• Archives of Pharmacal Research
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• v.3 no.1
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• pp.7-12
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• 1980

Ampliciline was synthesized from 6-amino-pencillanic acid (6-APA) and D-.alpha. phenylglycine methyl ester by using amplicilin synthesizing enzyme from Peudomonas melanogenum (IAM 1655). The whole cell enzyme was immobilized by entrapping it in the polyacrylamide gel lattices. The polymer used in the enzyme entrapment was made from 150 mg per ml of acrylamide monomer and 8 mg per ml of N, N'-methylenebisacrylamide. About 200 mg/whole cell enzyme was mixed in the polymer for entrapment. The maximal activity retention after immobilization was 56%. The optimal pH values for the whole cell enzyme and the immobilized whole cell enzyme were 6.0 and 5.9, respectively. The optimal temperature for the enzyme activity were the same for both type of preparations. The enzyme stabilities against pH and heat increased for immobilized whole cell enzyme. Immobilized cell was more stable especially in the acidic condition while both type were found to be very suceptible to thermal inactivation at a temperature above 4.deg.C. The kinetic constants obtained from Lineweaver-Burk plot based on two substate reaction mechanism showed somewhat higher value for immobilized whole cell enzyme as compared to the whole cell enzyme : the Km value for 6-APA were 7.0 mM and 12.5 mM while Km values for phenylglycine methyl ester were 4.5 mM and 8.2 mM, respectively. Using the immobilized whole cell enzyme packed in a column reactor, the productivity of ampiciline was studied by varying the flow rate of substrate solution. At the space velocity, SV, 0.14 hr$^{-1}$ the conversion was 45%. Operational stability found in terms of half life was 30 hr at SV = 0.2 hr.

• KSBB Journal
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• v.18 no.4
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• pp.306-311
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• 2003

Solid-phase refolding of immobilized proteins can be an effective way to reuse an immobilized enzyme column. Oriented immobilization methods are known to provide higher activity of the immobilized enzymes. In this study, using recombinant EK (enterokinase) as a model enzyme and a fusion protein, that consisted of recombinant human growth hormone and six His tag that was linked by the peptide of EK-specific recognition sequence, as a model substrate, we evaluated two oriented immobilization methods, i. e., reductive alkylation of N-terminus ${\alpha}$-amine and affinity interaction between poly-histidine tag and Ni-NTA (nickel-nitrilotriacetic acid). The immobilization yield, activity and cleavage of the immobilized enzymes, and the yield of solid-phase refolding were compared. The Ni affinity immobilization and the covalent immobilization yields were about 100％ and 65％, respectively. But the specific activities were the same, about 50％ of that of the soluble enzyme. The cleavage rate by the covalently immobilized EK was higher than the soluble enzyme and the side reaction of cryptic cleavage was significantly decreased. Covalently immobilized EK showed almost 100％ refolding yield but the affinity immobilized EK showed only 70％ yield, which suggested the covalent conjugation provided more rigid ‘reference structure’ for the solid-phase refolding. The monomeric hGH could be easily obtained by capturing the cleaved poly Histidine tag by the Ni affinity column.

• Journal of Microbiology and Biotechnology
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• v.29 no.6
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• pp.913-922
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• 2019

Magnetic $Ni_{0.7}Co_{0.3}Fe_2O_4$ nanoparticles that were prepared via the rapid combustion process were functionalized and modified to obtain magnetic $Ni_{0.7}Co_{0.3}Fe_2O_4@SiO_2-CHO$ nanocomposites, on which penicillin G acylase (PGA) was covalently immobilized. Selections of immobilization concentration and time of fixation were explored. Catalytic performance of immobilized PGA was characterized. The free PGA had greatest activity at pH 8.0 and $45^{\circ}C$ while immobilized PGA's activities peaked at pH 7.5 and $45^{\circ}C$. Immobilized PGA had better thermal stability than free PGA at the range of $30-50^{\circ}C$ for different time intervals. The activity of free PGA would be 0 and that of immobilized PGA still retained some activities at $60^{\circ}C$ after 2 h. $V_{max}$ and $K_m$ of immobilized PGA were 1.55 mol/min and 0.15 mol/l, respectively. Free PGA's $V_{max}$ and $K_m$ separately were 0.74 mol/min and 0.028 mol/l. Immobilized PGA displayed more than 50% activity after 10 successive cycles. We concluded that immobilized PGA with magnetic $Ni_{0.7}Co_{0.3}Fe_2O_4@SiO_2-CHO$ nanocomposites could become a novel example for the immobilization of other amidohydrolases.

• Journal of Microbiology and Biotechnology
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• v.6 no.2
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• pp.137-141
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• 1996

Acetoin and ammonia, the precursors of tetramethylpyrazine (TMP) having "meaty" or "roasted" flavors, were produced by the culture of Lactococcus lactis ssp. lactis biovar. diacetilactis FC1 in free and immobilized cell systems. Cells were immobilized using k-carrageenan and then were incubated at $34^{\circ}C$. The TMP productivity (0.34 g/l) and the conversion ratio (9.3%) of acetoin to TMP of the immobilized cell system were higher than those (0.24 g/l, 7.0%) of the free cell system. When the beads were activated for 12 h, the productivity of acetoin and TMP increased slightly.

• Journal of Environmental Science International
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• v.12 no.12
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• pp.1277-1284
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• 2003

The photocatalytic oxidation of Rhodamine B (RhB) was studied using immobilized TiO$_2$ and fluidized bed reactor. Immobilized TiO$_2$ onto GF/C was employed as the photocatalyst and a 30 W germicidal lamp was used as the light source and the reactor volume was 4.8 L. The effects of parameters such as the amounts of photocatalyst, initial concentration, initial pH, air flow rate and anion additives (NO$_3$$\^$-/, SO$_4$$\^$2-/, Cl$\^$-/, CO$_3$$\^$2-/) competing for reaction. The results showed that the optimum dosage of the immobilized TiO$_2$ was 40.0 g/L. Initial removal rate of immobilized TiO$_2$ was expressed Langmuir - Hinshelwood equation.

• KSBB Journal
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• v.15 no.3
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• pp.247-253
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• 2000

This study was aimed to enhance polycyclic aromatic hydrocarbon(PAHS) biodegradation rate by repeated-batch treatment using immobilized cells of Phanerochaete chrysosporium. In the repeated-batch operations with 30 mg/L of pyrene the maximum degradation rate was 6.58 mg/L day. As the number of batches increased the concentration of immobilized cells significantly decreased and the degradation rate and specific acitivity gradually increased to a maximum value and then decreased. To have PAH degradation activity and cell mass recovered one batch of cultivation using the growth medium instead of the PAH-degrading medium was carried in the course of repeated-batch operations. This maximum degradation rates of pyrene and anthracene were 4.29 and 4.46 mg/L$.$day respectively. Overall the rate of PAH degradation could be enhanced 2.5-30 folds by using immobilized cells compared to the case of using suspended cells.

• Microbiology and Biotechnology Letters
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• v.6 no.1
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• pp.33-40
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• 1978

Immobilization of alkaline protease was investigated by absorbing the enzyme on adsorbents. Alkaline protease was adsorbed on silica gel selected as a carrier to immobilize the enzyme. In this study, properties of the immobilized enzyme were compared with those of the soluble enzyme. 1) The optimum pH (10.0) of the enzyme was not changed, but the activity was increased at alkaline pH by immobilization. 2) The optimum temperature of the immobilized enzyme was shifted from 50$^{\circ}C$ to 45$^{\circ}C$, while the temperature-activity Profile became broader than those of the soluble enzyme. 3) The pH stability of the immobilized enzyme was significantely increased at pH 4.0, althouth it did not change in the neutral and alkaline pH region. 4) The heat stability of the enzyme was enhanced in the temperature range of 55$^{\circ}C$∼65$^{\circ}C$ by the immobilization. 5) The immobilized enzyme retained 40％ of its original activity after repetitive use for 6 times. 6) The enzyme stability was greately improved for a prolonged storage at 4$^{\circ}C$.

• Journal of Environmental Science International
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• v.4 no.3
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• pp.295-301
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• 1995

Continuous deodorization of malodorous sulfur compounds by Thiobaillus neapolitanusts R-10 immobilized onto a polypropylene pellet was studied using a column reactor at 30$^{\circ}$C. The maximum amounts of immobilized cells was 5.3 gall polypropylene with 5$\times$7.5mm in pellet size, and the amounts of immobilized cells in the higher part of the column was as twice as in the lower part. The optimum pH and temperature for removal of dimethyl sulfide were 6.0 and 30$^{\circ}$C, respectively. When 5-20 ${mu}ell$/l of hydrogen sulfide and methylmercaptan were employed 98% of removal efficiency were achieved. In contrast, lower concentrations of dimethyl sulfide and dimethyldisulfide should be supplied to meet satisfactory deodorization efficiency. The immobilized cell column was successfully operated for the deodorization of mixture of sulfur compounds over 15 days without significant loss of initial activity achieving high efficiency.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.35 no.5
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• pp.80-86
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• 2003

The wood-degrading fungus Trichophyton sp. LKY-7 (T. LKY-7) was immobilized in ca-alginate beads for laccase production and PCP bioremediation. The immobilized T. LKY-7 enabled the repeated use of this fungus for laccase production and produced high amount of laccase throughout 5 cycles incubation. As a laccase inducer. oak wood meal (Quercus variabilis) seemed to be effective laccase inducer for T. LKY-7, and the optimum addition amount was 1％ (W/W) in glucose-peptone medium. Bioremediation of pentachlorophenol by the immobilized T. LKY-7 reached an efficency of up to 90％ without toxic inhibition. The immobilized T. LKY-7 might thus be applicable for semicontinuous laccase production and bioremediation to serve inoculum for reactor system.

• Journal of Microbiology and Biotechnology
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• v.8 no.6
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• pp.696-699
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• 1998

Biological oxidation of D-sorbitol to L-sorbose using permeated and immobilized cells of Gluconobacter suboxydans was carried out to investigate the optimum reaction condition. The stabilization effect of cross-linking agents such as glutaraldehyde, tannic acid, and polyethylene imine to prevent the leakage of enzymes from beads containing permeated and immobilized cells of G. suboxydans was examined by the production of L-sorbose from the mixture of D-sorbitol and gluconic acid. The protein concentration effused from immobilized beads treated with only glutaraldehyde was $5.2\mug/m\ell$ after 20 h. The beads of G. suboxydans immobilized with alginate and cross-linked with 0.3% glutaraldehyde was the most useful for the oxidation of D-sorbitol to L-sorbose.