• Journal of Microbiology and Biotechnology
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• 제20권1호
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• pp.117-126
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• 2010

A chitinase (CHT) and a protease (PRO) were purified from the culture supernatant of Serratia sp. TKU017, with shrimp shell as the sole carbon/nitrogen source. The molecular masses of CHT and PRO determined by SDS-PAGE were approximately 65 kDa and 53 kDa, respectively. CHT was inhibited by $Mn^{2+}$ and $Cu^{2+}$, and PRO was inhibited by most tested divalent metals and EDTA. The optimum pH, optimum temperature, pH stability, and thermal stability of CHT and PRO were pH 5, $50^{\circ}C$, pH 5-7, and <$50^{\circ}C$, and pH 9, $40^{\circ}C$, pH 5-11, and <$40^{\circ}C$, respectively. PRO retained 95% of its protease activity in the presence of 0.5 mM SDS. The result demonstrates that PRO is an SDS-resistant protease and probably has a rigid structure. The $4^{th}$-day supernatant showed the strongest antioxidant activity (70%, DPPH scavenging ability) and the highest total phenolic content ($196{\pm}6.2\;{\mu}g$ of gallic acid equiv./ml). Significant associations between the antioxidant potency and the total phenolic content, as well as between the antioxidant potency and free amino groups, were found for the supernatant. With this method, we have shown that shrimp shell wastes can be utilized and it is effective in the production of enzymes and antioxidants, facilitating its potential use in industrial applications and functional foods.

• Applied Biological Chemistry
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• 제33권1호
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• pp.73-78
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• 1990

한국산 토마토로부터 endo-polygalacturonase를 gel filtration과 이온교환크로마토그라피에 의하여 약 24배 정제할 수 있었고, 최대 효소활성을 위한 PH는 5.0, 최적온도는 $50^{\circ}C$ 였으며 pH안정범위는 $4.0{\sim}5.0$, 열안정성은 $50^{\circ}C$에서 1시간 열처리 하였을 때 약 45 % 실활 되었다. 정제된 이 효소는 SDS-polyacrylamide gel 전기영동에 의하여 단일밴드로 확인되었으며, 그 분자량은 50,000정도였고, Km값은 $1.43{\times}10^{-1}\;mol/l이었다. 금속이온중 $Ag^+$, $Zn^{++}$이온이 효소의 활성을 촉진시켰으며, $Na^+$, $K^+$등의 이온에 의해서는 활성이 저해되었다.

• 생명과학회지
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• 제8권3호
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• pp.326-332
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• 1998

Cyclodextrin glucanotransferase from B. stearothermophilus KJ16 that can produce both cyclodextrin glucanotransferase and cyclodextrinase was purified by ammonium sulfate precipitation, DEAE-cellulose chromatography, Sephadex G-100 chromatography, and FPLC. The molecular weight of the purifice enzyme was about 65,000 dalton by SDS-PAGE. The optimal pH and temperature were 6.0 and $60^{\circ}C$, respectively. The enzyme was stable at $50^{\circ}C$ for 1 hr and in the pH range of 5.5 and 8.5. Mercaptoethanol and dithiothreitol inhibited the enzyme activity strongly. The enzyme produced 60% cyclodextrin(CD) from 5% soluble starch with the $^{\alpha}$, $^{\beta}$, $^{\gamma}$-CD ratio of 42:46:12. Amylopectin was the most suitable substrate with 67% conversion to CD.

• 환경위생공학
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• 제10권1호
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• pp.126-131
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• 1995

Chitinase, a potential pathogensis- related portein, was induced in the leaves of 30 day - old bean( phaseolus vulgaris ) plant with the treatment of 10 $\mu $ /mℓ ethylen for 30hrs. Chitinase was purified from the mature tissue of bean leaves( phaseolus vulgaris ) by ammonium sulfate precipitation followed by affinity chromatography on a regenerated chitin and sephadex G-75 chromatography. The purified chitinase gave a single band SDS- PAGE to be 32,000 Dalton. In order to elucidate the three- dimensional structure of chitinase and to shed light on the functional mechanism of this class of enzymes, the enzyme was tried to crystallize( Sitting Drop Method). Crystals grew at room temperature to their final size within two weeks(0.2mm $\times $0.2mm $\times $ 0.1 mm ), This enzyme are still continuing to crystallize for the study of X- ray.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권4호
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• pp.334-340
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• 2005

Lectin is a cell-agglutinating and carbohydrate-binding protein present in many plants. The lectin of Canavalia ensiformis shoot with specific affinity for D-glucose was purified by affinity chromatography using Sephadex G-100, and some of its biochemical characterizations were studied. Lectin was purified 8.87-fold and exhibited final specific activity of 225.74 units/mg protein with a $2.3\%$ yield. SDS-PAGE analysis demonstrated that the purified shoot lectin exists as a tetramer of 102 kD, composed of two subunits with molecular weight of 29 and 22 kD. The purified lectin was observed to agglutinate rabbit blood cell. The optimal temperature for the activity of this lectin was $40^{\circ}C$, and this lectin was relatively stable to heat with the highest activity at $50{\~}60^{\circ}C$. The maximal activity was observed at pH 7.2.

• Journal of Microbiology and Biotechnology
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• 제12권4호
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• pp.553-556
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• 2002

The inhibitory activity of fifty Bifidobacteria toward the infectivity of a rotavirus, which is the predominant cause of sporadic diarrhea in infants and young children, was investigated, and Bifidobacterium breve K-110 was found to have the most potent inhibitory activity. Accordingly, the rotavirus infection-inhibitory protein was purified, and its molecular weight was determined to be 76 kDa by SDS-PAGE. It was heat-labile and its 50% inhibitory concentration ($IC_{50}$) was 0.045 $\mu g/ml$.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.205-205
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• 1994

동물, 곤충, 해조류 등으로 부터 얻어진 독소는 해로운 것으로 알려져 있지만 의학적으로 유용한 화합물을 내포하고 있다. 본 연구는 이러한 점에 착안하여 독사, 또는 곤충의 독 30여종을 대상으로 위암 세포인 SNU-1에 대해서 MTT assay에 의한 세포 독성 시험을 실시하였다. 이 중 세포 독성 활성이 가장 놓은 코브라 계통인 Ophiphagus hannah의 독으로 부터 세포 독성 물질의 정제를 시도하였다. HPLC-GPC와 HPLC-lEX 또는 RP-HPLC를 이용하여 정제하였고 각 분획들을 암세포주에 대해서 시험하였다. TSK-2000SW로 부터 2개의 분획을 얻어 분획 I에서 $IC_{50}$/ 의 값이 0.5 - 3 $\mu\textrm{g}$/ml의 범위 내에서 확인되었다. 분자량이 작은 분획 II에서는 세포독성이 작게 나타났지만 농축하여 성분 분석을 시도하였다. 분획 I을 RP-HPLC를 이용하여 TFA와 acetonitrile의 linear gradient에 의해 더욱 분리를 하였고, phopholipase $A_2$ 활성의 가능성도 측정하였다. 분획 II는 Mono S 칼람을 이용하여 ammonium acetate농도에 따른 pH gradient에 의한 분리를 시도하여 순도를 SDS-PAGE에 의해서 확인하였다. 각각의 크로마토그리피로 부터 얻은 분획에 대해서 세포 독성을 실시중에 있으며 성질도 동정중에 있다.

• BMB Reports
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• 제30권6호
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• pp.448-452
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• 1997

Polyclonal antibody of the boar sperminogen was used to characterize the boar sperm proteinase sperminogen. Boar sperminogen was purified from the acid extracts of the washed epididymal spermatozoa by gel filtration through a Sephadex G-100 column. followed by preparative SDS-PAGE. The sperminogen band was sliced out and was eluted from the gel matrix. The purified sperminogen was used to produce the polyclonal antibody of the boar sperminogen. When characterized on a Western blot, the final preparation of sperminogen appeared as a homogenous protein with a molecular weight of 32 kDa. The relative migration of sperminogen was distinctly different from the major components of the proacrosin-acrosin system as well as all the observable proacrosin activation by-products detected on the Western blot. The sperminogen antibody, however. cross-reacted with the proacrosin-acrosin system.

• 한국축산식품학회:학술대회논문집
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• 한국축산식품학회 2004년도 제34차 추계 국제 학술대회
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• pp.345-348
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• 2004

젖소 초유중에 함유된 IGF-I 은 30kDa와 1kDa ultrafiltration(UF) membrane을 이용하여 IGF-I rich fraction을 효과적으로 분리하였으며 분획내의 IGF-I은 SDS-PAGE와 Western blot으로 확인하였다. 분획한 IGF-I rich fraction이 murine splenocyte의 면역 활성에 미치는 영향을 실험한 결과 $1{\mu}g$ 투여군의 경우 대조구에 대비하여 Bcell과 T cell의 증식능력은 각각 33%와 76%의 증식능력을 보였고, natural killer cell의 항암 능력은 42.3%의 세포 독성을 나타내었다.

• 한국축산식품학회:학술대회논문집
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• 한국축산식품학회 2004년도 제34차 추계 국제 학술대회
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• pp.340-344
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• 2004

젖소 초유 중에 함유된 IGF-I rich fraction을 30kDa와 1kDa ultrafiltration(UF) membrane을 이용하여 효과적으로 분리하였으며 분획내의 IGF-I은 SDS-PAGE와 Western blot으로 확인하였다. 분획한 IGF-I rich fraction이 murine macrophage의 면역 활성에 미치는 영향을 실험한 결과 $1{\mu}g$ 투여군의 경우 IL-6는 7.3ng/ml, NO는 $10.7{\mu}M$을 생성하였고 Phagocytosis는 대조구 대비하여 65% 증진시켰다. $TNF-{\alpha}$의 분비량은 L929세포의 증식 저해율로 측정한 결과 대조구에 대비하여 30% 더 높은 저해율을 나타내었고, $H_2O_2$는 대조구에 대비하여 6% 더 높은 과산화수소를 생산하였다.