• KSBB Journal
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• v.23 no.4
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• pp.350-354
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• 2008

Magnetically separable immobilized trypsin was developed and their biocatalytic activity was evaluated for the different immobilization media. The activity, recyclability, pH effect, and stability of immobilized enzymes were evaluated for the different supporting media. The biocatalytic activity of immobilized trypsin was highest with magnetically separable polyaniline (PAMP), and Vm and Km of PAMP were 0.169 mM/min and 0.263 mM respectively. With increasedpH, the biocatalytic activity increased for all supporting materials used. Immobilized enzymes were recycled and recycle activities were over 90% of their original activity after ten times reuse. The operational stabilities of enzymes were greatly improved with enzyme immobilization.

• KSBB Journal
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• v.16 no.5
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• pp.500-504
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• 2001

We scaled up a covalent immobilization system of urokinase to the activated Sepharose and used it repeatedly to cleava a fusion protein consisting of human growth hormone and GST fragment. After scale up from 6 ml to 250 ml. the column system still demonstrated basically the same performance in terms of urokinase immobilization and fusion protein cleavage. When the column was washed with 6 M guanidine HCI after the cleavage reaction, the immobilized urokinase showed no activity probably becasue it was fully unfoled. However, as the denaturant was gradually removed from the column the immobilized urokinase fully regained its bioactivity, which indicated it was properly refolded into is natie conformation as covalently attached to the solid matrix. After 20 cycles of this solid-phase unfolding/refolding. the immobilized urokinase maintained approx. 80% of the initial bioactivity. This method provides and efficient protocol to apply the solid-phase refolding technique to improve the longevity of immobilized enzyme columns.

• Korean Journal of Environmental Agriculture
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• v.29 no.2
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• pp.176-183
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• 2010

Endosulfan degrading ability of Klebsiella oxytoca KE-8 immobilized by entrapment with activated carbon was examined. Endosulfan degradation by the immobilized bacterial strains on several different activated carbon based support materials was investigated. Based on results, activated carbon ($8\times30$ mesh) was chosen as a support material. The immobilized Klebsiella oxytoca KE-8 with the cell density of 4 mg $g^{-1}$ (dry weight) degraded 22.18 ug $ml^{-1}$ endosulfan within 5 days at pH 7.0, $30^{\circ}C$ in batch shake flask cultures. Also, we an experimented recycle packed bed column mode and continuous packed bed column mode for endosulfan degradation. Under optimum operation condition, the immobilized cells in a laboratory scale pack bed column with support beads were able to degrade endosulfan completely in defined minimal salt medium at a maximum rate of 129.6 ug $ml^{-1}$ per day. Moreover, the endosulfan degradation activity could be demonstrated at $4^{\circ}C$ for one month without significant decrease in activity. Results of this study suggest that immobilized cells of Klebsiella oxytoca KE-8 might be applicable to endosulfan contaminated site.

• Journal of Microbiology and Biotechnology
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• v.13 no.1
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• pp.29-34
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• 2003

A Pseudomonas sp. strain NGK1 (NCIM 5120) capable of utilizing salicylate was immobilized in alginate and polyurethane foam (PUF). The degradation rate of salicylate by freely suspended cells was compared with the degradation rate by immobilized cells. In an initial 20 and 40 mM salicylate, free cells ($2{\times}10^{11}\;cfu\;ml^{-1}$) degraded to 16 and 14 mM, alginate-entrapped cells degraded to 18 and 26 mM, and PUF-entrapped cells degraded to 20 and 32 mM salicylate, respectively, in batch cultures. The alginate-and PUF-entrapped cells were used in repeated batch and continuous culture systems. The efficiency of both the immobilized systems f3r the degradation of salicylate was compared. It has been observed that the PUF-entrapped cells could be reused for more than 20 cycles whereas alginate-entrapped cells could be reused for a maximum of only 12 cycles, after which a decrease in degradation rat was observed with the initial 20 and 40 mM salicylate. The continuous degradation of sallcylate by freely suspended cells showed a negligible degradation rate of salicylate when compared with immobilized cells. With the immobilized cells in both alginate and polyurethane foam, the degradation rate increased with an increase in the dilution rate up to $2\;h^{-1}$ for 20 mM, and $1.5\;h^{-1}$ for 40 mM salicylate. The results revealed that PUF-entrapped cells were more efficient for the degradation of salicylate than alginate-entrapped cells and freely suspended cells.

• Microbiology and Biotechnology Letters
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• v.20 no.4
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• pp.459-469
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• 1992

The immobilization characteristics of Zymomonas mobilis for ethanol production were examined. Four different strains of Zymomonas mobilis have been used for ethanol production. Among those, Zymomonas mobilis KCTC 1534 has been selected as the best strain for the highest ethanol productivity from glucose and sucrose. The optimum temperature and pH of the selected strain for ethanol production were $37^{\circ}C$ and 5.0 respectively for both free and immobilized cells. When the cells were immobilized by the gel entrapment method, the immobilized cells could produce ethanol at a little higher temperature than free cells. Calcium alginate was selected as the best gel for immobilizing cells. The immobilized cells could maintain the viability of 80% in 10 weeks storage at $4^{\circ}C$ in the medium with 2% calcium chloride. 20-25 hours of preincubation in 10% glucose solution was required for the activation of immobilized cells entrapped within calcium alginate gel.

• Journal of Environmental Science International
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• v.6 no.1
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• pp.61-67
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• 1997

Heavy metal adsorption by microbial cells is an alternative to conventional methods of heavy metal removal and recovery from metal-bearing wastewater The waste Sac-chuomyces cerevisiae is an inexpensive, relatively available source of biomass for heavy metal biosorption. Biosorption was investigated by free and immobilized-S. cerevisiae. The order of biosorption capacity was Pb>Cu>Cd with batch system. The biosorption parameters had been determined for Pb with free , cells according to the Freundlich and Langmuir model. It was found that the data fitted reasonably well to the Freundlich model. The selective uptake of immobilized-S. cerevisiae was observed when all the metal ions were dissolved in a mixed metals solution(Pb, Cu, Cr and Cd). The biosorption of mixed metals solution by immobilized-cell was studied in packed bed reactor. The Pb uptake was Investigated in particular, as it represents one of the most widely distributed heavy metals in water. We also tested the desorption of Pb from immobilized-cell by us- ing HCI, $H_2SO_4$ and EDTA.

• Journal of the Korean Wood Science and Technology
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• v.29 no.3
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• pp.112-122
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• 2001

Laccase enzymes from Cerrena unicolor and Trametes versicolor were immobilized on the activated glass beads (CPG), silica gel (SG) and soil (SL). The heterogeneous matrices were activated by ${\gamma}$-aminopropyltriethoxysilane (APTES) and glutaraldehyde (GA), and their surfaces were coated by keratin (KER) on activated or non-activated CPG, SG and SL. The laccase activities were tested in the aqueous solution for the native and immobilized preparations using different pH and temperature conditions. By keratin coating on supports, in the cases of CPG-KER and SL-KER, the immobilization yield was increased from about 80% to 90%. Moreover, much less protein was immobilized in keratin coated matrices than in inorganic ones alone (e.g. on CPG-KER 57.6%, whereas on CPG alone 80.6%). Laccase immobilization on keratin coated inorganic matrices was generally more effective than that of non-coated matrices. Concerned to pH dependency, the optima pH for immobilized laccases generally shifted towards to higher values, 5.5-5.8 and even 5.9 in the case of keratin for C. unicolor and from 5.3 to 5.7 for T. versicolor, respectively, and decreased less gradually both in acidic and alkaline regions. The immobilized laccase was more stable against thermal denaturation. This seems particularly true at $75^{\circ}C$ in the case of C. unicolor, where the activity of immobilized enzyme is > 50% higher than that of the free enzyme. For T. versicolor the respective values were $65^{\circ}C$, and 50%.

• Microbiology and Biotechnology Letters
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• v.14 no.4
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• pp.285-291
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• 1986

Immobilized Candida lipolytica cells were prepared by entrapping the whole cells in calcium alginate gel. To enhance citric acid productivity, immobilized cells were Incubated with activation medium in fluidized-bed reactors. When the activation was done in batch operation, maximum citric acid productivity appeared in a much shorter time than in continuous operation. Activated immobilized cells were enhanced about 10-fold in citric acid production relative to non-activated immobilized cells. The productivity of citric acid was also influenced by bead size. When Immobilized cells were reacted in a fluidized-bed reactor with the same quantity of cells, the citric acid productivity was increased as the bead size was decreased.

• Microbiology and Biotechnology Letters
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• v.13 no.2
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• pp.115-118
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• 1985

Rifamycin B oxidase converts rifamycin B to rifamycin S using oxygen as cosubstrate. Humnicola spp. (ATCC 20620) was treated with acetone and the cell powder was immobilized with cellulose acetate. The properties of the immobilized enzyme was examined. The optimum pHs of the immobilized and the free enzymes were 7.2. The optimum temperature of the immobilized enzyme was at 50-55$^{\circ}C$, which was 5$^{\circ}C$ higher than that of the free enzyme. The activities of the immobilized enzyme appeared less sensistive with respect to the changes of temperature and pH as compared to those of the free enzyme. Twenty percent of the enzyme activity was recovered when the enzyme was immobilized in 3mm beads. The storage stability was good below 4$0^{\circ}C$, but the activity decreased very rapidly above 5$0^{\circ}C$. The physical strength of the beads was good and was suitable as packing material in a three-phase enzyme reactor.

• Journal of Microbiology and Biotechnology
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• v.23 no.4
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• pp.473-482
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• 2013

The desulfurization ability of Sphingomonas subarctica T7b was evaluated using resting and immobilized cells with dibenzothiophene (DBT), alkyl DBTs, and commercial light gas oil (LGO) as the substrates. The resting cells of S. subarctica T7b degraded 239.2 mg of the initial 250 mg of DBT/l (1.36 mM) within 24 h at $27^{\circ}C$, while 127.5 mg of 2-hydroxybiphenyl (2-HBP)/l (0.75 mM) was formed, representing a 55% conversion of the DBT. The DBT desulfurization activity was significantly affected by the aqueous-to-oil phase ratio. In addition, the resting cells of S. subarctica T7b were able to desulfurize alkyl DBTs with long alkyl chains, although the desulfurization rate decreased with an increase in the total carbon number of the alkylated DBTs. LGO with a total sulfur content of 280 mg/l was desulfurized to 152 mg/l after 24 h of reaction. Cells immobilized by entrapment with polyvinyl alcohol (PVA) exhibited a high DBT desulfurization activity, including repeated use for more than 8 batch cycles without loss of biodesulfurization activity. The stability of the immobilized cells was better than that of the resting cells at different initial pHs, higher temperatures, and for DBT biodesulfurization in successive degradation cycles. The immobilized cells were also easily separated from the oil and water phases, giving this method great potential for oil biodesulfurization.