• 제목/요약/키워드: enzyme adsorption

검색결과 96건 처리시간 0.025초

미생물 페니실린 아미다제에 관한 연구 (제 6 보) 흡착효소의 아크릴아마이드젤 포괄방법에 의한 Bacillus megaterium의 변이주가 생산하는 페니실린 아미다제의 고정화에 관한 연구 (Studies on Microbial Penicillin Amidase (Part 6) Immobilization of Penicillin Amidase from Bacillus megaterium by Adsorption and Acrylamide Gel Entrappment)

  • Seong, Baik-Lin;Son, Hyeung-Jin;Mheen, Tae-Ick;Moon H. Han
    • 한국미생물·생명공학회지
    • /
    • 제9권4호
    • /
    • pp.197-205
    • /
    • 1981
  • Bacillus megaterium의 발효액으로 부터 페니실린 아미다제를 셀라이트에 흡착시켜 분리한 후 이 흡착효소를 아크릴아마이드에 포괄시켜 고정화하였다. 관형식 반응조에서의 이 고정화효소의 안정도는 포괄시키지 않은 흡착효소에 비해 크게 증가하였으며 최적 반응 pH는 8.7, 그리고 최적 안정도는 7.5~8.0이었고 최적온도는 5$0^{\circ}C$ 였다. Km과 6-APA, 페닐초산에 의한 저해상수는 각각 4.55mM, 36.5mM, 그리고 10.5mM이었다. Effectiveness factor값은 0.95로 내부확산 효과는 무시할 수 있었다. pH8.0의 조건에서 관형식 반응조 내에서의 효소역가의 반감기는 4$0^{\circ}C$에서 6.8일 그리고 3$0^{\circ}C$ 에서는 47일로 포괄하지 않은 흡착효소에 비해 안정도가 각각 6.8배와 12배로 증가하였다. 이 고정화 효소에 의한 회분식 및 연녹식반응조에서의 6-APA의 생산성을 논의하였다. 실험결과로 미루어 보아 특히 흡착효소를 고정화효소로 사용하는 경우에 포괄방법을 이용함으로써 효소반응조의 안정도를 크게 증가시킬 수 있음을 시사하였다.

  • PDF

$\beta$-glucosidase의 고정화와 효소 반응특성 (Immobilization of $\beta$-glucosidase and properties of Immobilized Enzyme)

  • 정의준;이상호이용현
    • KSBB Journal
    • /
    • 제5권2호
    • /
    • pp.141-149
    • /
    • 1990
  • Aspergillus niger유래의 $\beta$-glucosidase를 (1)glutarald-ehyde를 가교제로 한 chitin과 chitosan담체에 covalent linkage (2)Amberite IRA93과 DEAE-cellulose담체에 succinylation시킨 후 흡착, 그리고 (3)alginarte, polyacry-lamide를 각종 가교제를 이용 entrapment등의 방법으로 고정화 하였다. Glutaraldehyde를 가교제로써 chitosan을 활성화시킨후 $\beta$-glucosidase를 고정화 시켰을때 효소활성 회수율이 31.5%로 가장 높았고, 또한 column형 반응기에서의 15일 경과 후 효소활성 유지도도 69%로서 가장 우수하였다. 또한 succinylation시킨 효소를 Amberite IRA93 담체에 흡착시켰을 때 효소활성 회수율은 24.7%였고 효소 활성유지도는 62%였다. 반면에 entra-pment 방법에 의한 $\beta$-glucosidase의 고정화는 효소의 계속적인 용출로 $\beta$-glucosidase의 고정화에는 적합하지 않았다. Chitosan담체에서의 고정화 최적조건을 조사한 결과 가교제인 glutaraldehyde의 최적농도는 0.4%였고, glutaraldehyde의 최적 반응 pH는 4.8이었다. 또한 column형 반응기를 이용하여 cellobiose로부터 glucose로의 전환율을 조사하여 고정화 효소의 효용성을 검토하였다.

  • PDF

재조합 Vibrio parahaemolyticus 콜라겐분해효소의 분리 및 특성 분석 (Isolation and Characterization of Recombinant Vibrio parahaemolyticus Collagenase)

  • 차재호;김수광;전인준;이재원
    • 생명과학회지
    • /
    • 제13권3호
    • /
    • pp.308-313
    • /
    • 2003
  • 식중독 병원균인 장염비브리오균 (V. parahaemolyticus)의 세포외 분비 효소 중 콜라겐분해효소를 발현벡터인 pET-29b에 클로닝시키고 대장균에서 발현시킨 다음, 부분정제하여 그 특성을 조사하였다. V. parahaemolyticus collagenase는 $(NH_4)_2So_4$침전, affinity adsorption, 그리고 Sephacryl S-100 gel filtration 과정을 통하여 부분정제 되었다. 이 collagenase는 73%의 회수율과 43.7의 정제도를 나타내었으며, 전기영동시 분자량은 약 35 kDa로 나타났다. 이 효소의 최적 pH 및 온도는 6~12와 $35^{\circ}C$이었고, 온도안정성 조사에서 $55^{\circ}C$까지는 90% 잔존 찬성을 유지하였으나 $60^{\circ}C$이상에서는 급격하게 효소활성이 실활되었다. 기질특이성조사에서 type I collagen과 콜라겐분해효소의 합성기질로 알려진 Z-GPGGPA에서 gelatin과 casein에 비해 높은 활성을 보이는 것으로 보아 이 효소가 진정한 콜라겐분해효소라는 것을 알 수 있었다.

2,4-Dichlorophenol Enzymatic Removal and Its Kinetic Study Using Horseradish Peroxidase Crosslinked to Nano Spray-Dried Poly(Lactic-Co-Glycolic Acid) Fine Particles

  • Dahili, Laura Amina;Nagy, Endre;Feczko, Tivadar
    • Journal of Microbiology and Biotechnology
    • /
    • 제27권4호
    • /
    • pp.768-774
    • /
    • 2017
  • Horseradish peroxidase (HRP) catalyzes the oxidation of aromatic compounds by hydrogen peroxide via insoluble polymer formation, which can be precipitated from the wastewater. For HRP immobilization, poly(lactic-co-glycolic acid) (PLGA) fine carrier supports were produced by using the Nano Spray Dryer B-90. Immobilized HRP was used to remove the persistent 2,4-dichlorophenol from model wastewater. Both extracted (9-16 U/g) and purified HRP (11-25 U/g) retained their activity to a high extent after crosslinking to the PLGA particles. The immobilized enzyme activity was substantially higher in both the acidic and the alkaline pH regions compared with the free enzyme. Optimally, 98% of the 2,4-dichlorophenol could be eliminated using immobilized HRP due to catalytic removal and partly to adsorption on the carrier supports. Immobilized enzyme kinetics for 2,4-dichlorophenol elimination was studied for the first time, and it could be concluded that competitive product inhibition took place.

Influence of Varying Degree of Salinity-Sodicity Stress on Enzyme Activities and Bacterial Populations of Coastal Soils of Yellow Sea, South Korea

  • Siddikee, Md. Ashaduzzaman;Tipayno, Sherlyn C.;Kim, Ki-Yoon;Chung, Jong-Bae;Sa, Tong-Min
    • Journal of Microbiology and Biotechnology
    • /
    • 제21권4호
    • /
    • pp.341-346
    • /
    • 2011
  • To study the effects of salinity-sodicity on bacterial population and enzyme activities, soil samples were collected from the Bay of Yellow Sea, Incheon, South Korea. In the soils nearest to the coastline, pH, electrical conductivity ($EC_e$), sodium adsorption ratio (SAR), and exchangeable sodium percentage (ESP) were greater than the criteria of saline-sodic soil, and soils collected from sites 1.5-2 km away from the coastline were not substantially affected by the intrusion and spray of seawater. Halotolerant bacteria showed similar trends, whereas non-tolerant bacteria and enzymatic activities had opposite trends. Significant positive correlations were found between EC, exchangeable $Na^+$, and pH with SAR and ESP. In contrast, $EC_e$, SAR, ESP, and exchangeable $Na^+$ exhibited significant negative correlations with bacterial populations and enzyme activities. The results of this study indicate that the soil chemical variables related with salinity-sodicity are significantly related with the sampling distance from the coastline and are the key stress factors, which greatly affect microbial and biochemical properties.

밀기울배지를 이용한 Bacillus macerans의 Cyclodextrin Glucanotransferase 생산과 효소특성 (Purification and Enzymatic Properties of Cyclodextrin Glucanotransferase from Bacillus macerans Cultivated in Wheat-bran Medium)

  • 선우양일;안태진
    • KSBB Journal
    • /
    • 제9권5호
    • /
    • pp.499-505
    • /
    • 1994
  • Bacillus macerans cyclodextrin glucanotrans­f ferase(CGTase)를 전분흡착법과 DEAE--cellulose 칼럼 크로마토그래피로 정제하였다. 효소의 분자량 은 67,000이였고 monomer였다. 정제된 효소는 전 분을${\alpha}$-, ${\beta}$-, ${\gamma}$-CD 로 전환시켰으며, CD생성비율은 각각 1 : 1.68: 0.32였다. ${\alpha}$-CD와 D-glucose의 coupling반응 초기에는 maltohexose가 주로 생성되었고, 그 후에 다른 oligosaccharide들이 생성되었다 .. a­C CD의 가수분해반응 초기에는 주로 maltotetrose가 생성되였고 그 이후에는 소량의 다른 이Igosa­C ccharide들이 생성되었다. 정제된 효소의 좋은 기질인 maltotriose 로부터 maltosyl 이나 D-glucopy ranosyl group이 전이될 수 있는데, 본 연구에서는 D정lucosyl transfer가 우세하였다.

  • PDF

Separation of Nattokinase from Bacillus subtilis Fermentation Broth by Expanded Bed Adsorption with Mixed-mode Adsorbent

  • Lu Miao-Hua;Lin Dong-Qiang;Wu Yuan-Chun;Yun Jun-Xian;Mei Le-He;Yao Shan-Jing
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • 제10권2호
    • /
    • pp.128-135
    • /
    • 2005
  • Mixed-mode hydrophobic/ionic matrices exhibit a salt-tolerant property for adsorbing target protein from high-ionic strength feedstock, which allows the application of undiluted feedstock via an expanded bed process. In the present work, a new type of mixed-mode adsorbent designed for expanded bed adsorption, Fastline $PRO^{\circledR}$, was challenged for the capture of nattokinase from the high ionic fermentation broth of Bacillus subtilis. Two important factors, pH and ion concentration, were investigated with regard to the performance of nattokinase ad-sorption. Under initial fermentation broth conditions (pH 6.6 and conductivity of 10 mS/cm) the adsorption capacity of nattokinase with Fastline PRO was high, with a maximum capacity of 5,350 U/mL adsorbent. The elution behaviors were investigated using packed bed adsorption experiments, which demonstrated that the effective desorption of nattokinase could be achieved by effecting a pH of 9.5. The biomass pulse response experiments were carried out in order to evaluate the biomass/adsorbent interactions between Bacillus subtilis cells and Fastline PRO, and to demonstrate a stable expanded bed in the feedstock containing Bacillus subtilis cells. Finally, an EBA process, utilizing mixed-mode Fastline PRO adsorbent, was optimized to capture nattokinase directly from the fermentation broth. The purification factor reached 12.3, thereby demonstrating the advantages of the mixed-mode EBA in enzyme separation.

Screening, Gene Cloning, and Characterizations of an Acid-Stable α-Amylase

  • Liu, Xinyu;Jia, Wei;An, Yi;Cheng, Kun;Wang, Mingdao;Yang, Sen;Chen, Hongge
    • Journal of Microbiology and Biotechnology
    • /
    • 제25권6호
    • /
    • pp.828-836
    • /
    • 2015
  • Based on its α-amylase activity at pH 5.0 and optimal pH of the crude enzyme, a strain (named B-5) with acid α-amylase production was screened. The B-5 strain was identified as Bacillus amyloliquefaciens through morphological, physiological, and biochemical characteristics analysis, as well as 16S rDNA phylogenetic analysis. Its α-amylase gene of GenBank Accession No. GU318401 was cloned and expressed in Escherichia coli. The purified recombinant α-amylase AMY-Ba showed the optimal pH of 5.0, and was stable at a pH range of 4.0-6.0. When hydrolyzing soluble starch, amylose, and amylopectin, AMY-Ba released glucose and maltose as major end products. The α-amylase AMY-Ba in this work was different from the well-investigated J01542-type α-amylase which also came from B. amyloliquefaciens. AMY-Ba exhibited notable adsorption and hydrolysis ability towards various raw starches. Structure analysis of AMY-Ba suggested the presence of a new starch-binding domain at its C-terminal region.

Fabrication of Sericin into Micro- and Macro Size Materials and its Application

  • Yang, Sejun;Kang, Yijin;Cho, Yejin;Shin, Bongseob;Lee, Ki Hoon
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • 제44권2호
    • /
    • pp.29-36
    • /
    • 2022
  • Over the past decades, silk sericin (SS) received increasing attention in the academic and industrial fields. In nature, SS acts as a glue that holds the two strands of silk fibrils together. However, recent works suggest that SS might have a more diverse role during the silk spinning process, such as stabilizing the SF in the silk gland. On the other hand, the sericulture industry has been trying to find novel applications for SS discarded from the silk fabric manufacturing process. Recovery and refining of SS would be the first step of the recycling of SS. Using a proper solvent SS could be shaped into various forms, such as spherical beads, microparticles, fibers, and films. Moreover, the applicability of these SS materials has been investigated in various fields such as cosmetics, templates for nanomaterials, drug delivery, heavy metal adsorption, and enzyme immobilization.

Application of Polymer Brush to Enzyme-Multilayered Porous Hollow-Fiber Membrane

  • Kawakita Hidetaka;Uezu Kazuya;Tsuneda Satoshi;Saito Kyoichi;Tamada Masao;Sugo Takanobu
    • 한국막학회:학술대회논문집
    • /
    • 한국막학회 2004년도 추계 총회 및 학술발표회
    • /
    • pp.142-145
    • /
    • 2004
  • Anion-exchange porous hollow-fiber membranes with a thickness of about 1.2 mm and a pore size of about $0.30{\mu}m$ were used as a supporting matrix to immobilize cycloisomaltooligosaccharide glucanotransferase (CITase). CITase was immobilized to the membrane via anion-exchange adsorption and by subsequent enzymatic cross-linking with transglutaminase, the amount of which ranged from 3 to 110 mg per g of the membrane. The degree of enzyme multilayer binding was equivalent to 0.3 to 9.8. Dextran, as the substrate, was converted into seven- to nine-glucose-membered cycloisomaltooligosaccharides (CI-7, -8, and -9) at a maxi mum yield of $28\%$ in weight at a space velocity of 10 per hour during the permeation of $2.0(w/w)\%$ dextran solution across the CITase-immobilized porous hollow-fiber membrane.

  • PDF