• Journal of Microbiology and Biotechnology
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• v.17 no.10
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• pp.1638-1644
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• 2007

The effect of carbon sources on tacrolimus production by a mutant strain of Streptomyces clavuligerus CKD 1119, an isolate from soil, was examined. Among the carbohydrates and oils tested in this work, a mixed carbon source of soluble starch and com oil was the best. An analysis of the culture kinetics also showed that, in contrast to the carbohydrates, the com oil was consumed later in the antibiotic production phase, implying that the oil substrate was the principal carbon source for the biosynthesis of tacrolimus, and this was directly proven by experiments using $^{14}C$-glucose and $^{14}C$-oleate substrates. Furthermore, com oil induced the formation of lipase by the mutant strain, whereas the addition of glucose significantly repressed lipase activity. The lipase activity exhibited by the FK-506-overproducing mutants was also observed to be directly proportional to their tacrolimus yield, indicating that a high lipase activity is itself a crucial factor for tacrolimus production. A feasibility study with a 200-1 pilot-scale fermentor and the best strain (Tc-XII-15322) identified in this work revealed a high volumetric and specific productivity of about 495 mg/l and 0.34 mg/mg dry mycelium, respectively.

• Journal of Microbiology and Biotechnology
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• v.5 no.5
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• pp.297-301
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• 1995

An anthracycline antibiotic, aclacinomycin A (aclarubicin), was produced from a mutant strain of Streptomyces lavendofoliae. The mutant strain which showed a 4-fold higher productivity of aclacinomycin A compared with the parent strain was also found to produce a significantly higher amount of aclacinomycin A than the reported production strain, Streptomyces galilaeus. The aclacinomycin A was produced up to 125 mg/l using potato starch and soybean meal as carbon and nitrogen sources, respectively, on a 3 liter scale fermentation in a 5 liter jar fermentor. The mutant strain also produced significant amount of aclacinomycins Band Y. Aclacinomycin A was isolated from the culture broth by solvent extractions and further purified by silica gel column chromatography. The yield of aclacinomycin A with over 99$%$ purity was found to be over 60$%$ starting from 3 liters of culture broth.

• Bulletin of the Korean Chemical Society
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• v.15 no.10
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• pp.832-835
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• 1994

The ${alpha}$-amylase gene of Bacillus licheniformis has been cloned and two mutant ${alpha}$-amylase genes of which histidine 235 was changed to glutamine (H235Q) and aspartic acid 328 to glutamic acid (D328E) have been produced by site-directed mutagenesis. The kinetic parameters, optimum pH and thermostability of wild type(WT) and these two mutant amylases expressed in E. coli MC1061 have been compared after purification. The $K_m$ values of WT, H235Q and D328E ${alpha}$-amylases were 0.22%, 0.73%, and 0.80% respectively, when using starch as the substrate. The $V_max$ values of wild type ${alpha}$ -amylase and mutant ${alpha}$-amylases were 0.6-0.7%/minute, and did not show any significant differences among them. The optimum pH of D328E ${alpha}$-amylase was shifted to more acidic pH. Also, the thermostability of H235Q ${alpha}$-amylase was increased compared to the wild type ${alpha}$-amylase.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.300-300
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• 2022

Wheat (Triticum spp.) is an important source of food worldwide and the focus of considerable efforts to identify new combinations of genetic diversity for crop improvement. In particular, wheat starch composition is a major target for changes that could benefit human health. Starches with increased levels of amylose are of interest because of the correlation between high amylose content and elevated levels of resistant starch, which has been shown to have beneficial effects on health for combating obesity and diabetes. In this study, high amylose wheat germplasms from other countries were collected and cultivated in Korea, and then the content of amylose was evaluated, we examined amylose content in 614 wheat germplasm. Furthermore, amylose content was validated using several milling processes such as roller, hammer, and grinding mill. As a result, the amylose content distribution was divided into five groups. The range of the amylose levels in whole wheat flour was 18.3% to 29.6%. In addition, the mutant lines were screened for high amylose, and two mutant lines (WX-1046 and WX-1074) exhibited a comparable amylose content to Keumkang whole wheat (19.6%). It has been established that high amylose indicated SS IIa null and necessitate GBSS. Based on these findings, it may be helpful to develop high amylose wheat germplasm and production techniques, particularly in Korea.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.6
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• pp.987-994
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• 2000

Aspergillus coreanus NR 15-1, Asp. oryzae NR 15-3 and Asp. oryzae NR 2-5 isolated from traditional Korean nuruk were screened as parental strains producing starch hydrolyzing enzymes. They were mutagenized by N-methyl -N'-nitro-N-nitrosoguanidine (NTG) and mutants were isolated for analysis of various amylase activities and the ability of acid production. Among them, the mutants harboring high saccharogenic activity, dextrinogenic activity, and the ability of acid production were selected. Fifteen, six, and five strains of mutants were isolated from Asp. coreanus NR 15-1, Asp. oryzae NR 2-5, and Asp. oryzae NR 15-3, respectively followed by NTG mutagenesis. Among these mutants, thirteen strains were identified as auxotrophic mutants. \ulcorner (Arg.￣) mutant from Asp. coreanus NR 15-1 showed high glucoamylase activity and total acid productivity. Z6 (Ade.￣) mutant from Asp. oryzae NR 2-5 showed the highest $\alpha$-amylase activity, therefore \ulcorner and Z6 mutant were selected.

• Korean Journal of Microbiology
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• v.38 no.1
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• pp.1-6
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• 2002

$\alpha$-Amylase producing bacteria were isolated from activated sludge of corn processing wastewater plant and paddy field soil samples and selected by the direct iodine reaction. The isolate was identified as Bacillus sp. after morphology, API system and fatty acid analyses. To enchance $\alpha$-amylase productivity, a successive mutation of Bacillus sp. AIV 19 was performed using the treatment of nitrosoguanidine(NTG).The mutant, Bacillus sp. AIV 1940, showed about 1.8-fold level of amylase activity compared with parental strain. The isolate was Gram-positive and rod (2.8-3.0 $\mu$m long, 0.5-0.6 $\mu$m wide) type. The strain increased the bacterial mass at 3000 mg/l starch concentration. Organic substance removal rate was 40.2, 72.3% respectively after 1 and 3 day reaction using starch synthetic wastewater (intial CODcr was 4,455 mg/l).

• Korean Journal of Food Science and Technology
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• v.32 no.2
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• pp.258-264
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• 2000

Starches from the eight varieties of rice were analyzed using scanning electron microscope(SEM), differential scanning calorimetry(DSC) and X-ray diffractometry, and tested on the starch-granule susceptibility to 15% $H_2SO_4$ and glucoamylase. The shape of starch granules from normal rice varieties and low-amylose mutants were polygonal while shrunken and floury mutants were globular. According to DSC, starches from Nampung CB243 showed higher onset temperature$(T_o)$, completion temperature$(T_c)$ and Punchilmi, Nampung EM90 showed higher enthalpy$({\triangle}H)$ of gelatinization than others. shr showed the highest hydrolysis rate to 15% $H_2SO_4$ while Nampung CB243 showed the lowest one. Eight varieties of rice starch granules showed A-type pattern on X-ray diffractograms. The lower amylose content rice varieties showed the higher hydrolysis rate treated with glucoamylase.

• Mycobiology
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• v.39 no.1
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• pp.20-25
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• 2011

Spores of Aspergillus sp. SU14 were treated repeatedly and sequentially with $Co^{60}$ ${\gamma}$-rays, ultraviolet irradiation, and N-methyl-N'-nitro-N-nitrosoguanidine. One selected mutant strain, Aspergillus sp. SU14-M15, produced cellulase in a yield 2.2-fold exceeding that of the wild type. Optimal conditions for the production of cellulase by the mutant fungal strain using solid-state fermentation were examined. The medium consisted of wheat-bran supplemented with 1% (w/w) urea or $NH_4Cl$, 1% (w/w) rice starch, 2.5 mM $MgCl_2$, and 0.05% (v/w) Tween 80. Optimal moisture content and initial pH was 50% (v/w) and 3.5, respectively, and optimal aeration area was 3/100 (inoculated wheat bran/container). The medium was inoculated with 25% 48 hr seeding culture and fermented at $35^{\circ}C$ for 3 days. The resulting cellulase yield was 8.5-fold more than that of the wild type strain grown on the basal wheat bran medium.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.230-233
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• 1999

A plasmid vector was constructed for the expression and secretion of Bacillus macerans cyclodextrin glucanotransferase (CGTase) in Bacillus subtilis. The vector, pUBACGT, was composed of the ribosome-binding sequence, signal sequence, and cgt gene from B. macerans under the control of amyR2, the promoter of amyE gene coding for $\alpha$-amylase from B. subtilis var. natto. Bacillus subtilis LKS88, a mutant strain lacking genes for an amylase and two proteases, was used as a host for the transformation of the plasmid vector. The transformants were selected on kanamycin-containing Luria-Bertani plates. The starch hydrolyzing activity was observed on the starch-containing plates by the iodine method and cyclodextrin-forming activity was detected in the culture medium. A SDS-PAGE analysis showed that most of the expressed CGTase in the recombinant B. subtilis was secreted into the medium at a high expression level.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.2
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• pp.214-219
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• 1997

Mutation rate of M$_2$ plants that were treated with three types of double treatments of chemical mutagens(1.5mol Na$N_2$ ＋ 0.75mol MNH, 0.75mol MNH ＋ 0.75mol MNH and 0.5mol MNH ＋ 0.5mol MNH) were estimated on the rate of chlorophyll mutant, changes of isozyme loci ; esterase (Est), glutamate oxaloacetate transaminase(GOT ; AAT) and leucyl aminopeptydase(LAP ; AMP). Rate of chlorophyll mutants (3.3％ =no. of seedling carrying mutant / all number of M$_2$ seedlings $\times$ 100) and rate of esterase isozyme loci mutants(3.5％ =no. of plant carrying mutant / all number of M$_2$ plant) in Dema were higher than one of Sacheon 6, but no significant differences in GOT, LAP. Among isozymes, most of mutants in M$_2$ plant of two varieties were found in esterase (73％ of total mutants were occurred in esterase loci). Although many of null bands were found in GOT 3, these were not repeatable and no real mutants. It might be due to qualities of starch, amount of extract buffer and degradation of isozyme during electrophoresis and staining.