• KSBB Journal
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• 제16권5호
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• pp.474-479
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• 2001

$_{L}$-Iysine 의 생산을 Corynebacterium glutamicum SH35을 이용하여 유가식 배양법과 연속 배양법으로 수행하였다. 유가시 배양에 의한 $_{L}$-Iysine 생산을 조사한 결과 최종 발효액 중의 lysine 농도는 129.2 g/L, lysine 수율 및 productivity는 가각 47%m 3.08g.$L^{-1}$.$h^{-1}$이였다. Single-stage 연속 배양에서 dilution rate에 따른 균의 생장 및 lysine 생산을 조사한 결과, critical diluation rate은 0.100 h$^{-1}$으로 이때 균체농동($OD_{610}$)은 67, lysine 농도는 44.0 g/L 그리고 lysine 수율 및 productivity 는 41%, 4.39 g.$L^{-1}$.$h^{-1}$이고 lysine 연속배양 시 최적의 dilution rate 이였다. 또한 lysine 생산을 위한 medium reservoir 내의 최적 탄소원 농도, 최적 질소원(ammonium sulfate) 농도 및 최적 pH는 각각 108 g/L, 25 g/L 및 6.9이었다. 유가식 배양법과 single-state 연속 배양법에 의한 lysine 생산에서, productivity 는 유가식 배양의 경우 3.08 g.$L^{-1}$.$h^{-1}$인 반면, 연속 배양에서는 4.39 g.$L^{-1}$.$h^{-1}$로 유가식 배양법에서보다 약 1.4배 정도 증가하였따. 연속 발효 공정을 통해 생산 역가의 향상은 있었으나, lysine 농도와 수율이 낮아 이에 대한 해결책으로 two-stage 연속 배양 방법을 수행하였다. 그 결과, 연속 배양 발효 수행보다 lysine 농도는 2배 정도 증가하였으며, lysine 수율은 46%로서 유가식 배양과 비슷한 정도로 얻어졌다. 이러한 실험 결과로 미루어 lysine 생산 역가 향상을 위한 방법으로 two-stage 연속배양의 산업적인 이용가능성이 있을 것으로 사료된다.

• 한국자기공명학회논문지
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• 제6권1호
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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.

• 한국미생물·생명공학회지
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• 제15권6호
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• pp.369-374
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• 1987

하상 오니 및 토양으로부터 D-$\alpha$-Amino-$\varepsilon$-Caprolactam(DAC)의 자화능이 우수한 균을 분리하여 Alcaligenes eutrophus로 동정하였다. Alcaligenes eutrophus A52의 생육을 위한 배양조건은 배양 초기 최적 pH가 6.0, 최적온도는 3$0^{\circ}C$, 최적 DAC 농도는 0.2%로서 50시간 배양으로 0.2% DAC를 거의 다 자화하여 정지기에 도달하였다. 본 균은 탄소원으로 glucose와 fructose, 질소원으로는 NH$_4$Cl, NH$_4$NO$_3$, (NH$_4$)$_2$SO$_4$ 및 NaNO$_3$를 잘 자화하였다. 탄소원 및 질소원으로 Iysine, glutamate를 잘 이용하였으나 그 밖의 아미노산 자화능은 낮았다. 균체로부터 추출한 3.1mg의 단백질에 상당하는 무세포 추출 액으로 0.5% DAC를 55%에서 12시간 반응시킨 생성물은 약 98% L-Iysine으로 확인되었다.

• KSBB Journal
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• 제7권3호
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• pp.209-215
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• 1992

당근 현탁배양세포에서 세포벽 합성 효소인 GSII의 활성에 다가양이온의 작용기작을 조사하였다. 다가양이온의 특성을 가진 poly-L-Iysine과 poly-L-ornithine은 GSII의 활성을 40~50% 정도 촉진 시켰으며, ATP도 NaF도 이와 비슷한 결과를 나타냈다. 한편 poly-L-Iysine과 poly-L-ornithine은 막투과성에는 영향을 미치지 못하였으며, protein kinase의 활성제인 TPA는 GSII의 활성을 대조구에 비해 35%정도 증가시켰고 억제제인 H-7은 30%정도 감소시켰다, 이러한 결과들은 다가양이온이 인산화 과정을 통해 GSII의 활성을 증가시켜 세포벽의 합성을 증진시키리라 사료된다.

• Journal of Plant Biotechnology
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• 제5권1호
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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.

• 센서학회지
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• 제6권6호
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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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• 제13권3호
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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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• 제7권5호
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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.

• 대한의용생체공학회:의공학회지
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• 제8권1호
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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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• 제17권2호
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• pp.153-157
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• 2002

Poly-Iysine이 tagging된 hEGF와 angiogenin(6L10ESA)의 융합단백질의 고체상 재접힘이 heparin-Sepharose colullln에서 수행되었을 때, untagging 단백질(E5h)의 기존의 액상 재접힘 방법과 비교하여 재접힘 수율은 약 13배 정도 증가하였다. 게다가 poly-Iysine tagging된angiogenin은 heparin에 친화도를 높여주므로 2.5배에서 3배 정도의 흡탁 수율이 증가한다. 재접힘 수율은 고체상 반응으로 인해 높은 재현성을 보였다. 재접힘 공정시간은 대략 8배 단축되었다. 고체상 재전힘된 단백질은 자신의 생물학적 역가를 유지하였다. 따라서 이 연구는 고체상 재접힘 방법이 분자간의 상호작용을 억제하여 응집현상을 현저히 줄였기 때문에 기인한 결과로 생각된다. 따라서 응집으로 인한 재접힘 수율이 낮은 단백질의 재접힘 긍정에 고체상 재접힘 공정을 사용하면 높은 재접힘 수율을 얻을 수 있다.