• Title/Summary/Keyword: enzyme solution

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The Extracellular Enzyme Activities in Culture Broth of Sparassis crispa. (꽃송이버섯(Sparassis crispa)의 세포외 효소활성)

  • Kim Ji-Young;Lim Chang-Soo;Kim Jae-Yong;Han Yeong-Hwan
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.230-231
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    • 2004
  • The mycelia of Sparassis crispa DSMZ 5201 were cultivated at $24^{\circ}C$ for 15 days in yeast-malt extract-glucose broth (pH 4.0) and the filtrate was used as crude enzyme solution to determined the extracellular enzyme activity. The specific activity of $\alpha$-amylase was 44.27 unit/protein. The specific activities of protease, CMCase, $\beta$-glucosidase, chitinase, exo-$\beta$-l,4-glucanase were relatively high. However, a very little activity of xylanase was found.

Simultaneous Dual-Enzyme Immunoassays in a Solid Phase

  • 백세환;박순재
    • Bulletin of the Korean Chemical Society
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    • v.18 no.1
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    • pp.44-50
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    • 1997
  • A method of dual-signal generation from two different enzymes was developed and utilized to simultaneously perform dual immunoassays in a single microwell. Two enzymes selected as tracers were horseradish peroxidase (HRP) and β-galactosidase (GAL). 3, 3', 5, 5'-Tetramethylbenzidine (TMB) and chlorophenolred-β-galactopyranoside (CPRG) as chromogenic substrates for the respective enzyme were used. Although the two enzymes showed their maximum activities at distinct pH conditions (pH 5.1 for HRP and 7.5 for GAL), the enzyme reactions were able to be concurrently carried out at pH 5.75 in a dual-substrate solution without signal loss. This performance was achieved by increasing TMB concentration two-fold, introducing potassium salt as activator of GAL reaction, and extending total reaction time 50%. The signal generation method was then used for dual-enzyme immunoassays to detect antibodies with co-immobilized Hepatitis C virus antigens (core and NS5) and a Hepatitis B virus antigen (PreS(2)) in a microwell. Dose-response curves of the assays revealed cooperativity between different antigen-antibody complex formation, which suggested that dual immunoassays can only be used for qualitative screening tests unless the antigens immobilized were spatially separated.

Potentiometric Homogeneous Enzyme-Linked Binding Assays for Riboflavin and Riboflavin Binding Protein

  • 김진목;김혜진;김미정;이동주;한상현;차근식
    • Bulletin of the Korean Chemical Society
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    • v.17 no.11
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    • pp.1018-1022
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    • 1996
  • Adenosine deaminase (ADA) has been utilized as the label in devising a potentiometric homogeneous assay for riboflavin and riboflavin binding protein (RBP). The proposed homogeneous assay method employs an ADA-biotin conjugate as the signal generator and an avidin-riboflavin conjugate as the signal modulator in the solution phase. The catalytic activity of the ADA-biotin conjugate is inhibited in the presence of an excess amount of the avidin-riboflavin conjugate, and the observed inhibition is reversed in an amount proportional to the concentration of RBP added. When the analyte riboflavin is added to this mixture of ADA-biotin, avidin-riboflavin and RBP, the activity of the enzyme conjugate is re-inhibited in an amount proportional to the concentration of riboflavin. Since the enzyme label used in this system is ADA, an ammonia-producing enzyme, a potentiometric rather than photometric detection scheme is used to monitor the enzymatic activity in the assay.

Catalytic Properties of Phospholipase D using Phosphatidic Acid as an Activator

  • Eun-hie Koh;Myung-Un Chol;Kwanyoung Jung
    • Bulletin of the Korean Chemical Society
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    • v.10 no.6
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    • pp.595-599
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    • 1989
  • The effects of phosphatidic acid(PA) on the activity of phospholipase D were examined in detail. The enzyme activity was examined in the liposome system containing phosphatidylcholine and PA, which was suspended in a desired buffer solution by ultrasonication. The substrate of large unilamella vesicle (LUV) state by ultrasonication was more effective on the enzyme activity than that of multilamella vesicle(MLV) by water-bath type sonication. The most effective molar ratio of PC-PA liposome for enzyme activity was found to be 1:0.7. The other optimum conditions were found 5 mM $Ca^{2+}$ ion, pH 6.6, and incubation temperature of $27^{\circ}C. K_m \;and \;V_{max}$ values were estimated to be 1.43 mM and 0.8 $nmole/min/{\mu}g$ protein respectively. These properties in a PC-PA liposome system were compared with those in a PC-SDS mixed micelle system. The effects of other phospholipids and organic phosphates on the enzyme activity were also examined.

Isolation of Protoplasts from Rhizopus nigricans (Rhizopus nigricans로부터 원형질체의 분리)

  • Kim, Myung-Hee;Kim, Mal-Nam
    • The Korean Journal of Mycology
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    • v.22 no.2
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    • pp.138-144
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    • 1994
  • Conditions for isolation of protoplasts from spores and mycelia of Rhizopus nigricans were studied. Larger amount of protoplasts was obtained from swollen spores in liquid medium contained with 5% of 2-deoxy-D-glucose for 4 hours than from mycelia. Enzyme mixture of Novozym 234(2%) and ${\beta}-glucuronidase(5000\;unit/ml)$ was most effective for the isolation of protoplasts from swollen spores and from mycelia. The solution of 0.6 M $MgSO_4$ or mannitol and pH 6.0 showed good results as the osmotic stabilizer and the optimal condition of pH of the enzyme solution for the isolation of protoplast from the swollen spores, respectively. At this condition, $8.0{\times}10^6\;cells/ml$ of protoplasts was obtained from swollen spores by digestion with lytic enzyme mixture for 2 hours.

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Structure-Activity Relationships Study of Angiotensin Converting Enzyme Inhibitor Captopril Derivatives: Importance of Solution Moleculnr Dynamics Study (Angiotensin 변환 효소 억제제인 Captopril 유도체들의 구조와 활성관계 연구: 수용액상의 분자동력학적 연구의 중요성)

  • 지명환;윤창노;진창배;박종세
    • Biomolecules & Therapeutics
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    • v.2 no.1
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    • pp.34-38
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    • 1994
  • In order to investigate the structure-activity relationships of the stereoisomers of angiotensin converting enzyme inhibitors, captopril and its derivatives were selected as model compounds. In vitro enzymatic activities of them depend on the symmetry at the asymmetric carbons. Especially, the alanyl carbon should have the S configuration to be biologically active. But the demethylated captopril having the achiral carbon also shows the activity although it is less active than captopril. Seven stereoisomers of captopril and its derivatives were chosen and their acidic and ionic forms were used for molecular dynamics simulations. Four computer simulations were practiced for each model compound in order to obtain the good condition for simulation to explain the experimental structure-activity relationships. From the computer simulation results, relativistic movements of three well-known pharmacophoric sites, carboxylate carbon, carbonyl oxygen, and sulfur atoms, were analyzed. Good results were obtained from the aqueous solution molecular dynamics simulation with ionic forms of model compounds. Active model compounds have the pharmacophoric areas of 6.08 to 6.38 $\AA$$^2$and the similarity in the geometrical data. But inactive ones have the largely deviated values of 4.51 to 4.87 $\AA$$^2$from those of active ones.

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Stiffness loss in enzyme-induced carbonate precipitated sand with stress scenarios

  • Song, Jun Young;Sim, Youngjong;Yeom, Sun;Jang, Jaewon;Yun, Tae Sup
    • Geomechanics and Engineering
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    • v.20 no.2
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    • pp.165-174
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    • 2020
  • The enzyme-induced carbonate precipitation (EICP) method has been investigated to improve the hydro-mechanical properties of natural soil deposits. This study was conducted to explore the stiffness evolution during various stress scenarios. First, the optimal concentration of urea, CaCl2, and urease for the maximum efficiency of calcite precipitation was identified. The results show that the optimal recipe is 0.5 g/L and 0.9 g/L of urease for 0.5 M CaCl2 and 1 M CaCl2 solutions with a urea-CaCl2 molar ratio of 1.5. The shear stiffness of EICP-treated sands remains constant up to debonding stresses, and further loading induces the reduction of S-wave velocity. It was also found that the debonding stress at which stiffness loss occurs depends on the void ratio, not on cementation solution. Repeated loading-unloading deteriorates the bonding quality, thereby reducing the debonding stress. Scanning electron microscopy and X-ray images reveal that higher concentrations of CaCl2 solution facilitate heterogeneous nucleation to form larger CaCO3 nodules and 11-12 % of CaCO3 forms at the interparticle contact as the main contributor to the evolution of shear stiffness.

DETERMINATION OF ANTIFUNGAL ABILITY OF DENTURE CLEANSING AGENTS TO CANDIDA ALBICANS (Candida albicans에 대한 의치 세정제의 항진균능 검사)

  • Chun, Sang-Sup;Chung, Chae-Heon;Lee, Zang-Hee
    • The Journal of Korean Academy of Prosthodontics
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    • v.31 no.1
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    • pp.28-38
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    • 1993
  • For the purpose of evaluating the cleansing efficiency against Candida albicans detected frequently in patients with denture stomatitis, two denture cleansers with or without enzymes were studied under the same conditions. The results were as fellows: 1. Enzyme-contain denture cleanser was showed more Candida albicans lytic ability than non-enzyme-contained denture cleanser. 2. It was observed that Candida albicans lytic activity in further diluted manufacturerers' recommended concentration was decreased. 3. In fungicidal test, the enzyme-contained denture cleanser sterilized Candida albicans, and the non-enzyme-contained denture cleanser did not sterilize Candida albicans. 4. Sterilizing time of Candid albicans was needed for at least 60 minutes in enzyme-contained denture cleanser solution which was diluted with manufacturerers' recommended concentrations., and was needed for more times with further diluted manufacturerers' recommended concentrations. 5. In vitro growth test of Candida albicans on acrylic resin surface, the only enzyme-contained denture cleanser inhibited growth of Candida albicans, and it was observed that inhibiton ability of growth of Candida albicans on arrylic resin surface was decreased in further diluted manufacturerers' recommended concentrations.

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Studies on the Cellulolytic Enzyme System of Rhizopus sp. G-211 Isolated from Rotting Ginseng (인삼부패란중 Rhizopus sp. G-211이 생성하는 Cellulase 에 관한 연구)

  • 노혜원;김상달;도재호;강성호
    • Microbiology and Biotechnology Letters
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    • v.10 no.1
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    • pp.1-7
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    • 1982
  • A Rhizopus sp. was selected for its strong cellulolytic activity among various strains of molds found in rotting ginseng roots. Studies were made on some properties of the cellyloiytic enzyme produced by the strain. The results obtained were summarized as follows: The optimum pH of the enzyme was 4.5 and the range of its stability to the pH was 3.0 to 7.0. The optimum temperature was 5$0^{\circ}C$, while the enzyme was instantly inactivated above 6$0^{\circ}C$. Mn$^{++}$ and Co$^{++}$ ions increased enzyme activity and the metal ions were found to increased the ther-mostability of the enzyme. This enzyme was inhibited by sodium dodecyl sulfate and 2,4-dinitrophenol. This enzyme had a strong cellulolytic enzyme activity on various native cellulose given a sufficient reaction time. The addition of 0.5% saponin solution into reaction mixture increased the enzyme activity.

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The Effect of Proteases on Contamination Removal (프로테아제의 오염 세정 효과)

  • Kim, Ju-Hye;Gwon, Mi-Yeon
    • Proceedings of the Korean Society of Dyers and Finishers Conference
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    • 2008.04a
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    • pp.181-183
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    • 2008
  • Four different subtilisins of protease were investigated to see their effects on the cleaning activity. The cleaning solution was formulated with various non-ionic surfactants and other additives such as propylene glycol, triethanolamine, pH balancer etc. to evaluate their effect on enzyme activity as well. Evaluation of formulated cleaning solution was carried under K0120 using pre-soiled textiles from EMPA. The results showed that the cleaning activity on soil removal was not severly influenced by surfactant but the enzyme mostly. In addition, the activity of enzymes was not much affected by the type of surfactants as long as the surfactants were non-ionic. Liquinase among the four enzymes used in this study showed the best performance on soil removal, especially blood stain.

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