• Microbiology and Biotechnology Letters
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• v.17 no.2
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• pp.136-141
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• 1989

A raw starch-digesting enzyme from Streptomyces sp. 4M-2 was purified by ammonium sulfate fractionation, DEAE-Sephadex A-50 column chromatography and Sephadex G-100 gel filtration. The specific activity of the purified enzyme was 51.22 RSU/mg protein and the yield was 4.5% of the total activity of the culture broth. The purified enzyme was found to be homogeneous by polyacrylamide gel electrophoresis and its molecular weight was estimated to be about 102, 000 daltons by SDS-polyacrylamide gel electrophoresis, The optimal temperature and pH for the enzyme activity were 42$^{\circ}C$ and PH 5.5, respectively. The enzyme had Km, value of 44.44mg/$m\ell$ for raw corn starch. The enzyme was activated by addition of calcium and barium ions. Corn amylose was degraded by the enzyme very easily and raw potato starch was also degraded easily. Main products of the enzymatic hydrolysis of raw corn starch were analyzed to be maltose and maltotriose. The enzyme was considered as $\alpha$-amylase.

• Korean Journal Plant Pathology
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• v.3 no.4
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• pp.239-244
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• 1987

Low molecular weight plant ribonucleic acids including potato spindle tuber viroid(PSTV) RNA were electrophoresed in 0M to 8M urea-gradient polyacrylamide gels. The electrophoresis was carried on in a urea - gradient gel system with 1/40 and 1/10 dilution of TBE buffer at three different temperatures, $17^{\circ}C,\;37^{\circ}C\;and\;57^{\circ}C$. The most effective separation of PSTV - RNA molecules into circular and linear forms was achieved at the highly denaturing temperature of $57^{\circ}C$ and at 1/40 dilution of TBE buffer. The electrophoretic mobility of the denatured circular viroid-RNA molecules is dependent mainly on the concentration of urea. In addition, a low concentration of TBE buffer would increase the separation distance between the circular and linear forms of PSTV-RNA molecules in the denaturing urea-gradient gel system

• Korean Journal of Microbiology
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• v.25 no.3
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• pp.180-183
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• 1987

Stu I, type II restriction endonuclease, has been purified to homogeneity from Streptomyces tubercidicus (ATCC 25502), and its catalytic properties have been studied. For the purification of Stu I endonuclease free of nonspecific nucleases, DEAE-Sephadex (A-50), QAE-Sephadex (A-50) and Heparin-agarose column chromatography have been performed after ammonium sulfate fractionation of the crude extract. The enzyme was further purified by gel filtration using Sephadex G-100 column to obtain homogeneous form of protein. The single polypeptide species of Stu I endonuclease has a subunit molecular weight of 34,000 $\pm$ 1,000 daltons as judged by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Stu I endonuclease requires $Mg^{2+}$ ion for its activity and is maximally active at neutral pH (7.0-8.0) in the absence of NaCl.

• Applied Biological Chemistry
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• v.37 no.4
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• pp.243-247
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• 1994

Endo-polygalacturonase was purified from Jujube. The purification procedures included DEAE-cellulose ion exchange chromatography and gel filtration on Sephdex G-100. Enzyme was purified as a single protein band and purification yield was about 6%. When the purified enzyme was applied to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular weight was estimated about 19,000. Purified enzyme formed hexagonal board type.

• Korean Journal of Microbiology
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• v.23 no.1
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• pp.13-19
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• 1985

Acc I endonuclease has been isolated from 300g (wet weight) cells of Acinetobacter calcoaceticus. The cells were broken by using French press at 20, 000p.s.i. After ammonium sulfate fractionation, the enzyme was further purified by heparin agarose, DEAE-sephades, Affi.-gel Blue, phosphocellulose, and hydroxylapatite column chromatography. The purified Acc I endonudlease has a single polypeptide species and its subunit molecular weight was 45,000 ${\pm}$ 1,000 daltons as judged by 10% SDS-polyacrylamide gel electrophoresis. The isolated enzyme was essentially free of contaminating nucleases as judged by homochromatography by using a $^{32}P-labeled$ oligonucleotide. The enzyme showed maximum activity at pH values between 8.0 and 11.0 and in the presence of $MgCl_2$. Acc I endonuclease was maximally active in the absence of NaCl and was completely inhibited at 200 mM NaCl.

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

• Journal of the Korean Society of Tobacco Science
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• v.12 no.1
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• pp.9-18
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• 1990

To study the determinants which are involved in the loss of pathogenicity in wilt-inducing Pseudomoms solamcewum, several physlologica I functions were compared in a virulent P. solanacearum strain and an avirulent, spontaneously derived mutant strain. The polyacrylamide gel electrophoresis showed the distinction between two strains in the patterns and the relative intensity of proteins produced intracellularly or extracellularly. Enzyme assays showed that the level of polygalacturonase activity in the culture filtrate of the avirulent mutant was markedly reduced, while carboxymethylcellulase(rondoglucanase) activity in both strains were nearly negligible. These results suggest that the loss of pathogenicity in mutant strain is attributed in part to the reduced production of polygalacturonase. In audit ion, comparative analyses by agarose gel electrophoresis of DNA molecules isolated from both strains show that the pathogenicity genes of p. solanaceerum are not located on plasmid but are on chromosome.

• Korean Journal of Food Science and Technology
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• v.18 no.4
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• pp.288-293
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• 1986

A raw starch-digesting enzyme from Rhizopus oryzae was purified by ammonium sulfate fractionation, DEAE-sephacel column chromatography and Sephadex G-150 gel filtration. The specific activity of purified enzyme was 45.2 Ulmg protein and the yield was 16.2%. The purified enzyme was found to be homogeneous bypolyacrylamide gel electrophoresis and its molecular weight was estimated to be 67,000 by SDS-polyacrylamide gel electrophoresis, and also the enzyme had Km value of 4.082 mg/ml for raw corn starch. The optimal temperature and pH for the enzyme activity were $50^{\circ}C$ and 4.0-5.0, respectively. Reaction product of raw corn starch by purified enzyme was glucose mainly.

• BMB Reports
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• v.28 no.6
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• pp.504-508
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• 1995

The electrophoretic patterns of plasma membrane surface proteins of normal rat liver cells and rat hepatomas were compared in 10% non-denaturing and 7-15% gradient non-denaturing gel. Chemical carcinogens, 2-Me DAB (2-methyl-4-dimethylaminoazobenzene) and DENA (diethylnitrosamine), were used to induce hepatoma in rats. One protein which disappeared in hepatoma was identified in normal rat liver by non-denaturing gel electrophoresis. Rabbit antisera were raised against this specific protein, and the protein was purified by Sephacryl S-200 column and immunoaffinity chromatography using the purified antibody. The purified protein showed two bands of molecular weights approximately 50 $kD_{\alpha}$ and 52 $kD_{\alpha}$ by SDS-polyacrylamide gel electrophoresis, which reacted specifically with the antibody. However only one band was observed in non-denaturing gel and also in isoelectric focusing with a pI value of 6.6. This study showed the existence of an unique protein on the plasma membrane surface of normal rat liver cells which disappeared in rat hepatomas induced by chemical carcinogens.

• Korean Journal Plant Pathology
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• v.2 no.2
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• pp.121-128
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• 1986

The biochemical properties of ribonucleic acid (RNA) and coat protein of the mild tobacco mosaic virus (TMV) mutant, Tw 333 are described. The molecular weight of the RNA calculated from polyacrylamide gel electrophoresis was $2.03\times10^6$ daltons. The molar ratio of the bases of the RNA was 25.4 guanine, 29.2 adenine, 17.5 cytosine and 27.9 uracil in moles. The hyperchromicity on Tw 333-RNA by thermal denaturation was $25.1\%$, indicating Tm value of $47^{\circ}C$. The virus coat protein migrated as a single component in SDS-polyacrylamide gel electrophoresis and had a molecular weight of 17,500 daltons. A total of 158 amino acid residues are present in the protein. Separation of the tryptic peptides by electrophoresis and chromatography yielded ninhydrin-positive compounds. The biochemical properties of RNA and coat protein of the mild mutant we very similar to those of wild type of TMV-OM strain, but some difference between the strains were observe in the base composition, hyperchromicity, amino acid composition and tryptic peptide map.