• 제목/요약/키워드: S100B protein

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Functional Characterization and Application of the HpOCH2 Gene, Encoding an Initiating $\alpha$l,6-Mannosyltransferase, for N-glycan Engineering in the Methylotrophic Yeast Hansenula polymorpha

  • Kim, Moo-Woong;Kim, Eun-Jung;Kim, Jeong-Yoon;Rhee, Sang-Ki;Kang, Hyun-Ah
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 2004년도 Annual Meeting BioExibition International Symposium
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    • pp.278-281
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    • 2004
  • The $\alpha$1,6-mannosyltransferase encoded by Saccharomyces cerevisiae OCH1 plays a key role for the outer chain initiation of the N-linked oligosaccharides. A search for Hansenula polymorpha genes homologous to S. cerevisiae OCHI (ScOCH1) has revealed seven open reading frames (ORF100, ORF142, ORF168, ORF288, ORF379, ORF576, ORF580). All of the seven ORFs are predicted to be a type II integral membrane protein containing a transmembrane domain near the amino-terminal region and has a DXD motif, which has been found in the active site of many glycosyltransferases. Among this seven-membered OCH1 gene family of H. polymorpha, we have carried out a functional analysis of H. polymorpha ORF168 (HpOCH2) showing the highest identity to ScOCH1. Inactivation of this protein by disruption of corresponding gene resulted in several phenotypes suggestive of cell wall defects, including hypersensitivity to hygromycin B and sodium deoxycholate. The structural analysis of N-glycans synthesized in HpOCH2-disrupted strain (Hpoch2Δ) and the in vitro $\alpha$1,6-mannosyltransferase activity assay strongly indicate that HpOch2p is a key enzyme adding the first $\alpha$1,6-mannose residue on the core glycan Man$_{8}$GlcNAc$_2$. The Hpoch2Δ was further genetically engineered to synthesize a recombinant glycoprotein with the human compatible N-linked oligosaccharide, Man$_{5}$GlcNAc$_2$, by overexpression of the Aspergillus saitoi $\alpha$1,2-mannosidase with the 'HDEL” ER retention signal.gnal.

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메주에서 분리된 Enterococcus faecium MJ-14가 생산하는 박테리오신의 부분정제 (Partial Purification of Bacteriocin Produced by Enterococcus faecium MJ-14 Isolated from Meju)

  • 이종갑;이군자;임성미
    • 한국식품위생안전성학회지
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    • 제20권4호
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    • pp.211-216
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    • 2005
  • E. faecium MJ-14 균주의 배양 상등액에 $50\%$ 황산암모늄을 처리한 결과, 박테리오신 활성은 3,840 BU/mL으로 배양 상등액에 비해 6배 증가되었고 회수율은 약 $60\%$에 이르렀다. 이온교환크로마토그래피에 의해 용출된 획분 중 $48\∼76$번에서 항균 활성이 나타났으며, 용출물의 비활성은 35.7배 증가하였고, 회수율은 약 $41\%$였다. 그리고 겔 크로마토그래피에 의해 비활성 127,293 BU/mg인 박테리오신 단백질을 정제하였으며, 이 때 정제도는 114배, 수율은 $36\%$였다. 정제된 박테리오신 단백질을 SDS-FACE한 결과. 항균 활성을 나타내는 4.3 kDa과 5.8 kDa의 분자량을 확인하였다.

작두콩분말 첨가에 따른 생면의 품질 특성 (Quality Characteristics of Wet Noodles added with Sword Bean Powder)

  • 박복희;고경미;전은례
    • 한국식생활문화학회지
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    • 제33권4호
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    • pp.374-381
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    • 2018
  • This study evaluated the quality characteristics of wet noodles added with sword bean powder(0, 5, 10, 15%) to determine the most preferred noodle recipe for consumer's desire. The proximate composition of the sword bean powder was as follows: Moisture contents were $13.4{\pm}0.08%$, protein $30.2{\pm}0.12%$, fat $0.3{\pm}0.09%$, protein $1.0{\pm}0.11%$ and carbohydrates $55.1{\pm}0.12%$, respectively. Total amino acid content of sword bean powder was measured 23,054.266 mg/100 g, especially in the order of histidine, glutamic acid and aspartic acid. The flavonoid content of sword bean powder was 17.9750 mg%. Water-binding capacity increased as the level of sword bean powder increased. When viscosity of wheat flours containing sword bean powder was measured by amylograph, the gelatinization point increased significantly as the level of sword bean powder increased, but peak viscosity decreased. As the level of sword bean powder increased, L value decreased, whereas a and b values increased. Weight, water absorption and volume of cooked noodles prepared with wheat flours containing sword bean powder decreased, whereas turbidity of soup increased. For textural properties, addition of sword bean powder increased hardness, springiness, chewiness of cooked noodles, whereas adhesiveness decreased. According to sensory evaluation such as appearance, color, flavor, taste, texture, overall preference, it was revealed noodles with 10% sword bean powder was the most preferred. According to the results, the addition of sword bean powder positively affects the overall sensory evaluation of wet noodle, and 10% is the optimal level for addition.

Energy Requirement of Rhode Island Red Hens for Maintenance by Slaughter Technique

  • Jadhao, S.B.;Tiwari, C.M.;Chandramoni, Chandramoni;Khan, M.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • 제12권7호
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    • pp.1085-1089
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    • 1999
  • Energy requirement of Rhode Island Red (RIR) hens was studied by comparative slaughter technique. Seventeen hens above 72 weeks of age were slaughtered in batches. Batch I consisted of 5 hens which were slaughtered initially. Batch II comprised of six hens, which were fed ad libitum broken rice (BR)-based diet for 18 days. Record of feed intake, number of eggs laid and egg weight during the period was kept. These hens were slaughtered and body energy content was determined. Egg energy was consisted as energy deposited. Batch III consisting of six hens which were fed varying quantity of diet for 15 days, were slaughtered similarly as hens of batch II. Regression equation (body weight to body energy) developed on batch I was applied to batch II and developed on batch II was applied to batch III hens, to find out initial body energy content of hens. Egg energy (EE) was calculated according to formula: EE (kcal) = -19.7 + 1.81 egg weight (g). Regressing metabolisable energy (ME) intake on energy balance (body energy change + egg energy), maintenance ME requirement of hens was found to be $119.8kcal/kg\;W^{0.75}/d$. Multiple regression of ME required for production on energy retained as protein and fat (body plus egg energy) indicated that RIR hens synthesize proteins with an efficiency of 85.5 and fat with an efficiency exceeding 100 percent on BR based diet.

Preparation and Characterization of a Polar Milk Lipid-enriched Component from Whey Powder

  • Lee, Kwanhyoung;Kim, Ara;Hong, Ki-Bae;Suh, Hyung Joo;Jo, Kyungae
    • 한국축산식품학회지
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    • 제40권2호
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    • pp.209-220
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    • 2020
  • Milk fat globule membrane (MFGM) is a lipid carrier in mammals including humans that consists mainly of polar lipids, like phospholipids and glycolipids. In this study, a process to enrich polar lipids in commercial butter and whey powder, including polar lipids of MFGM, was developed. WPC (whey protein concentrate) 60 was selected as the most suitable raw material based on the yield, phospholipid, protein, and lactose content of the polar lipid fraction obtained by ethanol extraction of two WPC (WPC60 and WPC70) and two buttermilk (A and B). After fractionation under optimum conditions, the polar-lipid enriched fraction from WPC60 contained 38.56% phospholipids. The content of glycolipids, cerebroside, lactosylceramide, ganglioside GM3, ganglioside GD3, was 0.97%, 0.55%, 0.09%, and 0.14%, respectively. Rancimat results showed that the oxidation stability of fish oil increased with an increase in the polar-lipid fraction by more than 30 times. In addition, the secretion of IL-6 and TNF-α decreased in a concentration-dependent manner after treatment of RAW 264.7 cells with 0.1 to 100 ppm of the polar lipid fraction. In this study, polar lipid concentrates with antioxidant and anti-inflammatory activity, were prepared from milk processing by-products. The MFGM polar lipid concentrates made from by-products are not only additives for infants, but are also likely to be used as antioxidants in cooking oils and as active ingredients for functional foods.

Biochemical Characterization and Genetic Diversity of Pongamia pinnata (L.) Pierre in Eastern India

  • Kumari, Kanchan;Sinha, Amrita;Singh, Sanjay;Divakara, B.N.
    • Journal of Forest and Environmental Science
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    • 제29권3호
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    • pp.200-210
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    • 2013
  • Biochemical characteristics of 24 Pongamia pinnata genotypes (candidate plus trees) from three agroclimatic zones were estimated and molecular characterization through RAPD markers was done. Various biochemical characters viz. seed oil, total carbohydrates, protein, acid value and Iodine number recorded significant variation among different genotypes. The highest seed oil content was 41.87% while seeds of 14 genotypes recorded above average (32.11%) for the trait. Seed oil and protein content exhibited a significant positive correlation and moderate heritability. Out of the initially selected twenty-five random primers, twenty-two RAPD primers were found to be highly reproducible and produced a total of 183 loci of which 147 (80.32%) loci were polymorphic. Percentage of polymorphism varied from 44% to 100% with an average of 80.62%. High level of genetic variation was found among different genotypes of P. pinnata. Both molecular and oil content (biochemical) markers appeared useful in analyzing the extent of genetic diversity in Pongamia and the result of these analyses will help to better understand the genetic diversity and relationship among populations. Overall, the Pongamia genotypes included in the study showed a correlation with their geographical origins such that genotypes from the same region tend to have higher genetic similarity as compared to those from different regions. However, in UPGMA based Nei's analysis, some genotypes were found not to be grouped based on geographical origins possibly due to the exchange of germplasm over time between farmers across the regions. The results from oil content analyses showed that several genotypes in 'Central and Western Plateau' agroclimatic zone of Jharkhand displayed a good potential for high oil content. The study provides insight about P. pinnata populations in Jharkhand (India) and constitutes a set of useful background information that can be used as a basis for future breeding strategy and improvement of the species.

피부(皮膚) 창상(創傷) 동물모델에서 흑삼(黑蔘) 열수 추출물 경구 투여의 효과 (Effects of Oral Administered Hot Water Extracts of Korean Black Ginseng on Wound Healing in Mice)

  • 김태령;김영준;우창훈
    • 한방재활의학과학회지
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    • 제32권1호
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    • pp.1-19
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    • 2022
  • Objectives This study aims to evaluate the wound healing effects of oral administered hot water extracts of Korean black ginseng (KBG). Methods 40 C57BL/6 mice were divided into five groups; normal, control, vitamin E 200 mg/kg, KBG 100 mg/kg, KBG 200 mg/kg, each n=8. Skin wounds were made in the back of all mice except normal group using biopsy punches. Wounds were observed on days 7 and 14 after injury. The anti-oxidant and inflammatory protein levels were evaluated using western blotting. Skin tissue was analyzed by hematoxylin & eosin and Masson's trichrome staining method. Results KBG significantly accelerated reducing wound area. KBG significantly decreased myeloperoxidase activity. KBG significantly decreased oxidative stress factors such as NADPH oxidase-4 and p22phox and increased antioxidant enzymes including nuclear factor erythroid 2-related factor2, kelch-like ECH-associated protein-1, heme oxygenase-1, superoxide dismutase, catalase and glutathione peroxidase-1/2. Moreover, KBG significantly decreased inflammation factors including nuclear factor-κB, phosphorylated inhibitor of κBα, cyclooxygenase-2, inducible nitric oxide synthase, tumor necrosis factor-α and interleukin (IL)-6 and increased anti-inflammation cytokine such as IL-4 and IL-10. In addition, KBG significantly increased tight junction proteins including claudin-1, claudin-3, claudin-4. In histopathologic, KBG made the epithelium thin and uniform, and accelerated the remodeling of collagen. Conclusions The results suggest that KBG has healing effects on skin wound in mice by anti-inflammatory and antioxidant activity.

Lipopolysaccharide로 자극된 RAW 264.7 세포와 마우스 귀부종 모델에 대한 참치 심장 Dichloromethane 분획물의 항염증 효과 (Anti-Inflammatory Activity of Dichloromethane Fraction from Katsuwonus pelamis Heart in LPS-Induced RAW 264.7 Cells and Mouse Ear Edema)

  • 김민지;배난영;최현덕;김꽃봉우리;박선희;성낙윤;변의홍;남희섭;안동현
    • 한국미생물·생명공학회지
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    • 제45권2호
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    • pp.101-109
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    • 2017
  • 본 실험에서는 참치 심장 70% ethanol 추출물을 dichloromethane으로 분획한 후 항염증 효과를 확인하기 위해 RAW 264.7 세포에 LPS로 염증을 유도시켜 염증 매개성 물질인 NO와 pro-inflammatory cytokine의 분비량의 변화를 확인하였다. 그 결과, 참치 심장 dichloromethane 분획물을 처리하였을 때, 농도 의존적으로 NO의 생성량을 감소시키는 것을 확인하였으며, 특히, $100{\mu}g/ml$에서 가장 높은 억제효과를 나타내었다. 따라서 dichloromethane 분획물의 억제 활성이 세포사멸에 의한 감소인지 알아보기 위해서 MTT assay를 하였을 때, 세포 생존율이 dichloromethane 분획물을 PBS 처리군과 비교하였을 때 유의적인 차이가 나타나지 않음을 확인하였고, 이를 통해 dichloromethane 분획물이 NO 및 전염증성 cytokine의 분비를 효과적으로 억제할 수 있는 물질임을 확인할 수 있었다. Dichloromethane 분획물을 처리하였을 때, 염증 관련 단백질 발현 정도를 western blot을 통해 확인한 결과, LPS에 의해 발현이 증가된 $NF-{\kappa}B$, iNOS 및 COX-2는 분획물을 처리함으로써 농도 의존적으로 감소되는 것을 확인할 수 있었다. 또한 dichloromethane 분획물의 처리가 인산화된 MAPKs의 발현을 저해함을 확인하여 참치 심장 dichloromethane 분획물이 $NF-{\kappa}B$와 MAPKs의 발현을 억제시킴으로써 NO 및 pro-inflammatory cytokine의 분비량을 감소시킴을 확인할 수 있었다. 동물 모델에서는, dichloromethane 분획물을 처리하였을 때 croton oil에 의한 귀 부종이 농도 의존적으로 감소함을 확인하였고, 특히, $250mg/kg{\cdot}body\;weight$ 농도로 투여시 시판 항염증제인 predinisolone을 $50mg/kg{\cdot}body\;weight$ 농도로 투여한 그룹과 유사한 효과를 나타내었다. 조직학적 변화를 확인한 결과에서는, 진피와 경피의 두께가 감소하였으며 진피내 mast cell 침윤이 감소되는 것을 확인할 수 있었다. 따라서 참치심장 dichloromethane 분획물이 효과적인 염증 예방 및 부종 완화를 위한 치료제로서 활용 가능성을 확인하였다.

Effect of Replacing Corn Silage with Whole Crop Rice Silage in Total Mixed Ration on Intake, Milk Yield and Its Composition in Holsteins

  • Ki, K.S.;Khan, M.A.;Lee, W.S.;Lee, H.J.;Kim, S.B.;Yang, S.H.;Baek, K.S.;Kim, J.G.;Kim, H.S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제22권4호
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    • pp.516-519
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    • 2009
  • This study was conducted to investigate the effects of replacing whole crop corn silage (WCCS) with whole crop rice silage (WCRS) in the total mixed ration (TMR) on nutrient intake, milk yield and its composition in Holstein cows. The Chucheong rice variety (Oryza sativa L. Japonica) and corn (Pioneer 32 P75) were harvested at yellow-ripe stage and were ensiled in round bales and in trench silos, respectively. Two TMR containing either WCCS or WCRS were prepared. These diets were randomly assigned to 16 midlactating Holstein cows (8 cows per treatment) and were fed for 120 days. The first 20 days were used for dietary adaptation and for the next 100 days daily feed intake, milk yield and its composition were recorded. The pH, lactic acid, NDF, ADF, CP, Ca and P contents were similar in WCRS and WCCS. The DM, ash and EE contents of WCRS were greater compared with WCCS. Nutrient (DM, NDF, TDN and CP) intakes were similar in cows fed WCCS- and WCRS-based TMR. Daily and 4% fat corrected milk yield were not affected by the treatments. Milk composition (percent milk fat, protein, lactose and total solids) was similar in cows fed either WCCS- or WCRSbased TMR. The concentration of milk urea N was greater in cows fed WCRS-based TMR than those fed WCCS-based TMR. In conclusion, round-baled WCRS can replace WCCS in the diet of mid- to late-lactating Holsteins without any deleterious effects on feed consumption, milk yield and its composition. The present findings raise the possibility that WCRS can be used as an alternative roughage source in the diets of dairy cows in countries with surplus rice production.

Streptozotocin으로 당뇨병을 유발시킨 쥐의 지방세포에 나타나는 $A_{1}$, Adenosine Receptor-Adenylyl Cyclase System의 변화 (Changes in $A_{1}$, Adenosine Receptor-Adenylyl Cyclase System of Rat Adipocytes Fellowing Induction of Experimental Diabetes by Streptozotocin Treatment)

  • 박경선;이명순;김경환
    • 대한약리학회지
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    • 제29권1호
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    • pp.97-105
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    • 1993
  • 흰쥐의 지방세포에 존재하는 아데노신 수용체는 $A_{1}$, subclass로 알려져 있다. 따라서 효현제 (Agonist)에 의해 자극되면 adenylyl cyclase가 억제되고 결과적으로 지방분해가 억제된다. 본 연구에서는 당뇨병으로 인하여 생기는 쥐의 지방세포의 $A_{1]$, 아데노신 수용체-adenylyl cyclase 시스템의 변화를 규명하고자 하였다. 흰쥐에 streptozotocin을 투여하여 당뇨병을 유발시킨 후 1주일이 되는 날, 굶기지 않은 쥐에 서 collagenase를 사용하여 지방세포를 분리하였다. 분리한 지방세포에 1 unit/ml adenosine deaminase와 $1\;{\mu}M$ isoproterenol을 가한 후 $37^{\circ}C$에서 1시간 동안 incubation하여 생성되는 glycerol을 측정하였을 때, 당뇨병군에서는 대조군에 비해 $A_{1}$, 아데노신 수용체의 효현제인 $(-)-N^{6}-(R-phenylisopropyl)$ adenosine (PIA)에 의해 지방분해가 약 두배 더 억제되었다. 그 기전을 밝히기 위하여 지방세포막의 $[{^3}H]PIA$ binding을 측정하였는데, 친화력이나 수용체의 수는 당뇨병군과 대조군 사이에 통계적으로 유의한 차이가 없었다 (대조군의 $K_{d}$$0.51{\pm}0.09\;nM$이었고 $B_{max}$$1.60{\pm}0.12\;pmoles/mg$ protein이었다. 당뇨병군의 $K_{d}$$0.54{\pm}0.21\;nM$이었고 $B_{max}$$1.72{\pm}10.31\;pmoles/mg$ protein이었다). 그러나 $100\;{\mu}M$ isoproterenol 자극에 의한 adenylyl cyclase activity의 PIA에 의한 억제를 비교하여 보았을 때 당뇨병군에서의 억제는 대조군의 1.9배로 나타났다. 이상의 결과로 보아 당뇨병군의 지방세포에 나타나는 변화, 즉 지방 분해가 PIA와 같은 $A_{1}$, 아데노신 수용체의 효현제에 의해 더욱 억제되는 것은 지방세포막의 수용체 자체의 변화때문이 아니라 수용체 이후의 신호전달 체계의 변화때문인 것으로 생각된다.

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