• Title/Summary/Keyword: n+ buffer

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Purification and Characterization of Oriental Pear(Niitaka, Pyrus pyrifolia Nak.) Protease (동양배(신고(新高)) Protease의 정제(精製) 및 성질(性質)에 관(關)하여)

  • Kim, Seung Yeol;Chung, Hai Jung;Kim, Seung Kyeom;Shin, Cheol Seung
    • Korean Journal of Agricultural Science
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    • v.16 no.2
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    • pp.225-238
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    • 1989
  • These studies were conducted to investigate the extraction, purification and characterization of oriental pear (Niitaka. Pyrus pyrifolia Nak.) protease, and the results obtained were as follows: 1. Oriental pear protease was effectively extracted by the method of homogenizing pear pulp with 0.7 volume of 0.1M-sodium phosphate buffer, pH 6.5 containing 5mM-cysteine, 40mM-2-mercaptoethanol and 2mM-EDTA at 10,000 rpm for 5 min. 2. The protease was purified by ammonium sulfate fractionation, Sephadex G-100 filtration and DEAE-Sephadex A-50 column chromatography, and the purified enzyme gave a single protein band on polyacrylamide gel electrophoresis. 3. The specific activity of purified enzyme was 29.65 unit/mg protein and the yield was 7.22%. 4. The moecular weight of the protease was estimated to be about 51,000 by SDS-polyacrylamide gel electrophoresis, and the enzyme had Km value of 54.5 mg/ml for casein. 5. The purified enzyme had a maximum activity at pH 6.0 and $50^{\circ}C$, and was stable from pH 5.5-6.5 and at temperatures below $50^{\circ}C$ 6. Casein was a better substrate for this protease compared to hemoglobin. 7. The enzyme activity was markedly inhibited by p-chloromercuribenzoic acid and heavy metal salts such as $HgCl_2$ and $MnSO_4$ also considerably inhibited the enzyme activity.

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Study on Reduction Effect of the Non-Point Pollutants through Riparian Buffer Zones (비점오염부하 저감을 위한 수변완충지대의 효율적 조성 연구)

  • Choi, I-Song;Kim, Sung-Won;Jung, Sang-Jun;Woo, Hyo-Seop;Oh, Jong-Min
    • Proceedings of the Korea Water Resources Association Conference
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    • 2007.05a
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    • pp.1793-1797
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    • 2007
  • 한강 "수변구역"에서 비점오염물질의 공공수역 유입을 억제하기 위한 다양한 방법들 중에서 보편적이고 자연친화적인 방법은 다양한 생물의 서식공간이며, 동시에 본류로 유입되는 과정에서 수질을 정화시키는 수질개선 공간인 수변완충구역, 또는 지대를 설정하여 관리하는 것이다. 그러나 이러한 수변완충지대 효과 분석 및 설정에 관한 연구는 국내에서 아직 수행되지 않았다. 본 연구의 목적은 수변구역의 자정능력을 높이는 것은 물론 그 밖의 하천 연안에서 비점오염물질의 차단과 처리능력을 증진시키고 수변 생태계의 서식처 보전 및 복원을 위해서 수변완충지대의 수질정화 기술개발과 생물다양성을 창출하는 수변완충지대 조성 기법을 개발하는데 있다. 본 연구에서는 기존 수변완충지의 추가적인 조성과 보완, 시험완충지 생태구조 및 기능 기초조사, 시험완충지 오염부하 저감효과의 실험 및 분석, 수변완충지대 설치 구상 등의 연구를 수행하였다. 수행 지역은 한강수계 지역으로 남한강 수변인 경기도 양평군 병산리에서 실시하였으며, 잔디와 갈대, 갯버들, 혼합지역, 자연그대로의 상태(대조지역)의 5 구역으로 구분하였고, 깊이별로 샘플을 채취하여 유입수와 표면유출, 하부유출을 비교해 보았다. 연구 결과, 5 가지 구역 중 잔디 구역의 SS, T-N, T-P, TOC의 제거 효율이 각각 76.7%, 85.2%, 97.6%, 83.3%로 가장 좋은 오염물질 제거 효율을 보였으며, 깊이 별 분석에서는 표면유출에서 하부유출로 갈수록 월등한 효율을 보였다. 따라서 본 연구를 통하여 비점오염원에 대한 한강수계의 수자원 보호 효과를 기대할 수 있고, 수변완충지대의 조성, 유지관리기술의 개발을 통한 수변완충지대의 계획과 설계에 직접적인 기여를 할 수 있으며, 수변구역에 설치 가능한 Riparian Buffer Zone의 중요성과 효율성을 알려 현재 하상 저니 준설 및 폭기 위주의 사업에서 생태 공학적 복원을 적극 고려한 정화사업으로 확대 추진하고자 한다.해결책을 얻어내는 상호보완적인 결과를 추구한다. 그가 디자인하는 작품은 전형적인 이미지를 내포하지 않는다. 즉 그의 작품은 기존의 가치와 이념적인 것은 배제하고, 창의적인 개념을 도출하였다.형모서리는 건물 특화 성격이 강하므로 불가피할 경우 소형 액센트 광고 위치를 미리 벽면으로 할애하는 것이 경관 및 입면계획에 유리한 것으로 분석되었다. 불확실도 해석모형 등의 새로운 기능을 추가하여 제시하였다. 모든 입출력자료는 프로젝트 단위별로 운영되어 data의 관리가 손쉽도록 하였으며 결과를 DB에 저장하여 다른 모형에서도 적용할 수 있도록 하였다. 그리고 HyGIS-HMS 및 HyGIS-RAS 모형에서 강우-유출-하도 수리해석-범람해석 등이 일괄되게 하나의 시스템 내에서 구현될 수 있도록 하였다. 따라서 HyGIS와 통합된 수리, 수문모형은 국내 하천 및 유역에 적합한 시스템으로서 향후 HydroInformatics 구현을 염두에 둔 특화된 국내 수자원 분야 소프트웨어의 개발에 기본 토대를 제공할 것으로 판단된다.았다. 또한 저자들의 임상병리학적 연구결과가 다른 문헌에서 보고된 소아 신증후군의 연구결과와 큰 차이를 보이지 않음을 알 수 있었다. 자극에 차이가 있지 않나 추측되며 이에 관한 추후 연구가 요망된다. 총대장통과시간의 단축은 결장 분절 모두에서 줄어들어 나타났으나 좌측결장 통과시간의 감소 및 이로 인한 이 부위의 통과시간 비율의 저하가 가장 주요하였다. 이러한 결과는 차가운 생수 섭취가 주로 결장 근위부를 자극하는 효과를 발휘하는 것이 아닌가 해석된다. 이와 같은 연구결과를 통해 생다시마를 주원료로 개발된 생다시마차와 생다시마 음료가 만성 기능성 변비 증세를 개선하는 효능이 잠재적으로 있음을 확인하였다. 그러나 생약제재의 변비약 수준으로 변비 개선 효능을 증대하기 위해서는 재료 배합비의 개선이나 대장 운동기능을 향상시키는 유효성분의 보강 등이 필요하다는 점도 알 수 있었다.더불어 산화물질 해독에 관여하는

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Effects of Saponin Contained Plant Extracts on Ruminal Fermentation Characteristics and Methane Production (Saponin 함유 식물 추출물의 첨가가 반추위 발효성상과 메탄생성에 미치는 영향)

  • Ok, Ji-Un;Baek, Youl-Chang;Kim, Kyoung-Hoon;Lee, Sang-Cheol;Seol, Yong-Joo;Lee, Kang-Yeon;Choi, Chang-Weon;Jeon, Che-Ok;Lee, Sang-Suk;Lee, Sung-Sil;Oh, Young-Kyoon
    • Journal of Animal Science and Technology
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    • v.53 no.2
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    • pp.147-154
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    • 2011
  • This study was conducted to evaluate the effects of saponin contained plant extracts on in vitro rumen fermentation characteristics and methane production. Ruminal fluid was collected from rumen cannulated Hanwoo steers fed rice straw and concentrate (5:5). Collected rumen fluids, corn starch and buffer including saponin contained plant extracts (ginseng, Ogapi, soapwort, tea plant and yucca; 0.5%/15 ml) were incubated at $39^{\circ}C$ for 24 h. All incubations were repeated five times. Rumen pH in all treatments was lower (p<0.05) compared with that of the control (no addition) during incubation time. The concentration of total VFA in all treatments was higher (p<0.05) than that of the control after 12h incubation. Compared with the control, the concentration of acetate and propionate in all treatments was lower and higher after 6h incubation, respectively. The concentration of $NH_3$-N in all treatments was lower (p<0.05) than that of the control except for Ogapi or yucca extracts supplementation. The number of protozoa in all treatments was significantly (p<0.05) lower than that of the control except for soapwort extract supplementation. The total gas production and methane production in all treatments was higher (p<0.05) and lower (p<0.05) compared with the control, except for ogapi or soapwort extracts supplementation after 12h incubation, respectively. Therefore, reduction in methane production by saponins may could be results from decreased protozoal population without any negative in vitro fermentation.

Studies on the analysis of phytin by the Chelatometric method (Chelate 법(法)에 의(依)한 Phytin 분석(分析)에 관(關)한 연구(硏究))

  • Shin, Jai-Doo
    • Applied Biological Chemistry
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    • v.10
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    • pp.1-13
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    • 1968
  • Phytin is a salt(mainly calcium and magnesium) of phytic acid and its purity and molecular formula can be determined by assaying the contents of phosporus, calcium and magnesium in phytin. In order to devise a new method for the quantitative analysis of the three elements in phytin, the chelatometric method was developed as follows: 1) As the pretreatment for phytin analysis, it was ashfied st $550{\sim}600^{\circ}C$ in the presence of concentrated nitric acid. This dry process is more accurate than the wet process. 2) Phosphorus, calcium and megnesium were analyzed by the conventional and the new method described here, for the phytin sample decomposed by the dry process. The ashfied phytin solution in hydrochloric acid was partitioned into cation and anion fractions by means of a ration exchange resin. A portion of the ration fraction was adjusted to pH 7.0, followed by readjustment to pH 10 and titrated with standard EDTA solution using the BT [Eriochrome black T] indicator to obtain the combined value of calcium and magnesium. Another portion of the ration fraction was made to pH 7.0, and a small volume of standard EDTA solution was added to it. pH was adjusted to $12{\sim}13$ with 8 N KOH and it was titrate by a standard EDTA solution in the presence of N-N[2-Hydroxy-1-(2-hydroxy-4-sulfo-1-naphytate)-3-naphthoic acid] diluted powder indicator in order to obtain the calcium content. Magnesium content was calculated from the difference between the two values. From the anion fraction the magnesium ammonium phosphate precipitate was obtained. The precipitate was dissolved in hydrochloric acid, and a standard EDTA solution was added to it. The solution was adjusted to pH 7.0 and then readjusted to pH 10.0 by a buffer solution and titrated with a standard magnesium sulfate solution in the presence of BT indicator to obtain the phosphorus content. The analytical data for phosphorus, calcium and magnesium were 98.9%, 97.1% and 99.1% respectively, in reference to the theoretical values for the formula $C_6H_6O_{24}P_6Mg_4CaNa_2{\cdot}5H_2O$. Statical analysis indicated a good coincidence of the theoretical and experimental values. On the other hand, the observed values for the three elements by the conventional method were 92.4%, 86.8% and 93.8%, respectively, revealing a remarkable difference from the theoretical. 3) When sodium phytate was admixed with starch and subjected to the analysis of phosphorus, calcium and magnesium by the chelatometric method, their recovery was almost 100% 4) In order to confirm the accuracy of this method, phytic acid was reacted with calcium chloride and magnesium chloride in the molar ratio of phytic: calcium chloride: magnesium chloride=1 : 5 : 20 to obtain sodium phytate containing one calcium atom and four magnesium atoms per molecule of sodium phytate. The analytical data for phosporus, calcium and magnesium were coincident with those as determine d by the aforementioned method. The new method employing the dry process, ion exchange resin and chelatometric assay of phosphorus, calcium and magnesium is considered accurate and rapid for the determination of phytin.

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The Ultrastructure of Leydig Cells in the Testis of Korean Native Chickens (한국 재래 닭 고환 간질세포의 미세구조에 관한 연구)

  • Jang, B.G.;Tae, H.J.;Choi, C.H.;Park, Y.J.;Yang, H.H.;Kim, N.S.;Park, S.Y.;Kang, H.S.;Park, B.Y.;Lee, Y.H.;Ahn, D.C.;Kim, I.S.
    • Korean Journal of Poultry Science
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    • v.33 no.3
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    • pp.181-188
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    • 2006
  • Changes in the fine structure of testicular Leydig cell from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The objective of this study were to elucidate Leydig cell ultrastructure during testicular development. Testes of chickens were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative(stereological) morphological studies were performed. The ultrastructural changes of the Leydig cell were investigated by ultrathin section with the transmission electron microscope. The stages of the Leydig cell development described focus on mitochondria, endoplasmic reticulum, and lipid droplets which are involved in androgens as fullows. 1) Approaching puberty. The closely packed Leydig cells and sparse intercellular space. The nucleus occupied a large portion of the Leydig cell volume. The population of Leydig cells contained two types of cells that differed in the appearance of their nuclei which were either highly electron-opaque or relatively electron-lucid. The cytoplasm was characterized by large amounts of lipid droplets, relatively few spherical mitochondria, and sparse smooth endoplasmic reticulum. 2) Puberty to adult. The Leydig cells which display features compatible with significant androgen synthesis: large volume of cytoplasm containing extended smooth endoplasmic reticulum, abundant mitochondria, and reduction of lipid droplets.

Studies on the Organo-mercury Residues in Rice Grain -I. Mercury residues in rice seeds treated with organo-mercury fungicide- (수도(水稻)에 처리(處理)된 유기수은제(有機水銀劑)의 잔류성(殘留性)에 관(關)한 연구(硏究) -제1보(第1報) 침지용유기수은제(浸漬用有機水銀劑)로 처리(處理)된 수도종자중(水稻種子中)의 수은잔류량(水銀殘留量)에 관(關)하여-)

  • Lee, Dong-Suk
    • Applied Biological Chemistry
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    • v.8
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    • pp.87-93
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    • 1967
  • 1) For the micro-analysis of mercury in plant materials, the method of Furutani was shown to be the simplest and most efficient way and the recovery of the assay was about 98%. 2) When the rice grain was soaked in 1/1000 diluted solution of organo-mercury fungicide for 8 hours at the end of March, the amounts of mercury residues in the brown rice and unhulled rice were 8.8 to $9.5\;{\mu}g/g$ seeds and 10.1 to $10.7\;{\mu}g/g$ seeds, respectively. 3) By washing the treated rice seeds with running water for three days, tile residual mercury concentration was reduced to 1/4 to 1/5; thus the mercury residues were 1.86 to $1.92\;{\mu}g/g$ for brown rice and 1.96 to $2.93\;{\mu}g/g$ for unhulled rice. 4) The residual mercury was present more in the unhulled rice than in the brown rice, either before or after washing of the treated seeds. 5) Among the different rice varieties, no difference was observed in mercury residues by seed treatment and washing.

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Characterization and Developmental Regulation of Polysialyltransferase from Embryos of Strongylocentrotus nudus (둥근성게, Strongylocentrotus nudus 배에 존재하는 Polysialyltransferase의 특성 및 발현 조절에 관한 연구)

  • 남지흔;김영대;박영제;조진원
    • Development and Reproduction
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    • v.2 no.2
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    • pp.149-155
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    • 1998
  • The polysialic acid (polySia) glycotope covalently modifies cell surface glycoconjugates on cells as evolutionarily diverse as microbes and human. The recent chemical identification of polysialylated glycoproteins in the jelly coat and on the cell surface of the sea urchin egg raises important questions about their biosynthesis and possible function. Using CMP-[$^{14}$ C]Neu5Ac as substrate and cell free preparations from eggs and embryos of the sea urchin Stronglylcentrotus nudus, we have identified a membrane associated CMP-Neu5Ac:poly-$\alpha$2, 8 sialosyl sialyltransferase (polyST) that transfers Neu5Ac to an endogenous acceptor. Optimal conditions for the polyST activity were found to be 2$0^{\circ}C$ in 20 mM MOPS buffer (pH 7.0). The polyST activity was increased 2.7 times by the addition of 10 mM $Mg^2$$^{+}$. The membrane-associated polyST also catalyzed the polysialylation of mammalian ganglioside GD3. Given that no structurally similar natural polysialylated gangliosides have been described, nor were observed in the present study, we conclude that a single polyST activity catalyzes sialylation of the endogenous acceptor and the gangliosides. Using an excess of GD3 as an exogenous acceptor, it was established that the expression of the polyST in S. nudus embryos increased rapidly at the mesenchyme blastula stage and reached at maximum at the gastrula stage. The finding that this polyST in the sea urchin embryo is developmentally regulated raises the possibility that it may play a role in the changing cell and tissue interactions that occur during gastrulation and the early stages of spicule formation.n.

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Effect of the Environmental and Nutritional Conditions on the Growth of Marine Microalga Isochrysis Galbana Parke (해양 미세조류 Isochrysis galbana Parke 성장에 대한 환경 및 영양 조건의 영향)

  • 오유관;박성훈
    • KSBB Journal
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    • v.11 no.3
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    • pp.303-310
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    • 1996
  • The marine microalga Isochrysis galbana Parke was studied to optimize its growth conditions in flask culture. Important medium components studied include nitrogen source, buffer, trace elements and vitamins. Environmental conditions include pH, temperature, light intensity, mixing extent and working volume. The medium prepared from natural sea-waters gave a higher final cell density than the medium prepared from synthetic sea-water Nitrate was a better source than ammonium. In the range of 0.4∼2mM, the final cell density was proportional to the initial nitrate concentration and the cell yield was estimated to be 8.5g dry cell wt/g N. For phosphate, optimal growth was observed in 0.1∼1.0mM but a considerable variation in pH was resulted. The addition of Tris at 5mM or 7mM could stabilize the medium pH, but this significantly reduced both growth rate and final cell density, The effect of trace elements and vitamins was negligible. Optimal temperature and initial pH were $20^{\circ}C$ and 8. When the intensity of incident light was varied in the range of 400∼2100 lux, the growth rate increased from 10mL to 70mL, the final cell density decreased although the initial growth rate did not change. Optimal agitation speed was 100rpm when working volume was 30mL. With optimal conditions, the maximum specific growth rate obtained was 0.021hr-1 and the final cell density was 1.1g/L.

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Molecular Cloning and Functional Expression of Extracellular Exo-β-(1,3)-Glucanase from Candida fermentati SI (Candida fermentati SI의 exo-β-(1,3)-glucanase유전자의 클로닝 및 그 특성)

  • Lim, Yu-Mi;Kim, Bong-Ki;Kim, Sang-Jun;So, Jai-Hyun;Kim, Won-Chan
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.317-323
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    • 2016
  • An isoflavone glucosidase that catalyzes the hydrolysis of isoflavone glucosides into glucose and corresponding aglycones was purified from Candida fermentati SI. The N-terminal sequence was determined to be GLNCDYCN. We designed degenerate primers on the basis of these amino acid sequences and successfully cloned the full structural gene sequence of the isoflavone glucosidase using inverse PCR. The exo-β-(1,3)-glucanase gene consists of 1227 base-pair nucleotides, encoding a 408-amino-acid sequence that shares 41–96% amino acid homology with other yeast exo-β-(1,3)-glucanases belonging to glycoside hydrolase family 5. The recombinant exo-β-(1,3)-glucanase was expressed in Pichia pastoris X-33, using a pPICZA vector system, and further characterized. The molecular mass of the purified exo-β-(1,3)-glucanase was estimated by SDS-PAGE to be 47 kDa. The optimal pH and temperature were pH 4.5 and 40℃, respectively. The Km values of the purified exo-β-(1,3)-glucanase for daidzin and genistin were 0.12 mM and 0.14 mM, respectively. The Vmax values of the purified isoflavone glucosidase were 945.03 U/mg for daidzin and 835.92 U/mg and for genistin.

Rumen fermentation and performance of Hanwoo steers fed total mixed ration with Korean rice wine residue

  • Jeong, Chang-Dae;Mamuad, Lovelia L.;Ko, Jong Youl;Sung, Ha Guyn;Park, Keun Kyu;Lee, Yoo Kyung;Lee, Sang-Suk
    • Journal of Animal Science and Technology
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    • v.58 no.1
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    • pp.4.1-4.7
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    • 2016
  • Background: This study was conducted to evaluate the effects of adding Korean rice wine residue (RWR) in total mixed ration (TMR) on in vitro ruminal fermentation and growth performance of growing Hanwoo steers. Methods: For in vitro fermentation, the experimental treatments were Control (Con: 0 % RWR + TMR), Treatment 1 (T1: 10 % RWR + TMR), and Treatment 2 (T2: 15 % RWR + TMR). The rumen fluid was collected from three Hanwoo steers and mixed with buffer solution, after which buffered rumen fluid was transferred into serum bottles containing 2 g dry matter (DM) of TMR added with or without RWR. The samples were then incubated for 0 h, 12 h, 24 h, or 48 h at $39^{\circ}C$ and 100 rpm. For the in vivo experiment, 27 Hanwoo steers (6 months old) with an average weight of $196{\pm}8.66kg$ were subjected to a 24-week feeding trial. The animals were randomly selected and equally distributed into three groups. After which the body weight, feed intake and blood characteristics of each group were investigated. Results: The pH of the treatments decreased significantly relative to the control during the 12 h of incubation. Total gas production and ammonia nitrogen ($NH_3-N$) was not affected by RWR addition. The total volatile fatty acid (VFA) was lower after 24 h of incubation but at other incubation times, the concentration was not affected by treatments. Feed cost was 8 % and 15 % lower in T1 and T2 compared to control. Blood alcohol was not detected and a significant increase in total weight gain and average daily gain were observed in Hanwoo steers fed with RWR. Conclusion: Overall, the results of this study suggest that TMR amended with 15 % RWR can be used as an alternative feed resource for ruminants to reduce feed cost.