• 한국미생물·생명공학회지
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• 제18권5호
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• pp.501-505
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• 1990

토양으로부터 분리한 Streptomyces 속 세균 KIS 13은 thiol 계통 단백질분해효소 활성을 특이적으로 저해하는 저분저량 저해물질을 생성하였다. 저해물질 생성은 세균체성장에 연관된 생성양상이 나타내었다. 배양액으로부토 butanol 추출, silicagel 60 column chromatography, Sephadex LH-2 gel-filtration chromatography, preparative HPLC 등의 과정을 통하여 단백질 분해효소 저해물질을 순수분리하였다. 이 저해물질은 Hammersten casein을 기질로 사용할때, papain에 대하여 non-competitive한 저해양상을 나타내었다.

• 한국미생물·생명공학회지
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• 제28권5호
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• pp.251-257
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• 2000

Serratia marcescens purine nucleoside phosphorylase (PNP) was purfied to homogeneity by streptomycin sulfate treatment, Sephacry HR S-200 gel filtration chromatography and AMP-agarose affinity chromatography. The specific activity of the enzyme was increased 49-fold during purification with an overall yield of 7.0%. The molecular weight was 168kD as estimated by Sephadex G-150 gel filtration chromatography. The S. marcescens enzyme was composed of six identical subunits with subunit molecular weight of 28kD, as estimated by SDS-PAGE. The Km values of S. marcescens enzyme for inosine and deoxyinsoine were 0.38 and 1.20 mM, respectively. The ph optimum was near 8.0, and the enzyme was relatively heat-stable protein. The enzyme was inactivated com-pletely by 0.5 mM of $Cu^{ 2+}$.

• Journal of Microbiology
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• 제34권1호
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• pp.82-89
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• 1996

Purine nucleoside phosphorylase (PNP) was purified in Micrococcus luteus (M. luteus) using streptomycin sulfate and amomonium sulfate fractionation, three times by a Sephadex G-100 gel filtration and a DEAE-Sephadex A-50 ion exchange chromatography. The enzyme was purified 72 folds with a 11% recovery and showed a single band in a nondenaturing gel electrophoresis. The M. W. of PNP turned out to be 1.35 * 10$^{5}$ delton in G-150 gel filtration chromatography. The stability of the enzyme was increased by treatment with both substrates, MgCI$_{2}$ or CaCI$_{2}$, but not significantly kcal/mol. M. luteus PNP catalyzed the phosphorolysis of inosine, deoxyinosine, guanosine and deoxyguanosine with the Km value of 1.5 * 10$^{-3}$ M, 3.0 * 10$^{-3}$ M, 5.0 * 10$^{-4}$ M, respectively. The enzyme was reacted with adenosine, 1-methylnosine and 1-methylguanosine as substrates, which were shown to be poor substrates for mammalian enzyme.

• 미생물학회지
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• 제37권1호
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• pp.21-27
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• 2001

Bacillus anthracis Sterne 34F$_2$균주로부터 PA를 생산하기 위해 RM배지를 변형하였다. NaHCO$_3$를 8 g/l에서 10 g/l로, glucose를 5 g/l로 첨가하여 새로운 배지조성에서 탄저균을 배양한 후, 배양액을 hydroxyapatite를 이용하여 농축하였다. 농축된 조단백질을 hydroxyapatite column chromatography, DEAE-Sepharose CL-4B column chromatography 및 Toyo-pearl gel filtration chromatography를 사용하여 PA를 정제하였다. 변형된 RM 배지를 사용해 얻은 PA 양은 8.6 mg/l 이었다.

• 한국양식학회지
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• 제21권4호
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• pp.285-293
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• 2008

본 연구에서는 동자개, Pseudobagrus fulvidraco의 난황단백질(Vitellin, Vg 난황단백전구체(Vitellogenin, Vg)를 분리하였고, 그 특성을 분석하였다. 난황단백질은 성성숙한 난소 난으로부터 gel filtration chromatography (Sephadex - G200)을 이용하여 분리되었다. 동자개의 난황단백질은 SDS-PAGE에서 분자량이 107 kDa인 1개의 subunit로 구성되어 있었고, 총 난황 단백질 분자량은 360 kDu으로_ 측정되었다. 난황단백전구체는 성숙한 수컷에 $E_2$를 삽입하여 유도하였고, anion exchange chromatography (Momo Q HR 5/5 column)을 이용하여 분리되었다. 분리된 난황단백전구체의 총 분자량은 450 kDa으로 계산되었고, 이는 SDS-PAGE하에서 110 KDa, 125 kDa와 147 kDa로 3개의 subunit로 구성되어져 있었다.

• Journal of Microbiology and Biotechnology
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• 제18권4호
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• pp.670-675
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• 2008

A laccase was isolated from the culture filtrate of the basidiomycete Fomitella fraxinea. The enzyme was purified to electrophoretical homogeneity using ammonium sulfate precipitation, anion-exchange chromatography, and gel-filtration chromatography. The enzyme was identified as a monomeric protein with a molecular mass of 47 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel-filtration chromatography, and had an isoelectric point of 3.8. The N-terminal amino acid sequence for the enzyme was ATXSNXKTLAAD, which had a very low similarity to the sequences previously reported for laccases from other basidiomycetes. The optimum pH and temperature for 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS) were 3.0 and $70^{\circ}C$, respectively. The enzyme also showed a much higher level of specific activity for ABTS and 2,6-dimethoxyphenol (DMP), where the $K_m$ values of the enzyme for ABTS and 2,6-DMP were 270 and $426{\mu}M$, respectively, and the $V_{max}$ values were 876 and $433.3{\mu}M/min$, respectively. The laccase activity was completely inhibited by L-cysteine, dithiothreitol (DTT), and sodium azide, significantly inhibited by $Ni^+,\;Mn^{2+}$, and $Ba^{2+}$, and slightly stimulated by $K^+$ and $Ca^{2+}$.

• International Journal of Industrial Entomology and Biomaterials
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• 제6권2호
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• pp.151-155
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• 2003

Using gel filtration chromatography, high molecular silk fibroin with high purity was obtained and silk flbroin microsphere particles (SFMP) could be simply made by spray dryer method. Also, some of the physicochemical properties of SFMP and morphology were investigated. The average molecular weight of pure silk fibroin protein dissolved in calcium chloride is about 61,500g/㏖ as measured by gel permeation chromatography. SFMP was spherical in shape, and particles, sized average of 2 ${\pm}$ 10 ${\mu}$, were observed by SEM and particle analyzer, respectively. Obtaining microspheres particles by spray dryer method accelerated the transition from the random coil to the $\beta$-sheet structure during spray dryer treatment. It was identified by the basic fourier transform infrared spectroscopy of SFMP. The swelling ratio of SFMP is majorly dependent on the pH of the solution, not on the occurred gelation. The characteristic structure, which might be applicable to immobilization of drugs is superior to other matrix materials for the use of biomaterials with skin affinity.

• 한국식품과학회지
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• 제31권5호
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• pp.1184-1187
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• 1999

치자황색소로부터 미생물에 의해 변환된 색소의 색조차이를 규명하고자 L. plantarum과 S. epidermidis에 의해 변환된 색소를 Amberlite XAD-4 column chromatography로 정제 한 다음 Sephadex LH-20 column의 gel filtration 용출양상과 분리된 색소의 분광학적 특성을 조사 하였다. L. plantarum에 의해 변환된 색소는 crocetin보다 큰 분자크기를 갖고 588 nm에서 홉수극대를 갖는 단일의 청색소를 생성하며, S. epidermidis에 의해 변환된 색소는 L. plantarum에 의해 생성되는 588 nm에 흡수극대를 갖는 청색소와 함께 crocetin 보다 작은 분자크기와, 413 nm에서 흡수극대를 갖는 황색소를 동시에 생성함으로서 청녹색을 띠는 것임이 밝혀졌다.

• Journal of Animal Science and Technology
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• 제46권1호
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• pp.77-82
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• 2004

본 연구는 우유의 angiogenin(ANG) 생리활성기능을 조사하기 위한 전단계로 ANG 정제 방법을 확립하기 위하여 실시하였다. 우유로부터 ANG을 CM-Sepharose ion exchange chromatography, HPLC, Gel-filtration의 3단계 방법을 이용하여 정제하였다. 1단계인 CM-Sepharose ion exchange chromatography를 수행 한 결과 0.6 M NaCl 농도의 36번 분획에서 ANG의 밴드를 볼 수 있었고 2단계인 Mono-S 컬럼을 이용하여 HPLC를 수행한 결과에서는 1.0 M에서 하나의 peak가 분리되었고 전기영동 결과는 LF와 ANG 두 가지 성분이 함유되어 있음을 확인하였다. 마지막 단계인 Gel-filtration을 수행한 결과 ANG이 단일 밴드로 분리되었다. 우유 10 $\ell$로부터 약 80 ${\mu}\ell$의 ANG를 얻었고 정제된 ANG을 N-말단 아미노산 배열을 분석한 결과 AQDDYRYIHFLTQHY로 밝혀졌다.

• KSBB Journal
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• 제13권5호
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• pp.535-539
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• 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%.