• Title/Summary/Keyword: oligosaccharide production

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Anti-Inflammatory Effect of Alginate Oligosaccharides Produced by an Alginate-Degrading Enzyme from Shewanella oneidensis PKA1008 on LPS-Induced RAW 264.7 Cells (Shewanella oneidensis PKA1008 유래 알긴산 분해 효소에 의해 제조된 알긴산 올리고당의 항염증 효과)

  • Kim, Min-Ji;Bae, Nan-Yong;Bark, Si-Woo;Kim, Koth-Bong-Woo-Ri;Park, Ji-Hye;Park, Sun-Hee;Ahn, Dong-Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.6
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    • pp.888-897
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    • 2015
  • The anti-inflammatory effect of alginate oligosaccharides on LPS-induced RAW 264.7 cells was investigated at different time points (0-60 h). The alginate oligosaccharides were produced by an alginate-degrading enzyme from Shewanella oneidensis PKA1008. The alginate oligosaccharides decreased the production of nitric oxide and proinflammatory cytokines [tumor necrosis factor-${\alpha}$, interleukin (IL)-$1{\beta}$, and IL-6] in a dose-dependent manner. The alginate oligosaccharides showed peak anti-inflammatory activity after 36 h of incubation; at that time point, reduced protein expression of NF-${\kappa}B$ p65, iNOS, and COX-2 was detected. Furthermore, the alginate oligosaccharide treatment reduced the formation of ear edema at 36 h compared to samples examined at 0 h when the oligosaccharides were administered at 50 and 250 mg/kg body weight, as well as dermal thickness and mast cell numbers in a histological analysis. These results suggest that alginate oligosaccharides are a promising anti-inflammatory agent.

Characterization of Mold Releasing Agent Obtained from Carbon Black Suspension in Natural Polymer Solution (카본 블랙과 천연 고분자를 이용하여 제조한 금속 주조용 이형제의 특성)

  • Lee, Soo;Jin, Seok-Hwan;Park, Jung-Hyun
    • Journal of the Korean Applied Science and Technology
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    • v.29 no.1
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    • pp.13-18
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    • 2012
  • A die casting mold releasing agent was prepared from aqueous mixture of powdery carbon black and water soluble natural polymeric materials such as xanthan gum(X-gum) and carboxymethyl cellulose(CMC), which were used as thickening agent as well as curing agent with aldehydes. The suitable concentration of natural polymers for stable dispersion of carbon black in water was 0.25 wt% of X-gum or 1.0 wt% of CMC. When CMC was used less than 1 wt%, the final carbon black disperion showed a rapid phase separation. The adhesion of carbon black releasing agent on glass plate was improved with the amount of crosslinking agent, glutaraldehyde and chain extender, oligosaccharide. However, the affinity of carbon black releasing agent prepared with X-gum was stronger than that with CMC on glass plate. The final carbon black mold releasing agents prepared under our mixing conditions can be applied to the production of castings of high quality with good workability and without worthening evironmental situations.

Effect of Different Commercial Oligosaccharides on the Fermentation Properties in Kefir during Fermentation

  • Oh, Nam Su;Lee, Hyun Ah;Myung, Jae Hee;Lee, Ji Young;Joung, Jae Yeon;Shin, Yong Kook;Baick, Seung Chun
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.325-330
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    • 2013
  • Kefir is traditional fermented milk produced by various lactic acid bacteria (LAB) and yeast, which produce lactic acid, ethanol, carbon dioxide, and other flavor compounds. The purpose of this study was to evaluate the effects of different commercial oligosaccharides, such as maltotriose, fructooligosaccharide (FOS), galactooligosaccharide (GOS), and isomaltooligosaccharide (IMO), on the fermentation properties of kefir. First, we determined the acidification kinetic parameters, such as $V_{max}$, $t_{max}$(h), $t_{pH5.0}$(h), and $t_f$(h) of fermented milk supplemented with 4% (w/w) of different oligosaccharides. The probiotic survival and chemical composition (pH, organic acids profile, and ethanol content) of kefir during fermentation were also measured. Compared to control fermentation, all oligosaccharides increased acidification rate and reduced the time to complete fermentation (pH 4.7). The addition of FOS, in particular, resulted in the lowest $t_f$(h) and the highest populations of LAB and yeast during fermentation. All oligosaccharides increased ethanol production during fermentation. Further, significant differences were observed in the formation rates of six organic acids during fermentation. This study provided comparative data on the properties of commercial oligosaccharides for kefir manufacturing. Consequently, FOS especially had the potential for adequate and effective oligosaccharides in commercial kefir for the improvement of cost- and time-effectiveness.

Biological Activity of Human Dimeric Hyperglycosylated Erythropoietin (dHGEPO) Fusion Proteins

  • Naidansuren, Purevjargal;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.34 no.4
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    • pp.289-297
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    • 2010
  • Erythropoietin (EPO) is a glycoprotein hormone secreted from primarily cells of the peritubular capillary endothelium of the kidney, and is responsible for the regulation of red blood cell production. We constructed and expressed dimeric cDNAs in Chinease hamster ovary (CHO) cells encoding a fusion protein consisting of 2 complete human EPO domains linked by a 2-amino acid linker (Ile-Asp). We described the activity of dimeric hyperglycosylated EPO (dHGEPO) mutants containing additional oligosaccharide chains and characterized the function of glycosylation. No dimeric proteins with mutation at the $105^{th}$ amino acid were found in the cell medium. Growth and differentiation of the human EPO-dependent leukemiae cell line (F36E) were used to measure cytokine dependency and in vitro bioactivity of dHGEPO proteins. MIT assay at 24 h increased due to the survival of F36E cells. The dHGEPO protein migrated as a broad band with an average molecular mass of 75 kDa. The mutant, dHGEPO, was slightly higher than the wild-type (WT) dimeri-EPO band. Enzymatic N-deglycosylation resulted in the formation of a narrow band with a molecular mass twice of that of of monomeric EPO digested with an N-glycosylation enzyme. Hematocrit values were remarkably increased in all treatment groups. Pharmacokinetic analysis was also affected when 2.5 IU of dHGEPO were intravenously injected into the tails of the mice. The biological activity and half-life of dHGEPO mutants were enhanced as compared to the corresponding items associated the WT dimeric EPO. These results suggest that recombinant dHGEPO may be attractive biological and therapeutic targets.

Characterization of Erwinia tasmaniensis Isolated from Nuruk Producing Alginate Lyase (누룩으로부터 분리한 알긴산 분해 효소 생산 균주인 Erwinia tasmaniensis의 특성)

  • Kim, Hyun Ji;Lee, Sung-Mok;Kim, Sung-Koo;Lee, Jae-Hwa
    • Applied Chemistry for Engineering
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    • v.23 no.1
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    • pp.100-104
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    • 2012
  • Oligosaccharides production showed various biological activities in vivo like functional foods and industrial materials utilized available within many practical applications which have obtained from the degradation of alginate. Alginate is rich in the main component of seaweeds especially the brown algae. We investigated what degrading alginate from seaweeds to make alginate oligosaccharides can utilize in various fields using enzyme secreting Erwinia tasmaniensis. In this study, we observed an optimal culture condition of E. tasmaniensis, and characteristics of alginate lyase secreting E. tasmaniensis. These bacteria, E. tasmaniensis, were isolated from Nuruk. In this case, a suitable growth factor for E. tasmaniensis was culture it for 36 h in broth media on concentration of 1.0% (w/v) alginate. The enzyme showed the highest level of alginate lyase activity when cultured on broth media containing 1.0% (w/v) sodium alginate for 72 h. Optimal condition of pH, temperature and duration time for alginate lyase activity were found to be pH 6.0, $20^{\circ}C$ and 60 min, respectively.

Systematic Review on Application of Whey Towards Production of Galacto-oligosaccharide Using β-Galactosidase Enzyme from Pichia pastoris

  • Ramachandran, C;Oh, Deog-Hwan
    • Journal of Food Hygiene and Safety
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    • v.35 no.4
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    • pp.304-311
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    • 2020
  • Galacto-oligosaccharides (GOS) are prebiotics that have a beneficial effect on human health by promoting the growth of probiotic bacteria in the gut, in addition to having various applications in the food industry. GOS are generally produced from lactose in a reaction catalyzed by β-galactosidase. Synthesis of GOS from whey permeate (WP) (ultrafiltration of whey, concentrated then spray dried) using surface engineered β-galactosidase in Pichia pastoris (P. pastoris) is a novel method to convert waste into a valuable product. Cell-surface display is the expression of peptides and proteins on the surface of living cells by fusing them to functional components of cells. Surface engineered cells have many potential uses. The Flo1p flocculation functional domain, thought to be located near the N terminus, recognizes and adheres non-covalently to cell-wall components such as α-mannan carbohydrates, causing reversible aggregation of cells into flocs.

Soy-oligosaccharide Production from Yeast Fermented Bean Cooking Water: Effects of Ultrafiltration and Reverse Osmosis (효모발효한 대두침출액으로부터의 대두올리고당 생산: 한외여과 및 역삼투 처리효과)

  • Ku, Kyung-Hyung;Park, Dong-June;Mok, Chul-Kyoon
    • Korean Journal of Food Science and Technology
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    • v.29 no.4
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    • pp.680-686
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    • 1997
  • Ultrafiltration and reverse osmosis were applied to produce soy-oligosaccharides from yeast fermented bean cooking water (BCW). The recovered total sugar by the ultrafiltration of bean cooking water on a cutoff molecular weight membrane of 20,000 and 5,000 was over 80%. The protein was removed up to 38% from the non-fermented BCW, 31% by S. cerevisiae KCTC 7039 and 21% by H. anomala KFRI 626. The recovery of oligosaccharides was above 70%, although fermented or non fermented bean cooking water was different on the recovery of oligosaccharides. The ultrafiltrated BCW was concentrated by reverse osmosis with respect to the volume concentration ratio (VCR). Total solid, total sugar, ash and protein contents increased up to VCR of 3.5 and then levelled off, showing that the optimum VCR was 3.5.

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Preparation of Oligosaccharides from Alginic Acid by Enzymic Hydrolysis (효소분해에 의한 알긴산 올리고당류의 제조)

  • Joo, Dong-Sik;Lee, Jung-Suck;Park, Jung-Je;Cho, Soon-Yeong;Kim, Hee-Kyung;Lee, Eung-Ho
    • Korean Journal of Food Science and Technology
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    • v.28 no.1
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    • pp.146-151
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    • 1996
  • For the purpose of production of oligosaccharides from alginates, a bacterium was isolated from seaweed, and then an enzyme which degraded alginates was obtained from the bacterium. A specific activity of the enzyme was shown in G-rich block and Na-alginate (Wako Co.) as a result of reaction between the enzyme and six types of alginates (G-rich block, M-rich block and 4 commercial Na-alginate). Degradation products were prepared from the Na-alginate (Wako Co.) by the enzyme. The oligosaccharides were fractioned by Sephadex G-25 and Bio-gel P-2 and identified on a thin layer chromatography (TLC). Degree of polymerization (DP) of the oligosaccharides was shown from 2.6 to 7.5.

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Anti-oxidative effects of exogenous ganglioside GD1a and GT1b on embryonic developmental competence in pigs

  • Kim, Jin-Woo;Park, Hyo-Jin;Yang, Seul-Gi;Koo, Deog-Bon
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.4
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    • pp.347-356
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    • 2020
  • Gangliosides are glycolipids in which oligosaccharide is combined with sialic acids. Our previous studies have suggested an interplay between ganglioside GD1a/GT1b and meiotic maturation capacity in porcine oocyte maturation. Furthermore, ganglioside GD1a and GT1b are known for its antioxidant activity, but it is still unclear whether possible antioxidant role of GD1a and GT1b is involved in porcine embryos development competence during in vitro culture (IVC). Here, the effects of ganglioside GD1a and GT1b on the embryonic developmental competence during in vitro culture of porcine were investigated. The effects of ganglioside GD1a and GT1b on the expression of ST3GAL2 were confirmed during embryos development (2-cell, 4-cell, 8-cell and blastocyst) using immunofluorescent staining (IF). As a result, the fluorescent expression of ST3GAl2 was higher in embryos at 4-8 cells stage than blastocysts. Blastocyst development rate significantly increased in only 0.1 μM GD1a and GT1b treated groups compared with control group. To investigate the cellular apoptosis, we analyzed TUNEL assay. In case of only 0.1 μM GD1a and GT1b treated groups, the total number of cells in blastocyst compared with control group, but there was no significant difference in the rate of apoptotic cells. We identified the intracellular ROS levels using DCF-DA staining. According to the result, ROS production significantly decreased in blastocysts derived from the 0.1 μM GD1a and GT1b treated groups. These results suggest that ganglioside GD1a and GT1b improve the developmental competence of porcine embryos via reduction of intracellular ROS during preimplantation stage.

Characterization of a Thermostable Lichenase from Bacillus subtilis B110 and Its Effects on β-Glucan Hydrolysis

  • Huang, Zhen;Ni, Guorong;Wang, Fei;Zhao, Xiaoyan;Chen, Yunda;Zhang, Lixia;Qu, Mingren
    • Journal of Microbiology and Biotechnology
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    • v.32 no.4
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    • pp.484-492
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    • 2022
  • Lichenase is an enzyme mainly implicated in the degradation of polysaccharides in the cell walls of grains. Emerging evidence shows that a highly efficient expression of a thermostable recombinant lichenase holds considerable promise for application in the beer-brewing and animal feed industries. Herein, we cloned a lichenase gene (CelA203) from Bacillus subtilis B110 and expressed it in E. coli. This gene contains an ORF of 729 bp, encoding a protein with 242 amino acids and a calculated molecular mass of 27.3 kDa. According to the zymogram results, purified CelA203 existed in two forms, a monomer, and a tetramer, but only the tetramer had potent enzymatic activity. CelA203 remained stable over a broad pH and temperature range and retained 40% activity at 70℃ for 1 h. The Km and Vmax of CelA203 towards barley β-glucan and lichenan were 3.98 mg/ml, 1017.17 U/mg, and 2.78 mg/ml, 198.24 U/mg, respectively. Furthermore, trisaccharide and tetrasaccharide were the main products obtained from CelA203-mediated hydrolysis of deactivated oat bran. These findings demonstrate a promising role for CelA203 in the production of oligosaccharides in animal feed and brewing industries.