• Title/Summary/Keyword: Hydrolysis rate

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The Brown-Rot Basidiomycete Fomitopsis palustris Has the Endo-Glucanases Capable of Degrading Microcrystalline Cellulose

  • Yoon, Jeong-Jun;Cha, Chang-Jun;Kim, Yeong-Suk;Son, Dong-Won;Kim, Young-Kyoon
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.800-805
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    • 2007
  • Two endoglucanases with processive cellulase activities, produced from Fomitopsis palustris grown on 2% microcrystalline cellulose(Avicel), were purified to homogeneity by anion-exchange and gel filtration column chromatography systems. SDS-PAGE analysis indicated that the molecular masses of the purified enzymes were 47 kDa and 35 kDa, respectively. The amino acid sequence analysis of the 47-kDa protein(EG47) showed a sequence similarity with fungal glycoside hydrolase family 5 endoglucanase from the white-rot fungus Phanerochaete chrysosporium. N-terminal and internal amino acid sequences of the 35-kDa protein(EG35), however, had no homology with any other glycosylhydrolases, although the enzyme had high specific activity against carboxymethyl cellulose, which is a typical substrate for endoglucanases. The initial rate of Avicel hydrolysis by EG35 was relatively fast for 48 h, and the amount of soluble reducing sugar released after 96 h was $100{\mu}g/ml$. Although EG47 also hydrolyzed Avicel, the hydrolysis rate was lower than that of EG35. Thin layer chromatography analysis of the hydrolysis products released from Avicel indicated that the main product was cellobiose, suggesting that the brown-rot fungus possesses processive EGs capable of degrading crystalline cellulose.

Development of Mathematical Model for the Hydrolysis Fish Oil (물고기 기름의 가수분해에 대한 수학적 모형개발)

  • Kim Won-Ho;Lee Yong-Hoon;Park Ji-Suk;Hur Byung-Ki
    • KSBB Journal
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    • v.20 no.2 s.91
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    • pp.106-111
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    • 2005
  • The functional relationship between the number of mole of an i-fatty acid (Si) included in fish oil and the hydrolysis time(t) was expressed as a mathematical model, $S_i=-{\alpha_i}1n(t)+\beta_i$. The average errors of calculated values on the basis of the measured values were distributed in the range of less than $5\%$ for all the 15 fatty aids composing of fish oil. The equation of hydrolysis rate of each fatty acid was deduced as $v_i={\gamma_i}exp(\frac{S_i}{\alpha_i})$ from the above-mentioned $S_i=-{\alpha_i}ln(t)+{\beta_i}$. Therefore the hydrolysis yields of fatty acids were analyzed using the equation of $S_i\;Vs.\;t.$. The 15 fatty acids were categorized into 4groups from the view point of hydrolysis yield. The hydrolysis yields of the first group, including C14:0, C16:0, C16:1, C18:0, C18:1 (n-7) and 1l8:1 (n-9), were higher than $70\%$ at 48 hr of hydrolysis. Those of the second group, C20:1, C22:1, C18:3, C20:4 and C20:5, were distributed from $40\%,\;to\;60\%$, and third group were around $30\%$. The final group containing only C22:6 was very hard to be hydrolyzed and the yield was less than $20\%$ at the same time.

Kinetics and Mechanism of the Hydrolysis of Imidoyl Halides (Imidoyl Halide의 가수분해 반응메카니즘과 그의 반응속도론적 연구)

  • Tae-Rin Kim;Jin-Hee Kim;Byung-Doo Chang;Kwang-Il Lee;Ung-Cho Kim
    • Journal of the Korean Chemical Society
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    • v.20 no.1
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    • pp.48-55
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    • 1976
  • The rate constants of the derivatives of N-(2,4-dinitrophenyl)-benzimidoyl chloride were determined at various pH and a rate equation which can be applied over wide pH range was obtained. The reaction mechanism of hydrolysis of N-(2,4-dinitrophenyl)-benzimidoyl chloride which has not been studied carefully earlier in acidic and basic solution can be fullly explained by the rate equation obtained. The rate equation reveals that, beow pH 7.00, the hydrolysis of benzimidoyl chloride proceeds through $S_N2$ reaction to form a carbonium ion intermediate.Above pH 8.5, however, the hydrolysis proceeds through the $S_N2$ type reaction which depends on hydroxide ion and imidoyl chloride concentration. At pH 7.0∼8.5, two reactions occur competitively.

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Aspergillus niger가 생성하는 생전분 분해효소의 정제와 특성

  • 정만재
    • Microbiology and Biotechnology Letters
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    • v.25 no.2
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    • pp.166-172
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    • 1997
  • Aspergillus niger was selected as a strain producing the potent raw starch hydorlyzing enzyme. These experiments were conducted to investigate the conditions of the glucoa- mylase production, the purification of the enzyme, some characteristics of the purified enzyme and hydrolysis rate on various raw starches such as com, rice, potato, glutinous rice, sweet potato, wheat and barley. The optimum cultural temperature and time for the enzyme production on wheat bran medium were $30^{\circ}C$ and 96hrs, respectively. The respective addition of yeast extract and nutrient broth on wheat bran medium increased slightly the enzyme production. The enzyme was purified by ammonium sulfate fractionation and DEAE-cellulose column chromatography. The specific activity of the purified enzyme was 30.7u/mg-protein and the yield of enzyme activity was 25.8%. The purified enzyme showed a single band on polyacrylamide disc gel electrophoresis and its molecular weight was estimated to be 56,000 by SDS-polyacrylamide disc gel electrophoresis. The isoelectric point for the purified enzyme was pH3.7. The optimum temperature and pH were $65^{\circ}C$ and pH 4.0, respectively. The purified enzyme was stable in the pH range of pH 3.0-9.5 and below $45^{\circ}C$, and its thermal stability was slightly increased by the addition of $Ca^{2+}$. The purified enzyme was activated by $Co^{2+},\;Sr^{2+},\;Mn^{2+},\;Fe^{2+},\;Cu^{2+}$. Raw rice starch, raw corn starch, raw glutinous rice starch, raw sweet potato starch, raw wheat starch and raw barley starch showed more than 90% hydrolysis rate in 48hrs incubation. Even raw potato starch, most difficult to be hydrolyzed, showed 80% hydrolysis rate. The purified enzyme was identified as glucoamylase.

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Optimization of fish oil extraction from Lophius litulon liver and fatty acid composition analysis

  • Hu, Zhiheng;Chin, Yaoxian;Liu, Jialin;Zhou, Jiaying;Li, Gaoshang;Hu, Lingping;Hu, Yaqin
    • Fisheries and Aquatic Sciences
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    • v.25 no.2
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    • pp.76-89
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    • 2022
  • The Lophius litulon liver was used as raw material for the extraction of fish oil via various extraction methods. The extraction rate by water extraction, potassium hydroxide (KOH) hydrolysis and protease hydrolysis were compared and the results revealed the protease hydrolysis extraction had a higher extraction rate with good protein-lipid separation as observed by optical microscope. Furthermore, subsequent experiments determined neutrase to be the best hydrolytic enzyme in terms of extraction rate and cost. The extraction conditions of neutrase hydrolysis were optimized by single-factor experiment and response surface analysis, and the optimal extraction rate was 58.40 ± 0.25% with the following conditions: enzyme concentration 2,000 IU/g, extraction time 1.0 h, liquid-solid ratio 1.95:1, extraction temperature 40.5℃ and pH 6.5. The fatty acids composition in fish oil from optimized extraction condition was composed of 19.75% saturated fatty acids and 80.25% unsaturated fatty acids. The content of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were 8.06% and 1.19%, respectively, with the ratio (6.77:1) surpassed to the recommendation in current researches (5:1). The results in this study suggest protease treatment is an efficient method for high-quality fish oil extraction from Lophius litulon liver with a satisfactory ratio of DHA and EPA.

Determination of Biodegradation Rate on Dichlorvos and Methidathion (Dichlorvos와 methidathion의 생분해율의 측정)

  • Min, Kyung-Jin;Cha, Chun-Geun
    • Journal of Environmental Health Sciences
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    • v.25 no.3
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    • pp.36-43
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    • 1999
  • The present study was performed to investigate biodegradation rate of dichlorvos and methidathion. In the biodegradation test of two pesticides by the modified river die-away method from June 17 to August 22, 1998, the biodegradation rate constants and half-life were determined in Nakdong(A) and Kumho River(B). Biodegradation rate of dichlorvos was 4.51% in A sampling point, 6.88% in B sampling point after 7 days. Biodegradation rate constants and half-life of dichlorvos were 0.0066 and 105 days in A sampling point, 0.0102 and 67.9 days in B sampling point, respectively. Biodegradation rate of methidathion was 23% in A sampling point, 36% in B sampling point after 7 days. Biodegradation rate constants and half-life of methidathion were 0.0377 and 18.4 days in A sampling point, 0.0641 and 10.8 days in B sampling point, respectively. Biodegradation rate of methidathion was faster than that of dichlorvos. This suggested that the difference in biodegradation of pesticides was due to difference in the water quality and standard plate counts in the Nackdong and Kumho Rivers. The result of correlation analysis between biodegradation rate constants of the pesticides and water quality(DO, BOD, SS, ABS, NH$_3$-N, and NO$_3$-N) showed significant correlation with BOD, SS and NH$_3$-N at the 5% significant level. A significant linear equation was obtained from regression analysis at the 5% significant level, whereas, dependent variables were BOD, SS and NH$_3$-N, and the biodegradation rate constant was independent variable. It is suggested that dichlorvos will be mainly degraded by hydrolysis, and for methidathion was both hydrolysis and biodegradation. A significant QSAR equation was obtained from regression analysis at the 10% significant level, whereas, dependent variable is biodegradation rate constants of BPMC, chlorothalonil, dichlorvos and methidathion, vapor pressures, partition coefficients and water solubilities of the pesticides are independent variables. Also, a significant linear equation was obtained from regression analysis at the 1% significant level, whereas, dependent variable is biodegradation rate constants of BPMC, chlorothalonil, dichlorvos and methidathion, hydrolysis rate constants of the pesticides are independent variables. It is suggested that the pesticides will be degraded by main degradation factor when the pesticides was affected both hydrolysis and biodegradation.

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Effect of Soil Water Contents on Urea Hydrolysis and Nitrification in a Newly Reclaimed Tidal Soils

  • Park, Mi-Suk;Kim, Hye-Jin;Chung, Doug-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.1
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    • pp.48-52
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    • 2011
  • The effect of soil water content on the transformation potential of N compounds derived from hydrolysis of urea applied in a reclaimed tidal soils which was saline-sodic was observed to evaluate nitrification rates of urea. Soil samples were collected from Moonpo series at the newly reclaimed area in Saemanguem. For the transformation potential of N compounds from urea (46% N), newly reclaimed tidal soils (RS) were amended with urea at the rates of 0, 10, and 20 kg $10a^{-1}$. With leachate obtained from the incubated RS in a leaching tube at $25^{\circ}C$, urea hydrolysis and nitrification were measured for a total of 30days. The cumulative amounts of $NO_3{^-}$-N in each of the four soils treated with urea was linear with time of incubation. Results showed that increase in pH occurred with increasing application rate of urea and volumetric water content due to hydrolysis of urea. The total N in the RS was decreased with incubation time, indicating that rates of urea hydrolysis was influenced by soil moisture conditions. Also, the cumulative amount of nitrate in RS gradually increased with increase in time of incubation.

Enzymatic Hydrolysis of Pretreated Chitin by Aspergillus carneus Chitinase

  • Mohamed, Abdel-Naby;Kwon, Dae-Young
    • Journal of Microbiology and Biotechnology
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    • v.2 no.3
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    • pp.197-203
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    • 1992
  • Studies of the pretreatment of chitin and its subsequent hydrolysis by Aspergillus carneus chitinase are reported. Ball milling was found to be the most effective way among the pretreatment methods tested. Data are presented describing the effect of enzyme and substrate concentrations on the rate and extent of the hydrolysis process. It was found that the successive addition of enzyme improved the saccharification yield. Significant product inhibition of the chitinase was observed when N-acetylglucosamine concentration was 3.6% or higher. Adsorption of enzymes to the substrate occurred during a 24 hr hydrolysis period. An initial rapid and extensive adsorption occurred, followed by gradual desorption which increased during the time of reaction. Intermediate removal of the hydrolyzate and continuation of the hydrolysis by adsorbed enzyme on the residual chitin was also investigated. A total of 75.4 g/l reducing sugars, corresponding to 69.2% saccharificaton yield (as N-acetylglucosamine) was obtained. In addition an increase in the amount of recoverable enzymes was observed under these conditions. Evidence presented here suggests that the technique, whereby the free enzymes in the recovered hydrolyzate are re-adsorbed onto the new substrate, may provide a means of recirculating the dissolved enzymes.

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Purification and Characteristics of Glucoamylase in Aspergillus oryzae NR 3-6 Isolated from Traditional Korean Nuruk

  • Yu, Tae-Shick;Kim, Tae-Hyoung;Joo, Chong-Yoon
    • Journal of Microbiology
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    • v.37 no.2
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    • pp.80-85
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    • 1999
  • The purification system of glucoamylase (glucan 1,4-${\alpha}$-glucosidase, EC 3. 2. 1. 3), some characteristics of the purified enzyme and hydrolysis rate of various raw starch were investigated through several experiments. The enzyme was produced on a solid, uncooked wheat bran medium of Aspergillus oryzae NR 3-6 isolated from traditional Korean Nuruk. The enzyme was homogeneously purified 6.8-fold with an overall yield of 28.3% by the criteria of disc- and SDS-polyacrylamide gel electrophoresis. The molecular weight was estimated to be 48 kDa by SDS-PAGE. The optimum temperature and pH were 55$^{\circ}C$ and 4.0, respectively. The enzyme was stable at a pH range of 3.0∼10.0 and below 45$^{\circ}C$. Enzyme activity was inhibited about 27% by 1mM Hg2+. The hydrolysis rate of raw wheat starch was shown to be 17.5-fold faster than the hydrolysis rate of soluble starch. The purified enzyme was identified as glucoamylase because the product of soluble starch by the purified enzyme was mainly glucose by thin layer chromatography.

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Conversion of water-insoluble components of the basidiocarps of ganoderma lucidum to water-soluble components by hydrolyzing with chitinase

  • Cheong, Jae-Yeon;Park, Won-Bong
    • Archives of Pharmacal Research
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    • v.19 no.4
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    • pp.326-334
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    • 1996
  • We investigated the optimum conditions for conversion of water-insoluble components of basidiocarps of Ganoderma lucidum to water-soluble components by hydrolyzing with chitinase. We also tried it with Ganoderma luciclum residue remaining after extracting hot water-soluble components of Ganoderma lucidum. After hydrolyzing under optimum conditions (20 ppm chitinase, 2% Ganoderma lucidum or 6% Ganoderma lucidum residue, at pH 3 and at $ 35^{\circ}C$), the contents of total water-soluble components (polysaccharide or protein) were measured, and it was found that the contents of water-soluble components increased to 1.5-2.7 fold. Michaelis constant, $K_m$ and maximum rate, $V_max$ calculated by Lineweaver-Burk plot for hydrolysis of Ganoderma lucidum were 1.75% and 0.02%/min respectively and those for hydrolysis of Ganoderma lucidum residue were 53.15% and 0.53%/min respectively The protein-bound polysaccharide was isolated after hydrolysis and molecular weights were measured by Sepharose CL-4B gel filtration and compared with the molecular weights of polysaccharide before hydrolysis.

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