• KSBB Journal
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• v.21 no.5
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• pp.394-400
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• 2006

When the papain, which is a sort of Cystein protease, is applied to the outer skin, it decomposes the protein which forms the peeled outer skin and speeds up metabolism. Therefore, it is one of the most important cosmetics compositic which keeps the function of skin normal. When the papain is used in cosmetics with surfactant, the activity of papain is reduced rapidly. In this study, the modified papain with extreme stability negative ionic environment was developed by directed evolution

• Korean Journal of Plant Resources
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• v.19 no.2
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• pp.348-354
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• 2006

Homogenized prothallus of 6 species (Cyrtomium falcatum, Cyrtomium caryoptideum var. coreanum, Dryopteris varia, Asplenium incisum, Camptosous sibiricus and Phyllitis scolopendrium) were treated with gamma radiation or by chemical mutagenesis with EMS, NMU, $NaN_3$ and colchicine to assess their sensitivities for each treatments and also with the aim of inducing mutations. Generally, decrease of proliferation ratio was dose-dependent and time-dependent. Based on proliferation ratio, optimum dose of gamma irradiation was $5{\sim}10krad$ except in D. varia with 20krad. Optimum condition of EMS treatment was considered as 50mM for 3h and for NMU as $5{\sim}10mM\;for\;1{\sim}3h$. Optimum condition of $NaN_3$ treatment was considered as $0.5{\sim}1mM\;for\;1{\sim}3h$. For colchicine, there were significant differences between species as to the proliferation ratios of prothallus for each treatment.

• Bulletin of the Korean Chemical Society
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• v.30 no.6
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• pp.1360-1364
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• 2009

The bacterium Sphingomonas chungbukensis DJ77 produces the extracellular polysaccharide gellan in high yield. Gellan produced by this bacterium is widely used as a gelling agent, and the enzyme UDP-glucose pyrophosphorylase (UGP) is thought to play a key role in the gellan biosynthetic pathway. The UGP gene has been successfully cloned and over-expressed in E. coli. The expressed enzyme was purified with a molecular weight of approximately 32 kDa, as determined by a SDS-polyacrylamide gel, but the enzyme appears as ca. 63 kDa on a native gel, suggesting that the enzyme is present in a homodimer. Kinetic analysis of UDP-glucose for UGP indicates $K_m$ = 1.14 mM and $V_{max}$ = 10.09 mM/min/mg at pH 8.0, which was determined to be the optimal pH for UGP catalytic activity. Amino acid sequence alignment against other bacteria suggests that the UGP contains two conserved domains: An activator binding site and a glucose-1-phosphate binding site. Site-directed mutagenesis of Lys194, located within the glucose-1-phosphate binding site, indicates that substitution of the charge-reversible residue Asp for Lys194 dramatically impairs the UGP activity, supporting the hypothesis that Lys194 plays a critical role in the catalysis.

• Bulletin of the Korean Chemical Society
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• v.30 no.7
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• pp.1547-1552
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• 2009

Sphingomonas chungbukensis DJ77 has the ability to produce large quantities of an extracellular polysaccharide that can be used as a gelling agent in the food and pharmaceutical industries. We identified, cloned and expressed the UDP-glucose dehydrogenase gene of S. chungbukensis DJ77, and characterized the resulting protein. The purified UDP-glucose dehydrogenase (UGDH), which catalyzes the reversible conversion of UDP-glucose to UDPglucuronic acid, formed a homodimer and the mass of the monomer was estimated to be 46 kDa. Kinetic analysis at the optimal pH of 8.5 indicated that the $K_m\;and\;V_{max}$ for UDP-glucose were 0.18 mM and 1.59 mM/min/mg, respectively. Inhibition assays showed that UDP-glucuronic acid strongly inhibits UGDH. Site-directed mutagenesis was performed on Gly9, Gly12 Thr127, Cys264, and Lys267. Substitutions of Cys264 with Ala and of Lys267 with Asp resulted in complete loss of enzymatic activity, suggesting that Cys264 and Lys267 are essential for the catalytic activity of UGDH.

• BMB Reports
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• v.45 no.8
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• pp.452-457
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• 2012

Diketoreductase (DKR) from Acinetobacter baylyi contains two tryptophan residues at positions 149 and 222. Trp-149 and Trp-222 are located along the entry path of substrate into active site and at the dimer interface of DKR, respectively. Single and double substitutions of these positions were generated to probe the roles of tryptophan residues. After replacing Trp with Ala and Phe, biochemical and biophysical characteristics of the mutants were thoroughly investigated. Enzyme activity and substrate binding affinity of W149A and W149F were remarkably decreased, suggesting that Trp-149 regulates the position of substrate at the binding site. Meanwhile, enzyme activity of W222F was increased by 1.7-fold while W222A was completely inactive. In addition to lower thermostability of Trp-222 mutants, molecular modeling of the mutants revealed that Trp-222 is vital to protein folding and dimerization of the enzyme.

• The Plant Pathology Journal
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• v.19 no.6
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• pp.301-304
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• 2003

Gamma-ray irradiation is known to induce various mutations in plants caused by chromosome alterations. This study investigated disease responses of selected gamma-ray induced rice mutants generated from seven Japonica-type rice cultivars against three plant diseases. Among the tested 22 mutants, three gain-of-function mutants and six loss-of-function mutants against rice blast were obtained, as well as three loss-of-function mutants against bacterial leaf blight (BLB). Two of the loss-of-function mutants were susceptible to both rice blast and BLB. Gain-of-function mutation has not been frequently observed in rice plants, thus, the mutants can be used to identify loci of novel genes for the regulation of disease resistant response.

• Korean Journal of Microbiology
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• v.20 no.2
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• pp.89-97
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• 1982

Mutants of plasmid pKM101 modified to enhance mutagenesis were induced and characterized in Salmonella typhimurium. The pKM101 mutant plasmid were transferred normally and stably maintained in cells. They had modified in their ability (i) to enhance the reversion of both point and frameshift mutations, (ii) to protect the cell against UV-irradiation and chemical mutagen treatment, (iii) of ampicillin resistance. A similar modification in enhancement of reversion was also observed in a $uvrB^-$ strains. These results indicated that mutator effect of pKM101 was coded by one plasmid gene.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.2
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• pp.247-253
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• 1997

Development of soybean cultivars with great nodulation and high nitrogen fixation activity, derived mostly from mutagenesis, may decrease inputs of chemical fertilizer nitrogen into the soil-plant system. Soybean seeds (cv. Jangyupkong, Hwanggeumkong, and Geomjungkong 1) were treated with three different levels of EMS (ethyl methanesulfonate) concentration(30, 50, and 70mM). Increasing the doses of EMS resulted in decreased field emergence rate of seeds, whereas it did not increase M$_2$ mutation frequencies. This indicated that the most efficient concentration of EMS was 30mM for generating mutants. Extensive mutagenesis of Sinpaldalkong 2 with 30mM EMS was undertaken to isolate soybean mutants with greater nodulation. Approximately 8, 200 M$_2$ families were screened for greater nodulation on 5 mM nitrate after inoculation with Bradyrhizobium japonicum strain YCK213-KFCC-10728. Mutant SS-2 nodulated more than the wild type. Comparison of supernodulation between SS-2 and two nts mutants(nts 1007 and nts 1116) revealed that SS-2 showed the supernodulation character at an earlier growth stage than the two nts mutants. Further studies should be needed to characterize the difference in timing of nodulation between SS-2 and nts mutants.

• Molecules and Cells
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• v.27 no.5
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• pp.547-556
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• 2009

Parathyroid hormone is the most important endocrine regulator of calcium concentration. Its N-terminal fragment (1-34) has sufficient activity for biological function. Recently, site-directed mutagenesis studies demonstrated that substitutions at several positions within shorter analogues (1-14) can enhance the bioactivity to greater than that of PTH (1-34). However, designing the optimal sequence combination is not simple due to complex combinatorial problems. In this study, support vector machines were introduced to predict the biological activity of modified PTH (1-14) analogues using mono-substituted experimental data and to analyze the key physicochemical properties at each position that correlated with bioactivity. This systematic approach can reduce the time and effort needed to obtain desirable molecules by bench experiments and provide useful information in the design of simpler activating molecules.

• Microbiology and Biotechnology Letters
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• v.19 no.2
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• pp.116-121
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• 1991

A bacterial strain KN, which highly produced a protease, was isolated from several soil samples and identified to to belong to the genus Bacillus. We selected mutant strain Bacillus sp. KN103N, which was hyperproducer of protease and was resistant to D-cyclowerine, from the strain KN by several steps of mutagenesis. Neutral protease productivity of mutant strain KN103N was about 55 times as much as that of the original strain KN. The optimum pH and temperature for the enzyme activity were 7.0 and 50$^{\circ}C$, respectively and the enzyme was relatively stable at pH6.0~8.0 and below 40$^{\circ}C$. The enzyme was inactivated by EDTA, but not by DFP. These results indicate that the enzyme from Bacillus sp. KN103N was a neutral (metallo-) protease.