• The Korean Journal of Mycology
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• v.12 no.1
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• pp.1-8
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• 1984

The mycelial growth of some mushrooms was inhibited by benomyl treatment. The $ED_{50}$ of benomyl to that of Pleurotus spp., Agaricus bisporus and Flammulina velutipes was 25ppm, 50ppm and 200ppm, respectively, which indicates the former was the most sensitive to the fungicide. The mycelial growth of mushrooms growing on artificial media amended by benomyl was increased when they were cultured successively 5 times and 10 times on the media. The increasing rate of that of each mushroom was the highest at the concentration of $ED_{50}$ of benomyl. The mycelial growth of P. ostreatus was increased progressively as the number of successive culturing increased, while that of P. florida and A. bisporus was increased until they were cultured successively up to 5 times and 7 times, respectively, but they were decreased after that. Mutant sectors of mycelia were induced by successive treatment of benomyl. Mutant sectors of P. ostreatus appeared earlier than those of P. florida and all of them were induced earlier on the media of low contration of benomyl than on that of high concentration. The mycelia of mutant sectors induced by benomyl treatment grow faster than those of mother colony treated with benomyl successively, but there was no difference in resistance against the fungicide between them. The increase of mycelial growth of the mushrooms culturing successively on media containing benomyl indicated that they might obtain the resistance against benomyl.

• Applied Biological Chemistry
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• v.4
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• pp.11-16
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• 1963

(1) The 30 strains of Penicillia and the 52 strains of Aspergilli have been screened for their producibility of 5'-Phosphodiesterase, and Penicillium sclerotiorum 7321, Penicillium sp M-11 and Penicillium citrinum UV-mutant 2032-72 were selected as having high 5‘-Phosphodiesterase activity. (2) Using the wheat bran medium the 5‘-Phosphodiesterase production was reached at maximum state by the plate culture for 10 days at $30^{\circ}C$ (3) The optimum conditions of the 5'-Phosphodiesterase activity of Penicillium sclerotiorum 7321 and Penicillium sp M-11 were pH 4.0 at $62.5^{\circ}C$, while the optimum condition of the 5'-Phosphodiesterase activity of Penicillium citrinum UV-mutant 2032-72 was pH 5.0 at $50^{\circ}C$.

• Biomolecules & Therapeutics
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• v.21 no.2
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• pp.114-120
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• 2013

Most patients with mutant B-Raf melanomas respond to inhibitors of oncogenic B-Raf but resistance eventually emerges. To better understand the mechanisms that determine the long-term responses of mutant B-Raf melanoma cells to B-Raf inhibitor, we used chronic selection to establish B-Raf (V600E) melanoma clones with acquired resistance to the new oncogenic B-Raf inhibitor UI-152. Whereas the parental A375P cells were highly sensitive to UI-152 ($IC_{50}$ < $0.5{\mu}M$), the resistant sub-line (A375P/Mdr) displayed strong resistance to UI-152 ($IC_{50}$ < $20{\mu}M$). Immunofluorescence analysis indicated the absence of an increase in the levels of P-glycoprotein multidrug resistance (MDR) transporter in A375P/Mdr cells, suggesting that resistance was not attributable to P-glycoprotein overexpression. In UI-152-sensitive A375P cells, the anti-proliferative activity of UI-152 appeared to be due to cell-cycle arrest at $G_0/G_1$ with the induction of apoptosis. However, we found that A375P/Mdr cells were resistant to the apoptosis induced by UI-152. Interestingly, UI-152 preferentially induced autophagy in A375P/Mdr cells but not in A375P cells, as determined by GFP-LC3 puncta/cell counts. Further, autophagy inhibition with 3-methyladenine (3-MA) partially augmented growth inhibition of A375P/Mdr cells by UI-152, which implies that a high level of autophagy may protect UI-152-treated cells from undergoing growth inhibition. Together, our data implicate high rates of autophagy as a key mechanism of acquired resistance to the oncogenic B-Raf inhibitor, in support of clinical studies in which combination therapy with autophagy targeted drugs is being designed to overcome resistance.

• Korean Journal of Environmental Agriculture
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• v.25 no.2
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• pp.124-128
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• 2006

To reducing offensive odor form compost UIW-6 mutant obtained by UV treatment from sulfur-oxidizing bacteria, Thiobacillus sp. IW. The UIW-6 mutant was found 1.6 times faster in growth than the parent strain on thiosulfate medium (TM) at 36 h after incubation. Initial pH, temperature and agitation for the optimum growth of UIW-6 were 6.5, $35^{\circ}C$ and 200 rpm, respectively. The UIW-6 mutant growth was two times higher than parent strain at 6 h culture in TM liquid medium containing 50 mM sodium thiosulfate. The UIW-6 mutant used fructose and sucrose as carbon sources and yeast extract> tryptone> peptone as nitrogen ones. It was found that the growth of UIW-6 was increased in addition of 0.2% yeast extract.

• The Journal of Natural Sciences
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• v.8 no.1
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• pp.33-39
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• 1995

Klebsiella sp. L-10 was treated with physical and chemical mutagens, and one of the NTG mutant which increased hyaluronic acid complex yield 2.5 folds was selected. The yield of hyaluronic acid complex from Klebsiella sp. L-10 NTG 50 mutant reached maximum level I the YPD medium containing 0.1% yeast extract, 3% Bacto-tryptone, 3% dextrose, each 30mM of $K_2HPO_4$ and $KH_2PO_4$ (pH 6.0-6.5) with shaking culture at $37^{\circ}C$ for 24 hrs, and 2900mg of hyaluronic acid complex per litre of culture was produced under the above condition.

• Korean Journal of Microbiology
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• v.20 no.3
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• pp.125-133
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• 1982

Mutational experiments were performed to imporve the cellulase productivity of Aspergillus phoenicis KU175, isolated from the southern part of Korea, as a high cellulase producer. By treatment ultra-violet light nad 4-NQO(4-Nitroquinoline-N-Oxide), mutation waas induced, and treatment ultra-violet light and 4-NQO (4-Nitroquinoline-N-Oxide), mutation was induced, and A.phoenicis KU175-115 was finally selected for its highest avicelase production. Avicelase production of the mutant was increased about 2 times compared with those of the wild strain. However, activities of other hydrolytic enzymes, such as amylase, protease and nuclease, of the mutant strain didn't show a marked difference compared with those of the nuclease, of the mutant strain didn't show a marked difference compared with the wild strain, except slight increase in ribonuclease activity and slight decrease in glucoamylase activity. Avicelases from the mutant strain selected were purified from wheat bran culture by successive salting out, followed by dialysis and column chromatography, and their charcteristics were compared with thosw of the wild strain. Avicelase was separated into three peaks in the mutant strain as well as in the case of wild strain. Avicelase II activity of the mutant strain was prominently higher than that of the wild strain, while avicelase I and III activities of those were equivalent. The optimal pH ranges and stability of avicelase II from the mutant strain were pH4-5 and pH3.5-6.0, respectively, as well as in the case of the wild strain. The optimal temperature and thermal stability of avicelase II from the mutant strain were $40{\sim}50^{\circ}C\;and\;20{\sim}55^{\circ}C$, respectively. These results were same as those of the wild strain. By the using of Eadie-Hofastee plot, $K_m\;and\;V_{max}$ of avicelase II from the mutant and the wild strain were calculated to be 2.29mg/ml and $4.84{\mu}g$ reducing sugar as glucose per min equally, from the line fitted to the data by the least square method. Activity of avicelase II from the mutant strain was slightly activated by $Mg^{++}\;but\;inhibited\;by\;Cu^{++}, \;Mn^{++}\;and\;Zn^{++}$, as well as in the case of the wild strain. Therefore, it was concluded that the mutant didn't induce the formation of another avicelase isozyme, or the changes in the properties of avicelase, but induce the changes in the productively of the same avicelase II by the action of regulatory gane.

• Bulletin of Food Technology
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• v.17 no.4
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• pp.98-106
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• 2004

Two mutant enzymes of $\gamma$-glutamylcysteine synthetase ($\gamma$-GCS) which catalyzed the synthesis of $\gamma$-glutamylcysteine from L-glutamic acid and L-cysteine in the presence of ATP, were prepared bypoint mutation of $\gamma$-GCS gene with site-directed mutagensis in E. coli. Conformational structuresand catalytic reaction kinetics of mutant enzymes were compared with wild type $\gamma$-GCS afterpurification. The S495F mutant enzyme (serine at 495 residue was substituted with phenylalanine),which had no catalytic activity for $\gamma$-glutamylcysteine synthesis, rarely folded even in neutral pH.However, the mutant A494V (alanine of 494 residue was replaced by valnine) which showed 50 %increase of activity, had a high folding structure. The folding structure of A494V also more stable athigh temperature and extreme pH compared to wild type and S495F. Reaction kinetics of wild typeand A494V were also investigated, Km value of A494V was smaller than that of wild type, while itshowed a little difference at Vmax values. This result evolved that alanine at 494 may be involved inbinding site of substrate rather than catalytic site. In addition, change of catalytic activity by onepoint mutation was highly correlated with the folding structure of enzyme.

• Journal of Microbiology and Biotechnology
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• v.17 no.9
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• pp.1533-1537
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• 2007

In this study, in order to develop a continuous production process of lactosucrose in a packed-bed reactor, Sterigmatomyces elviae ATCC 18894 was selected and mutated. The mutant strain of S. elviae showed 54.3% higher lactosucrose production than the wild type. Reaction conditions such as temperature, pH, substrate concentration and flow rate were also optimized. Under optimized reaction conditions ($50^{\circ}C$, pH 6.0, 25% sucrose and 25% lactose as substrate, flow rate 1.2 ml/min), the maximum concentration of lactosucrose (192 g/l) was obtained. In a packed-bed reactor, continuous production of lactosucrose was performed using S. elviae mutant immobilized in calcium alginate, and about 180 g/l of lactosucrose production was achieved for 48 days.

• Biomedical Science Letters
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• v.2 no.1
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• pp.21-30
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• 1996

MAP kinases are a family of serine/threonine specific protein kinases becoming activated in response to different proliferative stimuli by phosphorylation at both threonine and tyrosine residue. Present study shows that MAP kinase was purified from P388 murine leukema cells by SP sephadex C-50, phenyl superose and Mono Q column chromatography and identified with anti-ERKl antibody by western blotting. Immnublotting analysis to the crude extract of P388 cell lysate shows 44 kD and other minor bands but partial purified fraction eluted from phenyl supherose column have 44kD and 66 kD isoform. Subcloned GST-fusion protein from N-terminal of $p56^{kk}$ was tested as a substrate for MAP kinase phosphorylation. It was showed that the wild type and mutant forms(S42A) were fully phosporylated by purified MAP kinase fraction as com-pare with the other mutant form(S59A). This finding suggest that those GST-fusion proteins may be used as substrate for the in vitro test of MAP kinase.

• Korean Journal of Microbiology
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• v.40 no.2
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• pp.160-166
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• 2004

The optimal conditions for production of Monascus sp. KM100l cell mass on submerged culture and production of monacolin K on rice solid culture were investigated. An overproducing mutant of Monascus pigments, KM 1001 mutant, from Monascus purpureus KCCM60016 was selected by NTG treatment. The optimal medium for the production of KM100l mutant cell mass is instructed to be composed of 3% glucose, 2% yeast extract, 0.1 % KH$_2$PO$_4$, 0.05% The optimal conditions for production of Monascus sp. KM100l cell mass on submerged culture and production of monacolin K on rice solid culture were investigated. An overproducing mutant of Monascus pigments, KM 1001 mutant, from Monascus purpureus KCCM60016 was selected by NTG treatment. The optimal medium for the production of KM100l mutant cell mass is instructed to be composed of 3% glucose, 2% yeast extract, 0.1 % KH$_2$The optimal conditions for production of Monascus sp. KM100l cell mass on submerged culture and production of monacolin K on rice solid culture were investigated. An overproducing mutant of Monascus pigments, KM 1001 mutant, from Monascus purpureus KCCM60016 was selected by NTG treatment. The optimal medium for the production of KM100l mutant cell mass is instructed to be composed of 3% glucose, 2% yeast extract, 0.1 % $(KH_2PO_4$, 0.05% $MgSO_4{\cdot}7H_2O$, 0.2% L-asparagine, pH 4.5, and the optimal inoculum size and shaking speed were $1.5{\times}10^6$ spores/50 m1 medium and 150 rpm, respectively. On optimal conditions, 4.1 g/l of the cell mass was obtained at 28$^{\circ}C$ for 3 days. The mycelium were inoculated on 500 g of steamed rice using vinyl bag ($30.6{\times}44$ cm) and incubated at $30^{\circ}C$, 85% humidity for 21 days. Lactone form monacolin K was rapidly increased for 2 days and reached highest concentration of monacolin K (2,930 mg/kg) for 15 days, and monacolin K was decreased after 15 days.