• Title/Summary/Keyword: enzymatic hydrolysis rate

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pH-Controlled Synthesis of Cephalexin by a Purified Acetobacter turbidans Ampicillin Acylase

  • Nam, Doo-Hyun;Ryu, Yeon-Woo;Dewey D.Y Ryu
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.329-332
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    • 2001
  • It has been known that, in enzymatic synthesis of cephalexin, the conversion yield was reduced by high loading of ampicillin acylase. In order to elucidate this phenomena, pH-controlled synthesis of cephalexin was examined using a purified Acetobacter turbidans acylase. When the pH of the reaction mixture was maintained at $6.20{\pm}0.04$, the reduction of the maximal conversion rate was not observed even with high enzyme loading. The kinetic parameters also suggest that pH drop during the enzymatic synthesis of cephalexin was mainly attributed to the rapid hydrolysis of D-${\alpha}$-phenylglycine methyl ester to D-${\alpha}$-phenylglycine, rather than the disappearance of 7-amino-3-deacetoxycephalosporanic acid for cephalexin synthesis. At higher molar ratio of two substrates, [D-${\alpha}$-phenylglycine methyl ester]/[7-amino-3-deacetoxycephalosporanic acid], the conversion rate was also elevated under pH-controlled enzymatic synthesis, which implies that the main reason for the pH drop is due to the production of D-${\alpha}$-phenylglycine methyl easter, the effect of a water-methanol cosolvent system on the ester, the effect of a water-methanol cosolvent system on the conversion profile was also examined. Even the though the conversion rate was increased in 10% methanol solution, a higher than 16% methanol in the reaction mixture caused an inactivation of enzyme.

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Reaction Optimization for Enzymatic Synthesis of Astragalin (효소를 이용한 아스트라갈린 합성 반응의 최적화)

  • Lee, Seul Bi;Chung, Dae-won
    • Applied Chemistry for Engineering
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    • v.23 no.4
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    • pp.394-398
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    • 2012
  • Astragalin (AS), kaempferol monoglycoside, is classified as a polyphenol, and a minute quantity of AS is known to be present in several plants. Recently, it was reported that AS can be prepared by the partial hydrolysis of camelliaside A (CamA) and camelliaside B (CamB) in the tea seed extract (TSE) in the presence of a commercial enzyme complex such as Mash. In this paper, the effects of reaction temperature, amount of enzyme, and the substrate concentration on the reactivity were investigated. As the reaction temperature or the amount of enzyme increased, the reaction rate to produce AS increased, however, the hydrolysis of AS into KR was also enhanced. As a conclusion, the reaction, when 2 mL of Mash to 1 g of TSE was applied with a substrate concentration of 15% at $50^{\circ}C$, was found to be optimum, based on the reaction rate and the selectivity to AS.

Effects of Surfactant Tween 80 on Enzymatic Accessibility and Degradation of Orchardgrass (Dactylis glomerata L.) at Different Growth Stages

  • Goto, M.;Bae, Hee-Dong;Yahaya, M.S.;Karita, S.;Wanjae, K.;Baah, J.;Sugawara, K.;Cheng, K.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.1
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    • pp.83-87
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    • 2003
  • The study evaluates the enzymatic dry matter (DM) degradability and water holding capacity of leaf and stem fractions of orchardgrass (Dactylis glomerata L.) at different growth stages with or without the presence of surfactant Tween 80. While Tween 80 significantly (p<0.05) increased water and enzyme holding capacities in the leaf blades fraction, less was observed in the fraction of leaf sheath and stem of orchardgrass. The enzyme holding capacity in the leaves was also altered more than that for water holding capacity. This resulted in the increased rate and extent of enzymatic hydrolysis of the leaf blade fractions at two growth stages, whereas little was with leaf sheath and stem fractions. It was also observed that at 0.005% concentrations of Tween 80 the enzymatic DM degradability of young leaf blades was higher (p<0.05) by 20-30% compared to that of the control, as well as for water and enzyme holding capacity. For matured leaf blades the DM degradability were increased with over 0.01% concentrations of the surfactant, but the increase was less than leaf blades of young orchardgrass. This result suggests the possibility of using the surfactant Tween 80 to improve forage digestibility in the rumen.

Development and Evaluation of the Attrition Coupled Bioreactors for Enzymatic Hydrolysis of Biomass ; Tumbling-Drum Type Bioreactor for Enzymatic Hydrolysis of Cellulose (Biomass의 고효율 효소당화에 적합한 Attrition Coupled Bioreactor 개발에 관한 연구 ; Tumbling Drum Type Bioreactor를 활용한 섬유소 당화)

  • 이용현;조구형;박진서
    • KSBB Journal
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    • v.4 no.2
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    • pp.87-93
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    • 1989
  • To develop high dfficiency-low energy consumption attrition coupled bioreactor for enhanced enzymatic hyerolysis of insoluble biomass, a tumbling drum type bioreactor was installed, and its efficiency was evaluated. The effects of drum structure and poerational conditions were investigated. The optimal saccharification at 3L drum was obtained at 8 baffled drum, drum diameter to baffle height ratio of 1:0.05, 100rpm, and addition of 600g of 3mm glass bead per liter. The consumed power for rolling of drum and energy consumption for half digestion of cellulose were measured, and compared with enhanced rate and yield to predict the economic prospect of the process. The tumbling drum type bioreactor seems to have appropriated structure for industrial scale operation, and further investigation for scale-up need to be conducted.

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Effect of External and Intramolecular Nucleophiles on Nature of Products of Carboxypeptidase A-Catalyzed Hydrolysis of Esters. Attempted Trapping of Acyl-Enzyme Intermediate (카르복시펩티다제A의 에스테르 가수분해 반응생성물의 종류에 대한 외부 및 분자내 친핵체의 영향. 아실-효소중간체의 포획시도)

  • Junghun Suh;Emil Thomas Kaiser
    • Journal of the Korean Chemical Society
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    • v.22 no.3
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    • pp.164-172
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    • 1978
  • Carboxypeptidase A-catalyzed hydrolysis of ester substrates was carried out at room temperature in the presence of a number of external reagents. If the acyl-enzyme intermediate, an anhydride, is attacked by the external reagents, products formed by trapping at the acyl portion or at the enzyme portion of the anhydride group can be obtained. Examination of the uv/vis spectral properties of the reaction products and of changes in enzyme activity indicated that such trapping reactions did not occur. Also performed was evaluation of enzymatic rate parameters for the the hydrolysis of O-(o-hydroxyphenylacetyl)-L-${\beta}$-phenyllactate. Detection of 2-coumaranone possibly formed by attack of the o-hydroxy group as an intramolecular trapping group at the acyl-enzyme intermediate was tried, but no evidences for the intramolecular trapping reaction were obtained. Failure to trap the intermediate was discussed in terms of steric hindrance imposed on the approach of the trapping reagents to the anhydride group of the acyl-enzyme intermediate and of the fast enzymatic breakdown of the intermediate.

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Enzymatic Hydrolysis of Crystalline Chitin in an Agitated Bead Reaction System and Its Reaction Characteristics

  • Lee, Yong-Hyun;Bae, Young-Ki;Jeong, Eui-Jun
    • Journal of Microbiology and Biotechnology
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    • v.6 no.6
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    • pp.432-438
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    • 1996
  • Native crystalline chitin was hydrolyzed in an agitated bead reaction system using crude chitinase excreted from Aspergillus fumigatus JC-19. The reaction was enhanced significantly, and the concentration and yield of reducing sugar after 48 hours were measured to be 35.42 g/I (w/v) and 0.64, respectively, around 1.86 times higher than those of the conventional system that was carried out without glass beads. The effect of reaction conditions, such as the amounts of chitin, chitinase and glass beads, and the size of glass bead, were examined. Ball milled chitin was also hydrolyzed in the agitated bead reaction system, the conversion yield and reaction rate of ball milled chitin for 24 hours increased up to 0.87 and 48.02 g/I, respectively. Chitinase showed relatively high stability in the agitated bead reaction system, particularly in the presence of enzyme stabilizer, $Ca^{++}$, which played a critical role in preventing the deactivation of chitinase by the physical impact of glass beads. The variations of the structural features of chitin during the reaction were followed by SEM and X-ray diffraction, and the enhanced hydrolysis reaction was caused by both the fragmentation of chitin particles and the destruction of the crystalline structure owing to the synergic effects of the attrition of glass beads and the hydrolytic action of chitinase.

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Design of Recycle Bubble Column Reactor for Continuous Enzymatic Hydrolysis of Cellulose (섬유소의 연속 효소 가수분해를 위한 순환식 기포탑 반응기의 설계)

  • 김춘영;홍석표정봉우이태원
    • KSBB Journal
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    • v.5 no.1
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    • pp.59-67
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    • 1990
  • Enzymatic hydrolysis of insoluble cellulose was performed in a bubble column with tangential flow ulrafiltration membrane unit. The reactor was operated in a batch mode as well as semi-continuous and continuous with continuous removal of products through the tangential flow ultrafiltration membrane. The optimum superficial gas velocity was 1-3cm / sec so as to avoid bubble coalescence and enzyme denaturation. In continuous and selni-cotinuous process, the conversion was gradually increased but the total reduced sugar concentration was drcastically dereased with the dilution rate. It was concluded that the bubble column attaching tangential flow ultrafiltration membrane unit was effective on continuous hydrolysis of cellulose and recovery of enzyme.

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Kinetic Studies on Enzymatic Hydrolysis of Cellulose(II) - Evaluation of Several Factors for Enzyme Adsorption and Initial Hydrolysis - (섬유소 가수분해반응에 관한 연구(II) - 효소흡착과 가수분해반응에 관여하는 여러인자의 영향 -)

  • Lee, Yong-Hun;Kim, Chul
    • KSBB Journal
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    • v.6 no.2
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    • pp.167-174
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    • 1991
  • Enzymatic cellulose hydrolysis depends on the several factors such as the structural features (CrI, particle size and surface area, etc.), the nature of cellulase enzyme system, the inhibitory effects of products, and enzyme deactivation. At the presence of products on the initial hydro- lysis rate of cellulose, cellobiose has more severe inhibitory effect than glucose. Othewise, the inhibition effect of products for adsorbed enzyme is related to the glucose and cellobiose conentration hyperbolically. Enzyme deactivation of FPA and ${\beta}-glucosidase$ were expressed by exponential decay profile.

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Effects of Lactobacillus helveticus Fermentation on the Ca2+ Release and Antioxidative Properties of Sheep Bone Hydrolysate

  • Han, Keguang;Cao, Jing;Wang, Jinghui;Chen, Jing;Yuan, Kai;Pang, Fengping;Gu, Shaopeng;Huo, Nairui
    • Food Science of Animal Resources
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    • v.38 no.6
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    • pp.1144-1154
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    • 2018
  • Both the calcium and collagen in bone powder are hard to be absorbed by the body. Although enzymatic hydrolysis by protease increased the bio-availability of bone powder, it was a meaningful try to further increase $Ca^{2+}$ release, oligopeptide formation and antioxidant activity of the sheep bone hydrolysate (SBH) by lactic acid bacteria (LAB) fermentation. Lactobacillus helveticus was selected as the starter for its highest protease-producing ability among 5 tested LAB strains. The content of liberated $Ca^{2+}$ was measured as the responsive value in the response surface methodology (RSM) for optimizing the fermenting parameters. When SBH (adjusted to pH 6.1) supplemented with 1.0% glucose was inoculated 3.0% L. helveticus and incubated for 29.4 h at $36^{\circ}C$, $Ca^{2+}$ content in the fermented SBH significantly increased (p<0.01), and so did the degree of hydrolysis and the obtaining rate of oligopeptide. The viable counts of L. helveticus reached to $1.1{\times}10^{10}CFU/mL$. Results of Pearson correlation analysis demonstrated that LAB viable counts, $Ca^{2+}$ levels, obtaining rates of oligopeptide and the yield of polypeptide were positively correlated with each other (p<0.01). The abilities of SBH to scavenge the free radicals of DPPH, OH and ABTS were also markedly enhanced after fermentation. In conclusion, L. helveticus fermentation can further boost the release of free $Ca^{2+}$ and oligopeptide, enhance the antioxidant ability of SBH. The L. helveticus fermented SBH can be developed as a novel functional dietary supplement product.

Microbial Conversion of Woody Waste into Sugars and Feedstuff (II) - Production of Cellulolytic Enzymes from Aspergillus fumigatus and Saccharification of Popla Wood (미생물(微生物)에 의한 목질자원(木質資源)의 당화(糖化) 및 사료화(飼料化)에 관(關)한 연구(硏究) (II) - Aspergillus fumigatus KC-1으로부터 섬유소 분해 효소의 생산 및 현사시나무의 효소가수분해)

  • Chung, Ki-Chul;Huh, Jeong-Weon;Myung, Kyu-Ho;Kim, Yoon-Soo
    • Journal of the Korean Wood Science and Technology
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    • v.15 no.4
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    • pp.18-25
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    • 1987
  • The cellulolytic activities of Aspergillus fumigatus KC-1 was investigated, which showed the most active producer of cellulase among the 256 strains of cellulose-decomposing microorganisms screened in our laboratory. All the examined cellulolytic activities (filter paper-, Avicel-, cotton-, CMC-, salicin- and xylansaccharifying activity) in a culture of A. fumigatus KC-1 grown on 1% popular sawdust pretreated with peroxide alkaline reached a maximum within 4-5 days. The optimum pH and temperature for the enzymatic activity was found to be pH 4.5 and $60^{\circ}C$ respectively. The sawdust of poplar wood delignified with 1% NaOH and 20% peracetic acid succesively recorded the highest hydrolysis rate in the tests of enzymatic saccharification. The major end product of hydrolysis of poplar wood with the cellulolytic enzymes obtained from A. fumigatus KC-1 was glucose with small amount of cellobiose and xylose. It can be concluded from these results that A. fumigatus KC-1 is an advantagous source of a cellulase that is capable of hydrolyzing cellulose to glucose rapidly. The influence of degree of delignification, substrate size and its concentration on the rate of hydrolysis of poplar wood was also discussed.

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