• KSBB Journal
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• v.8 no.4
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• pp.328-335
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• 1993

A flow injection analysis(FIA) system was developed for the determination of cholesterol using immobilized cholesterol oxidase and cholesterol ester hydrolase. The enzymes were immobilized on controlled pore galas(CPG) by the glutaraldehyde method. The glass colunms packed with immobilized enzymes were found to contain 3-5 I.U. for each enzyme. A hydrogen peroxide sensitive electrode was contructed and applied to the FIA system. The operational conditions for FIA response were investigated and optimized with variation of sampling volume, flow rate and composition of carrier solution. The FIA response were linear upto 60 and 400mg/m1 for free cholesterol and cholesterol ester, respectively. All samples were analyzed with a good precision (<2.5% CV) and accuracy. 23 samples were mea sured succesively within about an hour. Intermittent assays of more than 500 times caused 50% decrease in response sensitivity.

• Microbiology and Biotechnology Letters
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• v.46 no.3
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• pp.234-243
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• 2018

The Antarctic Ocean contains numerous microorganisms that produce novel biocatalysts that can have applications in various industries. We screened various psychrophilic bacterial strains isolated from the Ross Sea and found that a Croceibacter atlanticus strain (Stock No. 40-F12) showed high lipolytic activity on a tributyrin plate. We isolated the corresponding lipase gene (lipCA) by shotgun cloning and expressed the LipCA enzyme in Escherichia coli cells. Homology modeling of LipCA was carried out using the Spain Arreo lake metagenome alpha/beta hydrolase as a template. According to the model, LipCA has an ${\alpha}/{\beta}$ hydrolase fold, Gly-X-Ser-X-Glymotif, and lid sequence, indicating that LipCA is a typical lipase enzyme. Active LipCA enzyme was purified fromthe cell-free extract by ammonium sulfate precipitation and gel filtration chromatography. We determined its enzymatic properties including optimum temperature and pH, stability, substrate specificity, and organic solvent stability. LipCA was immobilized by the cross-linked enzyme aggregate (CLEA) method and its enzymatic properties were compared to those of free LipCA. After cross-linking, temperature, pH, and organic solvent stability increased considerably, whereas substrate specificities did not changed. The LipCA CLEA was recovered by centrifugation and showed approximately 40% activity after 4th recovery. This is the first report of the expression, characterization, and immobilization of a C. atlanticus lipase, and this lipase could have potential industrial application.

• KSBB Journal
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• v.7 no.2
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• pp.144-148
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• 1992

The purpose of this study is to develop a continuous process for sorbitol production using dried Zymomonas mobilis immobilized in K-carrageenan. The methods of glularaldehyde cross-linking of enzymes in CTAB (celyltrimetylammoniumbromide) treated cells immobilized in K-carrageenan showed stability for the production of sorbitol for 30 days of operation. K-carrageenan beads entrapping permeabilized cells were dried to Improve bead rigidity and storage stability. A semi-batch process with dry beads was carried out and only a small loss of enzyme activity (less than 8%) was observed during 72h. The value of Vmax for the dry K-carrageenan beads was found to be one half or that for free cells. It was shown that the productivities of the continuous process with wet K-carrageenan beads and dry beads at a dilution rate 0.1h-1 were 3.4g/L-h and 2.88h/L-h, respectively.

• Korean Journal of Microbiology
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• v.37 no.2
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• pp.164-169
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• 2001

The conversion of cephalosporin C to 7$\alpha$-hydroxycephalosporin C was examined with the cell-free extract of several cephamycin producing strains. Streptomyces clavuligerus ATCC 27064 was the most potent strain for the activity of cephalosporin 7$\alpha$-hydroxylase. Partially purified and immobilized cephalosporin 7$\alpha$-hydroxylase with resins were used to synthesize 7$\alpha$-hydroxycephalosporin C from the substrate, cephalosporin C. The molecular weight of the product isolated from the reaction mixture were determined to be 431 by ESI-Mass. $^1H$ NMR also support the conversion of cephalosporin C to 7$\alpha$-hydroxycephalosporin C by immobilized enzyme.

• Korean Chemical Engineering Research
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• v.59 no.3
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• pp.373-378
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• 2021

The study aimed to develop a high-power enzymatic electrode for a wearable fuel cell that generates electricity utilizing lactate present in a sweat as fuel. Anode was fabricated by immobilizing lactate oxidase (LOx) on flexible carbon paper. As the lactate concentration in the electrolyte solution increased, the amount of current generated by catalysis of lactate oxidase increased. The immobilized LOx generated 1.5-times greater oxidation current density in the presence of gold nanoparticles than carbon paper only. Bilirubin oxidase (BOD)-immobilized cathode generated a larger amount of reduction current in the electrolyte saturated with oxygen than purged with nitrogen. A fuel cell composed of two electrodes was fabricated and cell voltage was measured under different discharge current. At the discharge current density of 66.7 ㎂/cm², the cell voltage was 0.5±0.0 V leading to maximum cell power density of 33.8±2.5 ㎼/cm².

• Korean Chemical Engineering Research
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• v.61 no.4
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• pp.576-583
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• 2023

This study aimed to investigate the impact of enhancing the electrode conductivity and mitigating the production of hydrogen peroxide - a by-product arising from lactate oxidation - on the performance of lactate electrodes. The electrical conductivity of the electrode was improved by modifying the surface of carbon paper with single-walled carbon nanotubes. Catalase was introduced to effectively eliminate the hydrogen peroxide produced during the lactate oxidation reaction. The carbon paper electrode, with simultaneous immobilization of both lactate oxidase and catalase, yielded a current 1.7 times greater than the electrode where only lactate oxidase was immobilized. The electrode in which lactate oxidase and catalase were co-immobilized on the surface of carbon paper modified with single-walled carbon nanotubes, produced a current of 171 µA, which was more than twice as much current as the carbon paper with only lactate oxidase immobilized. The optimized electrode showed a linear response up to lactate concentration of 20 mM, confirming that it can be used as a sensor electrode.

• Applied Biological Chemistry
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• v.28 no.4
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• pp.233-238
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• 1985

Glucoamylases catalyze a stepwise hydrolysis of starch with the production of glucose. In order to make an efficient conversion of starch into glucose, glucoamylases prepared from Rhizopus spp. (Sigma Co.) were attached to a porous glass and immobilized by glutaraldehyde-induced crosslinking. The porous glass used in this study was $ZrO_2$ coated, $40{\sim}80$ mesh, 550 A pore diameter. Using the forgoing glass, we could couple as much as 50mg of protein per gram of carrier. Substrate for the glucoamylase was an enzyrne-modified thin-toiling 30% cornstarch solution used where greater solubility and low viscosity are desired. Immobilized glucoamylase had an optimum pH 7.0 to the alkaline side of soluble enzyme. Km values of immobilized and soluble enzyme were 1.04 mM and 1.25mM, respectively. The thermal stability of glucoamylase was increased by immobilization and the immobilized enzyme showed an optimum temperature at $40{\sim}60^{\circ}C$. The continuous conversion of cornstarch to glucose by use of immobilized glucoamylase resulted in the production of a more than 90 DE product.

• Proceedings of the KIEE Conference
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• 2002.11a
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• pp.248-250
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• 2002

본 논문은 당뇨병의 지표물질인 glucose의 농도를 극미량의 시료를 사용하여 정량 할 수 있는 방법을 개발하기 위하여 효소 고정화 전극을 제작하였다. 전극은 실리콘 웨이퍼상에 마이크로 크기의 전극을 반도체 공정을 이용하여 제작하였고, 전기 화학적 방법으로 마이크로 전극에 전도성 고분자 Polypyrrole(PPy) 및 glucose oxidase(GOx)를 고정화한 고감도의 전기화학 전극을 개발하였다. 도전성 고분자의 전기 화학적 중합은 순환 전압 전류법으로 하였으며, 용액의 액성에 따른 효소의 표면 전하를 이용하여, 도전성 고분자를 코팅한 전극에 일정한 전압을 인가하고 GOx를 도우핑 하였다. 제작된 전극은 시간대 전류법으로 glucose의 농도에 따른 감도 측정결과 마이크로 리터의 시료에 $5{\mu}A$/decade를 얻었다. 전극의 표면분석은 Scanning electron microscopy(SEM), Energy dispersive X-ray spectroscopy(EDX)를 이용하였다.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2001.10a
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• pp.51-51
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• 2001

• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.504-511
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• 2016

Lactulose, a synthetic disaccharide, has received increasing interest because of its role as a prebiotic that can increase the proliferation of Bifidobacterium and Lactobacillus spp. and enhance the absorption of calcium and magnesium. While the industrial production of lactulose is still mainly achieved by the chemical isomerization of lactose in alkaline media, this process has drawbacks including the need to remove catalysts and by-products, as well as high energy requirements. Recently, the use of cellobiose 2-epimerase (CE) has been considered an interesting alternative for industrial lactulose production. In this study, to develop a process for enzymatic lactulose production using CE, we screened improved mutant enzymes ($CS-H^RC^E$) from a library generated by an error-prone PCR technique. The thermostability of one mutant was enhanced, conferring stability up to $75^{\circ}C$, and its lactulose conversion yield was increased by 1.3-fold compared with that of wild-type CE. Using a recombinant Escherichia coli strain harboring a CS35 $H^RC^E$-expressing plasmid, we prepared cell beads immobilized on a Ca-alginate substrate and optimized their reaction conditions. In a batch reaction with 200 g/l lactose solution and the immobilized cell beads, lactose was converted into lactulose with a conversion yield of 43% in 2 h. In a repeated 38-plex batch reaction, the immobilized cell beads were relatively stable, and 80% of the original enzyme activity was retained after 4 cycles. In conclusion, we developed a reasonable method for lactulose production by immobilizing cells expressing thermostable CE. Further development is required to apply this approach at an industrial scale.