• Journal of Life Science
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• v.15 no.4 s.71
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• pp.553-560
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• 2005

The stf (Salmonella typhimurium fimbriae) operon consisting of stfA(CDEFG assumes to encode putative fimbriae. The complete stf operon is existed in S. typhimurium and S. choleraesuis, whereas it is absent in S. typhi. Analyses of the amino acid residues between major subunit StfA of the Stf fimbriae and those of known other fimbriaes suggested that Stf belongs to class I type fimbriae. Through comparison of StfD chaperone with the other fimbrial chaperones, and of C-terminus in subunits of Stf fimbriae, it belongs to FGS (with a short Fl-G1 loop) subfamily. In order to investigate the expression of stf operon, we have constructed a Salmonella strain containing a chromosomal stfA::lacZYA transcriptional fusion, resulting in S. typhimurium $_X8532$. The strain $_X8532$ lacked the expression of \beta-galactosidase$ under normal culture conditions. However, with longer incubation time of the S. typhimurium $_X8532$, we have isolated 21 individual strains exhibiting $Lac^+$ phenotype. $Lac^+$ phenotype was appeared as approximately 0.03 frequency per generation. All isolates expressed lacZ constitutively in the various environmental conditions. Various global regulatory proteins including RpoS, OmpR, and CpxR were not involved in the regulation of the stf operon. A S. typhimurium $_X8661$ mutant lacking stfAC function attenuated 6.7 folds more than that of wild type $_X3761$ in the mouse virulence test, suggesting in the somehow involved in the Salmonella pathogenesis.

• Journal of Life Science
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• v.32 no.3
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• pp.202-209
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• 2022

The green alga Chlamydomonas reinhardtii, a unicellular haploid eukaryote, has long been used by researchers and industries as a cell factory to produce high value-added microalgae substances using genetic modification. Microalga K01, presumed to be Chlamydomonas, was isolated from 12 freshwater samples from the Chungcheong and Jeolla regions to replace C. reinhardtii, an introduced species currently used in most basic and industrial research. The isolated K01 strain was identified as C. reinhardtii through morphological and phylogenetic studies of the 18S rDNA gene sequence (NCBI accession number KC166137). The growth and lipid content of the isolated C. reinhardtii K01 were compared with three wild and four mutant strains in TAP medium, and it was found that the K01 strain could produce 1.74×10⁷ cells/ml by the third day of culture. The growth rate of C. reinhardtii K01 was 1.5 times faster than UTEX2244, which showed the highest number of cells (1.20×10⁷ cells/ml) among the compared strains. The lipid content of the isolated C. reinhardtii K01 (20.67%) was similar to those of the wild strains, although the fatty acid oleate C18:1 was not detected in the isolated strain but was identified in the seven others. The cell density of the isolated strain increased to 0.87 g/l during a six-day culture in BG11 medium, where nitrate (NaNO₃) was introduced as a nitrogen source, while the seven acquired strains showed almost no cell proliferation.

• Microbiology and Biotechnology Letters
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• v.48 no.3
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• pp.344-357
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• 2020

Strain improvement and bioprocess development were undertaken to enhance hyaluronic acid(HA) production by Streptococcus zooepidemicus cells. Using a high-yielding mutant strain, statistical medium optimization was carried out in shake flask cultures, resulting in 52% increase in HA production (5.38 g/l) at the optimal medium composition relative to the parallel control cultures. For sufficient supply of dissolved oxygen (DO), which turned out to be crucial for enhanced production of HA, agitation system and speed were intensively investigated in 5 L bioreactor cultures. Increase in oxygen mass transfer coefficient (k_La) through increment of agitation speed (rpm) and 35% expansion of diameter of the newly-designed impellers showed significantly positive effects on HA production. By installing an expanded Rushton-turbine impeller for efficient break-down of sparged air, and an extended marine impeller above the Rushton-turbine impeller for efficient mixing of the air-born viscous fermentation broth, maximum amount of HA (9.79 g/l) was obtained at 450 rpm, 1.8 times higher level than that of the corresponding flask culture. Subsequently, the possibility of bioprocess scale-up to a 50 L bioreactor was investigated. Despite almost identical maximum HA production (9.11 vs 9.25 g/l), the average HA volumetric productivity (r_p) of the 50 L culture turned out only 74% compared to the corresponding 5 L culture during the exponential phase, possibly caused by shear damages imposed on the producing cells at the high stirring in the 50 L culture. The scale-up process could be successfully achieved if a scale-up criterion of constant oxygen mass transfer coefficient (k_La) is applied to the 50 L pilot-scale bioreactor system.

• Korean Journal of Food Science and Technology
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• v.40 no.4
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• pp.424-429
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• 2008

To investigate the acid tolerance characteristics of the acid-resistant mutant, Leuconostoc paramesenteroides P-200, as a kimchi starter, this study examine proton permeability, ATPase activity, glycolysis activity, $Mg^{2+}$ release, and membrane fatty acid composition, and compared the data to that of its wild-type, L. paramesenteroides LP-W. In the proton permeability experiment, the LP-W and P-200 strains' average maximum half-time $(t_{1/2})$ values for pH equilibration through the cell membrane were approximately 5.7 and 9.3 min in 150mM KCl solution, and 4.2 and 8.3 min in 3% NaCl solution, respectively. Their values and pH levels for maximal specific ATPase activity showed that P-200 had greater activity than LPW. And the results of pH-dependent glycolysis activity showed that P-200 had greater activity than LP-W. Furthermore, after 2 hr at pH 4.0, LP-W and P-200 had percent magnesium release values of approximately 12% and 34%, respectively. A comparison of their membrane fatty acid compositions indicated that C18 and cyclo-C19 were the major different fatty acids between the two strains, and their contents of C18 and cyclo-C19 were 2.5% and not detected, respectively, in LP-W, and 6.4% and 11.4%, respectively, in P-200. These results indicate that the P-200 strain has significantly improved acid tolerance as compared to its wild type, LP-W.

• Korean Journal of Microbiology
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• v.46 no.3
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• pp.233-239
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• 2010

Four genes (yqfR, yfmL, ydbR, deaD) were identified as putative DEAD-box RNA helicase genes in the genomic sequence of Bacillus subtilis by homology search. To understand the function of these genes, each of the genes was deleted and the constructed strains were tested for their growth charateristics at different temperatures. The growth rate of ydbR deletion mutant ($T_d$=53 min) was a little bit reduced at $37^{\circ}C$ as compared to that of wild type strain (CU1065). But the growth rate of other three (yqfR, yfmL, deaD) deletion mutants ($T_d$=30-40 min) is nearly equal to the growth rate of wild type ($T_d$=32 min). On the other hands, the growth rate of deletion mutants were reduced at $22^{\circ}C$ in order of yqfR ($T_d$=151 min), yfmL ($T_d$=214 min), ydbR ($T_d$=343 min), which showed cold-sensitive phenotype. The deletion mutant of deaD ($T_d$=109 min) grew equally as compared to the growth rate ($T_d$=102 min) of the wild type at $22^{\circ}C$ and did not show cold-sensitive growth. Double, triple and quadruple deletion mutants of these genes were constructed, and growth rate of these mutants were measured at various temperature conditions ($22^{\circ}C$, $37^{\circ}C$, $42^{\circ}C$) using LB broth. Multiple deletion mutations showed more severe cold-sensitive growth than single deletion mutations, and double deletion of ydbR and yfmL ($T_d$=984 min) showed most cold-sensitive growth than any other double mutants. Such a cold-sensitive growth of these mutations is quite similar to the result of csdA or srmB deletion in E. coli and suggested that physiological role of ydbR and yfmL is related with ribosome assembly.

• Korean Journal of Soil Science and Fertilizer
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• v.27 no.2
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• pp.136-146
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• 1994

To research the effect of chemotaxis of Rhizobia toward the root exudate on nitrogen fixing ability in soybean Rhizobia symbiosis system. Root exudate from seedlings of Glycine max. L was collected aseptic conditions. B. japonicum KCTC 2422 induced the formation of symbiotic nitrogen fixing nodules on the root of soybean plant and possessed motility and chemotaxis toward the 2mM proline. LPN-100 mutant was $Nod^-$, $Che^+$, and LPN-101 was $Che^-$, $Nod^+$ strains. Physiological properties of mutants were similar to parent strain. The crude root exudate was tested for its chemotactic ability using the capillary tube method. Chemotactic responses of RCR 3407 toward crude root exudate were 2.2, 2.6, 2.9, those of KCTC 2422 were 2.3, 2.9, 3.0, respectively. The crude root exudate was fractionated into neutral, cationic and anionic fractions. Chemotactic responses of KCTC 2422 was least with anionic fraction, most with neutral and intermediate with cationic fraction. B. japonicum KCTC 2422 was attracted by carbohydrates, amino acids and carboxylic acid. Carbohydrates and amino acids were good chemoattractants and carboxylic acids were intermediate chemoattractants. The peak concentration was $10^{-3}M$ for ribose, glucose, glutamine, aspartic acid and carboxylic acids, with exception of xylose, arabinose, tryptophan, which elicited maximum responses at $10^{-4}M$. The formation of nodules and nitrogenase activity of soybean inoculated with KCTC 2422 was determined in 7days after inoculation, and those of LPN-101 was detected in 15days after inoculation, but LPN-100 didn't form of nodules in soybean plants.

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.140-146
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• 2009

Cyclosporin, an immunosuppressant, is a representative group of biologically active secondary metabolites produced by the fungus Tolypocladium inflatum. The amount and ratio of cyclosporin derivatives in the culture broth are an important factors for the production of cyclosporin A and the purification in the industrial process. Therefore, we studied the effect of amino acids and complex organic nitrogen sources using Tolypocladium inflatum mutants on the productivity of cyclosporin A and the ratio of cyclosporin derivatives. Overproducing mutant YHC-004 having seven times higher productivity than mother strain's could be obtained through the artificial mutation by UV irradiation. The concentration and kind of organic nitrogens and amino acids shows the profound effect on the productivity of cyclosporin A and ratio of cyclosporin derivatives. As a result, it was possible to raise the productivity and the ratio of cyclosporin A up to 3,430 mg/L and 93% respectively, but on the other hand the other cyclosporin derivatives decreased less than 2% in the culture broth.

• Korean Journal of Microbiology
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• v.31 no.2
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• pp.141-147
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• 1993

Plasmid DNAs were detected from the mitochondrial fraction of four strains of whiterot fungus, Pleurotus ostreatus. The size of the plasmids were 10.2 and 7.2 kb in strain NFFA 2, 10.2 kb in NFFA 4001, 11.2 kb in NFFA 4501, and 10.2 and 11.2 kb in KFCC 11635. The two strains,NFFA 2ml and NFFA 2m2, which are mutant derivatives of NFFA 2, did not contain any plasmids. The cleavage by proteinase K indicated that these plasmids have DNA ends associated with proteins. In digestion with proteinase K all the plasm ids remained resistant to lambda exonuclease which hydrolyzes DNA from 5' ends and were sensitive to exonuclease III which hydrolyzes DNA from 3' ends. This suggests that the plasmids are linear double-stranded DNA and the terminal proteins are covalently linked to 5' ends of plasm ids. In order to find relationship between these plasmids, hybridization of plasm ids by each separate plasmid DNA was done. The result indicated that the plasmids can be classified into at least 3 groups. Plasmids of group I were present in all the P ostreatus. More mitochondrial plasmids were detected in P cornucopiae. P ,florida, P pulmonarius, P sajor-caju, and P spodoleucus. The size of plasmids ranged between 7.2 kb and 14 kb. All the species except P cornucopiae contained plasmids of approximately 10 kb which hybridized with the 10.2 kb plasmid (group I) of P ostreatus NFFA 2.

• Journal of Microbiology and Biotechnology
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• v.16 no.11
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• pp.1690-1698
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• 2006

In order to investigate the effect of dissolved oxygen (DO) level on AVM $B_{1a}$ production by a high yielding mutant of Streptomyces avermitilis, five sets of bioreactor cultures were performed under variously controlled DO levels. Using an online computer control system, the agitation speed and aeration rate were automatically controlled in an adaptive manner, responding timely to the oxygen requirement of the producer microorganism. In the two cultures of DO limitation, the onset of AVM $B_{1a}$ biosynthesis was observed to casually coincide with the fermentation time when oxygen-limited conditions were overcome by the producing microorganism. In contrast, this phenomenon did not occur in the parallel fermentations with DO levels controlled at around 30% and 40% throughout the entire fermentation period, showing an almost growth-associated mode of AVM $B_{1a}$ production: AVM $B_{1a}$ biosynthesis under the environments of high DO levels started much earlier than the corresponding oxygen-limited cultures, leading to a significant enhancement of AVM $B_{1a}$ production during the exponential stage. Consequently, approximately 6-fold and 9-fold increases in the final AVM $B_{1a}$ production were obtained in 30% and 40% DO-controlled fermentations, respectively, especially when compared with the culture of severe DO limitation (the culture with 0% DO level during the exponential phase). The production yield ($Y_{p/x}$), volumetric production rate (Qp), and specific production rate (${\bar{q}}_p$) of the 40% DO-controlled culture were observed to be 14%, 15%, and 15% higher, respectively, than those of the parallel cultures that were performed under an excessive agitation speed (350 rpm) and aeration rate (1 vvm) to maintain sufficiently high DO levels throughout the entire fermentation period. These results suggest that high shear damage of the high-yielding strain due to an excessive agitation speed is the primary reason for the reduction of the AVM $B_{1a}$ biosynthetic capability of the producer. As for the cell growth, exponential growth patterns during the initial 3 days were observed in the fermentations of sufficient DO levels, whereas almost linear patterns of cell growth were observed in the other two cultures of DO limitation during the identical period, resulting in apparently lower amounts of DCW. These results led us to conclude that maintenance of optimum DO levels, but not too high to cause potential shear damage on the producer, was crucial not only for the cell growth, but also for the enhanced production of AVM $B_{1a}$ by the filamentous mycelial cells of Streptomyces avermitilis.

• Microbiology and Biotechnology Letters
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• v.32 no.1
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• pp.52-59
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• 2004

When both fructose and galactose were added to a production medium as carbon sources, the productivity of PAFS (Psedomonas Antifungal Substance) biosynthesized by Pseudomonas aeruginosa was observed to be reduced significantly due to the well-known phenomenon of catabolite repression. In order to overcome this phenomenon by use of fermentation bioprocess, fed-batch cultivation method was examined. In addition, a high producer mutant strain, AP-20 obtained by a rational screening method was tested for its productivity of PAFS in both batch and fed-batch fermentation processes. Notably fed-batch operation showed approximately 4 fold higher PAFS productivity than traditional batch operation process. It was appeared that galactose was utilized principally for the cell growth of Pseudomonas aeruginosa whereas large portion of fructose was used for the biosynthesis of PAFS. Furthermore it was observed that composition and feeding rate of production media should be optimized even in the fed-batch fermentation bioprocess. As an example, very slow feeding of carbon sources gave rather negative effect on the production of PAFS due to significant limitation of carbon and energy sources available for the producer microorganism.