• Korean Journal of Food Science and Technology
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• v.27 no.5
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• pp.753-756
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• 1995

Optimum conditions for immobilization of thermolysin, a metalloendopeptidase catalyzing synthesis of aspartame precursors, were investigated with using Amberlie XAD-7 as carrier and glutaraldehyde as cross-linking agent. Adsorption of thermolysin onto the carrier was rapid at the initial stage and 96% of the enzyme was adsorbed after 24 hours at $5^{\circ}C$. There was a linear relationship between amount of thermolysin adsorbed and thermolysin loaded upto 300g per liter of carrier. The effective range of cross-linking time, concentration of glutaraldehyde and pH for immobilization of the enzyme were $3{\sim}7\;hours,\;6{\sim}12.5%\;and\;pH\;6.0{\sim}7.0$, respectively. Degree of cross-linking and residual enzyme activity were high when cross-linked for 7 hours with 6% glutaraldehyde or for 3 hours with 12.5% glutaraldehyde. The residual enzyme activity was over 30% under these conditions.

• Journal of Microbiology and Biotechnology
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• v.7 no.2
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• pp.107-113
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• 1997

A strain producing a high amount of chitinase was isolated from soil, identified as Pseudomonas sp., and tentatively named Pseudomonas sp. YHS-A2. An extracellular chitinase of Pseudomonas sp. YHS-A2 was purified according to the procedure of ammonium sulfate saturation, affinity adsorption, Sephadex G-100 gel filtration and Phenyl-sepharose CL-4B hydrophobic interaction column chromatography. The molecular weight of the purified enzyme was estimated to be 55 kDa on SDS-PAGE was confirmed by active staining. Optimal pH and temperature of the enzyme are pH 7.0 and $50^{\circ}C$, respectively, and the enzyme is stable between pH 5.0 and 8.0 and below $50^{\circ}C$. The main products of colloidal chitin by the chitinase were N-acetyl-D-glucosamine and N,N'-diacetylchitobiose both of which were detected by HPLC analysis. The enzyme is supposed to be a random-type endochitinase which can degrade any position of ${\beta}$-l,4-linkages of chitin and chitooligosaccharides. The chitinase inhibited the growth of some phytopathogenic fungi, Fusarium oxysporum, Botrytis cineria, and Mucor rouxii and these antifungal effects were thought to be due to the characteristics of endochitinase.

• Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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• 2006.06a
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• pp.43-49
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• 2006

The surface and pore structure of cellulose fibers have a significant impact on the properties and performance in applications. Cellulase enzymatic hydrolysis of cellulose fibers can result in changes to the surface and pore structure thus providing a useful tool for fiber modification. This research characterizes these changes using various test methods such as fiber dimension, water retention value, hard-to-remove water content, freezing and non-freezing bound water content, polymer adsorption, and crystallinity index. For a high-dosage enzyme treatment (0.10 g/g), the fiber length was significantly decreased and the fibers were 'cut' in the cross direction, not in the axial direction. The swelling capacities as measured by the WRV and HR water content increased for the high-dosage treatment. Three independent measurements (non-freezing bound water, polymer adsorption, and crystallinity index) are in good agreement with the statement that the amorphous regions of cellulose fibers are a more readily available substrate relative to crystalline regions. Based on the experimental results obtained herein, a model was proposed to explain surface and pore structure modification of cellulose fibers via enzymatic treatment.

• Journal of the East Asian Society of Dietary Life
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• v.23 no.6
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• pp.789-798
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• 2013

In the present study, optimization on ${\beta}$-glucanase-assisted extraction was made in order to isolate waxy barley starch from domestic cultivar using the D-optimal design suitable for response surface methodology (RSM). The results demonstrated that the amount of enzyme was found to be a major influencing factor on the extraction yield, which was substantially increased by increasing the amount of enzyme. It was also influenced by the reaction time and amount of water addition; however, the two factors were less influential than the amount of enzyme. The optimized condition by RSM for the reaction time was found to be 2.63 hours and amount of enzyme 1.7%, and amount of water addition 4.38 times the weight of raw material. With the enzyme treatment, the starch content in residues (R), particularly in R1 and R5, was reduced considerably, resulting in an increase in the extraction yield and therefore primarily and effectively releasing B-type starch small granule confirmed by scanning electronic microscopy. In addition, the study determined the physicochemical properties of isolated waxy starch (i.e., purity, water adsorption capacity, thermal properties, rheology and starch morphology) and compared them with those from the enzyme-not treated sample. It was found that they were almost similar to each other, except for the purity of starch, which was lower in the enzyme-treated sample than in the enzyme-not treated one.

• Journal of Life Science
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• v.13 no.1
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• pp.90-98
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• 2003

An allium rhizobacterium Serratia plymuthica AL-1 was previously selected as a biocontrol agent of allium white rot. The chitinase from S. plymuthica AL-1 produced in medium containing colloidal chitin was purified by ammonium sulfate precipitation (40~70%), affinity adsorption, column chromatography on DEAE-sephadex A-50 and sephadex C-200 gel filtration. The enzyme was purified 10.8-fold with a yield of 7.3% from the starting culture broth. The purified chtinase gave a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis, it's molecular weight was estimated to be 55 kDa. The optimum pH and temperature of the purified enzyme were pH 5.5 and $55^{\circ}C$, respectively and it is stable up to $50^{\circ}C$ and maintains around 90% of its activity for 60min. The enzyme were activated by $Ca^{2+}$, $Mn^{2+}$ and $Mg^{2+}$ and inhibited by $Cu^{2+}$, SDS, $\rho$-CMB, MIA, respectively. The purified chitinase showed broad spectrum of antifungal activities against plant pathogenic fungi Sclerotium cepivoruin, Alternana alternnta, Colletotrichum glceosporioidrs, Phoma sp., Sclerotinia sclerotiorum, Stemphylium solani, Fusarium oxysporium f. sp. niveum but rarely inhibited Phytophthora capsici and Pythium ultimum.. The purified chitinase from S. plymuthica AL-1 caused swelling, lysis, deceleration and degradation of the hyphal tips of S. sczerotiorum causing allium white rot. It suggest that S. prymuthica AL-1 chitinase play an important part in the bifunctional chitinase / lysozyme activity.

• KSBB Journal
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• v.24 no.5
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• pp.415-419
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• 2009

In this study, the detoxification methods were evaluated for the removal of fermentation inhibitors from synthetic solution containing the composition similar to the lignocellulosic hydrolysate. The enzyme peroxidase and laccase were used as a biological treatment method. The physico-chemical methods such as adsorption and ion exchange were applied by using activated charcoal and ion exchange resins. The enzyme peroxidase showed a excellent removal of phenolic compounds. The 5-HMF and furfural were completely removed by activated charcoal. The anion exchange resin showed a good result for detoxification of acetic acid. The activated charcoal and ion exchange resins lead to a loss of sugars more or less. The choice of detoxification method must be made after considering the composition and inhibitors in hydrolysates.

• Korean Journal of Food Science and Technology
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• v.12 no.3
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• pp.176-181
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• 1980

Yeast invertase was immobilized on the 2 kinds of matrices : one is an indirectly coupled enzyme to the cyanogen bromide activated Sepharose by using ${\omega}-aminohexyl$ group as an extension arm, and the other is a tightly adsorbed enzyme on the modified hydrophobic cellulose derivative which has a phenoxyacetyl group as a linkage. The enzyme preparation coupled on Sepharose retained 26.0% of the original activity against sucrose as a substrate, while the preparation immobilized on phenoxyacetyl cellulose retained 72.9% . The immobilized invertase preparation on ${\omega}-aminohexyl$ Sepharose showed the optimal pH 4.5, optimal temperature $60^{\circ}C$, activation energy $5,941\;cal/mole{\cdot}deg$ and Km' 22.2 mM against sucrose, while the preparation adsorbed on phenoxyacetyl cellulose showed the optimal pH 4.0, optimal temperature $60^{\circ}C$, activation energy $7,769\;cal/mole{\cdot}deg$ and Km' 69.9 mM.

• Journal of Life Science
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• v.23 no.11
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• pp.1365-1370
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• 2013

Immobilized cellulase on weak ion exchange resin showed a typical Langmuir adsorption isotherm. Immobilized cellulase had better stability with respect to pH and temperature than free cellulase. Kinetics of thermal inactivation on free and immobilized cellulase followed first order rate, and immobilized cellulase had a longer half-life than free cellulase. The initial rate method was used to characterize the kinetic parameters of free and immobilized enzyme. The Michaelis-Menten constant $K_m$ was higher for the immobilized enzyme than it was for the free enzyme. The effect of the recirculation rate on cellulose degradation was studied in a recycling packed-bed reactor. In a continuous packed-bed reactor, the increasing flow rate of cellulose decreased the conversion efficiency of cellulose at different input lactose concentrations. Continuous operation for five days was conducted to investigate the stability of long term operation. The retained activity of the immobilized enzymes was 48% after seven days of operation.

• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.491-505
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• 2020

A newly isolated strain, Proteus vulgaris OR34, from olive mill waste was found to secrete an alkaline extracellular lipase at 11 U·ml-1 when cultivated on an optimized liquid medium. This lipase was purified 94.64-fold with a total yield of 9.11% and its maximal specific activity was shown to be 3232.58 and 1777.92 U·mg-1 when evaluated using the pH-stat technique at 55℃ and pH 9 and Tributyrin TC4 or olive oil as the substrate. The molecular mass of the pure OR34 lipase was estimated to be around 31 kDa, as revealed by SDS-PAGE and its substrate specificity was investigated using a variety of triglycerides. This assay revealed that OR34 lipase preferred short and medium chain fatty acids. In addition, this lipase was stable in the presence of high concentrations of bile salt (NaDC) and calcium ions appear not to be necessary for its activity. This lipase was inhibited by THL (Orlistat) which confirmed its identity as a serine enzyme. In addition, the immobilization of OR34 lipase by adsorption onto calcium carbonate increased its stability at higher temperatures and within a larger pH range. The immobilized lipase exhibited a high tolerance to organic solvents and retained 60% of its activity after 10 months of storage at 4℃. Finally, the OR34 lipase was applied in biodiesel synthesis via oleic acid mediated esterification of methanol when using hexane as solvent. The best conversion yield (67%) was obtained at 12 h and 40℃ using the immobilized enzyme and this enzyme could be reused for six cycles with the same efficiency.

• The Korean Journal of Pesticide Science
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• v.2 no.3
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• pp.70-78
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• 1998

Dissipation, adsorption and desorption of oxadiazon were examined in two soils containing different amounts of soil organic matter. In addition, reactivity of oxadiazon with humic monomers was searched to clarify binding mechanism of oxadiazon to soil organic matter in the presence of a laccase of Myceliophthera thermophila. Half lives of oxadiazon were 38 days in Soil I and 45 days in Soil II. Freundlich constant, k values of fresh soils were higher than those of oxidized soils. Adsorption rates of oxadiazon were increased 17.1% in Soil I and 9.3% in Soil II in the presence of a laccase but no significant increase was observed in oxidized soils. Desorption rates of oxadiazon in fresh soils were lower than those in oxidized soils. Desorption rates of adsorbed oxadiazon in soils addes with the enzyme were not changed in oxidized soils but decreased in fresh soils. The herbicide oxadiazon alone underwent no transformation by a laccase but in the presence of catechol, guaiacol and gallic acid as humic monomer, transformation rates of it were from 20% to 24%.