• Title/Summary/Keyword: enzymatic method

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Acute toxicity response caused by mixture or tank mix of several insecticides (몇 가지 살충제의 혼용 및 혼합 시 독성반응)

  • Lee, Je-Bong;Jeong, Mi-Hye;Sung, Ha-Jung;Lee, Hae-Keun;Yang, Jae-Sul
    • The Korean Journal of Pesticide Science
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    • v.5 no.4
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    • pp.57-61
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    • 2001
  • Tank mixing application of pesticides has been used to reduce labor and to control wide spectrum of pests, but it may cause significant pesticide poisoning on human and animals due to carelessness. The toxic response of pesticides for the tank mixtures and mixtures was investigated to determine acute toxicity and enzymatic change using experimental animals. Acute oral toxicity and acute dermal toxicity were tested by RDA test guideline. The $LD_{50}$ was calculated by probit analysis method and cholinesterase was measured with automatic analyzer. The toxicities were generally higher than estimated toxicities in tank mixing and mixture. Serum cholinesterase activity was inhibited more than expected at the dose levels of 1/5, 1/10 and 1/20 of $LD_{50}$. Therefore, the results of this study showed that acute toxicity caused by the pesticide mixtures should be considered before the tank mixing method is applied.

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Enzymatic Analysis of Organophosphorus Pesticides Using Cholinesterase Inhibition Activities (Cholinesterase 저해 활성을 이용한 유기인계 농약의 효소적 분석)

  • Kim, Jung-Ho
    • The Korean Journal of Pesticide Science
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    • v.5 no.1
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    • pp.12-18
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    • 2001
  • The effects of organophosphorus were examined with inhibition of the cholinesterase activity on tile chicken plasma in vivo and in vitro. The cholinesterase activity in chicken plasma determined by tile Ellman mettled was $23{\mu}mol$/min/g protein. After oral administration with 0.2 and 0.5 times of organophosphorus terbufos $LD_{50}$(1.81 mg/kg), cholinesterase activity were inhibited to 36% and 96% of control after 15min in vivo, respectively. After oral administration with 0.2 and 0.5 times of terbufos $LD_{50}$(1.81 mg/kg), then the recovery of cholinesterase activity followed to 99% and 56% of control after 11hr, respectively. Ki of phosphorodithioate and phosphorothioate with P=S was $74{\sim}322\;mole^{-1}min^{-1}$ in vitro. Ki of phosphate and phosphorothiolate with P=O was $13898{\sim}79610\;mole^{-1}min^{-1}$. Toxicology of organophosphorus with P=S was higher than that of organophosphorus with P=S by oxidation. $pI_{50}$ of phosphorodithioate and phosphorothioate with P=S was $21{\sim}102$ mg/L. $pI_{50}$ of phosphate and phosphorothiolate with P=O was $0.519{\sim}0.071$ mg/L. Enzyme-Inhibition method with cholinesterase was the rapid bioassay method to detect the organohpophorus pesticides in vitro.

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The Correlation of $CO_2$ Content with Non-Enzymatic Browning Color in Non-Fat Dried Milk (탈지분유(脫脂粉乳)에서 $CO_2$함량(含量)과 비효소적(非酵素的) 갈변색소(褐變色素)와의 상관관계(相關關係))

  • Chang, Kyu-Seob;Min, David B.
    • Applied Biological Chemistry
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    • v.29 no.2
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    • pp.190-197
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    • 1986
  • In order to develop a simple and effective method for determining the rate of the Maillard reaction in non-fat dry milk, the carbon dioxide content of the headspace as an indicator were used and the amount of correlation between $CO_2$ content and brown color development were determined by the gas chromatograph. There is a high correlation between brown color and $CO_2$ content. The use of gas chromatography to analyze the $CO_2$ in the headspace of samples is a quick, simple and effective method of monitoring the Maillard reaction. Volatile concentration increases with storage time and varies inversely with oxygen content. Lysine is more effective than glucose in catalyzing the Maillard reaction. Product samples can be stored at $55^{\circ}C$ and $68^{\circ}C$ to accelerate the rate of the Maillard reaction and shorten testing period, but product stored at $75^{\circ}C$ is degraded too rapidly to be of any real use.

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Principles of Simulated Moving Bed Reactor(SMBR) (Simulated Moving Bed Reactor(SMBR)의 원리)

  • Song, Jae-Ryong;Kim, Jin-Il;Koo, Yoon-Mo
    • Korean Chemical Engineering Research
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    • v.49 no.2
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    • pp.129-136
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    • 2011
  • Simulated Moving Bed(SMB) process consists of multiple chromatographic columns, which are usually partitioned into four zones. Such a process characteristic allows a continuous binary separations those are impracticable in conventional batch chromatographic processes. Compared with batch chromatography, SMB has advantages of continuity, high purity and productivity. Various researches have been reported for the integration of reaction and recovery during process operation on the purpose of economics and effectiveness. Simulated Moving Bed Reactor(SMBR) is introduced to combine SMB as a continuous separation process and reactor. Several cases of SMBR have been reported for diverse reactions with catalytic, enzymatic and chemical reaction on ion exchange resin as main streams. With an early type of fixed bed using catalyst, SMBR has been developed as SMB using fluidized enzyme, SMB with immobilized enzyme and SMB with discrete reaction region. For simple modeling and optimization of SMBR, a method considering convection only is possible. A complex method considering axial dispersion and mass transfer resistance is needed to explain the real behavior of solutes in SMBR. By combining reaction and separation, SMBR has benefits of lower installation cost by minimizing equipment use, higher purity and yield by avoiding the equilibrium restriction in case of reversible reaction.

Production of Biodiesel Using Immobilized Lipase from Proteus vulgaris (Proteus vulgaris에서 유래한 리파아제의 고정화 및 바이오디젤 생산)

  • Yoon, Shin-Ah;Han, Jin-Yee;Kim, Hyung-Kwoun
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.238-244
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    • 2011
  • Biodiesel, mono-alkyl esters of long chain fatty acids, is one of the alternative fuels derived from renewable lipid feedstock, such as vegetable oils or animal fats. For decade, various lipases have been used for the production of biodiesel. However, the production of biodiesel by enzymatic catalyst has profound restriction in industry application due to high cost. To overcome these problems, many research groups have studied extensively on the selection of cheap oil sources, the screening of suitable lipases, and development of lipase immobilization methods. In this study, we produced biodiesel from plant oil using Proteus vulgaris lipase K80 expressed in Escherichia coli cells. The recombinant lipase K80 was not only expressed in high level but also had high specific lipase activity and high stability in various organic solvents. Lipase K80 could produce biodiesel from olive oil by 3-stepwise methanol feeding method. The immobilized lipase K80 also produced biodiesel using the same 3-stepwise method. The immobilized lipase could produce biodiesel efficiently from various plant oils and waste oils.

Production of Inulo-oligosaccharides from Chicory(Cichorium intybus, L.) with Endo-inulinase from Arthrobacter sp.537 (새로운 endo-inulinase를 이용한 치커리 추출물로부터 Inulo올리고당의 생산)

  • Kang, Su-Il;Kim, Su-Il
    • Applied Biological Chemistry
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    • v.40 no.1
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    • pp.34-38
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    • 1997
  • For the effective production of functional oligosaccharides(DP 3-5) from inulin in chicory extracts, the acid hydrolysis and enzymatic endo-inulinase reaction were compared. Acid hydrolysis was unfavorable ; the content of oligosacharides in total sugar increased to 26.0% for 12 min at $55^{\circ}C$ and 24.6% at 6 min at $65^{\circ}C$ and showed little change for 30 min. The content of high DP(DP 6) decreased from 83.5 to 49.5% and 23.0% for 30 min, repectively. Glucose, fructose and sucrose increased to 24.6% and 50.3%, respectively. Hydrolysis of chicory extracts with purified endo-inulinase from Arthrobacter sp. S37 was carried out at $40^{\circ}C$ and pH 7.5 for 44 hrs. The content of high DP($DP{\geq}6$) in total sugar decreased from 83.5 to 23.0% and that of inulobiose(F2) and DP 3-5 increased to 66.1%. Glucose, fructose and sucrose were not produced. The hydrolysis of chicory extracts without DP 1 and DP 2 with crude or with purified enzyme were also carried out. In contrast to the hydrolysate of crude enzyme, that of purified endo-inulinase did not contain glucose, fructose, sucrose, F2 and 1-kestose(GF2). The content of oligosaccharides in the hydrolysate of the purified endo-inulinase were 79.2%, composed mainly of inulotriose(F3), inulotetraose(F4) and inulopentaose(F5), which shows that the enzymatic hydrolysis using purified endo-inulinase from Arthrobacter sp. S37 is the best method for oligosaccharides production from inulin in chicory extracts.

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Characteristics of mushroom Phellinus baumii extracts with enzyme pretreatment (효소 전처리에 의한 상황버섯 β-glucan 추출물의 특성)

  • Son, Eun Ji;Ryu, Eun-Ah;Lee, Sang-Han;Kim, Young-Chan;Hwang, In-Wook;Chung, Shin-Kyo
    • Journal of Applied Biological Chemistry
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    • v.61 no.1
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    • pp.101-108
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    • 2018
  • This study was conducted to establish optimized ${\beta}-glucan$ extraction method through enzymatic hydrolysis from Phellinus baumii and investigate ${\beta}-glucan$ contents and physicochemical properties. The optimal condition was obtained with the enzyme concentration of 0.66% (v/v), reaction time of 6.08 h ($R^2=0.9245$) and the ${\beta}-glucan$ contents from the Phellinus baumii extracts under the optimized condition was 1.9594 g/100 g. ${\beta}-Glucan$ yield (0.76-16.40%) of enzyme beta-glucan extract (EBE) was three fold higher than that of non-enzyme beta-glucan extract (NEBE). ${\beta}-Glucan$ purity (11.15-59.05%) of non-enzyme beta-glucan (NEB) and that of enzyme beta-glucan (EB) were higher than that of NEBE and that of EBE. ${\beta}-Glucan$ purity of EB (59.05%) and ${\beta}-glucan$ contents of EB (3.38 g/100 g) showed higher than those of others. Total sugar contents (0.61-1.17 mg/mL) showed that NEB and EB were higher than that of NEBE and EBE, EB had the highest total sugar content as 1.17 mg/mL, respectively. Protein contents (0.44-11.73 mg/mL) of NEBE and that of EBE were higher than that of NEB, that of EB. In FT-IR spectrum, the band at $890cm^{-1}$ of microcapsule was attributed to a ${\beta}-1,3-glucan$. The toxicities of ${\beta}-glucan$ from Phellinus baumii in both melanoma cell lines was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoli um bromide assay and ${\beta}-glucan$ from Phellinus baumii has no toxicity until $30{\mu}g/mL$. The effects of ${\beta}-glucan$ from Phellinus baumii on inhibition of cancer cell proliferation were detected by using a wound healing assay. The effect of NEB and EB were higher than NEBE and EBE, especially $30{\mu}g/mL$ of EB had the highest in both melanoma cell lines.

Development of Optimum Process for Continuous Hydrolysis of Fish Skin Gelatin Using a Three-Step Recycle Membrane Reactor (재순환 3단계 막반응기를 이용한 어피젤라틴의 연속적 가수분해 최적화 공정 개발)

  • Kim, Se-Kwon;Byun, Hee-Guk
    • Applied Chemistry for Engineering
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    • v.5 no.4
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    • pp.681-697
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    • 1994
  • The enzymatic hydrolysate of gelatin extracted from fish skin was fractionated and recycled through the membrane reactor according to the molecular weight for the purpose of using as functional material. In addition, the enzymatic hydrolysis conditions of gelatin, enzyme stability by membrane and mechanical shear, and effect on the long-term operational stability of the recycle membrane reactor were investigated. Using the pH-drop technique, Alcalase, pronase E and collagenase were identified as the most suitable enzymes for the hydrolysis of fish skin gelatin. The optimum hydrolysis conditions in the 1st-step membrane reactor(1st-SMR) by Alcalase were enzyme concentration 0.2mg/ml, substrate-to-enzyme ratio(S/E) 50(w/w), $50^{\circ}C$, pH 8.0, reaction volume 600ml and flow rate 6.14ml/min. In the 2nd-SMR by pronase E were enzyme concentration 0.3mg/ml, S/E 33(w/w), $50^{\circ}C$, pH 8.0, reaction volume 600ml and flow rate 6.14ml/min. In the case of 3rd-SMR, enzyme concentration 0.1mg/ml, S/E 100(w/w), $37^{\circ}C$, pH 7.5, reaction volume 600ml and flow rate 10ml/min. Decreased enzyme activities by mechanical shear and membrane were 30% and 15% in the 1st-SMR, were 14% and 5% in the 2nd-SMR, and 18% and 8% in the 3rd-SMR, respectively. Under the optimum conditions, the degree of hydrolysis in the 1st, 2nd and 3rd-SMR were 3.5%(Kjeldahl method, 87%), 3.1%(77%) and 2.7%(70%), respectively. The productivity of hydrolysate in the continuous three-step membrane reactor was 430mg per enzyme(mg) for 10 times of volume replacements.

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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.

Hydrolysis of Rice Syrup Meal Using Various Commercial Proteases (쌀 시럽박의 단백질 가수분해 특성)

  • Kim, Chang-Won;Park, Jin-Woo;Choi, Hyuk-Joon;Han, Bok-Kyung;Yoo, Seung-Seok;Kim, Byung-Yong;Baik, Moo-Yeol;Kim, Young-Rok
    • Journal of Life Science
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    • v.21 no.2
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    • pp.309-315
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    • 2011
  • Rice syrup meal (RSM) was enzymatically hydrolyzed using eight commercial proteases (Protamex, Neutrase, Flavourzyme, Alcalase, Protease M, Protease N, Protease A, Molsin F) for 4 hr at optimum pH and temperature. Proteolytic hydrolysates were examined in supernatant and precipitate using Lowry protein assay, semimicro Kjeldahl method and gravimetric method using weight difference before and after enzymatic hydrolysis. Although RSM contains a high amount of protein (71.2%), only a very small amount of protein was hydrolyzed. Two proteases (Protease M and Protease N) were found to be the most effective in the hydrolysis of RSM protein. In Lowry method, 57.5 and 59.0 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments, respectively. In gravimetric method, 80.0 and 85.4 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments. In Kjeldahl method, 67.43 and 70.43 mg protein/g RSM were hydrolyzed after Protamex and Protease N treatments, respectively. For synergistic effect, two or three effective commercial proteases (Protease M, Protease N and Protease A) were applied to RSM at one time. The highest hydrolysis of RSM protein was observed in both Lowry protein assay (80.3 mg protein/g RSM) and gravimetric methods (153.2 mg protein/g RSM) when three commercial proteases were applied at one time, suggesting the synergistic effect of those proteases.