitm-1) has been reported to have an important role in primordial germ cell formation, and it has expressed in female reproductive organ. In the present study, Ifitm-1 gene expression was identified in testes and all part of epididymis using western immunoblot and immunohistochemistry. Interestingly, Ifitm-1 expression was observed on the head of spermatozoa. To investigate the role of Ifitm-1 gene expression in behavior of spermatozoa after acrosome reaction, fresh sperm was incubated with calcium ionophore to induce acrosome reaction, whereas the expression of Ifitm-1 was not altered after the acrosome reaction. Then to identify the effect of Ifitm-1 in sperm motility and other seminal parameters, different concentration of Ifitm-1 antibody was incubated with spermatozoa, and seminal parameters were assessed using computer-assisted semen analysis (CASA). Interestingly, motility, progressive, and VAP were increased in the sperm with Ifitm-1 antibody treated compared to rabbit serum, however other parameters such as straightness were not changed. In order to identify the functional significance of Ifitm-1 in fertilization, capacitated spermatozoa were pre-incubated with anti-Ifitm-1 antibody and subsequently examined the ability to adhere to mouse oocytes. However, any defection or alteration in sperm-egg fusion was not found, Ifitm-1 antibody treated or non-treated spermatozoa showed a normal penetration. Although the precise role of Ifitm-1 in sperm motility and following fertilization need to be elucidated, this study suggests that the activation of Ifitm-1 on the sperm may enhance the motility of spermatozoa in mice.

Genome-wide Association Study to Identify Quantitative Trait Loci for Meat and Carcass Quality Traits in Berkshire

  • Iqbal, Asif;Kim, You-Sam;Kang, Jun-Mo;Lee, Yun-Mi;Rai, Rajani;Jung, Jong-Hyun;Oh, Dong-Yup;Nam, Ki-Chang;Lee, Hak-Kyo;Kim, Jong-Joo
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1537-1544
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    • 2015
  • Meat and carcass quality attributes are of crucial importance influencing consumer preference and profitability in the pork industry. A set of 400 Berkshire pigs were collected from Dasan breeding farm, Namwon, Chonbuk province, Korea that were born between 2012 and 2013. To perform genome wide association studies (GWAS), eleven meat and carcass quality traits were considered, including carcass weight, backfat thickness, pH value after 24 hours (pH24), Commission Internationale de l'Eclairage lightness in meat color (CIE L), redness in meat color (CIE a), yellowness in meat color (CIE b), filtering, drip loss, heat loss, shear force and marbling score. All of the 400 animals were genotyped with the Porcine 62K SNP BeadChips (Illumina Inc., USA). A SAS general linear model procedure (SAS version 9.2) was used to pre-adjust the animal phenotypes before GWAS with sire and sex effects as fixed effects and slaughter age as a covariate. After fitting the fixed and covariate factors in the model, the residuals of the phenotype regressed on additive effects of each single nucleotide polymorphism (SNP) under a linear regression model (PLINK version 1.07). The significant SNPs after permutation testing at a chromosome-wise level were subjected to stepwise regression analysis to determine the best set of SNP markers. A total of 55 significant (p<0.05) SNPs or quantitative trait loci (QTL) were detected on various chromosomes. The QTLs explained from 5.06% to 8.28% of the total phenotypic variation of the traits. Some QTLs with pleiotropic effect were also identified. A pair of significant QTL for pH24 was also found to affect both CIE L and drip loss percentage. The significant QTL after characterization of the functional candidate genes on the QTL or around the QTL region may be effectively and efficiently used in marker assisted selection to achieve enhanced genetic improvement of the trait considered.

Preparation and characterization of protein isolate from Yellowfin tuna Thunnus albacares roe by isoelectric solubilization/precipitation process

  • Lee, Hyun Ji;Lee, Gyoon-Woo;Yoon, In Seong;Park, Sung Hwan;Park, Sun Young;Kim, Jin-Soo;Heu, Min Soo
    • Fisheries and Aquatic Sciences
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    • v.19 no.3
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    • pp.14.1-14.10
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    • 2016
  • Isoelectric solubilization/precipitation (ISP) processing allows selective, pH-induced water solubility of proteins with concurrent separation of lipids and removal of materials not intended for human consumption such as bone, scales, skin, etc. Recovered proteins retain functional properties and nutritional value. Four roe protein isolates (RPIs) from yellowfin tuna roe were prepared under different solubilization and precipitation condition (pH 11/4.5, pH 11/5.5, pH 12/4.5 and pH 12/5.5). RPIs contained 2.3-5.0 % moisture, 79.1-87.8 % protein, 5.6-7. 4 % lipid and 3.0-3.8 % ash. Protein content of RPI-1 and RPI-2 precipitated at pH 4.5 and 5.5 after alkaline solubilization at pH 11, was higher than those of RPI-3 and RPI-4 after alkaline solubilization at pH 12 (P < 0.05). Lipid content (5.6-7.4 %) of RPIs was lower than that of freeze-dried concentrate (10.6 %). And leucine and lysine of RPIs were the most abundant amino acids (8.8-9.4 and 8.5-8.9 g/100 g protein, respectively). S, Na, P, K as minerals were the major elements in RPIs. SDS-PAGE of RPIs showed bands at 100, 45, 25 and 15 K. Moisture and protein contents of process water as a 2'nd byproduct were 98.9-99.0 and 1.3-1.8 %, respectively. Therefore, yellowfin tuna roe isolate could be a promising source of valuable nutrients for human food and animal feeds.