• Title/Summary/Keyword: Enzyme Conversion

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Production and Characterization of Multi-Polysaccharide Degrading Enzymes from Aspergillus aculeatus BCC199 for Saccharification of Agricultural Residues

  • Suwannarangsee, Surisa;Arnthong, Jantima;Eurwilaichitr, Lily;Champreda, Verawat
    • Journal of Microbiology and Biotechnology
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    • v.24 no.10
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    • pp.1427-1437
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    • 2014
  • Enzymatic hydrolysis of lignocellulosic biomass into fermentable sugars is a key step in the conversion of agricultural by-products to biofuels and value-added chemicals. Utilization of a robust microorganism for on-site production of biomass-degrading enzymes has gained increasing interest as an economical approach for supplying enzymes to biorefinery processes. In this study, production of multi-polysaccharide-degrading enzymes from Aspergillus aculeatus BCC199 by solid-state fermentation was improved through the statistical design approach. Among the operational parameters, yeast extract and soybean meal as well as the nonionic surfactant Tween 20 and initial pH were found as key parameters for maximizing production of cellulolytic and hemicellulolytic enzymes. Under the optimized condition, the production of FPase, endoglucanase, ${\beta}$-glucosidase, xylanase, and ${\beta}$-xylosidase was achieved at 23, 663, 88, 1,633, and 90 units/g of dry substrate, respectively. The multi-enzyme extract was highly efficient in the saccharification of alkaline-pretreated rice straw, corn cob, and corn stover. In comparison with commercial cellulase preparations, the BCC199 enzyme mixture was able to produce remarkable yields of glucose and xylose, as it contained higher relative activities of ${\beta}$-glucosidase and core hemicellulases (xylanase and ${\beta}$-xylosidase). These results suggested that the crude enzyme extract from A. aculeatus BCC199 possesses balanced cellulolytic and xylanolytic activities required for the efficient saccharification of lignocellulosic biomass feedstocks, and supplementation of external ${\beta}$-glucosidase or xylanase was dispensable. The work thus demonstrates the high potential of A. aculeatus BCC199 as a promising producer of lignocellulose-degrading enzymes for the biomass conversion industry.

Optimum Conditions for Glycoside Conversion to Aglycone by ${\gamma}-Galactosidase$ (유당분해효소에 의한 Glycoside의 Aglycone으로 전환을 위한 최적 조건 확립)

  • Kim, Nam-Chul;Jeon, Byung-Ju;Ahn, Joung-Jwa;Kwak, Hae-Soo
    • Food Science of Animal Resources
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    • v.27 no.3
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    • pp.329-336
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    • 2007
  • This study was designed to find the optimum conditions for converting isoflavone glycoside to aglycone by ${\beta}-galactosidase$. Three different forms of the enzyme were tested and the optimum enzyme concentration, incubation temperature, pH, and incubation time were determined. Before treatment with enzyme, isoflavone contained 89.4% glycoside including daidzin, glycitin and genistin, and only 10.6% aglycone including daidzein, glycitein and genistein. Among the enzymes tested, the highest rate of isoflavone hydrolysis to aglycone, 35%, was observed when 3 unit/g Fungal Lactase (Amano Enzyme) was used. Higher incubation temperatures resulted in a higher rate of hydrolysis along with a greater loss of isoflavone mass. Therefore, body temperature $(37^{\circ}C)$ may be adequate for isoflavone conversion, with 44.9% hydrolysis and less than 10% loss of mass. As expected, a higher amount of aglycone was produced at pH 7 compared with other pH values. During 5hr of incubation, the conversion of glycoside to aglycone increased dramatically from 0 to 1hr, and plateaued thereafter. In addition, commercial soy-based milk was hydrolyzed more effectively with ${\beta}-galactosidase$ when incubated for 5hr. Based on the above results, the optimum conditions for isoflavone hydrolysis by ${\beta}-galactosidase$ were for 3 hr at $37^{\circ}C$, pH 7 with 3 unit/g Fungal Lactase (Amano Lactase), yielding an average total amount of aglycone ranging from 40 to 47%.

Characteristics of Lactose Hydrolysis by Immobilized β-Galactosidase on Chitosan Bead (Chitosan 담체에 고정화된 β-galactosidase에 의한 유당 분해 특성)

  • Kang, Byung-Chul
    • Journal of Life Science
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    • v.21 no.1
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    • pp.127-133
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    • 2011
  • ${\beta}$-Galactosidase was immobilized on chitosan bead by covalent bonding using glutaraldehyde. The characteristics of the immobilized enzyme were investigated. Maximum immobilization yield of 75% was obtained on chitosan bead. Optimum pH and temperature for the immobilized enzyme was 7.0 and $50^{\circ}C$, respectively. The immobilized enzyme showed a broader range of pH and temperature compared to a free one. A mathematical model for the operation of the immobilized enzyme in a packed-bed reactor was established and solved numerically. Under different inlet lactose concentrations and feed flow rate conditions, lactose conversion was measured in a packed-bed reactor. The experimental results of continuous operation in a packed-bed reactor were compared to theoretic results using Michaelis-Menten kinetics with competitive product inhibition and external mass transfer resistance. The model predicted the experimental data with errors less than 5%. Process optimization of continuous operation in a packed-bed reactor was also conducted. In a recirculation packed-bed operation, conversion of lactose was 97% in 3 hours. In a continuous packed-bed operation, the effect of flow rate and initial lactose concentration was investigated. Increasing flow rates and initial lactose concentration decreased the conversion of substrate.

Performance of Broiler Chicks Fed Normal and Low Viscosity Rye or Barley with or without Enzyme Supplementation

  • He, T.;Thacker, P.A.;McLeod, J.G.;Campbell, G.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.2
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    • pp.234-238
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    • 2003
  • This experiment was conducted to measure nutrient digestibility and performance in broiler chicks fed diets based on normal and low viscosity rye or barley fed with and without enzyme (pentosanase and $\beta$-glucanase) during a 17 day growth trial. A total of 150 one-day old, male broiler chicks (5 birds per pen and 5 pens per treatment) were randomly assigned to one of six dietary treatments in a $3{\times}3$ factorial design experiment (3 cereals${\times}$2 enzyme levels). Digestibility coefficients were determined using chromic oxide. Digestibility coefficients for dry matter and crude protein were significantly (p=0.0001) higher for the barley-based diets than for any of the rye-based diets. Digestibility coefficients for gross energy did not differ (p>0.05) due to cereal grain. There were no differences in the digestibility coefficients for dry matter and gross energy between chicks fed normal and low viscosity rye. However, the digestibility coefficient for crude protein was higher (p=0.01) for the low viscosity rye compared with the normal viscosity rye. Addition of enzyme to the diet significantly (p=0.0001) increased digestibility coefficients for dry matter, crude protein and energy. There were no significant differences in weight gain, feed intake or feed conversion between birds fed barley or rye or between birds fed normal or low viscosity rye. Enzyme supplementation significantly improved (p=0.0001) weight gain, intake and feed conversion. The overall results of this experiment indicate that unsupplemented barley and rye do not support adequate growth rates in poultry. Enzyme supplementation dramatically improved broiler performance. In addition, genetic selection to reduce the viscosity of rye had only a modest effect on the nutritive value of rye for broilers.

Energy Consumption of Biodiesel Production Process by Supercritical and Immobilized Lipase Method (초임계와 Lipase 고정화에 의한 바이오디젤 생산 공정의 에너지소비량)

  • Min, Eung-Jae;Lee, Euy-Soo
    • Korean Chemical Engineering Research
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    • v.50 no.2
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    • pp.257-263
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    • 2012
  • Biodiesel is a renewable energy which is nontoxic and acting as a replacement for conventional diesel which derived from fossil fuel. Classified biodiesel producing way such as acid, base, supercritical and enzyme methods, this study focused on eco-friendly production of biodiesel using supercritical and immobilized enzyme process. Assuming a plant with a production rate of 10,000 tons a year, a PRO II simulator program was used to simulate the product conversion rate and total energy consumption. The product conversion in supercritical process and immobilized enzyme was found to be 91.17% (including 0.9% glycerol) and 93.18% (including 1.0% glycerol) respectively. The result shows that the efficiency of immobilized enzyme process is higher compared to supercritical process but having lower end product purity. From the energy consumption point of view, supercritical process consume about 8.9 MW while immobilized enzyme process consume much lower energy which is 3.9 MW. Consequently, this study certifies that energy consumption of supercritical process is 2.3 times higher than immobilized enzyme process.

Prediction of Continuous Reactors Performance Based on Batch Reactor Deactivation Kinetics Data of Immobilized Lipase

  • Murty, V.Ramachandra;Bhat, Jayadev;Muniswaran, P.K.A.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.4
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    • pp.225-230
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    • 2002
  • Experiments on deactivation kinetics of immobilized lipase enzyme from Candida cyl-indracea were performed in stirred bath reactor using rice bran oil as the substrate and temperature as the deactivation parameter. The data were fitted In first order deactivation model. The effect of temperature on deactivation rate was represented by Arrhenius equation. Theoretical equations were developed based on pseudo-steady state approximation and Michaelis -Menten rate expression to predict the time course of conversion due to enzyme deactivation and apparent half-life of the immobilized enzyme activity in PFR and CSTH under constant feed rate polity for no diffusion limitation and diffusion limitation of first order. Stability of enzyme in these continuous reactors was predicted and factors affecting the stability were analyzed.

Purification of the Convertible Enzyme of Ginseng Saponin from Rhizopus japonicus (Rhizopus japonicus가 생산하는 인삼 Saponin 전환효소의 정제)

  • 김상달;서정훈
    • Microbiology and Biotechnology Letters
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    • v.16 no.6
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    • pp.438-442
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    • 1988
  • The enzyme produced by Rhizopus japonicus was able to convert selectively ginsenoside-Rb$_1$which is the most abundant ginseng saponin, into ginsenoside-Rd which was known to be superior to ginsenoside-Rb$_1$pharmaceutically. The convertible enzyme was purified homogeneous from wheat bran culture of Rhizopus japonicus by ammonium sulfate fractionation and column chromatography of TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150, Sepharose 2B. Specific activity of the purified enzyme was increased to a bent 96 folds and yield was appeared to be 11% of culture extract. Evidence for homogenity was obtained from polyacrylamide and SDS-polyacrylamide gel electrophoresis. Molecular weight of the enzyme was estimated about 88, 000 daltons by Sephadex G-l50 gel filtration and SDS-polyacrylamide gel electrophoresis, and it did not consist of any subunit.

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Intestinal Structure and Function of Broiler Chickens on Wheat-Based Diets Supplemented With a Microbial Enzyme

  • Iji, P.A.;Hughes, R.J.;Choct, M.;Tivey, D.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.1
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    • pp.54-60
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    • 2001
  • A study was conducted to assess the nutritive value of two diets based on a low-energy variety of wheat, RAC C1 and their effects on intestinal mucosal structure and function in broiler chickens. The diets were fed with or without microbial enzyme supplement to male and female broiler chickens. The digesta viscosity was reduced (p<0.001) through supplementation with a microbial enzyme in male and female chicks. Enzyme supplementation also improved the dietary apparent metabolizable energy content (p<0.001) and had slight but non-significant positive effects on chick growth and feed conversion ratio. Intestinal mucosal structure and enzyme function were not affected by microbial enzyme supplement. Male chicks consumed more feeds (p<0.001), attained higher final body weight (p<0.001) and were more efficient at feed utilization (p<0.01) than the female chicks. Except for duodenal villus surface area and ileal protein content, intestinal mucosal structure and enzyme activities were similar between the two sexes and dietary treatment groups. The study showed an improvement in the nutritive value of the diets in the presence of the microbial enzyme supplement.

Influence of enzyme mixture supplementation on growth performance, nutrient digestibility, and fecal score in growing pigs

  • Thamaraikannan, Mohankumar;Kim, In Ho
    • Korean Journal of Agricultural Science
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    • v.48 no.2
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    • pp.201-207
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    • 2021
  • This study evaluated the influence of enzyme mixture supplementation on the growth performance, nutrient digestibility, and fecal score of growing pigs. A total of 72 pigs with an initial body weight of 20.23 ± 1.46 kg were randomly assigned to two treatments consisting of a basal diet and the basal diet supplemented with 0.5% enzyme mixture. During a 19-day trial, no significant difference was observed in the body weight (BW) and average daily feed intake (ADFI) of the pigs. However, a gradual increase in the average daily gain (ADG) was observed during the period from day 14 to day 19 and the overall period in pigs fed a diet supplemented with the 0.5% enzyme mixture (p < 0.10) as compared to the pigs that were fed the control diet. From days 4 to 14 and in the overall experiment, a gradual increase in the feed conversion ratio (FCR) (p < 0.10) was observed with the inclusion of 0.5% enzyme mixture supplementation. The nutrient digestibility of dry matter (DM), nitrogen (N), and energy were not affected by enzyme mixture supplementation. In addition, dietary supplementation with the enzyme mixture had no significant effects on the fecal score of growing pigs. In summary, supplementation with the enzyme mixture had beneficial effects on the ADG performance but failed to have a significant effect on growth performance (BW), nutrient digestibility, and fecal score.

Characterization of NAD-Dependent Formate Dehydrogenase from Trametes versicolor Using a Cell-Free Protein Expression System

  • LEE, Su-Yeon;JANG, Seokyoon;LEE, Soo-Min
    • Journal of the Korean Wood Science and Technology
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    • v.50 no.3
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    • pp.159-166
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    • 2022
  • CO2 emissions are the primary reason for global warming; hence, biological and chemical technologies for converting CO2 into useful compounds are being actively studied. Biological methods using enzymes can convert CO2 under mild conditions. Formate dehydrogenase (FDH) is a representative CO2 conversion enzyme. Its function was revealed after isolation from bacteria, yeast, and plants. In this study, we evaluated the CO2 conversion potential of FDH isolated from wood-rotting fungi. After isolating the FDH gene (TvFDH) from Trametes versicolor, we cloned the full-length FDH from T. versicolor and expressed it in a cell-free expression system. The gene encoding TvFDH was identified as 1,200 bp open reading frame (ORF) and the expected molecular weight of the protein was approximately 42 kDa. Overexpression of the recombinant crude protein including TvFDH was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Enzyme activities and metabolite analyses confirmed the efficiency of TvFDH for CO2 reduction.