• Title/Summary/Keyword: CMCase

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Microbial Population Diversity of the Mud Flat in Suncheon Bay Based on 16S rDNA Sequences and Extracellular Enzyme Activities (남해안 갯벌 미생물의 세포외효소 활성 및 16S rDNA 분석에 의한 다양성 조사)

  • Kim, Yu-Jeong;Kim, Sung-Kyum;Kwon, Eun-Ju;Baik, Keun-Sik;Kim, Jung-Ho;Kim, Hoon
    • Applied Biological Chemistry
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    • v.50 no.4
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    • pp.268-275
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    • 2007
  • Diversity of the mud flat microbial population in Suncheon Bay was investigated by studying extracellular enzyme activities and 16S rDNA sequences. Four culturable bacterial strains with CMCase, xylanase and protease activities were isolated from the wetland and the mud flat. All the strains produced more xylanase activity than CMCase or protease activity, and the properties of the isolate enzymes from the wetland were similar to those from the mud flat. About 2,000 clones were obtained with the 16S rDNA amplified from the metagenomic DNA isolated from the mud samples. Based on the restriction pattern(s), seventeen clones were selected for base sequence analysis. Of the 17 clones, only 35% (6 clones) were found to be cultured strains and 65% (11 clones) to be uncultured strains. The similarities in the base sequences of the clones ranged from 91.0% to 99.9% with an average similarity of 97.3%. The clones could be divided into 7 groups, Proteobacteria (9 clones, 52.9%), Firmicutes (3 clones, 17.6%), Bacteroidetes (1 clone), Flavobacteria (1 clone), Verrucomicrobia (1 clone), Acidobacteria (1 clone), and Chloroflexi (1 clone). Most of the Proteobacteria clones were gamma Proteobacteria associated with oxidation-reduction of sulfur.

Comparison of Endo-, Exo-Cellular Enzyme Activity for New Strains of Hypsizygus marmoreus (느티만가닥버섯의 신품종에 대한 endo-, exo-cellular 효소 활성도의 비교)

  • Lee, Chang-Yun;Song, Ho-Sung;Ro, Hyeon-Su;Woo, Ju-Ri;You, Young-Hyun;Kim, Jong-Guk
    • Journal of Life Science
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    • v.22 no.6
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    • pp.837-843
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    • 2012
  • This study was carried out to investigate the morphological and physiological characteristics of six new cultivars of Hypsizygus marmoreus (Hm) and measure endo-, exo-cellular enzyme-specific activity. The domestic wild stain (Hm3-10) and commercial strain in Japan (Hm1-1) were mated by crossing monokaryon mycelia. We gained 58 strains from one of 400 crosses through the $1^{st}$ cultivation experiment, and selected six strains from one of 58 strains through the $2^{nd}$ cultivation experiment. When six of the selected new strains were grown during several spawn culture periods (60, 70, 80, 90, and 100 days), a spawn culture period of more 80 days was considered to be excellent as being shorter than 19~20 days. Therefore, we determined the period of spawn culture as 80 days. Three strains such as Hm15-3, Hm15-4, and Hm17-5 showed an excellent result. When endo-cellular enzyme activity measured eight strains, we obtained a result of that specific activity of ${\alpha}$-amylase at the highest as 73.9~102.2 unit/mg protein, and chitinase is lower than ${\alpha}$-amylase at 8.1~13.1 unit/mg protein. When exo-cellular enzyme activity measured eight strains, we determined the result of that specific activity of ${\alpha}$-amylase is the highest at 5,292~1,184 unit/mg protein, and CMCase and xylanase were 1,140~245 unit/mg protein, 94~575 unit/mg protein, compared to each other. However, the enzyme activity of ${\beta}$-glucosidase and chitinase is low.

Isolation and Characterization of Cellulolytic Anaerobic Fungi from the Guts of the Hanwoo Cattle and the Korean Native Goat (한우 및 산양의 장내 섬유소 분해 혐기 곰팡이의 분리 및 특성 구명)

  • Kim, C.H.;Lee, S.S.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.1019-1030
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    • 2003
  • The study was conducted to isolate and identify highly fibrolytic anaerobic fungi from the guts of a Hanwoo steer and a Korean native goat, and then investigate the characterization of cellulolytic activity of an anaerobic fungus. Twenty-one anaerobic fungal colonies were isolated in the study, in which 16 colonies were isolated from the rumen contents of the Hanwoo steer and 5 colonies from the duodenal fluids of the Korean native goat. Four anaerobic fungi were selected based on higher cellulolytic enzyme activities to identify under a optical microscope. NLRI-M003 and -T004 belong to Neocallimastix genus and NLRI-M014 belongs to Piromyces genus based on the morphology of their thallus, sporangia, rhizoid and the number of flagella. NLRI-M001 appeared to be an unknown strain of anaerobic fungi due to its different morphology from existing types of anaerobic fungi, though the morpholgoy is similar to Orpinomyces sp. Supplementation of 2% anaerobic fungal culture(NLRI-M003) in rumen-mixed microorganisms increased in vitro DM degradability of rice straw and filter paper up to 4 and 11%, respectively, compared with non-supplementation(control). CMCase and xylanase activities in in vitro culture were also higher in 2% fungal supplementation than controls in both rice straw and filter paper substrates.

Partial Purification and Some Properties of Carboxymethyl Cellulases from Alkalophilic Cephalosporium sp. RYM-202 (호알칼리성 Cephalosporium sp. RYM-202가 생산하는 carboxymethyl cellulase의 부분정제 및 특성)

  • Kang, Myoung-Kyu;Park, Hee-Moon;Rhee, Young-Ha;Kim, Yun-Seog;Kim, Yeo-Kyung
    • The Korean Journal of Mycology
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    • v.21 no.4
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    • pp.301-309
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    • 1993
  • An alkalophilic Cephalosporium sp. RYM-202 capable of producing cellulase components was isolated from soil. This organism grew best at an initial pH 9.0 and produced cellulase maximal at an initial pH 9.5-10.0. Three carboxymethyl cellulases(CMCases), P-I-I, P-I-II and P-II-I, were partially purified by DEAE-Sephadex A-50 ion exchange column followed by Sephadex G-150 gel filtration. The optimum pH values for activity were 7.5 for P-I-I, 8.0-9.5 for P-I-II and 7.5-10.0 for P-II-I. All CMCases were stable between pH 4.5 and 12.0. Temperature optima for activity ranged between 40 and $60^{\circ}C$ and more than 50% of the maximum activity was observed at $20^{\circ}C$ for both of P-I-I and P-II-I. The activity of CMCases was significantly stable in the presence of various laundry components, such as, surfactants, chelating agents and alkaline proteinases.

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성스테로이드 호르몬과 해산 태생 경골어류의 성분화

  • Gwon, Jun-Yeong;Han, Hyeong-Gyun;Im, Han-Gyu;Kim, Dae-Jung;Lee, Chan-Hui;Gwon, Hyeok-Chu
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2004.05a
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    • pp.355-355
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    • 2004
  • 난생 경골어류의 성분화는 내인성 및 외인성 성스테로이드 호르몬의 영향을 받는 것으로 알려져 있지만, 해산 태생 경골어류의 성분화 과정에서 성스테로이드 호르몬의 작용은 아직 명확하게 밝혀져 있지 않다. 본 연구에서는 유용 양식어종의 하나인 해산 태생어류 조피볼락 Sebastes schlegeli를 대상으로 이 종의 성분화에 미치는 성스테로이드 호르몬의 영향을 조사하였다. 실험에 사용한 조피볼락은 출산후 일령 45일 전후의 성이 미분화한 개체들이었다. 실험어는 3개 실험군으로 나누어 (각각 3반복), 1개월간 각각 estradiol-l7${\beta}$ 또는 methyltestosterone을 처리하였고, 하나의 실험군은 호르몬 처리를 하지 않았다 처리기간 동안에는 각 실험군으로부터 무작위 sampling을 실시하여, 호르몬 처리에 따른 체내 성스테로이드 호르몬 변화를 조사하였으며, 각 실험군의생식소 분화과정을 조직학적으로 조사하였다. 성스테로이드 호르몬은 diethylether로 추출한후, rndioirmmunoassay 방법 (RIA)으로 분석하였다. 실험결과 대조구의 실험어는 1:1의 자연성비에 부합하는 성분화 결과를 나타내었지만, estradiol-17${\beta}$ 처리구는 자성화, methyltestosterone 처리구는 웅성화하는 경향을 나타내었으며, 이러한 성분화 shift는 외부에서 투여한 호르몬이 실재 내부호르몬양의 변화를 초래하여 이루어졌음이 RIA 분석에 의해 부분적으로 확인되었다. 본 연구의 결과는 해산 태생 경골어류의 성분화 역시 성스테로이 드호르몬의 작용에 의하여 매개 또는 조절되어진다는 사실을 제시한다. 하지만 태생어류에서 이 호르몬의 실질적 작용 메카니즘 및 명확한 작용시기에 대해서는 보다 많은 연구가 요구되어 진다.을 설계하는 것이 가능하였다.적(最適) 온도(溫度)는 $30^{\circ}C$, avicelase와 ${\beta}-glucosidase$의 최적(最適) pH는 5.0, CMCase는 pH 5.5 이었으며, 균사(菌絲) 생육(生育)은 pH 5.0에서 양호(良好)하였다. 배양(培養) 기간(期間)은 avicelase가 8일(日), CMCase가 10일(日), ${\beta}-glucosidase$는 16 일간(日間) 배양(培養)하였을 때 최대치(最大値)를 보였고, 균사(菌絲) 생육(生育)은 12일(日) 배양(培養)했을 때 가장 양호(良好)하였다.가한 반면, 중국인들은 고소한 향의 강도, 고소한 향의 기호도, 전체적인 맛에서 뚜렷하게 일본참기름을 우수하게 평가하였다.s의 항체(抗體)로 반응(反應)시킨 후 protein-A gold(15 nm)로 표식(標識)시킨 바 제일 바깥 상층(上層)의 keratinocyte에 있어서 세포막표면(細胞膜表面)을 따라 표식(標識)되어 세포막항원(細胞膜抗元)을 나타내었으며, 이와 같은 소견(所見)으로 미루어 정상피부(正常皮膚) 중층편평상피세포(重層扁平上皮細胞)에서도 동일(同一)한 소견(所見)을 관찰(觀察)할 수 있다고 본다.al remnants, Resorption of fetus로 관찰된 것이다. Fetal death는 수정후 $14{\sim}18$일까지의 사망으로써 Maceration of fetus로 관찰되는 것이다. 통계학적 분석은 각 Group의 착상 을과 자궁 내 사망 율을 산출할 때에는 각 임신마우스에 따라 발생빈도가 크게 다르기 때문에 통계처리에는 Non parametric 검정인 Kluskal Wallis 검정을 사용하여 분석하였다. 또한 개체 Level 영향인 착상을, 태아사망,

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Comparison of in vitro digestibility and chemical composition among four crop straws treated by Pleurotus ostreatus

  • Nie, Haitao;Wang, Ziyu;You, Jihao;Zhu, Gang;Wang, Hengchang;Wang, Feng
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.1
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    • pp.24-34
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    • 2020
  • Objective: The effects of Pleurotus ostreatus on the feed utilization of broad bean stalks (BBS), rape straw (RS), paddy straw (PS), and corn stalk (CS) was examined. Methods: The four roughages were co-cultured with Pleurotus ostreatus. The chemical composition; enzyme activities of laccase, carboxymethylcellulase (CMCase) and xylanase; carbohydrate and protein fractions (based on The Cornell Net Carbohydrate and Protein System [CNCPS]) were assessed at different days after inoculation (7, 14, 21, 28 d) and un-inoculated roughages (control, 0 d). The digestibility of nutrient components and the gas production of roughage with various incubation times were monitored at 0, 2, 4, 6, 9, 12, 24, 36, 48, 60, and 72 h using an in vitro ruminal fermentation method. Results: A higher CMCase activity (0.1039 U/mL) and earlier time to peak (14 d) were detected in Pleurotus ostreatus cultured with CS (p<0.05). Significantly, the incubation length-dependent responses of cumulative gas production were observed from 24 to 72 hours post fermentation (p<0.05), and these incubation length-dependent effects on cumulative gas production of PS and CS appeared earlier (24 h) for PS and CS than those (48 h) for BBS and RS (p<0.05). The fast-degradable carbohydrate (CA) content for all four roughages significantly increased over time (p<0.05). Nonetheless, increased degradation efficiency for CA treated with Pleurotus ostreatus was detected at both 21 and 28 days of incubation (p<0.05). With the exception of PS (p<0.05), there were no significant difference among the roughages (p>0.05) in slowly-degradable carbohydrate (CB2) at different incubation times (p<0.05). Conclusion: Assessment of the alterations in chemical composition, CNCPS system fractions, and the fermentation kinetics after biological pretreatment may yield a valuable database for evaluating the biological pretreatment of Pleurotus ostreatus in ruminant feed.

Effects of Types of TMR on Rumen Fermentation Characteristics and Nutrients Digestibility in Sheep (유형별 완전혼합사료 급여가 반추위내의 발효성상 및 영양소 소화율에 미치는 영향)

  • Li, D.Y.;Ko, J.Y.;Choi, N.J.;Lee, Sang S.;Song, J.Y.;Lee, S.Y.;Park, S.H.;Sung, H.G.;Ha, J.K.
    • Journal of Animal Science and Technology
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    • v.45 no.5
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    • pp.805-812
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    • 2003
  • This study was conducted to examine effects of feeding dry TMR(DTMR), wet TMR(WTMR) and fermented TMR(FTMR) on rumen fermentation, enzyme activity and digestibility in the total tract of sheep. Three rumen cannulated sheep were used in a 3${\times}$3 latin square design. The present results showed that pH, NH3-N, total and individual VFA, A/P ratio and enzymes (CMCase, Xylanase and Protease) activity in the rumen were higher in WTMR and FTMR compared with DTMR. In addition, dry matter, organic matter, crude protein, ether extract, NDF and ADF digestibility in the total tract were also higher in WTMR and FTMR compared with DTMR. Therefore, the present results showed that WTMR and FTMR are better than DTMR for rumen fermentation and nutrients digestibility.

Characterization of Bacillus licheniformis KJ-9 Isolated from Soil (토양으로부터 분리한 Bacillus licheniformis KJ 9의 특성)

  • Seo, Dong-Cheol;Ko, Jeong-Ae;Gal, Sang-Won;Lee, Sang-Won
    • Journal of Life Science
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    • v.20 no.3
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    • pp.403-410
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    • 2010
  • In order to produce high-quality fermenting composts, a microorganism was isolated from the natural world. The bacterium has not only in high enzyme activities but also had good antimicrobial activities against phytopathogenic microorganisms. Its cultivating characteristics were then investigated. Bacterium KJ-9, which contains high CMCase, protease and chitinase activities and excellent antimicrobial activities against phytopathogenic microorganisms, was separated from leaf mold and identified as Bacillus licheniformis by two methods: Bergey's Manual of Systematic Bacteriology and API 50 CHL Carbohydrate Test Kit (Bio Merieux, France) using an ATB (Automated Identification) computer system (Bio Merieux, France). Optimal medium for cultivation of B. licheniformis was 2% soluble starch as a carbon source, 0.5% yeast extract as a nitrogen source and 0.05% $MgSO_4{\cdot}7H_2O$. Optimal growth conditions of pH, temperature and shake speed were pH 7.0, $50^{\circ}C$ and 180 rpm, respectively. Culture broth of B. licheniformis KJ-9 cultured for 36~60 hr was effective in fungicidal activities against plant pathogens including Botrytis cinerea, Corynespora cassicola, Fusarium oxysporum, and Rhizoctonia solani.

Studies on the Production of Fermented Feeds from Agricultural Waste Products (Part IV) -On the Production of Cellulase by Aspergillus niger and Trichoderma viride- (농산폐기물(農産廢棄物)에서 발효사료(醱酵飼料)의 생산(生産)에 관(關)한 연구(硏究)[제4보(第四報)] -Aspergillus niger와 Trichoderma viride에 의(依)한 Cellulase의 생산성(生産性)에 관(關)하여-)

  • Lee, Ke-Ho;Koh, Jeong-Sam;Lee, Kang-Hup
    • Applied Biological Chemistry
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    • v.19 no.3
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    • pp.139-144
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    • 1976
  • In order to investigate the properties of enzymes from two strains of mold, reported in the previous paper, (1) studies have been made concerning the characteristics of cellulase of Aspergillus niger-SM6 and Trichoderma viride-SM10, and summarized as follows. 1. In the semi-purification the recovery of ${\beta}-glucosidase$ was the highest when 80-90% ethanol was used and 0.8 saturation of $(NH_4)_2SO_4$. 2. The characteristics of the semi-purified enzyme were as follows. Aspergillus niger-SM6 Trichoderma viride-SM10 Optimum pH 3.5 4.0 pH stability 3.0-6.0 3.0-6.0 Optimum temperature $60^{\circ}C$ $60^{\circ}C$ Heat stability below $60^{\circ}C$ below $50^{\circ}C$ Optimum reaction time 30 min. 60 min. Optimum CMC concentration 3% 3% 3. The Km values of CMCase were 0.8% and 1.01 for Aspergillus niger-SM6 and Trichoderma viride-SM10, respectively. 4. In the strain of Aspergillus niger-SM6, there were high activity of xylanase and pectinase.

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Biosynthetic Regulation and Enzymatic Properties of $\beta$-Glucosidase from Cellulomonas sp. CS 1-1 (Cellulomonas sp. CS1-1으로 부터의 $\beta$-Glucosidase의 합성조절과 그의 효소학적 성질)

  • Lee, Hee-Soon;Min, Kyung-Hee;Bae, Moo
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.119-125
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    • 1988
  • $\beta$-Glucosidase of Cellulomonas sp. CS1-1 in cellular compartment was localized with cell-bound form while Avicelase and carboxymethylcellulase (CMCase) were appeared with extracellular enzyme. Cell growth on cellulose or CMC minimal broth was increased by glucose addition. $\beta$-Glucosidase production on cellobiose or CMC minimal broth was repressed by the addition of glucose. However, on CMC minimal broth, the enzyme production was specially stimulated by cellobiose addition. $\beta$-Glucosidase production was also induced by CMC, starcth and maltose compared with glycerol, arabinose, xylose and trehalose. From the above results, it was concluded that glucose effect on $\beta$-glucosidase biosynthesis showed catabolite repression, but enzyme production was induced by cellobiose, CMC, and starch, indicating that $\beta$-glucosidase is inducible enzyme. Yeast extract stimulated $\beta$-glucosidase production more than peptone and ammonium sulfate. $\beta$-Glucosidase activity was increased with 50mM MgCl$_2$in 10mM potassium phosphate buffer (pH 7.0). Optimum conditions for enzyme activities were pH 6.0 and 42$^{\circ}C$, Km value of $\beta$-glucosidase for p-nitrophenyl-$\beta$-D-glucosidase was 0.256mM and Ki for $\beta$-D(+)-glucose was 9.0mM.

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