• 미생물학회지
• /
• 제28권3호
• /
• pp.278-282
• /
• 1990

Diffusion and partition of progesterone into the polyacrylamide gel was examined. Diffusion coefficient of progesterone decreased down to an asymptotic value as the concentration of the organic solvents in the diffusing medium increased. However the partition coefficient diminished steadily. Crosslinking density in the gel didn't affected the diffusion coefficient considerably but lowered the partition coefficient due to the contraction of pore volume of the gel. Progesterone showed higher diffusion coefficient as well as partition coefficient in the polyurethane than in the polyacrylamide gel, which seems to be ascribed to the difference in hydrophobicity, pore volume and pore size of the polymer matrix.

• Journal of Microbiology
• /
• 제34권4호
• /
• pp.384-386
• /
• 1996

Application of polyacrylamide gel (PAG) instead of agar to solid cultures of Streptomyces spp. was studied. The improved media were prepared by 1) gelling 20 ml of 5% acrylamide in a glass petri dish at room temperature, 2) washing by running water for more than 8 hr to remove residual reaction reagents, 3) drying at 5$0^{\circ}C$ for 12 hr to make a gel film, 4) autoclaving at 121$^{\circ}C$ for 15 min, and 5) swelling gel for about 4 hr by adding sterile liquid medium. In PAG media there were no differences from the observation of morphological characteristics showing during the cellular differentiation on agar media, whereas the ability to utilize carbohydrates differed somewhat from agar media. Agar media thus were little favorable for biochemical tests which the growth was determined depending on the formation of colony, but washed PAG was superior to serve as a solidifying agent.

• Applied Biological Chemistry
• /
• 제27권2호
• /
• pp.73-78
• /
• 1984

Pseudomonas stutzeri IAM 12097의 trypticase 배지(培地)에서 36시간(時間), initial pH는 6.3일때 Exo-maltotetraohydrolase가 최대로 생산되었다. Exo-maltotetraohydrolas, 황산(黃酸)암모니아분획(分劃)과 2회(回)의 DEAE-cellulose column chromatography에 의하여 정제(精製)하였으며 정제효소(精製酵素)의 specific activity는 108.6 u/mg protein, 수율(收率)은 9.4%이었다. 본정제효소(本精製酵素)는 polyacrylamide gel electrophoresis와 SDS-polyacrylamide gel electrophoresis에 의하여 각각(各各) 단일(單一) band를 나타내었다.

• BMB Reports
• /
• 제57권2호
• /
• pp.116-121
• /
• 2024

We investigated the therapeutic potential of bone marrow-derived mesenchymal stem cell-conditioned medium (BMSC-CM) on immortalized renal proximal tubule epithelial cells (RPTEC/TERT1) in a fibrotic environment. To replicate the increased stiffness characteristic of kidneys in chronic kidney disease, we utilized polyacrylamide gel platforms. A stiff matrix was shown to increase α-smooth muscle actin (α-SMA) levels, indicating fibrogenic activation in RPTEC/TERT1 cells. Interestingly, treatment with BMSC-CM resulted in significant reductions in the levels of fibrotic markers (α-SMA and vimentin) and increases in the levels of the epithelial marker E-cadherin and aquaporin 7, particularly under stiff conditions. Furthermore, BMSC-CM modified microRNA (miRNA) expression and reduced oxidative stress levels in these cells. Our findings suggest that BMSC-CM can modulate cellular morphology, miRNA expression, and oxidative stress in RPTEC/TERT1 cells, highlighting its therapeutic potential in fibrotic kidney disease.

• KSBB Journal
• /
• 제19권5호
• /
• pp.368-373
• /
• 2004

The optimal condition for the production of a glucan and a fructan synthesizing enzymes from Leuconostoc mesenteroides NRRL B-1149 were studied based on the different medium compositions. Response surface methodology was applied to find the optimistic condition showing the relationship between the fermentation response (enzyme activities) and the fermentation variable concentrations of yeast extract, peptone concentration, K2HP04 concentration and sucrose. Optimum medium composition for both enzymes production was $0.75\%$ yeast extract, $0.72\%$ peptone, $1\%$ K2HP04 and $2.17\%$ sucrose. Using this medium, the activities produced in culture was 0.90 U/m~ for glucosyltransferase (GTase) and 0.96 U/ml for fructosyltransferase (FTase). After purification of 1149FTase by consecutive chromatographies using Sephadex G-150 and DEAE-Sepharose, a 1149FTase of 210 kDa on $7\%$ polyacrylamide gel was isolated and it synthesized soluble fructan. The 1149GTase showed a band of 180 kDa on $8\%$ polyacrylamide gel after purification using Bio-Gel P-100 gel chromatography and DEAE-Sepharose ion exchange chromatography and it synthesized insoluble glucan. The linkages of polymers were determined by methylation using Hakomori reagent and following NMR analysis. The glucan was composed of a(1~6) and a(1~3) linkages and the fructan was levan.

• KSBB Journal
• /
• 제13권5호
• /
• pp.535-539
• /
• 1998

The monellin, a sweet-taste protein, was expressed and secreted in Saccharomyces cerevisiae. The secreted menellin was concentrated using an ultrafiltration membrane with a nominal molecular weight cut off of 3,000 or by ammonium sulfate precipitation. The monellin was purified by G-25 gel filtration chromatography, followed by CM-Sepharose ion exchange chromatography. The purified monellin was characterized by SDS-PAGE (SDS-Polyacrylamide Gel Electrophoresis) and PHLC. The molecular weight of monellin was found to be 10,700 dalton, and its purity was over 95%.

• 한국미생물·생명공학회지
• /
• 제20권4호
• /
• pp.409-416
• /
• 1992

Bacillus cereus에 의한 pullulanase 생산의 최적 배양온도 및 배양시간은 각각 $15^{\circ}C$, 72시간이고, 기본배지에 casein, nutrient broth, egg albumin의 첨가는 효소의 생산을 크게 증가시켰다. 황산암모늄분획, CM-cellulose와 DEAE-cellulose column chromatography에 의하여 효소를 정제하였고 정제효소의 specific activity는 29.09U/mg protein, 수율은 17.1이었다. 정제효소는 polyacrylamide disc gel electrophoresis에 의하여 single band를 나타내었고, SDS-polyacrylamide disc gel electrophoresis에 의하여 추정된 분자량은 61,000, 등전점은 pH7.0, 최적온도는 $40^{\circ}C$, 최적 pH는 6.5, $35^{\circ}C$ 이하에서 안정하였고, pH 안정 범위는 6.5-11.0, $Ag^{+}$, $Hg^{2+}$, $Zn^{2+}$에 의하여 크게 저해되었고, $Ca^{2+}$은 효소의 내열성을 증가시켰다. 정제효소는 공시기질중 pullulan을 잘 분해시켰으며 pullulan에 대한 분해산물은 maltoriose이었다.

• 한국미생물·생명공학회지
• /
• 제23권6호
• /
• pp.695-701
• /
• 1995

An alkaline lipase producing bacteria was isolated from soil and identified as Serratia liquefaciens AL-11. from the results of analysis of its morphological, biochemical and physiological properties. This strain showed the highest productivity of alkaline lipase when grown at pH 9.0 and 30$\circ$C for 42 hours in the medium of 1% peptone, 0.5% tryptone, 0.9% yeast extract, 1% starch, 1% tween 80, 0.05% CaCl$_{2}$ and 0.05% NaCl. The enzyme was purified by ammonium sulfate treatment, Sephadex G-100 gel filtration and DEAE-Sephadex A-50 column chromatography. The specific activity of the purified enzyme was 27 unit/mg protein and the yield of enzyme activity was 61.3%. The homogeneity of the purified enzyme was verified by polyacrylamide gel disc electrophoresis. Molecular weight of the purified enzyme was estimated about 53,000 by sodium dodecyl sulfate- polyacrylamide gel electrophoresis. This enzyme is composed of 17 amino acids of which glycine, proline and glutamic acid were three miajor acids.

• 한국식품영양과학회지
• /
• 제24권4호
• /
• pp.570-577
• /
• 1995

Aspergillus sp. SB-2704 was selected for its strong polygalacturonase activity among various strain of mold found in soil. It was found that production of polygalacturonase reached to maximum when the wheat bran medium containing 1% polypepton, 1% glucose, and 0.2% FeSO4 were cultured for 3 days at 35$^{\circ}C$. Polygalacturonase was purified 20.90 fold from Aspergillus SB-2704. The purification procedures include ammonium sulfate treatment, gel filtration on Sephdex G-150 and DEAE-cellulose ion exchange chromatography. Yield of the enzyme purification was 4.34%. Purified enzyme was confirmed as a single band by the polyacrylamide gel electrophoresis. When the purified enzyme was applied to SDS-polyacrylamide gel electrophoresis, the molecular weight was estimated to be 36,000. The optimum pH for the enzyme activity was 5.5 and optimum temperature was 5$0^{\circ}C$. The enzyme is stable in acidic condition. The activity of purified enzyme was inhibited by Pb2+, Hg2+ and Ba2+, whereas activated by Cu2+, Mn2+, Mg2+ and Fe2+. The activity of polygalacturonase was inhibited by the treament wit maleic anhydride, iodine, and EDTA. The result indicate the possible involvement of histidine and metal ion at active site.

• 한국미생물·생명공학회지
• /
• 제18권3호
• /
• pp.251-259
• /
• 1990

생전분 분해력이 강력한 glucoamylase를 생산하는 균주로서 Asp.usamii IAM 2185를 선정하였다. 밀기울배지에서의 효소생산의 최적 initial pH는 6.0-8.0, 최적 배양온도는 25-$30^{\circ}C$, 최적 배양시간은 72시간이고, 밀기울배지에 ammonuim nitrate와 albumin의 첨가는 효소의 생산을 약간 증가시켰다. 황산암모늄분획, CM-cellulose와 DEAE-cellulose column chromatography에 의하여 효소를 정제하였고, 정제효소의 specific activity는 34.3U/mg.protein, 수율은 10.3 이었다.