• Microbiology and Biotechnology Letters
• /
• v.18 no.4
• /
• pp.344-351
• /
• 1990

An alkalophilic microoganism producing a detergent-resistant alkaline protease was isolated from soil and identified as Baeiltus sp. The alkaline protease has been partially purified by ammonium sulfate fractionation, DEAE-Cellulose, CM-Cellulose and Sephdex G-100 column chromatography. The purified alkaline protease was highly active at pH 12-13 toward casein and stable at pH values from 6 to ll. The optimum temperature for the enzyme reaction was $55^{\circ}C$. The enzyme was completely inactivated by diisopropyl fluorophosphate (DFP) indicating that the enzyme was serine protease, but considerabiy stable in the presence of surface active agents.

• The Korean Journal of Zoology
• /
• v.31 no.4
• /
• pp.300-308
• /
• 1988

Human placental alkaline phosphatase (PLAP), one of the oncofetal antigen was purified from placentas through the procedures including butanol extraction, concanavalin A-Sephar-ose, DEAE-cellulose and Sephadex G-200 gel chromatography. Monoclonal antibodies (fibs) against human PMP were produced by hybridizing SP 210-Ag 14 mouse myeloma cells with spleen ceils of Balblc mice immunized with PLAP. Six stable monoclones uvere obtained by cloning tuvice in serial dilutions, and the monoclonal speclfidty of these MAbs was confirmed by biochemical and immunonogical criteria. Tumor marker의 하나인 태반형 alkaline phosphatase(PLAP)에 대한 단일 클론항체의 생산과 분석을 위하여, 태반조직을 재료로 butanol 추출법 및 concanavaline A-Sepharose, DEAE-cellulose, Sephadex G-200 gel 크로마토그라피법에 의하여 PLAP를 순수 분리하였다. 이를 항원으로 하여 하이브리도마 방법에 의해 항-PLAP 단일 클론항체를 생산 분비하는 안정된 6클론세포를 얻었으며 생화학적 및 면역학적 분석방법으로 이들의 단일 클론성을 확인하였다.

• The Korean Journal of Zoology
• /
• v.29 no.4
• /
• pp.283-293
• /
• 1986

For the preliminary step to make and characterize the monoclonal antibodies of human alpha-fetoprotein (AFP) was purified from 534g of human fetal tissues through the procedures of tissue extraction, DEAE-cellulose, concanavalin A-Sepharose, Cibacron Blue F3GA-agarose and immunoadsorbent affinity chromatography. The isolated AFP preparation showed a single band on polyacrylamide gel electrophoresis and a single precipitin are against rabbit anti-human cord serum and anti-human AFP on immunoelectrophoresis. Our AFP also displayed a single band on SDS-polyacrylamide gel electrophoresis. The recovery of AFP was 8.76mg total.

• Applied Biological Chemistry
• /
• v.27 no.2
• /
• pp.73-78
• /
• 1984

The optimum culture time and initial pH, for the production of exo-maltotetraohydrolase from Pseudomonas stutzeri IAM 12097, in the trypticase medium were 36 hrs and pH 6.3, respectively. Exo-maltotetraohydrolase was purified by $(NH_4)_{2}SO_4$ and two times of column chromatography on DEAE-cellulose. Specific activity of the purified enzyme was 108.6U/mg protein and yield of the enzyme activity was 9.4%. The purified enzyme showed a single band on polyacrylamide gel electrophoresis and SDS-polyacrylamide gel electrophoresis.

• Applied Biological Chemistry
• /
• v.36 no.1
• /
• pp.1-6
• /
• 1993

Chitinase was induced in rice cell suspension culture with treatment of chitooligosaccharides mixture. Among eleven isozymes found in 10% polysacrylamide gel electropherogram, four isozymes were identified as induced enzymes. Acidic chitinase fraction separated in DEAE-cellulose column chromatography, includes three induced chitinase, while basic fraction contains only one induced isozyme. Treatment of chitooligosaccharides mixture enhanced the contents in both protein and chitinase activity in cell suspension culture media, but increase in chitinase activity was much higher than in protein.

• Microbiology and Biotechnology Letters
• /
• v.20 no.4
• /
• pp.401-408
• /
• 1992

The extracellular cholesterol oxidase produced from a soil microorganism HSL613 was purified and partially characterized. Through a series of purification procedures including concentration with CH2 concentrator, DEAE-cellulose column chromatography and gel filtration on Superose12, the purified enzyme was shown to have a specific activity of 108 units/mg protein giving 30.8-fold purification and final yield of 66%. The molecular weight of the enzyme was estimated to be 59,500 daltons by SDS-PAGE. The optimum temperature and pH for this enzyme were $50^{\circ}C$ and 6.0, respectively. The activity of the purified cholesterol oxidase was inhibited by $Ag^{2+}$, $Hg^{2+}$ and SDS.

• Journal of Life Science
• /
• v.8 no.5
• /
• pp.598-603
• /
• 1998

A marine microbe, Vibrio anguillarum was isolated from fish and studied for its concerning pathogenic substance of hemolysin. Purification of hemolysin was achieved by the procedure of ammonium sulfate precipitation from cul-ture filtrate, DEAE-cellulose chromatography, and G-200 gel filtration with 36 fold of purification and 2.3% yield. The molecular weight of the purified hemolysin was 38,000 dalton by SDS-PAGE. The purified hemolysin was stable at pH 6-9, below 45$^{\circ}C$, and up to 1% of NaCl, respectively. $Ca^{2+}, Cu^{2+}, Zn^{2+}, Fe^{2+}$ inhibited the hemolytic activity whereas EDTA and $Mg^{2+}$ did not.

• The Korean Journal of Food And Nutrition
• /
• v.16 no.2
• /
• pp.152-157
• /
• 2003

Serratia marcescens ATCC 25419 protease was purified to homogeneity by ammonium sulfate treatment, and DEAE-cellulose anion exchange chromatography. The specific activity of the enzyme was increased 448-fold during purification with an overall yield of 43.0%. Metal reactivation on the purified protease from S. marcescens was studied. S. marcescens protease was a metalloenzyme to be completely inhibited its activity by EDTA and the enzyme outstandingly inhibited by Hg, Fe, Cu, but the activity was increased approximately 20% by Co. The reactivation of the apoenzyme was effective with Mn, Co, Zn in pH range from 6 to 8. Among metalloenzymes prepared to the addition of Mn, Co, Zn to restore the degree of activity of native enzyme, Zn-enzyme was similar to the native enzyme in respects with enzyme activity, alkali-inactivation, thermo-stability.

• Microbiology and Biotechnology Letters
• /
• v.17 no.3
• /
• pp.198-201
• /
• 1989

Steptomyces sp. GI 32 with high production of glucose isomerase was isolated from soil. The maximum enzyme production was observed in the culture medium containing 1% sorbitol, 0.6% tryptone, 0.4% yeast extract, 1mM Fe$_2$(SO$_4$)$_3$ with initial pH 7.0 when the cell was cultured at 35$^{\circ}C$ about 18 hours with shaking. The enzyme was partially purified by ammonium sulfate fractionation and DEAE cellulose chromatography. The enzyme was also appeared to be relatively thermostable, and no apopreciable inactivation was observed after incubation at 7$0^{\circ}C$ for 1 hour. The optimal pH and temperature of the enzyme were pH 8.0 and 7$0^{\circ}C$, respectively.

• Microbiology and Biotechnology Letters
• /
• v.15 no.6
• /
• pp.425-429
• /
• 1987

The extra-cellular hemolysin from Bacillus thuringiensis subsp. israelensis was purified in the process of suiting with (NH$_4$)$_2$SO$_4$, Sephadex G-200 gel filtration, and DEAE-cellulose column chromatography. The purified hemolysin had molecular weight of approximately 47,000 dalton on SDS-polyacrylamide gel electrophoresis. The activity of purified hemolysin on human red blood cells was increased by thiol agents, but that was Inhibited by cholesterol, protease treatment, and metal salts such as CuSO$_4$, and FeSO$_4$, respectively.