• Title/Summary/Keyword: enzyme hydrolysis

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Real Time Scale Measurement of Inorganic Phosphate Release by Fluorophore Labeled Phosphate Binding Protein (형광단이 붙어 있는 인산결합 단백질에 의한 인산 배출의 실시간 측정)

  • Jeong Yong-Joo
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.935-940
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    • 2005
  • Fluorescence change of coumarin labeled phosphate binding protein (PBP-MDCC) was monitored to measure the amount of released inorganic phosphate ($P_{i}$) during nucleoside triphosphate (NTP) hydrolysis reaction. After purification of PBP-MDCC, fluorescence emission spectra showed that fluorescence responded linearly to $P_{i}$ up to about 0.7 molar ratio of $P_{i}$ to protein. The correlation of fluorescence signal and $P_{i}$ standard was measured to obtain [$P_{i}$] - fluorescence intensity standard curve on the stopped-flow instrument. When T7 bacteriophage helicase, double-stranded DNA unwinding enzyme using dTTP hydrolysis as an energy source, reacted with dTTP, the change of fluorescence was able to be converted to the amount of released $P_{i}$ by the $P_{i}$ standard curve. $P_{i}$ release results showed that single-stranded Ml3 DNA stimulated dTTP hydrolysis reaction several folds by T7 helicase. Instead of end point assay in NTP hydrolysis reaction, real time $P_{i}$-release assay by PBP-MDCC was proven to be very easy and convenient to measure released $P_{i}$.

Enzymatic Hydrolysis of Pre-treated Ulva pertusa with Alkaline Peroxide (구멍갈파래의 알칼리 과산화수소 전처리 및 효소 가수분해 특성)

  • Yoon, Byung-Tae;Kim, Young-Wun;Chung, Keun-Wo;Kim, Jin-Seog
    • Applied Chemistry for Engineering
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    • v.22 no.3
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    • pp.336-339
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    • 2011
  • Algae is an abundant and potential fermentation substrate. The enzymatic hydrolysis of algae was investigated by pre-treating an alkaline hydrogen peroxide with commercial cellulase and viscozyme. Algae used in this study was the Ulva pertusa. The evaluated response was the yield of released glucose after the enzymatic hydrolysis. Alkaline hydrogen peroxide containing mixtures of 1 wt% hydrogen peroxide and 1~1.75 wt% sodium hydroxide was also used. The results show that the highest glucose conversion was obtained for Ulva pertusa using 5 wt% hydrogen peroxide at $60^{\circ}C$ for 3 h. The required amount of enzymes after the pre-treatment with alkaline hydrogen peroxide were reduced by far compared to that of untreated Ulva pertusa. Also, the amount of glucose that is released during the enzymatic hydrolysis was increased.

Hydrolysis Methods for the Efficient Manufacture of Sugar Solutions from the Freshwater Alga Water-net (Hydrodictyon reticulatum) (담수조류 그물말로부터 당 용액의 효율적 제조를 위한 가수분해 방법)

  • Kim, Ji-Hyun;Kim, Sul Ki;Ko, Eun Hye;Kim, Jin-Cheol;Kim, Jin-Seog
    • Weed & Turfgrass Science
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    • v.2 no.2
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    • pp.176-183
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    • 2013
  • To explore hydrolysis methods for the efficient manufacture of sugar solutions from the freshwater alga Water-net (Hydrodictyon reticulatum, HR), acid hydrolysis, enzymatic hydrolysis, and combined hydrolysis (acid followed by enzymatic hydrolysis) were investigated. In the one-step acid hydrolysis, the reaction of 8% solids content using 2% sulfuric acid at $120^{\circ}C$ for 1 hour was desirable. In this case, glucose 27.44 g 100 g $DM^{-1}$ could be obtained from the HR-d13 samples. In the two-step acid hydrolysis, the primary hydrolysis (HR powder : 72% sulfuric acid = 1 g : 1.5 mL) was carried out for 1 hour at $60^{\circ}C$, and then the secondary hydrolysis was done for 1 hour at $120^{\circ}C$ after addition of distilled water 23.5 mL. In this case, glucose 35.11 g/100 g DM could be obtained from the HR-d13 samples. In the combined hydrolysis, 25% solids content using 2% hydrochloric acid were reacted for 1 hour at $120^{\circ}C$, and then citrate buffer and hydrolysis enzyme complexes (E1 1.0 mL+E2 0.2 mL $g^{-1}$ dried matter) were added and reacted for 1 - 2 days at $50^{\circ}C$. In this case, glucose 33.5 g 100 g $DM^{-1}$ could be obtained from the HR-d23+26 samples. In conclusion, combined hydrolysis was likely to be more useful saccharification method of HR biomass at a practical level, considering the glucose productivity, generation of fermentation-inhibiting substances (hydroxyl methyl furfural, furfural), and limited use of strong acid.

Enzymatic Deinking of Old Newsprit with Alkalophilic Enzymes from Coprinus cinereus 2249

  • Lee, Jung-Myong;Eom, Tae-Jin
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.31 no.5
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    • pp.12-17
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    • 1999
  • This study was to evaluate applications of the alkalophilic enzymes from Coprinus cinereus 2249 with old newsprint(ONP). A enzymatic deinking process based on alkalopholic enzymes was investigated . It was found that alkalophilic enzymes could effectively deink old newsprint. When applied on deinking of the old newsprint, it increases the freeness and brightness due to effect of hydrolysis at 0.1% enzyme concentration . Also, the physical properties of deinked pulp were improved.

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Novel Liposome Immunoassay for Detection of Ultratrace Amount of Bioactive Substances : an Assay for Insulin

  • Lim, Soo-Jeong;Kim, Chong-Kook
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.281-281
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    • 1996
  • The immunoassay method is frequently used for the identification and quantitation of ultratrace amount of bioactive substances. Homogeneous liposome immunoassays, which can avoid the use of radioisotopes and separation steps, have recently been reported in many publications. Cytolysin-mediated liposome immunoassay using melittin ever been studied but showed limited applications. Here, we designed a homogeneous liposome immunoassay using Clostridium perfringens phospholipase C (PLC), an enzyme which catalyzes the hydrolysis of phosphatidylcholine in biological membranes, as a cytolysin.

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Inhibitory Effect of Trans-N-p-Coumaroyl Tryamine from the Twigs of Celtis chinensis on the Acetylcholinesterase

  • Kim, Dae-Keun;Lee, Kie-Seung
    • Archives of Pharmacal Research
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    • v.26 no.9
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    • pp.735-738
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    • 2003
  • The methanolic extract of the twigs of Celtis chinensis was found to show inhibitory activity on acetylcholinesterase (AChE), an enzyme that plays a role in the metabolic hydrolysis of ACh. Bioassay-guided fractionation of the methanolic extract resulted in the isolation of N-p-coumaroyl tyramine. as an inhibitor on AChE. This compound inhibited AChE activity in a dose-dependent manner, and the $IC_50$ value of trans-N-p-coumaroyl tyramine was 34.5 $\mu$g/mL (122 $\mu$M).

Cooper ions and hypochlorite are mainly responsible for oxidative inactivation of paraoxon-hydrolyzing activity in human high density lipoprotein

  • Su, Nguyen-Duy;Kim, Ju-Ryung;Eun, Sok-Dai
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.141.1-141.1
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    • 2003
  • Paraoxonase, an antioxidant enzyme, exclusively located on HDL is well known for both hydrolysis of organophosphate and prevention against LDL oxidation. It have been reported that PON1 decrease its activity under oxidative stress and that PON1 activity is lower in subject with higher vulnerability to organophosphate poisoning. The aim of our study is to examine the effect of oxidative system on paraoxon-hydrolzing activity and to elucidate the plausible mechanisms responsible for the decline of HDL-asscociated PON1 activity in vivo system. (omitted)

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Enzymatic Hydrolysis Optimization of a Snow Crab Processing By-product (홍게 가공부산물의 효소적 단백질 가수분해 최적화)

  • Jang, Jong-Tae;Seo, Won-Ho;Baek, Hyung-Hee
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.622-627
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    • 2009
  • The objectives of this study were to evaluate a protease suitable for the enzymatic hydrolysis of a snow crab processing by-product (SPB) and to optimize the hydrolysis conditions using response surface methodology (RSM). The SPB was hydrolyzed at $50^{\circ}C$ and pH 7.0-7.2 to obtain various degree of hydrolysis (DH) using Flavourzyme at an enzyme/substrate (E/S) ratio of 3.0%. The reaction progress curve exhibited an initial fast reaction rate followed by a slowing of the rate. The DH was increased to 30% at 90 min with a final DH 32 to 36%. A central composite experimental design having three independent variables (reaction temperature, reaction time, and E/S ratio) with five levels was used to optimize the enzymatic hydrolysis conditions. Based on the DH data, the optimum reaction conditions for the enzymatic hydrolysis of the SPB were a temperature of $51.8^{\circ}C$, reaction time of 4 hr 45 min, and an E/S ratio of 3.8%. It was demonstrated that the enzymatic hydrolysate of SPB could be used as a flavoring agent or a source of precursors for the production of reaction flavors.