• 제목/요약/키워드: cellulase C

검색결과 476건 처리시간 0.03초

Effect of Dilute Alkali on Structural Features and Enzymatic Hydrolysis of Barley Straw (Hordeum vulgare) at Boiling Temperature with Low Residence Time

  • Haque, Md. Azizul;Barman, Dhirendra Nath;Kang, Tae Ho;Kim, Min Keun;Kim, Jungho;Kim, Hoon;Yun, Han Dae
    • Journal of Microbiology and Biotechnology
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    • 제22권12호
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    • pp.1681-1691
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    • 2012
  • This work was conducted to evaluate the effect of dilute sodium hydroxide (NaOH) on barley straw at boiling temperature and fractionation of its biomass components into lignin, hemicellulose, and reducing sugars. To this end, various concentrations of NaOH (0.5% to 2%) were applied for pretreatment of barley straw at $105^{\circ}C$ for 10 min. Scanning electron microscopy (SEM), atomic force microscopy (AFM), and Fourier transform infrared (FTIR) spectroscopy studies revealed that 2% NaOH-pretreated barley straw exposed cellulose fibers on which surface granules were abolished due to comprehensive removal of lignin and hemicellulose. The X-ray diffractometer (XRD) result showed that the crystalline index was increased with increased concentration of NaOH and found a maximum 71.5% for 2% NaOH-pretreated sample. The maximum removal of lignin and hemicellulose was 84.8% and 79.5% from 2% NaOH-pretreated liquor, respectively. Reducing sugar yield was 86.5% from 2% NaOH-pretreated sample using an enzyme dose containing 20 FPU of cellulase, 40 IU of ${\beta}$-glucosidase, and 4 FXU of xylanase/g substrate. The results of this study suggest that it is possible to produce the bioethanol precursor from barley straw using 2% NaOH at boiling temperature.

큰느타리버섯 수확후배지 발효사료 급여가 비육한우에 미치는 영향 (Effect of dietary supplementation of fermented spent mushroom substrates from Pleurotus eryngii on Hanwoo Steers)

  • 문여황;김혜수;김철환;조웅기;유영복;신평균;조수정
    • 한국버섯학회지
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    • 제13권2호
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    • pp.108-113
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    • 2015
  • 본 연구는 큰느타리버섯 수확후배지 발효사료에 적합한 생균제를 개발하고 큰느타리버섯 수확후배지 발효사료 급여가 비육한우에 미치는 영향을 조사하기 위하여 수행되었다. 큰느타리버섯 수확후배지 발효사료를 제조하기 위하여 큰느타리버섯 수확후배지로부터 cellulase, xylanase 활성이 우수하면서 Asp. flavus에 대해 항균활성을 나타내는 균주를 선발하여 Bacillus subtilis CS21로 명명하고 버섯수확후배지 발효사료용 생균제로 사용하였다. 사양시험은 13개월령 비육한우 20두를 공시하여 29-30개월령까지 수행하였다. 공시동물은 성장단계(육성기, 비육전기, 비육후기)에 따라 배합비를 조절한 TMR 사료를 급여한 대조구와 30%의 큰느타리버섯 수확후배지 발효사료가 첨가된 TMR 사료(30% F-SMS TMR)를 급여한 처리구로 나누어 수용하였다. 시험기간 동안 총 증체량과 일당 사료섭취량은 대조구보다 30% F-SMS TMR 급여구에서 약간 높게 나타났다. 육질등급에서는 대조구와 30% F-SMS TMR 급여구 모두 1등급 이상 출현율이 100%를 나타내었으나 육량등급의 A등급 출현율은 대조구(57%)가 30% F-SMS TMR 급여구(41.67%) 보다 높은 경향을 나타내었으며 C등급 출현율은 대조구(0%)보다 30% F-SMS TMR 첨가구(25%)에서 높은 경향을 나타내었다. 이상의 결과를 종합하면 발효공정을 거쳐 기호성과 저장성이 개선된 큰느타리버섯 수확후배지 발효사료 급여는 육질등급에서는 대조구와 비슷하였지만 증체량은 대조구 보다 30% F-SMS TMR 급여구가 높은 반면 육량등급의 A등급 출현율은 낮고 C등급 출현율은 높은 경향을 나타내었으므로 육량등급을 높일 수 있도록 배합사료 조성비를 개선한다면 큰느타리버섯 수확후배지는 비육한우의 대체사료자원으로 이용될 수 있을 뿐만 아니라 사료비절감 효과를 기대할 수 있을 것으로 사료된다.

고추역병균 Phytophthora capsici를 방제하는 길항균주 Bacillus megaterium KL39의 선발과 길항물질 (Purification and Characteriztion of an Antifungal Antibiotic from Bacillus megaterium KL 39, a Biocontrol Agent of Red-Papper Phytophtora Blight Disease.)

  • 정희경;김상달
    • 한국미생물·생명공학회지
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    • 제31권3호
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    • pp.235-241
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    • 2003
  • 항진균성 항생물질에 의한 고추역병의 생물방제에 관한 연구를 위하여 지역 경작지에서 고추역병균 P. capsici 생육을 저해는 다기능의 강력한 길항세균을 선발하였다 선발된 균주는 배양학적, 생리학적, 생화학적 실험과 AEI(CHB50) 및 Biolog(Microlog 4.01C) 시스템을 이용하여 동정한 결과 B. megaterium와 97%이상 일치하여 최종적으로 선발된 길항세균을 B. megaterium KL 39로 명명하였다. B.megaterium KL 39의 P. capsic에 대한 길항은 열에 안정하고 n-BuOH에 추출이 되는 저분자성 항생물질과 용매에 추출되지 않은 열에 약한 고분자성의 식물병원균 세포벽 가수분해효소인 cellulase에 의한 것으로 확인되었다. 길항균주 B.megaterium KL 39가 생산하는 항진균성 항생물질은 P.capsici의 포자발아와 균사성장에 큰 영향을 미쳤으며 0.4% fructose, 0.3% yeast extract, 5 mM KCI (pH 8.0)을 포함한 배지에서 3$0^{\circ}C$, 40시간 배양하였을 때 그 생산성이 최대치를 나타내었다. Diaion HP-20 column, silica gel column, Sephadex LH-20 column과 HPLC에 의해 항생물질을 정제하였으며 정제된 항생물질은 TLC plate상에서 Ethanol:Ammonia:Wate.(8:1:1)로 전개한 결과 RF value가 0.32를 나타내었으며, 단일물질로 정제된 항생물질을 KL39로 명명하였다. 선발된 길항미생물 E. megaterium KL 39와 정제된 항생물질 KL39를 가지고 고추를 기주식물로 하여 in vivo pot시험 결과 고추역병균 P.capsic게 의해 유발되는 고추역병에 대한 길항력으로 두 가지 모두 고추역병 방제력이 있음을 식물실험에서 확인할 수 있었다.

Characterization of a Multimodular Endo-β-1,4-Glucanase (Cel9K) from Paenibacillus sp. X4 with a Potential Additive for Saccharification

  • Lee, Jae Pil;Kim, Yoon A;Kim, Sung Kyum;Kim, Hoon
    • Journal of Microbiology and Biotechnology
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    • 제28권4호
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    • pp.588-596
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    • 2018
  • An endo-${\beta}$-1,4-glucanase gene, cel9K, was cloned using the shot-gun method from Paenibacillus sp. X4, which was isolated from alpine soil. The gene was 2,994 bp in length, encoding a protein of 997 amino acid residues with a predicted signal peptide composed of 32 amino acid residues. Cel9K was a multimodular enzyme, and the molecular mass and theoretical pI of the mature Cel9K were 103.5 kDa and 4.81, respectively. Cel9K contains the GGxxDAGD, PHHR, GAxxGG, YxDDI, and EVxxDYN motifs found in most glycoside hydrolase family 9 (GH9) members. The protein sequence showed the highest similarity (88%) with the cellulase of Bacillus sp. BP23 in comparison with the enzymes with reported properties. The enzyme was purified by chromatography using HiTrap Q, CHT-II, and HiTrap Butyl HP. Using SDS-PAGE/activity staining, the molecular mass of Cel9K was estimated to be 93 kDa, which is a truncated form produced by the proteolytic cleavage of its C-terminus. Cel9K was optimally active at pH 5.5 and $50^{\circ}C$ and showed a half-life of 59.2 min at $50^{\circ}C$. The CMCase activity was increased to more than 150% in the presence of 2 mM $Na^+$, $K^+$, and $Ba^{2+}$, but decreased significantly to less than 50% by $Mn^{2+}$ and $Co^{2+}$. The addition of Cel9K to a commercial enzyme set (Celluclast 1.5L + Novozym 188) increased the saccharification of the pretreated reed and rice straw powders by 30.4% and 15.9%, respectively. The results suggest that Cel9K can be used to enhance the enzymatic conversion of lignocellulosic biomass to reducing sugars as an additive.

Production and Characterization of Ethanol- and Protease-Tolerant and Xylooligosaccharides-Producing Endoxylanase from Humicola sp. Ly01

  • Zhou, Junpei;Wu, Qian;Zhang, Rui;Yang, Yuying;Tang, Xianghua;Li, Junjun;Ding, Junmei;Dong, Yanyan;Huang, Zunxi
    • Journal of Microbiology and Biotechnology
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    • 제23권6호
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    • pp.794-801
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    • 2013
  • This paper reports the production and characterization of crude xylanase from the newly isolated Humicola sp. Ly01. The highest (41.8 U/ml) production of the crude xylanase was obtained under the optimized conditions (w/v): 0.5% wheat bran, 0.2% $KH_2PO_4$, and 0.5% peptone; initial pH 7.0; incubation time 72 h; $30^{\circ}C$; and 150 rpm. A considerable amount of the crude xylanase was induced using hulless barley bran or soybean meal as the carbon source, but a small amount of the enzyme was produced when supplementary urea was used as the nitrogen source to wheat bran. The crude xylanase showed apparent optimal cellulase-free xylanase activity at $60^{\circ}C$ and pH 6.0, more than 71.8% of the maximum xylanase activity in 3.0-30.0% (v/v) ethanol and more than 82.3% of the initial xylanase activity after incubation in 3.0-30.0% (v/v) ethanol at $30^{\circ}C$ for 2 h. The crude xylanase was moderately resistant to both acid and neutral protease digestion, and released 7.9 and 10.9 ${\mu}mol/ml$ reducing sugar from xylan in the simulated gastric and intestinal fluids, respectively. The xylooligosaccharides were the main products of the hydrolysis of xylan by the crude xylanase. These properties suggested the potential of the crude enzyme for being applied in the animal feed industry, xylooligosaccharides production, and high-alcohol conditions such as ethanol production and brewing.

STUDIES ON THE TISSUE CULTURE OF PANAX GINSENG

  • Harn C
    • 고려인삼학회:학술대회논문집
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    • 고려인삼학회 1974년도 학술대회지
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    • pp.9-22
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    • 1974
  • Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

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도심공원으로부터 산내성 xylanase를 생산하는 박테리아 분리 및 효소학적 특성 (Isolation and biochemical characterization of acid tolerance xylanase producing Bacteria, Bacillus sp. GJY from city park soil)

  • 장민영;박화랑;이총규;추갑철;조현서;박삼봉;오기철;김봉규
    • Journal of Applied Biological Chemistry
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    • 제60권1호
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    • pp.79-86
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    • 2017
  • 토양 내에 존재하는 미생물은 다양한 종류의 다당류 분해 효소들을 생산함으로써 토양의 비옥도 증진뿐만 아니라 토양내의 생태계를 건전하게 유지하게 위해 중요한 역할을 한다. 본 연구는 경남과학기술대학교에 위치한 쥬라기숲에서 0.4 % carboxymethyl cellulose와 0.01 % trypan blue가 첨가된 LB agar plate를 이용하여 CMCase와 xylanase를 생산하는 박테리아를 분리하였다. 16S rRNA 유전자 염기서열 분석과 API kit 분석을 바탕으로 분리된 박테리아는 Bacillus 종에 속하는 것으로 동정되었으며, Bacillus sp. GJY으로 명명하였다. Bacillus sp. GJY에서 CMCase와 xylanase의 활성을 책임지고 있는 단백질을 알아보기 위하여 Zymogram 분석을 실시하였다. 그 결과 CMCase의 경우 약 28 kDa 크기에 xylanase의 경우 약 25 kDa 크기에 활성밴드가 하나씩 존재하였다. Bacillus sp. GJY의 최적 생장온도는 $37^{\circ}C$이었으며, CMCase와 xylanase의 활성은 배양 후 12시간에 최고에 달하였다. CMCase의 경우 pH 5.0, 온도 $40^{\circ}C$에서 최적의 활성을 보인 반면, xylanase는 pH 4.0, $40^{\circ}C$에서 최적의 활성을 보였다. CMCase와 xylanase 모두 40, $50^{\circ}C$에서는 열 안정성을 보였지만, $60^{\circ}C$ 이상에서는 두 효소의 열 안정성이 급격하게 감소하는 경향을 보였다.

갯지렁이(Perinereis aibuhitensis)에서 분리한 Bacillus spp.의 생리생화학적 특성 분석 (Physiological and Biochemical Characterization of Bacillus spp. from Polychaete, Perinereis aibuhitensis)

  • 신세연;;이상석;강경호;강형일
    • 생명과학회지
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    • 제23권3호
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    • pp.415-425
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    • 2013
  • 본 연구에서는 연안 갯벌에서 유기물 분해능이 매우 우수한 것으로 알려진 갯지렁이(Perinereis aibuhitensis)에 내생하고 있는 균주 중 호기성과 혐기성 조건에서 생장 가능한 5 종류의 Bacillus 균주 CBW3, CBW4, CBW9, CBW14 그리고 EBW10를 선별하여 그 특성을 비교 분석하였다. 16S rRNA 염기서열에 기초하여 동정한 결과, CBW3과 CBW14는 B. nanhaiensis, B. arsenicus 그리고 B. barbaricus와 99.8% 이상의 높은 상동성을, CBW4, CBW9 그리고 EBW10 균주는 B. anthracis, B. algicoa 그리고 B. thuringiensis와 각각 92.7%, 99.8% 그리고 99.8%의 상동성을 보였다. 이들 대부분 균주들의 생장온도 범위는 $4-45^{\circ}C$, 염도는 0-17%, pH는 5-9 범위로 매우 다양하게 나타났다. 모든 균주들이 casein, starch 분해 효소를 가지고 있었으며 특히 균주 EBW10은 시험한 모든 고분자 물질을 분해할 수 있는 protease, amylase, cellulase. lipase 등의 효소 활성을 가지고 있을 가능성이 높음을 제시해주었다. 대상 균주 5 종 모두 alkaline phosphatase 활성을 가지며, CBW3, CBW4 그리고 EBW10은 acid phospatase 활성을 동시에 가지고 있었으며, CBW3, CBW14, EBW10은 esterase (C4) 활성을, CBW3, CBW9, EBW10은 CBW4는 esterase lipase (C8) 활성을 나타냈다. 지방산 분석 결과 CBW3, CBW9, CBW14, EBW10 균주에서는 anteiso $C_{15:0}$이 균주에 따라 차이가 있었지만 약 42.8%에서 55.7%까지 가장 많은 비율을 차지하는 지방산으로 분석되었고, 오직 CBW4에서만 iso $C_{15:0}$이 전체 지방산의 46.9%로 가장 많은 비율로 나타났다.

Lactobacillus bulgaricus와 Lactobacillus plantarum 균주에서 Clostridium thermocellum 유래 endoglucanase의 발현과 발현 유산균의 in vitro 생존 특성 (Expression of Clostridium thermocellum Endoglucanase Gene in Lactobacillus bulgaricus and Lactobacillus plantarum and in vitro Survival Characteristics of the Transformed Lactobacilli)

  • 조재순;강승하;이홍구;이현준;우정희;문양수;양철주;최윤재
    • Journal of Animal Science and Technology
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    • 제45권4호
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    • pp.659-666
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    • 2003
  • 다양한 미생물들로부터 유래한 cellulase 중에서, 특히 장내 단백질 가수분해효소에 안정한 Clostridium thermocellum 균주 유래의 endoglucanase를 선별하였다. 그 후 그 유전자의 자체 프로모터에 의해 발현되는 재조합 Lactobacillus용 발현벡터를 구축하였고, 그 발현벡터를 pSD1이라 명명하였다. 이 발현벡터를 L. bulgaricus와 L. plantarum 균주에 각각 전기천공법을 이용하여 형질전환시키는데 성공하였으며 그 재조합 균주들로부터 endoglucanase 효소역가를 조사한 결과 각각 배지 상층액에서 0.12, 0.144 U/ml로 조사되었다. 한편 이들 균주들의 생균제로 갖추어야할 특성인 내산성, 내담즙성 및 항생제내성 여부를 조사한 결과, 이들 균주들은 모두 pH3과 같은 산성 조건하에서도 안정하였으며, 내담즙성에 있어서는 특히 L. plantarum 균주의 경우 0.3, 1% 의 oxgall에서도 안정하였다. 또한 항생제 내성을 조사한 결과 두 균주 모두 amikacin, gentamicin, kanamycin, colistin에 저항성이 높은 것으로 나타났다.

Saccharification of Foodwastes Using Cellulolytic and Amylolytic Enzymes from Trichoderma harzianum FJ1 and Its Kinetics

  • Kim Kyoung-Cheol;Kim Si-Wouk;Kim Myong-Jun;Kim Seong-Jun
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권1호
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    • pp.52-59
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    • 2005
  • The study was targeted to saccharify foodwastes with the cellulolytic and amylolytic enzymes obtained from culture supernatant of Trichoderma harzianum FJ1 and analyze the kinetics of the saccharification in order to enlarge the utilization in industrial application. T. harzianum FJ1 highly produced various cellulolytic (filter paperase 0.9, carboxymethyl cellulase 22.0, ${\beta}$-glucosidase 1.2, Avicelase 0.4, xylanase 30.8, as U/mL-supernatant) and amylolytic (${alpha}$-amylase 5.6, ${\beta}$-amylase 3.1, glucoamylase 2.6, as U/mL-supernatant) enzymes. The $23{\sim}98\;g/L$ of reducing sugars were obtained under various experimental conditions by changing FPase to between $0.2{\sim}0.6\;U/mL$ and foodwastes between $5{\sim}20\%$ (w/v), with fixed conditions at $50^{\circ}C$, pH 5.0, and 100 rpm for 24 h. As the enzymatic hydrolysis of foodwastes were performed in a heterogeneous solid-liquid reaction system, it was significantly influenced by enzyme and substrate concentrations used, where the pH and temperature were fixed at their experimental optima of 5.0 and $50^{\circ}C$, respectively. An empirical model was employed to simplify the kinetics of the saccharification reaction. The reducing sugars concentration (X, g/L) in the saccharification reaction was expressed by a power curve ($X=K{\cdot}t^n$) for the reaction time (t), where the coefficient, K and n. were related to functions of the enzymes concentrations (E) and foodwastes concentrations (S), as follow: $K=10.894{\cdot}Ln(E{\cdot}S^2)-56.768,\;n=0.0608{\cdot}(E/S)^{-0.2130}$. The kinetic developed to analyze the effective saccharification of foodwastes composed of complex organic compounds could adequately explain the cases under various saccharification conditions. The kinetics results would be available for reducing sugars production processes, with the reducing sugars obtained at a lower cost can be used as carbon and energy sources in various fermentation industries.