• Microbiology and Biotechnology Letters
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• v.26 no.4
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• pp.345-351
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• 1998

In an effort to develop a potent system for the production of various dp (degree of polymerization) chitooligosaccharides, 32 enzymes or microbial systems were screened for chitosanolytic acitivity using chitosan as a substrate. The efficiency of each enzyme system was evaluated by the changes of turbidity and viscosity of chitosan solution, the amount of precipitate and the reducing sugar-producing activity in the enzymatic reaction mixture. Based on these assay methods for the chitosanase activity, Bacillus sp. P2l out of 32 screened systems showed highly potent endochitosanase, which was comparable with a commercially available enzyme (E7). Chitooligosaccharides of dp 3-7 were separated by TLC as major enzymatic reaction products, suggesting that the chitosanase from Bacillus sp. P2l be endo-splitting type.

• Microbiology and Biotechnology Letters
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• v.27 no.4
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• pp.304-310
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• 1999

We have developed a new process for the production of new structure oligosaccharides using the mixed-culture fementation of Lipomyces starkeyi KSM22 and leuconostoc mesenteroides B-512FMCM.L.starkeyi KSM22 produces a novel DXAMase(an enzyme containing both dextranase and amylase activities). It hydrolyzes the soluble starch and dextran. The hydrolyzates were used as acceptors for dextransucrase of L.mesenteroides to synthesize the new oligosaccharides(NOS). In fermentation, as the concentration of sucrose was increased from 9%(w/v) to 15%(w/v), the yields of dextran(sum of dextran I, MW=66kD, and dextran II, MW=21kD) was increased from 12.7% to 42.5%, and NOS was increased from 3.9% to 5.2% of the theoretical, respectively. The NOS of dp(degree of polymerization) 5 and over was increased from 33.1% to 58.3% of the total NOS. The NOS showed heat resistant up to 12$0^{\circ}C$ and was stable at pHs ranged from 2 to 6. The NOS decreased the pH changes in the culture of S. mutans, and also showed inhibitory effects on the growth of S. aureus or S. typhimurium.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.563-566
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• 2000

In order to obtain microbial endochitosanase for enzymatic production of chitooligosaccharides from chitosan, we screened four microbes from soil and selected. Bacillus sp. HSB-21 which showed highest activity. Chitosanase, produced from isolating microbe, was endo-type and molecular mass of the enzyme was estimated as 21,000 by active staining. Its optimum pH and temperature were 5.5 and $50^{\circ}C$, respectively. It was stable in the pH range of 3.0 to 8.0 and up to $40^{\circ}C$. It did not produce chitomonosaccharide and produced chitooligosaccharide ranging from chitobiose to chitooctaose as major end-products from chitosan. The chitosanase from Bacillus sp. HSB-21 can be applicable to enzymatic production of chitooligosaccharide which has high degree of polymerization .

• Preventive Nutrition and Food Science
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• v.11 no.2
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• pp.146-154
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• 2006

Structural modification of corn starch by gamma irradiation was evaluated for under dry conditions at varied intensities from 0 to 40 kGy. Under scanning electron microscopy, the granule shape of corn starch was not significantly affected by the irradiation up to 40 kGy. In addition, X-ray diffraction and melting patterns of the irradiated starches were similar to those of the native starch, indicating that crystalline regions in the starch granules were not changed by irradiation. However, the pattern of gel permeation column chromatography showed a significant increase in partial hydrolysis of gamma irradiated starch samples. The degree of polymerization and the paste viscosity of irradiated starch samples dose-dependently decreased significantly with irradiation, and increased solubility and clarity were observed in the irradiated starch solution. In addition, the degree of retrogradation decreased as irradiation dose increased. Irradiation of corn starch has advantages over the ordinary acid or the enzyme hydrolysis modification methods. It does not affect the granular shape and crystalline phase of starch during hydrolysis, and the process can be carried out in dry state.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1513-1520
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• 2010

A bacterium capable of producing melanin pigment in the presence of L-tyrosine was isolated from a crop field soil sample and identified as Klebsiella sp. GSK based on morphological, biochemical, and 16S rDNA sequencing. The polymerization of this pigment occurs outside the cell wall, which has a granular structure as melanin ghosts. Chemical characterization of the pigment particles showed then to be acid resistant, alkali soluble, and insoluble in most of the organic solvents and water. The pigment got bleached when subjected to the action of oxidants as well as reductants. This pigment was precipitated with $FeCl_3$, ammoniacal silver nitrate, and potassium ferricynide. The pigment showed high absorbance in the UV region and decreased absorbance when shifted towards the visible region. The melanin pigment was further charecterized by FT-IR and EPR spectroscopies. A key enzyme, 4-hydroxyphenylacetic acid hydroxylase, that catalyzes the formation of melanin pigment by hydroxylation of L-tyrosine was detected in this bacterium. Inhibition studies with specific inhibitors, kojic acid and KCN, proved that melanin is synthesized by the DOPA-melanin pathway.

• Journal of Electrochemical Science and Technology
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• v.4 no.1
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• pp.41-45
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• 2013

Platinum is a well known element which shows a significant electrocatalytic activity in many important applications. In glucose sensor, because of the poisoning effect of reaction intermediates and the low surface area, the electrocatalytic activity towards the glucose oxidation is low which cause the low sensitivity. So, we fabricate a nanoporous PtZn alloy electrode by deposition-dissolution method. It provides a high active surface and a large enzyme encapsulating space per unit area when it used for an enzymatic glucose sensor. Glucose oxidase was immobilized on the electrode surface by capping with PEDOT composite and PPDA. The composite and PPDA also can exclude the interference ion such as ascorbic acid and uric acid to improve the selectivity. The surface area was determined by cyclic voltametry method and the surface structure and the element were analyzed by Scanning Electron Microscope (SEM) and Energy Dispersive X-ray spectroscopy (EDX), respectively. The sensitivity is $13.5{\mu}A/mM\;cm^2$. It is a remarkable value with such simply prepared senor has high selectivity.

• Proceedings of the KIEE Conference
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• 2003.07c
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• pp.1935-1937
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• 2003

최근 콜레스테롤 센서는 전극 상에 효소를 고정화 하는 방식을 이용하여 센서의 집적도를 높이는 시도가 이루어지고 있다. 이러한 전극 상의 효소고정화 방식으로 entrapment, cross liking, covalently binding 등이 있다. 본 논문에서는 이러한 효소 고정화 방식-전도성 고분자인 P3MT를 사용하여 entrap시키는 방법과 silanization을 이용한 covalent bonding 시키는 방법-에 따른 전기화학 센서의 감도 특성에 관한 연구를 수행하였다. 전도성 고분자를 사용한 고정화 방법은 cyclic voltammograms으로 scan rate 10 mA/s, potential 0.5-1.3V의 조건하에서 P3MT를 Polymerization하고, 효소 고정화를 위해 chromoampermeter로 potential 0.6V에서 900초 동안 수행하였다. silanization을 이용한 covalent bonding 시키는 방법은 nitric acid로 Pt 전극표면을 산화시키고, APTER로 silanization 공정을 시행하였다. 효소 고정화를 위해 전해질로는 0.1M Phosphate buffer solution을 사용하여 cyclic voltammograms으로 scan rate 50 mA/s 전위 0.0-0.7V의 조건 하에서 수행하였다. 이 결과 전도성 고분자를 이용한 고정화 방법에서의 senstivity가 0.89 ${\mu}A/mM{\cdot}cm^2$이고, silanization을 이용한 효소 고정화 방법에서는 1.51 ${\mu}A/mM{\cdot}cm^2$였다. 이처럼 후자의 방법에서 더 좋은 감도 특성이 나타났다. 따라서, silanization을 이용한 고정화 방법이 센서 제작 방식으로 더 적합하다고 사료된다.

• Journal of Microbiology and Biotechnology
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• v.25 no.8
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• pp.1361-1370
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• 2015

A blue laccase was purified from a white rot fungus of Trametes trogii, which was a monomeric protein of 64 kDa as determined by SDS-PAGE. The enzyme acted optimally at a pH of 2.2 to 4.5 and a temperature of 70℃ and showed high thermal stability, with a half-life of 1.6 h at 60℃. A broad range of substrates, including the non-phenolic azo dye methyl red, was oxidized by the laccase, and the laccase exhibited high affinity towards ABTS and syringaldazine. Moreover, the laccase was fairly metal-tolerant. A high-molecular-weight kraft lignin was effectively polymerized by the laccase, with a maximum of 6.4-fold increase in weight-average molecular weight, as demonstrated by gel permeation chromatography. Notable structural changes in the polymerized lignin were detected by Fourier transform infrared spectroscopy and ¹H NMR spectroscopy. This revealed an increase in condensed structures as well as carbonyl and aliphatic hydroxyl groups. Simultaneously, phenolic hydroxyl and methoxy groups decreased. These results suggested the potential use of the laccase in lignin modification.

• Korean Journal of Food Science and Technology
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• v.12 no.1
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• pp.45-52
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• 1980

The nylon (poly 6, 10) microcapsules containing ${\beta}-galactosidase$ were obtained by the interfacial polymerization of 1, 6-diaminohexane and sebacoyl chloride with ${\beta}-galactosidase$ from Escherichia coli. They were generally spherical and had a mean diameter of $80{\mu}$ with 45 % of the activity recovery. In particular, there was no transport hamper of lactose through the membrane of microcapsules. The characteristics of the microencapsulated enzyme were similar to those of soluble enzyme optimal pHs, $7.0{\sim}7.2$ for the soluble and $7.3{\sim}7.5$ for the microencapsulated ; optimal temperatures, $50^{\circ}C$ for both ; apparent $K_m,\;3.33{\times}10^{-4}(on ONPG),$ $2.86{\times}10^{-3}$ M(on lactose) for the soluble and $5.28{\times}10^{-4}$ (on ONPG), $4.25{\times}10^{-3}$ M (on lactose) for the microencapsulated ; activation energies, 8.94 for the soluble and 9.78 Kcal/mole for the microencapsulated enzyme. Using this microencapsulated ${\beta}-galactosidase$, hydrolyses of lactose and milk lactose were carried out and 80 % of 5 % lactose solution and 70 % of lactose in skim milk were hydrolyzed in 40 hr at $27^{\circ}C$. The reusability and operational stability showed that the remaining activity was 50 % of the original activity after 5 runs and 120 hr of total operating time at $27^{\circ}C$.

• Journal of Food Hygiene and Safety
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• v.35 no.1
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• pp.83-92
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• 2020

This study investigated the optimal conditions of enzymatic reaction for production of isomaltooligosaccharides (IMO) using rice flour. To manufacture IMO, commercial enzymes (Termamyl 2X, Maltogenase L, Promozyme D2, Fungamyl 800L and Transglucosidase L) were used. The sugar composition and amount of IMO were examined by HPLC with charged aerosol detector (HPLC-CAD) in each manufacturing process. Liquefaction reaction was performed according to different Termamyl 2X concentrations (0.025%, 0.05%, 0.075%, 0.1%) and reaction times (1 h, 2 h). As a result, the reducing sugar content was the highest at 138.26 g/L when 0.075% Termamyl 2X was added for 2 hours. In order to optimize simultaneous saccharification and transglucosylation, experiments on enzyme selection, enzyme concentration and enzyme reaction time were conducted. Reaction with 0.0015% Maltogenase L, 0.05-0.1% Promozyme D2 and 0.1% Tansglucosidase L was effective in decreasing glucose content and increasing content of IMO with a high degree of polymerization. A change in sugar content was observed every 6 hours to determine the optimal reaction time, and the highest IMO was produced after 36 hours of reaction (75.36 g/L). The IMO prepared under optimal conditions showed isomaltose, 35.11 g/L; panose, 11.97 g/L; isomaltotriose, 19.95 g/L; isomaltotetraose, 7.46 g/L; isomaltopentaose, 1.05 g/L at 18 brix and the ratio of IMO in the total sugar was 56.37%.