• 제목/요약/키워드: Enzyme production

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Production and Characterization of Multi-Polysaccharide Degrading Enzymes from Aspergillus aculeatus BCC199 for Saccharification of Agricultural Residues

  • Suwannarangsee, Surisa;Arnthong, Jantima;Eurwilaichitr, Lily;Champreda, Verawat
    • Journal of Microbiology and Biotechnology
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    • 제24권10호
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    • pp.1427-1437
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    • 2014
  • Enzymatic hydrolysis of lignocellulosic biomass into fermentable sugars is a key step in the conversion of agricultural by-products to biofuels and value-added chemicals. Utilization of a robust microorganism for on-site production of biomass-degrading enzymes has gained increasing interest as an economical approach for supplying enzymes to biorefinery processes. In this study, production of multi-polysaccharide-degrading enzymes from Aspergillus aculeatus BCC199 by solid-state fermentation was improved through the statistical design approach. Among the operational parameters, yeast extract and soybean meal as well as the nonionic surfactant Tween 20 and initial pH were found as key parameters for maximizing production of cellulolytic and hemicellulolytic enzymes. Under the optimized condition, the production of FPase, endoglucanase, ${\beta}$-glucosidase, xylanase, and ${\beta}$-xylosidase was achieved at 23, 663, 88, 1,633, and 90 units/g of dry substrate, respectively. The multi-enzyme extract was highly efficient in the saccharification of alkaline-pretreated rice straw, corn cob, and corn stover. In comparison with commercial cellulase preparations, the BCC199 enzyme mixture was able to produce remarkable yields of glucose and xylose, as it contained higher relative activities of ${\beta}$-glucosidase and core hemicellulases (xylanase and ${\beta}$-xylosidase). These results suggested that the crude enzyme extract from A. aculeatus BCC199 possesses balanced cellulolytic and xylanolytic activities required for the efficient saccharification of lignocellulosic biomass feedstocks, and supplementation of external ${\beta}$-glucosidase or xylanase was dispensable. The work thus demonstrates the high potential of A. aculeatus BCC199 as a promising producer of lignocellulose-degrading enzymes for the biomass conversion industry.

수송용 바이오 부탄올 생산을 위한 미강발효의 최적화 (Fermentation of rice bran and defatted rice bran for butanol production using Clostridium beijerinckii NCIMB 8052)

  • 이지은;서은종;박기문;진용수
    • 한국신재생에너지학회:학술대회논문집
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    • 한국신재생에너지학회 2008년도 춘계학술대회 논문집
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    • pp.235-238
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    • 2008
  • We examined butanol fermentation by Clostridium beijerinckii NCIMB 8052 using various hydrolyzates obtained from rice bran which is one of the most abundant agricultural by-products in Korea and Japan. In order to increase the amount of fermentable sugars in the hydrolyzates of rice bran, various hydrolysis procedures were applied. Total eight different hydrolyzates were prepared using rice bran (RB) and defatted rice bran (DRB) with enzyme or acid treatment and both. Each hydrolyzate was evaluated in terms of total sugar concentration and butanol production after fermentation by C. beijerinckii NCIMB 8052. Acid treatment yielded more sugar than enzyme treatment and combined treatment with enzyme and acid yielded even more sugars as compared to single treatment with enzyme or acid. As a result, the highest sugar concentration (33 g/L) was observed from the hydrolyzate from DRB (100 g/L) with combined treatment using enzyme and acid. Prior to perform fermentation of the hydrolyzates, we examined the effect of P2 solution containing yeast extract, buffer, minerals, and vitamins on production of butanol during the fermentation. Fermentation of the hydrolyzates with or without additionof P2 was performed using C. beijerinckii NCIMB 8052 in a 1 L anaerobic bioreactor. Although the hydrolyzates RB were able to support growth and butanol production, addition of P2 solution into the hydrolyzates significantly improved cell growth and butanol production. Highest butanol production (12.24 g/L) was observed from the hydrolyzate of DRB with acid and enzyme treatment after supplementation of P2 solution.

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Fermentation of Rice Bran and Defatted Rice Bran for Butanol Production Using Clostridium beijerinckii NCIMB 8052

  • Lee, Ji-Eun;Seo, Eun-Jong;Kweon, Dae-Hyuk;Park, Ki-Moon;Jin, Yong-Su
    • Journal of Microbiology and Biotechnology
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    • 제19권5호
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    • pp.482-490
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    • 2009
  • We examined butanol fermentation by Clostridium beijerinckii NCIMB 8052 using various hydrolyzates obtained from rice bran, which is one of the most abundant agricultural by-products in Korea and Japan. In order to increase the amount of fermentable sugars in the hydrolyzates of rice bran, various hydrolysis procedures were applied. Eight different hydrolyzates were prepared using rice bran (RB) and defatted rice bran (DRB) with enzyme or acid treatment or both. Each hydrolyzate was evaluated in terms of total sugar concentration and butanol production after fermentation by C. beijerinckii NCIMB 8052. Acid treatment yielded more sugar than enzyme treatment, and combined treatment with enzyme and acid yielded even more sugars as compared with single treatment with enzyme or acid. As a result, the highest sugar concentration (33 g/l) was observed from the hydrolyzate from DRB (100 g/l) with combined treatment using enzyme and acid. Prior to fermentation of the hydrolyzates, we examined the effect of P2 solution containing yeast extract, buffer, minerals, and vitamins on production of butanol during the fermentation. Fermentation of the hydrolyzates with or without addition of P2 was performed using C. beijerinckii NCIMB 8052 in a 1-1 anaerobic bioreactor. Although the RB hydrolyzates were able to support growth and butanol production, addition of P2 solution into the hydrolyzates significantly improved cell growth and butanol production. The highest butanol production (12.24 g/l) was observed from the hydrolyzate of DRB with acid and enzyme treatment after supplementation of P2 solution.

Streptomyces sp.가 생산하는 Penicillinase 에 관한 연구 (제1보) Streptomyces sp. YS-40에 의한 Penicillinase의 생산조건 (Studies on the Penicillinase Produced by a Streptomyces sp. (Part I). Optimal Conditions for the Penicillinase Production by Streptomyces sp. YS-40.)

  • 도재호;김상달;이동의
    • 한국미생물·생명공학회지
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    • 제10권3호
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    • pp.177-184
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    • 1982
  • 토양에서 분리, 선별한 Streptomyces 속의 한 균주가 생산하는 Penicillinase의 최적생산조건을 조사한 결과를 요약하면 다음과 같다. 탄소원으로서는 glucose, 실소원으로는 L-as-paragine이 가장 우수했으며 금속감중에서 Mn C $l_2$를 첨가했을때 효소생성을 37% 증가시켰다. Amino 산중에서 L-leucine이 본 효소생산을 약간 증가시켰으며 L-histidine L-methionine은 크게 저하시켰다. Riboflavine, i-inositol, hesperidine, niacinamide, biotin, DL-$\alpha$- lipoic acid, folic acid가 효소생산을 증가시켰으며 항생물질중에서 cephradine, cephalexin, ampicillin, cloxacillin의 첨가가 효소생성을 크게 증가시켰다. 본 효소생산의 최고 pH는 7.0이며 최적온도는 28$^{\circ}C$였다. 또 500$m\ell$ Erlenmeyer flask에서 175$m\ell$의 배지를 가하여 3 일간 배양했을 때 효소생성이 최고에 도달했다.

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Continuous Production of Fructooligosaccharides Using Fructosyltransferase Immobilized on Ion Exchange Resin

  • Yun, Jong-Won;Song, Seung-Koo
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제1권1호
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    • pp.18-21
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    • 1996
  • A continuous production of fructooligosaccharides from sucrose was investigated by fructosyltransferase immobilized on a high porous resin, Diaion HPA25. The optimum pH(5.5) and temperature(55$^{\circ}C$) of the enzyme for activity was unaltered by immobilization, and the immobilized enzyme became less sensitive to the pH change. The optimal operation conditions of the immobilized enzyme column for maximizing the productivity were as follows: 600g/L of sucrose feed concentration, flow rate of superficial space velocity 2.7h-1. When the enzyme column was run at 50$^{\circ}C$, about 8% loss of the initial activity of immobilized enzyme was observed after 30 days of continuous operation, during which high productivity of 1174g/L$.$h was achieved. The kinds of products obtained using the immobilized enzyme were almost the same as those using soluble enzymes or free cells.

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Partial Purification and Characterization of Thermostable Alkaline $\beta$-Mannanase from Bacillus sp. JB-99 Suitable for Pulp Bleaching

  • VIRUPAKSHI S.;BABU K. GlREESH;NAIK GAJANAN R.
    • Journal of Microbiology and Biotechnology
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    • 제15권4호
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    • pp.689-693
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    • 2005
  • Bacillus sp. JB-99, when grown in a chemically defined medium containing lactose as a carbon source, yielded 3,860 U/ml extracellular $\beta$-mannanase, which was high compared to other examined carbon sources. Among the nitrogen sources, yeast extract enhanced the enzyme activity. The enzyme production was growth-associated. The enzyme was optimally active at $65^{\circ}C$, pH 10, and had a half-life of 190 min at $65^{\circ}C$. N-Bromosuccinamide and $AgNO_3,\;CuSO_4$, and $HgCl_2$ strongly inhibited the enzyme, whereas $Ca^{2+}$ stimulated the enzyme activity. The $\alpha$-galactosidase enzyme production was not found in any of the enzyme assays.

세균에 의한 생전분 분해효소의 생성 및 특성 (Production and Characterization of Raw Starch Hydrolyzing Enzyme from Bacteria)

  • 박인식;남인;고선옥;김기남;서경순
    • 한국미생물·생명공학회지
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    • 제18권3호
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    • pp.244-250
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    • 1990
  • 토양으로부터 생전분을 분해하는 세균을 분리하였으며, Bacillus sp.로 동정하였다. 분리한 세균으로부터 생전분 분해효소의 생성을 위한 최적조건을 검토하였다. 세균은 탄소원으로 wheat 및 soluble starch를, 질소원으로 polypeptone을 사용했을 때 최대 효소생성을 얻을 수 있었다. 그리고 탄소원을 0.5, 질소원을 0.5 수준으로 사용했을 때 효소 생성을 극대화 하였다. 그리고 배지의 초기 pH를 6.5, 배양온도를 $35^{\circ}C$로 유지했을 때 효소생성이 높았다. Seepharose CL-6B 젤 여과 및 DEAE-Sephacel 이온교환수지를 사용하여 부분정제한 효소활성의 최적조건은 pH6.5, 온도 $70^{\circ}C$ 였다.

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새우젓에서 혈전용해효소 생산균주의 분리, 동정 및 효소생산 배지의 최적화 (Isolation and Identification of Fibrinolytic Enzyme Producing Strain from Shrimp Jeot-Gal, a Tiny Salted Shrimps, and Medium Optimization for Enzyme Production)

  • 장순애;김명희;이명선;이명자;지옥화;오태광;손천배
    • 한국식품과학회지
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    • 제31권6호
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    • pp.1648-1653
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    • 1999
  • 새우젓으로부터 혈전용해효소를 생산하는 균주를 분리하여 Bacillus sp.로 동정하였다. Bacillus sp. S19는 screening 배지에서는 18 U/mL의 효소를 생산하였다. 그러나 2% soluble starch, 2% skim milk, 3% NaCl(pH 7.5)의 최적화된 배지로 $37^{\circ}C$에서 72시간 동안 배양하였을 경우 효소활성이 약 3배 증가된 62 U/mL의 효소 생산을 결과하였다.

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도축 폐혈액 단백질로부터의 Angiotensin I Converting Enzyme 저해 펩타이드의 생산 (Production of Angiotensin I Converting Enzyme Inhibitory Peptides from Bovine Blood Plasma Proteins)

  • 현창기;신현길
    • KSBB Journal
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    • 제14권5호
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    • pp.600-605
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    • 1999
  • 본 연구는 도축 폐기물인 가축혈액을 이용하여 항고혈압 기능성 식품소재로서의 angiotensin I converting enzyme 저해 펩타이드 분획을 생산하기 위한 조건과 가능성을 조사하기 위하여 수행되었다. 산업적으로 이용 가능한 단백분해효소 중 Alcalase가 혈장 원액 및 그로부터 분리된 albumin에 대하여 가장 높은 활성의 가수분해물을 생성하였다. 특히 albumin의 Alcalase 가수분해물과 이를 gel chromatography를 통새 분획하여 얻은 고활성 분획의 $IC_50$값은 각각 0.5 및 0.02 mg/mL로서 지금까지 보고된 식품단백질 유래 펩타이드 혼합물들과 비교할 때 활성이 매우 높은 것에 속함을 알 수 있었다. 또한 이 고활성 펩타이드 분획은 혈장 원액으로부터 단순한 한외여과만을 거쳐도 얻을 수 있음을 확인함으로써 산업적 실용화 가능성이 높은 공정임을 알게 되었다.

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내열성 Amylase의 생산에 관한 연구 (제1보) 최적배양조건과 효소의 정제 (Studies on the Production of Thermostable Amylase. Part 1. Optimal Culture Conditions and Purification of Enzyme.)

  • 오두환;이강표;변유량;유주현
    • 한국미생물·생명공학회지
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    • 제9권2호
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    • pp.91-97
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    • 1981
  • 고온균중에서 amylase의 생산능이 우수한 Bacillus sp. Y-127을 토양에서 분리, 선정하고 다음과 같은 실험결과를 얻었다. 1. Amylase 생산의 최적배양조건은 nutrient broth 0.8% (w/v), soluble starch 2 % (w/v), urea 0.2 % (w/v, N기준), MgSO$_4$.7$H_2O$ 0.02 % (w/v, Mg기준), $K_2$HPO$_4$ 0.02 % (w/v, P기준), yeast extract 0.2 % (w/v), 6$0^{\circ}C$, pH7.0이었다. 2. Amylase를 (NH$_4$)$_2$SO$_4$침전, 투석, Sephadex G-150 column chromatography 및 Sephadex G-150 column rechromatography를 통해 정제한 결과 123배의 비활성 증가를 볼 수 있었다. 3. Amylase의 pH 안정성은 pH 4.0에서 7.0 사이였으며, 온도에 따른 효소의 불활성도는 온도가 증가함에 따라 증대되었다.

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