• Title/Summary/Keyword: enzyme 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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    • v.19 no.5
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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.

Flavobacterium meningosepticum이 생산하는 Nucleoside Oxidase의 효소학적 특성

  • 최양문;조홍연;양한철
    • Microbiology and Biotechnology Letters
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    • v.24 no.5
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    • pp.579-584
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    • 1996
  • The molecular weight of the purified nucleoside oxidase estimated by gel filtration column chromatography was 480,000 and the enzyme protein was composed of four nonidentical subunits (81,000, 69,000, 32,000 and 16,000). On the basis of the visible absorption spectra and the enzymatic determination of the purified enzyme, the enzyme was supposed as a hemoprotein and also a flavoprotein containing 3 moles of FAD per I mole of enzyme. The isoelectric point of the enzyme was pH 5.1. Addition of metal salts such as 1 mM SnCl$_{2}$ and PbCl$_{2}$ into an enzyme reaction solution inhibited the enzyme activity by 94 and 90%, respectively. The enzyme activity was also lost significantly by hemoenzyme inhibitors such as NaCN and NaN$_{3}$ and flavoenzyme inhibitor, acriflavine and quinacrine. The maximal nucleoside oxidase activity was observed at pH 7.0 and 55$\circ$C. The nucleoside oxidase was relatively stable in the range of pH 5.5-9.0 and below 55$\circ$C.

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A Sensing of Glucose Solution and Diabetic Serum using Polypyrrole Nanotubules Enzyme Electrode Immobilized Glucose Oxidase (포도당 산화효소를 고정화한 Polypyrrole 나노튜뷸 효소전극의 포도당 용액 및 당뇨병 혈청에 대한 감응특성)

  • Kim, Hyun-Cheol;Gu, Hal-Bon
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2001.05a
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    • pp.6-10
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    • 2001
  • We synthesized polypyrrole (PPy) nanotubules by oxidative polymerization of the pyrrole monomer on the pore of a polycarbonate membrane. The electrochemical behavior was investigated using cyclic voltammetry and AC impedance. The redox potential was about -0.5 V vs. Ag/AgCl reference electrode, while the potential was about 0 V for electro-synthesized PPy film. It is considered as the backbone grows according to the pore wall. Therefore, it is possible to be arranged regularly. That leads to improvement in the electron hopping. The AC impedance plot gave a hint of betterment of mass transport. PPy nanotubules have improved in mass transport, or diffusion. That is because the diffusion occurs through a thin pore wall of PPy nanotubules. The kinetic parameter of PPy nanotubules enzyme electrode with glucose solution was evaluated. The formal Michaelis constant and maximum current calculated by computer were about 23.8 mmol $dm^{-3}$ and $440\;{\mu}A$ respectively. Obviously, an affinity for the substrate and current response of the PPy nanotubules enzyme electrode are rather good, comparing with that of PPy film. What is more, the enzyme electrode is sensitive to blood sugar of a diabetic serum despite an obstruction of ascorbic acid, oxygen, some protein and/or hormone.

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Isolation of psychrotrophic microorganism producing soymilk-clotting enzyme from marine fish (생선으로부터 분리한 두유 응고 효소 생산 호냉성 미생물)

  • Kim, Jung-Ho
    • Applied Biological Chemistry
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    • v.36 no.1
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    • pp.45-50
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    • 1993
  • A psychrotrophic microorganism isolated from Alaska pollack (Theragra chalcoramma) produced soymilk-clotting enzyme(s) with relatively low proteolytic activity. The isolate No. 268 was tentatively identified as Pseudomonas sp. Soymilk-clotting activity of the crude enzyme solution was observed at temperatures ranging from 20 to $60^{\circ}C$ and the optimum temperature was $40^{\circ}C$. When the crude enzyme solution was preincubated for 30 minutes, the clotting activity was stable at temperatures up to $30^{\circ}C$ and 75% of the activity was retained at $40^{\circ}C$. The clotting activity was decreased as the pH of soymilk was increased from 5.8 to 7.3.

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Improved Sensitivity of a Glucose Sensor by Encapsulation of Free GOx in Conducting Polymer Micropillar Structure

  • Jung, Shin-Hwan;Lee, Young-Kwan;Son, Yong-Keun
    • Journal of Electrochemical Science and Technology
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    • v.2 no.2
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    • pp.124-129
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    • 2011
  • A simple process of fabricating micropillar structure and its influence upon enhancing electrochemical biosensor response were studied in this work. Conducting polymer PEDOT was used as a base material in formulating a composite with PVA. Micro porous PC membrane filter was used as a template for the micropillar of the composite on ITO electrode. This structure could provide plenty of encapsulating space for enzyme species. After dosing enzyme solution into this space, Nafion film tent was cast over the pillar structure to complete the micropillar cavity structure. In this way, the encapsulation of enzyme could be accomplished without any chemical modification. The amount of enzyme species was easily controllable by varying the concentration of the dosing solution. The more amount of enzyme is stored in the sensor, the higher the electrochemical response is produced. One more reason for the sensitivity improvement comes from the large surface area of the micropillar structure. Application of 0.7 V produced the best current response under the condition of pH 7.4. This biosensor showed linear response to the glucose in 0.1~1 mM range with the average sensitivity of $14.06{\mu}A/mMcm^2$. Detection limit was 0.01 mM based on S/N = 3.

Degradation and Preparation of Blend Films Using Natural Polymers Chitosan and Algin (키토산과 알긴을 이용한 블랜드필름의 제조와 분해)

  • 류정욱;이홍열;오세영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.2
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    • pp.417-422
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    • 1999
  • Algin and chitosan are known as biodegradable natural polymers. PVA is useful for the production of water soluble packaging, paper, textile sizes. PVA/Algin and PVA/chitosan films were prepared by solution blends method in the weight ratio of chitosan, algin for the purpose of useful biodegradable films. Thermal and mechanical properties of blend films such as DSC, impact strength, tensile strength and morphology by SEM were determined. As a result, The ratio of 10.0wt% PVA/chitosan films were similar to PVA at thermal and mechanical properties. PVA/Algin films were found that phase separation was occured as more than 25wt% increasing the blend ratio of algin. PVA/Algin films were observed to be less partially compatibility than 10wt% increasing the blend ratio of algin by DSC, mechanical properties and SEM. Blend films were completely degraded pH 4.0 better than 7.0, 10.0 in the buffer solution. Also, they were rapidly degraded in the enzyme( glucosidase) solution better than pH solution by enzymolysis.

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Study of the Antioxidant and Alcohol-degrading Enzyme Activities of Soybean Sprout Sugar Solutions (콩나물 당 침지액의 항산화 효능 및 알코올 분해 효소 활성 연구)

  • Kim, Kyoung Mi;Jung, Hyun Jung;Sung, Hea Mi;Wee, Ji-Hyang;Kim, Tae Yong;Kim, Ki Myong
    • Korean Journal of Food Science and Technology
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    • v.46 no.5
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    • pp.581-587
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    • 2014
  • The antioxidant and alcohol-degrading enzyme activities of soybean sprout sugar solutions (oligosaccharide and sucrose, $50^{\circ}Bx$) were characterized under different soaking conditions. The ratio of sugar solution to sprout content was 25%, 50%, and 75% (w/w), and the soaking times studied were 1, 3, 6, 12, and 24 h. Higher 1,1-diphenyl-2-pycrylhydrazyl (DPPH) radical-scavenging activity was detected in case of the oligosaccharide solution, compared to the sucrose solution. A similar tendency was observed for alcohol-degrading enzyme activity. When the ratio of sugar solution to sprout content was 50% (w/w), the total phenol and flavonoid contents were found to be higher, compared to those observed at 20% (w/w). However, we did not observe a significant difference between 50% and 75% (w/w). Soaking time did not significantly affect the antioxidant and alcohol-degrading enzyme activities of the solutions. As a result, when oligosaccharide solution was used for soaking soy sprouts at a ratio of >50% (w/w), higher antioxidant and alcoholdegrading enzyme activities were observed.

Enzyme Reactions in Organic Solvents on the Biosurfactant (미생물 계면활성제에 있어 유기용매중의 효소반응)

  • Nam, Ki-Dae;Kim, Sang-Chun;Choi, Jae-Hyo
    • Journal of the Korean Applied Science and Technology
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    • v.10 no.1
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    • pp.9-22
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    • 1993
  • Recent studies on enzyme reactions in organic solvents are revived. The reactions are classified into three categories: heterogeneous, biphasic and homogeneous systems. The following subjects are described and discussed about the heterogeneous system. 1) The maximal expression of enzyme activity in organic solvents in terms of water content, hydration of enzyme, and equilibriun of water between enzyme and substrate solution. 2) Solvent effect on the catalytic power of enzyme. 3) Thermostability and thermoreactivity. 4) Applications of the enzyme reactions to synthetic chemistry.

NMR peak assignment for the elucidation of the solution structure of T4 Endonuclease V

  • Im, Hoo-Kang;Hyungmi Lihm;Yu, Jun-Suk;Lee, Bong-Jin
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.183-183
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    • 1996
  • Bacteriophage T4 endonuclease V initiates the repair of ultraviolet (UV)-induced pyrimidine dimer photoproducts in duplex DNA. The mechanism of DNA strand cleavage involves four sequential stens: linear diffusion along dsDNA, pyrimidine dimer-specific binding,l pyrimidine dimer-DNA glycosylase activity, and Af lyase activity. Although crystal structure is known for this enzyme, solution structure has not been yet known. In order to elucidate the solution structure of this enzyme NMR spectroscopy was used. As a basis for the NMR peak assignment of the protein, HSQC spectrum was obtained on the uniformly $\^$15/N-labeled T4 endonuclease V. Each amide peak of the spectrum were classified according to amino acid spin systems by interpreting the spectrum of $\^$15/N amino acid-specific labeled T4 endonuclease V. The assignment was mainly obtained from three-dimensional NMR spectra such as 3D NOESY-HMQC, 3D TOCSY-HMQC. These experiments were carried out will uniformly $\^$15/N-labeled sample. In order to assign tile resonance of backbon atom, triple-resonance theree-dimensional NMR experiments were also performed using double labeled($\^$15/N$\^$13/C) sample. 3D HNCA, HN(CO)CA, HNCO, HN(CA)HA spectra were recorded for this purpose. The results of assignments were used to interpret the interaction of this enzyme with DNA. HSQC spectrum was obtained for T4 endonuclease V with specific $\^$15/N-labeled amino acids that have been known for important residue in catalysis. By comparing the spectrum of enzyme*DNA complex with that of the enzyme, we could confirm the important role of some residues of Thr, Arg, Tyr in activity. The results of assignments were also used to predict the secondary structure by chemical shift index (CSI).

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Spectro-electrochemical Analyses of Immobilization of Glucose Oxidase (Glucose Oxidase 고정화에 대한 전기화학적/광학적 분석)

  • Kim, Hyun-Cheol;Cho, Young-Jai;Gu, Hal-Bon;SaGon, Geon
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2000.05b
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    • pp.316-319
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    • 2000
  • In the case of immobilizing of glucose oxidase into polypyrrole (PPy) using electrosynthesis, the glucose oxidase (GOx) forms a coordinate bond with the polymer's backbone. However, because of intrinsic insulation and net-chain of the enzyme, the charge transfer and mass transport are obstructed during the film growth. Therefore, the film growth is dull. We synthesized the enzyme electrode by electropolymerization added some organic solvent, A formative seeds of film growth is delayed by adding the solvent. The delay is induced by radical transfer between the solvent and pyrrole monomer. In the case of adding ethanol, the radical transfer shares the contribution of dopant between electrolyte anion and GOx polyanion. This may lead to increase amount of immobilized the enzyme in ppy. However, adding tetrahydrofuran (THF), the radical transfer is more brisk, resulting in short chained polymer. Therefore, the doping level is lowered and then amount of immobilized of enzyme is decreased. For the UV absorption spectra of synthetic solution before synthesis and after, in the case of ethanol added, the optical density was slightly decreased for the GOx peaks. It suggests amount of GOx in the solution was decreased and amount of GOx in the film was increased. We established qualitatively that amount of immobilization can be improved by adding a little ethanol in the synthetic solution. It is due to radical transfer reaction. The radical transfer shares the contribution of dopant between small and fast electrolyte anion and big and slow GOx polyanion.

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