• Title/Summary/Keyword: Enzyme I

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Development of an Enzyme Electrode Biosensor for Lactic Acid Bacteria (효소 전극을 이용한 유산균 측정 바이오센서 개발)

  • Park T. S.;Cho S. I.
    • Journal of Biosystems Engineering
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    • v.30 no.4 s.111
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    • pp.249-253
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    • 2005
  • This study was carried out to develop enzyme biosensor for lactic acid bacteria. Lactic acids produced by lactic acid bacteria (LAB) was measured and good correlation $R^2=0.98$ between LAB count and lactic acids concentration was found. Hydrogen ion produced by L-lactate dehydrogenase (L-LDH) was measured by a potentiometer. Glutamic-pyruvic transminase (GPT) was used for eliminating inhibitor in the reaction. Polyacrylamide gel was used for immobilizing matrix of the sensor. The biosensor was tested and showed good feasibility with $R^2=0.99$ on validation.

DEINKING OF COLORED OFFSET NEWSPRINT WITH ENZYME TREA TMENT IN COOPERATION WITH ULTRASONIC WAVE

  • Yimin XIE;U, Hong-W;Yanming LAI
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 1999.11b
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    • pp.149-152
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    • 1999
  • A new process for deinking of colored offset newsprint, i.e. enzyme treatment in cooperation with ultrasonic wave was developed in the present study. The physical characteristics such as fiber length, coarseness, crystallinity index of the deinked pulps were investigated and the sugar residues released from the treatment were analyzed. It was found that colored offset newsprint could be deinked effectively by cellulase treatment when ultrasonic wave was applied. The brightness increased by 5% ISO over that of control experiment and the pigment content was reduced markedly. Though the ultrasonic wave had little effect on the strength and crystallinity of the pulp, the treatment of enzyme combined with ultrasonic wave reduced the coarseness and fiber length to some extent. It was also found that ultrasonic wave could accelerate the hydrolysis of cellulose and hemicellulose during the cellulase treatment.

Development of an Enzyme-Linked Immunosorbent Assay for the Organophosphorus Insecticide Cyanophos

  • Park, Jae-Hyun;Park, Won-Chul;Kim, Yoo-Jung;Lee, Yong-Tae
    • Bulletin of the Korean Chemical Society
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    • v.23 no.4
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    • pp.605-609
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    • 2002
  • A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of organophosphorus insecticide cyanophos. An analogue (hapten) of cyanophos was synthesized and was coupled to BSA to produce polyclonal antibodi es from rabbits. The antisera were screened against another hapten coupled to ovalbumin (OVA). Using the sera of highest specificity, an antigen-coated ELISA was developed, which showed an I50 of 310 ng/mL with the detection limit of 20 ng/mL. The antibodies showed negligible cross-reactivities with other organophosphorus pesticides except for parathion-methyl, which makes the assay suitable for the selective detection of cyanophos.

Production of the Bacteriocin from the Tofu-Residue (두부비지를 이용한 박테리오신 생산)

  • 이명숙;이원재;김동수;박지현;강지희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.28 no.1
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    • pp.74-80
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    • 1999
  • Growth and bacteriocin production by Lactobacillus sp. GM7311 in tofu residue treated with two commercial amylases were investigated. The optimal condition of amylase Ⅰ(liquefied enzyme for sauce) and Ⅱ(multienzyme 2,000) for the enzyme reaction was showed at pH 6.0 and 4.0, respectively. The optimal temperature was 40oC both. At the enzyme dosage 4% and 3% and reaction time 1hr, about 2% of reduced sugar needed bacteriocin production was obtained. The enzymatic treatment of tofu residue enhanced bacteriocin production by lactic acid bacteria, particularly in the tofu residues added 2.0% yeast extract. But, we couldn't see the increment of bacteriocin activity in the tofu residues added other nitrogen sources such as proteose peptone No. 3 and lab lemco powder. Also, in the comparision of amylase I and Ⅱ, bacteriocin activity in the tofu residue treated with amylase Ⅰ was better than that of amylase Ⅱ.

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Chemical Modification of Yeast Farnesyl Protein Transferase Expressed in E. coli

  • Kim, Hyun-Kyung;Yang, Chul-Hak
    • Bulletin of the Korean Chemical Society
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    • v.27 no.4
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    • pp.529-534
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    • 2006
  • Chemical modification of the S. cerevisiae farnesyl protein transferase (FPT) with CMC, phenylglyoxal and DEPC resulted in enzyme inactivation, depending upon the reagent concentration. The peptide substrate GST-PEP-I, a GST-fused undecapeptide mimicking the C-terminus of $p21^{Ki-ras}$, protected the enzyme against inactivation by CMC which is specific to either aspartate or glutamate, while the other substrate farnesyl pyrophosphate (FPP) showed protection against phenylglyoxal which is the specific modifier of arginine residues, dependent on the substrate concentrations. Neither of the two substrates protected the enzyme against histidine inactivation by DEPC. It is suggested that there is at least one aspartate or glutamate residue at the peptide substrate binding site, and that at least one arginine residue is located at the binding site of FPP. There also seems to be at least one histidine residue which is critical for enzymic activity and is exposed toward the bulk solution, excluded from the substrate binding sites.

Molecular cloning of phospho-$\beta$-galactosidase gene of lactobacillus casei in escherichia coli (Lactobacillus casei의 phospho-$\beta$-galactosidase 유전자의 대장균내 분자클로닝)

  • 문경희;박정희;최순영;이유미;김태한;김연수;민경희
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.188-193
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    • 1989
  • Gene for lactose catabolism in Lactobacillus casei SW-M1 was encoded by a 60Kb metabolic plasmid. A derivative of only 10kb, pPlac 15 of recombinant plasmid, was constructed by introducing into pBR322 and was cloned into E. coli using restriction endonuclease Pst I. A 10kb insery DNA in plasmid pBR322 was identified as a gene encoded phospho-$\beta$-galactosidase by the determination of enzyme activity. Phospho-$\beta$-galactosidase was apparently expressed in E. coli. The enzyme activities of cell-free extract from transformant E. coli HB101 carrying pPLac 15 DNA were not different from that of L. casei as a donor strain on the basis of enzyme properites. However, specific activity of phospho-$\beta$-galactosidase in the cloned strain with Lac $Y^{-}$ phenotype of E. coli HB101 was lower than that in L. casei strain.

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Studies on the Digestive Enzyme of Cynthia roretzi V. Drasche. I . Some Enzymatic properties of Hmylase. (우릉쉥이(Cynthia roretzi v. Drasche)의 소화효소에 대하여 (제1보) Amylase의 효소적 성질)

  • 서석구;양한술
    • YAKHAK HOEJI
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    • v.5 no.1
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    • pp.45-50
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    • 1960
  • Some enzymatic properties of Cynthia roretzi v. (Drasche Korean : U-Rung-Shei) was studied by author and obtained the following results; 1. The optimum pH of the digestive gland amylase was 6.8-7.0 2. Activity of metallic ion on the amylase showed the following order; 10$^{-3}$ M M $n^{++}$>10$^{-3}$ M $Co^{++}$>10$^{-4}$ M $Mg^{++}$>10$^{-4}$ M $Ca^{++}$>10$^{-2}$ M Z $n^{++}$>10$^{-2}$ M P $b^{++}$ 3. The digestive gland enzyme inactivated at 70.deg. C. 4. When the enzyme concentration increase 2 times, the enzymatic activity also increase, but not propertionally. 5. The digestine gland amylase showed remarkably higher enzymatic activity than the intestinal amylase. 6. The digestive gland amylase from the ascidian showed remarkably higher enzymatic activity than the heptancreatic amylase from shell fish (Turbo (Batillus) Cornutus Solander).nder).nder).

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Activity and stability of purified amylase produced by streptomyces aureofaciens 77

  • Ibrahim, A.N.;Ahmed, F.H.;Ibrahim, M.M.K.;Arafa, M.A.I.
    • Archives of Pharmacal Research
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    • v.13 no.1
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    • pp.33-37
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    • 1990
  • The effects of pH values, temperature and some elements on the amylolytic activity and stability of the purified S. aureofacienc 77 amylase were studied in this investigation. The purified enzyme showed its maximum activity at pH 6 within 8 min incubation at $40^{\circ}C$. None of the tested 6 metals showed on stimulatory effect on the enzymatic activity, $Fe^{+++}$, $Cu^{++}$ and $Hg^{++}$ at high dose inhibited the enzyme activity to great extent as compared with $Zn^{++}$, $Mn^{++}$ and $Fe^{++}$ whih gave less effect in this respect. The enzyme liquor was found to be thermolabile, since it lost completely its activity after 4 days incubation under room temperature and showed maximum activity during this period as a result of additions of $Ca^{++}$and NaCl, Gradual reduction was however recorded until activity reached 30% after 60 days of incubation.

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Precepitation and purification of amylase enzyme produced by streptomyces aureofaciens 77

  • Ibrahim, A.N.;Ahmed, F.H.;Ibrahim, M.M.K.;Arafa, M.A.I.
    • Archives of Pharmacal Research
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    • v.13 no.1
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    • pp.28-32
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    • 1990
  • Precipitation and purification of amylase secreted by Streptomyces aureofaciens 77 in liquid inorganic salts-starch medium under the optimum conditions were carried out. Ammonium sulphate fractionation was used to precipitate amylase in cell free culture filtrate. (NH/sub 4/)/sub 2/ SO/sub 4/ at a concentration of 50-70% saturation gave the highest enzyme yield. The obtained precipitates were redissolved in phosphate buffer (pH 7.0) and subjected to dialysis. The dialyzed enzyme preparation was applied to DEAE-cellulose column chromatography which resulted in an increase of purification up to 59.48 fold. A further step of purification was done by applying the obtained purified sample to Sephadex-G200 column chromatography which resulted in ann increase of purification up to 73. 92 fold. The results clearly indicated that the isolated amylase from S. aureofaciens 77 was only on type.

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Analyses of single nucleotide polymorphisms and haplotypes of BoLA-DRB3 gene in Holstein and Hanwoo (홀스타인종과 한우에 있어서 BoLA-DRB3 유전자의 단일염기다형과 반수체 분석)

  • Jeong, Hang-Jin;Yu, Seong-Lan;Hoque, M.R.;Lee, Jun-Heon;Do, Chang-Hee;Ryoo, Seung-Heui;Sang, Byung-Chan
    • Korean Journal of Agricultural Science
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    • v.38 no.1
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    • pp.51-63
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    • 2011
  • BoLA (bovine leukocyte antigens) have been known as gene complex related with bovine diseases and immunological traits. This study was conducted to find out the characteristics of BoLA-DRB3 gene related to mastitis and BL(bovine leukocyte) from 280 cattle [193 animals of Holstein cattle and 87 animals of Hanwoo]. As a result, five PCR-RFLP types (b, d, e, f and g) using HaeIII restriction enzyme, three BstYI restriction patterns (b, d and e) and eight RsaI restriction types(b, d, f, I, j, n, o and w) were identified. Moreover, we identified new d' type ($197{\rightarrow}175$/22), having one more cutting site by BstYI enzyme than d type allele and n' type ($180{\rightarrow}169$/11) having one more cutting site by RsaI enzyme than n allele was additionally identified. Next, we identified 53 SNPs in BoLA-DRB3 exon2 from 280 cattle. SNP frequency and heterozygosity of Holstein and Hanwoo were investigated in all the SNP genotype. These results might be based on research for identifying marker associated with bovine diseases.