• KSBB Journal
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• v.16 no.5
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• pp.474-479
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• 2001

Fed-batch culture, single stage and two stage continuous cultures of Corynebacterium glutamicum SH 35 for the production of $_{L}$-Iysine were performed and compared. In the case of fed batch culture, $_{L}$-Iysine concentration, $_{L}$-Iysine yield and $_{L}$-Iysine productivity was 129.2 g/L, 47.0% and 3.08 g/L/h, respectively. In a single-stage continuous culture, optimum dilution rate and pH was 0.1 h$^{-1}$ and 6.9, respectively, and optimum concentration of sugar and ammonium sulfate in a medium reservoir was 108 g/L and 25 g/L, respectively. Under the optimized conditions, 67 of cell concentration($OD_{610}$), 44.2 g/L of lysine concentration, 41% of $_{L}$-Iysine yield and 4.39 g$L^{-1}$ of $_{L}$-Iysine productivity were obtained. In a two-stage continuous culture, optimum dilution rate was 0.075 $h^{-1}$. Under the conditions, 103 of cell concentration($OD_{610}$) 84.0g/L of $_{L}$-Iysine concentration and 46% of $_{L}$-Iysine yield were obtained.

• Journal of the Korean Magnetic Resonance Society
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• v.6 no.1
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• pp.69-77
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• 2002

During the screening of material which has the antimicrobial activity against aminoglycoside-resistant bacteria, A new material $\varepsilon$-(L-$\beta$-Iysine) polypeptide from a culture medium of Streptomyces sp.(DWGS2) was isolated, and the structure and the physicochemical properties of the new material were elucidated. The new material was separated by column chromatography of the culture medium using Dowex1$\times$2, Silica gel, and Sephadex LH20 etc. The chemical structure and molecular weight were determined with the data of various NMR experiments, MALDI mass, and ESI mass experiments. The antimicrobial activity of $\varepsilon$-(L-$\beta$-Iysine) polypeptide is not only better than equal to the activity of known aminoglycoside type of antibiotics(MIC=3.125 - 6.25ug/mL) but also effective against aminoglycoside-resistant bacteria and fungi. If the mechanism of antimicrobial activity against aminoglycoside- resistant bacteria is figured out, the $\varepsilon$-(L-$\beta$-Iysine) polypeptide can be utilized for the treatment of diseases caused by aminoglycoside-resistant bacteria.

• Microbiology and Biotechnology Letters
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• v.15 no.6
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• pp.369-374
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• 1987

A bacterium which grows on D-$\alpha$-Amino-$\varepsilon$-Caprolactam as sole carbon, energy and nitrogen source was isolated from the sludge of industrial areas in Taegu, and identified as Alcaligenes eutrophus. The optimum pH, temperature and concentration of D-$\alpha$-Amino-$\varepsilon$-Caprolactam for the growth were 6.0, 3$0^{\circ}C$ and 0.2% respectively. The bacteria could utilize glucose and fructose as a carbon source, and utilize ammonium chloride, ammonium nitrate, ammonium sulfate and sodium nitrate as a nitrogen source, and utilize L-Iysine and L-glutamate as a carbon and nitrogen source. It was found with thin layer chromatography and polarimeter that D-$\alpha$-Amino-$\varepsilon$-Caprolactam was converted to L-Iysine by the cell-free extracts of Alcaligenes eutrophus A52.

• KSBB Journal
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• v.7 no.3
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• pp.209-215
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• 1992

The aims of this sutdy was to investgate the action mechanism of polycation on the $\beta$-glucan synthetase II (GS II) related to cell wall synthesis in suspension cultured carrot cells. In the suspension cultured cells treated with poly-L-Iysine($12{\mu}M$) and poly-L-ornithine ($12{\mu}M$) having ploycationic nature, GS II activity increased about 40% and 50% than that of the control respectively. And similar response was observed when ATP and NaF were treated. On the other hand, ploy-L-lysine and ploy-L-ornithine did nor affect the membrane permeability. Phorbol-12-myrlstate-13-acetate (TPA), activator of protein klnase, increased about 35% and 1-(5-isoquinolinyl sulfonyl)-2-methyl-piperrazine (H-7), inhibitor of protein kinase, decreased about 30% of GSII activity than that of control. These results suggest that polycation plays a role in the cell wall synthesis by increasing GS II activity through phosphorylation.

• Journal of Plant Biotechnology
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• v.5 no.1
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• pp.33-41
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• 2003

Microspore-derived cell lines resistant to 5-methyltryptophan (5MT, a tryptophan analog) or S-(2-aminoethyl)-L-cysteine (AEC, a Iysine analog) were selected in rice by in vitro mutagenesis. For selection of 5MT or AEC resistant cell lines, suspension-cultured cells were irradiated with gamma rays. Thirteen 5MT resistant cell lines were selected and they were able to grow stably at 2 times higher 5MT concentration. A feedback insensitive form of anthranilate synthesis, the pathway specific control enzyme for tryptophan synthesis, was detected from the 5MT resistant lines. Contents of the free amino acids in five resistant lines (MR12-1 to MR12-5) showed a 7.4 to 46.6 times greater level than that in the control culture. Tryptophan, phenylalanine, and tyrosine levels in the shikimate pathway were 28.1 and 22.5 times higher in MR12-3 and MR12 4, respectively, than that measured in the control cells. Four AEC resistant cell lines were isolated from cultures grown on medium containing 1 mM AEC, They were able to grow stably with 2 mM AEC, while sensitive calli were inhibited by 0.5 mM AEC. Aspartate kinase activities of the resistant lines were insensitive to the natural inhibitor, Iysine, and accumulated 2.2 to 12.9-fold higher levels of free Iysine than that of the control cells. Especially, the levels of aspartate, asparagine, and methionine in the aspartate pathway showed higher accumulation in the AEC resistant lines than that in the control cells.

• Journal of Sensor Science and Technology
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• v.6 no.6
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• pp.437-444
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• 1997

A potentiometric L-lysine-selective sensor is described for the direct determination of lysine. The sensor system is based on a carbon dioxide gas sensing electrode and an L-lysine decarboxylase immobilized to CNBr-activated sepharose 4B. A highly selective L-lysine sensor has been prepared with immobilizing enzyme slurry put into reaction buffer solution. The optimum conditions for the measurement were evaluated by various experiments. This sensor exhibits a linear response to L-lysine concentrations from $10^{-4}M$ to $10^{-1}M$. Response time of this lysine sensor is shorter than 30secs and the immobilized enzyme slurry is stable over one year.

• Macromolecular Research
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• v.13 no.3
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• pp.257-264
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• 2005

Polyurethanes containing z-Iysine segments in the main chain (PULL) were synthesized from 4,4'-diphe-nylmethyl diisocyanate, poly(tetramethylene glycol), and z-Iysine oligomer as a chain extender. The PULL film was treated first with a $10\%$ HBr-acetic acid solution and subsequently with a saturated sodium bicarbonate aqueous solution to produce a primary amine group on the surface (PULL-N). Lactobionic acid (LA)-immobilized PULL (PULL-L) was prepared by the coupling reaction of the PULL surface amine groups and the LA carboxylic acid groups. The surface-modified PULLs were then characterized by attenuated total reflection-Fourier transform infra-red spectroscopy, electron spectroscopy for chemical analysis, atomic force microscopy, and contact angle goniometry. In the hepatocytes adhesion experiment, the cells poorly adhered to the PULL surface, although they adhered moderately well to the PULL-N surface. On the other hand, the cells adhered well to the PULL-L surface, suggesting the good affinity of the surface $\beta$-galactose moieties for hepatocytes. When hepatocytes were cultured in the presence of epidermal growth factor for 48 h, the cells rapidly aggregated on the PULL-L surface, whereas they aggregated only slowly on the other surfaces. The PULL prepared in this study has the potential to be used as a coating material for the enhancement of hepatocyte adhesion.

• Journal of Microbiology and Biotechnology
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• v.7 no.5
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• pp.333-340
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• 1997

An actinomycetes, KT-10 isolated from ginseng field in Kyongpook, Korea was selected based on its ability to produce a lysosomal cathepsin B inhibitor. The inhibitor purified from the culture supernatant of the isolate KT-10 showed strong inhibitory effects against cathepsin B as well as against papain when the activities were measured using synthetic substrate, ${\alpha}$-N-benzyloxycarbonyl-L-Iysine p-nitrophenyl ester (CLN) or ${\alpha}$-N-benzoyl-D,L-arginine 2-naphthylamide (BANA). The isolate KT-10 was identified as a species of Streptomyces based on its morphological characteristics and chemotaxonomic data. The TAXON program of Ward was used to identify Streptomyces sp. KT-10 as a strain of Streptomyces luteogriseus belong to cluster 18 of the genus Streptomyces with a Willcox probability 0.999388. The cathepsin B inhibitor was presumed to a novel material composed of a polyhydroxylamine.

• Journal of Biomedical Engineering Research
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• v.8 no.1
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• pp.87-92
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• 1987

In order to evaluate the artificial skin for burn would covering materials, copoly(N. carbobenzoxy-L-Iysine-L-leucine)s were prepared by Ipolymerization of N - carbobenzoxy-L- I sine anhydride and L-leucine anhydride in homogeneous solvents using triethylamine as an initiator. The synthetic polypeptides and the oxter type polyurethane(PV)of medical grade were used as the sheet type membranes were prepared ; monolayer membrances were composed of only the polypeptides, bilayer membranes and blend membranes were controlled by composition of the polypeptides and PU. Test of the swelling degree, mechanical tensile strength, elongation, oxygen permeability, water-vapor loss and In vitro degradation treated by pretense TV of samples of artificial skin were measured by adequate methods so as to mechanical, physincal characterization and biodegradation. As a result, all the values of samples were found to be similar to desired value of skin which was nature. The Artificial skin based on polypeptides can be considered as ideal burn wound covering materials.

• KSBB Journal
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• v.17 no.2
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• pp.153-157
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• 2002

A fusion protein, consisting of a human epidermal growth factor as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as an inclusion body in recombinant E. coli, yet when the conventional solution-phase refolding process was used the refolding yield was very low due to severe aggregation, probably because of the opposite surface charge resulting from the vastly different pl values of each domain. Accordingly the solid-phase refolding process, which exploits the ionic interactions between a solid matrix and the protein, was tried, however the ionic binding yield was also very low regardless of the resins and pH conditions used. Therefore, to provide a higher affinity toward the solid matrix, six Iysine residues were tagged to the N-terminus of the hEGF domain. When cation exchange resins, such as heparin- or CM-Sepharose, were used as the matrix, the adsorption capacity increased 2.5~3-fold and the subsequent refolding yield increased nearly 15-fold compared to the conventional process. A similat result was also obtained when an Ni-NTA metal affinity resin was used.