• Journal of Microbiology and Biotechnology
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• v.24 no.12
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• pp.1609-1621
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• 2014

The plant pathogen Burkholderia gladioli, which has a broad host range that includes rice and onion, causes bacterial panicle blight and sheath rot. Based on the complete genome sequence of B. gladioli BSR3 isolated from infected rice sheaths, the genome of B. gladioli BSR3 contains the luxI/luxR family of genes. Members of this family encode N-acyl-homoserine lactone (AHL) quorum sensing (QS) signal synthase and the LuxR-family AHL signal receptor, which are similar to B. glumae BGR1. In B. glumae, QS has been shown to play pivotal roles in many bacterial behaviors. In this study, we compared the QS-dependent gene expression between B. gladioli BSR3 and a QS-defective B. gladioli BSR3 mutant in two different culture states (10 and 24 h after incubation, corresponding to an exponential phase and a stationary phase) using RNA sequencing (RNA-seq). RNA-seq analyses including gene ontology and pathway enrichment revealed that the B. gladioli BSR3 QS system regulates genes related to motility, toxin production, and oxalogenesis, which were previously reported in B. glumae. Moreover, the uncharacterized polyketide biosynthesis is activated by QS, which was not detected in B. glumae. Thus, we observed not only common QS-dependent genes between B. glumae BGR1 and B. gladioli BSR3, but also unique QS-dependent genes in B. gladioli BSR3.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.579-582
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• 2000

(R,S)-3-amino-n-butanoic acid$(DL-\;{\beta}\;-homoalanine)$ has been kinetically resolved using Alcaligenes denitrificans Y2k-2 as a biocatalyst, which was isolated from soil by enrichment culture, which was carried out with minimal media containing (R,S)-3-amino-n-butanoic acid as a sole nitrogen source. The enzyme which peformed this kinetic resolution assumed to belong to the ${\omega}-transaminase$ family, because A. denitrificans used pyruvate as amino acceptor and its transaminase activity was inhibited by gabaculine, aminooxy acetic acid and hydroxylamine. In whole cell reaction, (R,S)-3-amino-n-butanoic acid was kinetically resolved to the corresponding (R)-3-amino-n-butanoic acid with excellent E (>100) in the presence of pyruvate as an amino acceptor at $37^{\circ}C$. (S-specific) We observed the substrate inhibition for pyruvate at 100mM. In this study, characteristics of transaminase activity of Alcaligenes denitrificans Y2k-2, such as substrate specificity and thermostability, are carried out for the development of (R)-3- amino-n-butanoic acid production system.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.2
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• pp.154-160
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• 2016

Several our studies have focused recently on the mass production of the freshwater Cladoceran Moina macrocopa which can substitute Artemia nauplii for the culture of larval marin fish. A 6 weeks experiment was conducted to investigate the effects of enrichment on the fatty acid composition of Moina macrocopa through feeding Schizochytrium sp. containing highly unsaturated fatty acids and to study the impacts of n-3-HUFA enriched Moina on improving survival rate and fatty acid compostion of larval rockfish Sebastes schlegeli. After feeding for 6 weeks, the Moina-fed fry resulted in a higher survival rate of 99.2% compared to the Artemia-fed fry 12.8%. In addition, the Moina-fed fry had the fast growth rate 45.6mm compare to the Artemia-fed fry 25.7 mm at the end of the experiment. The Moina-fed fry showed significantly higher level of 16.47% DHA than their Artemia-fed fry counterparts of the level of 3.97% with respect to DHA. PL, the cell membrane components in living food organisms, constituted 63.8% of the Moina, which was significantly higher than in the 40.1% of the Artemia. The present study indicate that Moina macrocopa can be used as Artemia substitute and improving the survival rate rockfish larvae through enchriment Schizochytrium sp.

• Korean Journal of Plant Tissue Culture
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• v.24 no.4
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• pp.197-201
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• 1997

Plant seed oil-bodies or oleosomes ate the repository of the neutral lipid stored in seeds. These organelles in many oilseeds may comprise half of the total cellular volume. Oleosomes are surrounded by a half-unit membrane of phospholipid into which are embedded proteins called oleosins. Oleosins are present at high density on the oil-body surface and after storage proteins comprise the most abundant proteins in oilseeds. Oleosins are specifically targeted and anchored to oil-bodies after co-translation on the ER. It has been shown that the amino-acid sequences responsible for this unique targeting reside primarily in the central hydrophobic tore of the oleosin polypeptide. In addition, a signal-like sequence is found near the junction of the hydrophobic domain and ann N-terminal hydrophilic / amphipathic domain. This "signal" which is uncleaved is also essential for correct targeting. Oil-bodies and their associated oleosins may be recovered by floatation centrifugation of aqueous seed extracts. This simple partitioning step results in a dramatic enrichment for oleosins in the oil-body fraction. In the light of these properties, we reasoned that it would be feasible to create fusion proteins on oil-bodies comprising oleosins and an additional valuable protein of pharmaceutical or industrial interest. It was further postulated that if these proteins were displayed on the outer surface of oil-bodies, it would be possible to release them from the purified oil-bodies using chemical or proteolytic cleavage. This could result in a simple means of recovering high-value protein from seeds at a significant (i.e. commercial) scale. This procedure has been successfully reduced to practice for a wide variety of proteins of therapeutic, industrial and food no. The utillity of the method will be discussed using a blood anticoagulant, hirudin, and industrial enzymes as key examples.

• KSBB Journal
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• v.11 no.1
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• pp.92-98
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• 1996

A chitinase-producing bacterium was isolated from seashore mud around Beobseongpo in Chunnam province by selective enrichment culture, and among it, one isolate which was the best in producing of chitinase was selected. Nutrient or MacConkey medium was confirmed with secreting of prodigiosin pigment by Serratia sp. JM, and it was performed by the production of clear zone on medium containing chitin. Serratia sp. JM was almost same compared with Serratia marcescens ATCC 27117 in respect of its morphological, physiological and biochemical characteristics except succinic, urea and pyruvic acid. Serratia sp. JM was resistant to tetracycline but was not resistant to kanamycin and chloramphenicol. The optimal temperature and pH for the production of chitinase from Serratia sp. JM were $30^{\circ}C$ and 7.5, respectively. Production of chitinase and pH in the medium increased until the cultivation of 120 hours, but after 120 hours, they were decreased due to the acetic acid accumulated from degradation of chitin by Serratia sp. JM.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.4
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• pp.467-473
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• 2016

Several of our recent studies have focused on mass production of the marine Cladoceran Diaphanosoma celebensis, which can serve as a feed substitute for Artemia nauplii in the culture of larval marine fish. We compared the growth and survival rates of tiger puffer Takifugu rubripes fed D. celebensis with those fed Artemia. The group fed D. celebensis showed an increase in growth from 0.10 g to 1.78 g, while the group fed enriched Artemia had a growth from 0.10 g to 0.92 g. The survival rate of fish fed D. celebensis was 86.7% while that of fish fed enriched Artemia was 51.1%, indicating that the former group was superior to the latter both in growth and survival rate. Fatty acid analysis revealed that D. celebensis fed Nannochloropsis oculata and Schizochytrium sp. had an eicosapentaenoic acid (EPA) content of 16.42% and a docosahexaenoic acid (DHA) content of 3.93%. Meanwhile, juvenile tiger puffer fed D. celebensis had an EPA content of 9.12% and a DHA content of 10.69%. Juvenile tiger puffer fed D. celebensis had a similar n-3 highly unsaturated fatty acid content to that of tiger puffer fed enriched Artemia which had an EPA content of 5.82% and a DHA content of 15.90%, indicating that no additional enrichment was required.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.847-853
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• 1999

A Pseudomonas sp. strain that is capable of utilizing dicarboxylic acids as a sole carbon source was isolated from activated sludge by using the enrichment culture technique. This organism accumulated polyhydroxyalkanoates (PHAs) with an unusual pattern of monomer units that depends on the carbon sources used. Polyhydroxybutyrate (PHB) homopolyester was synthesized from glucose or small $C_{-even}$ alkanoic acids, such as butyric acid and hexanoic acid. Accumulation of PHB homopolyester was also observed in the cells grown on $C_{-odd}$ dicarboxylic acids, such as heptanedioic acid and nonanedioic acid as the sole carbon sources. In contrast, a copolyester consisting of 6 mol% 3-hydroxybutyrate (3HB) and 94 mol% 3-hydroxyvalerate (3HV) was produced with a PHA content of as much as 36% of the cellular dry matter. This strain produced PHAs consisting both of the short-chain-length (SCL) and the medium-chain-length (MCL) 3-hydroxyacid units when heptanoic acid to undecanoic acid were fed as the sole carbon sources. Most interestingly, polyester consisting of significant amount of relevant fractions, 3HB, 3HV, and 3-hydroxyheptanoate (3HHp), was accumulated from heptanoic acid. According to solvent fractionation experiments, the polymer produced from heptanoic acid was a blend of poly(3HHp) and of a copolyester of 3HB, 3HV, and 3HHp units. The hexane soluble fractions contained only 3HHp units while the hexane-insoluble fractions contained 3HB and 3HV units with a small amount of 3HHp unit. The copolyester was an elastomer with unusual mechanical properties. The maximum elongation ratio of the copolyester was 460% with an ultimate strength of 10 MPa, which was very different from those of poly(3HB-co-3HV) copolyesters having similar compositions produced from other microorganisms.

• Korean Journal of Microbiology
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• v.16 no.4
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• pp.170-179
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• 1978

By the successive enrichment culture, more than 250 methanol-utilizing bacteria were isolated from various samples such as soil, waste water and sewage. Two strains of which were selected and tentatively identified as Acinetobacter sp. and Pseudomonas sp. experiments were carried out to determine the growth conditions for the higher biomass yield and to demonstrate the difference to protein composition dependent upon carbon sources of these two species. the results were as follows ; 1. the optimum pH was determined as 8 in the both species. The optimum temperature in Acinetobacter sp. was $25^{\circ}C{\sim}30^{\circ}C$ and pseudomonas sp. was $30^{\circ}C-35^{\circ}C$. The optimum initial concentration of mthanol was determined as 1-2% in Acinetobacter sp. and 2-3% in pseudomonas sp. 2. The optimum concnetrations of nitrogen source, micro-elements, and vitamins such as biotin and thiamine-HCl in Acnetobactar sp. were 1g $(NH_4)_3SO4,\;1{\sim}3mg\;Mn^{++},\;4mg\;Fe^{++},\;10{\mu}g\;biotin,\;and\;100{\mu}g$ thiamine-HCl per liter medium. In the Pseudomonas sp., 2g $(NH_4)_3SO4,\;1mg\;Mn^{++},\;trace\;amounts\;of\;Fe^{++},\;5{\mu}g\;biotin,\;and\;100{\mu}g$ thiamine HCl per liter were effective. Maximum biomass yield was 2.5g/l in Acinetobacter sp. and 4.8g/l in Pseudomonas sp. 3. Protein composition of the two strains exhibited that alkai-labile protein was higher than alkali-stable protein. In Pseudomonas sp., the contents of acid soluble fraction and alkali-stable protein of the cells grown in the methanol medium were higher than in sucrose medium. On the other hand, in Acinetobacter sp., alkalilabile protein of the cells grown in sucrose medium was higher than in methanol medium.

• KSBB Journal
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• v.15 no.2
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• pp.208-213
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• 2000

Using three types of porcelain clays such as White, Blue, and Yellow clays, which were used as raw materials for Bae씨a, C Chungja, and common porcelains, the biological refinement by an enrichment culture of iron reducing bacteria was studied. | In the biological clay refining, amounts of leached iron increased as increasing sucrose $\infty$ncentration, which was s supplemented as a carbon and electron donor source for cell growth and iron reduction. Total amounts of the leached iron a and specific rate of iron reduction were dependent on the types of the clay. Strength and chromaticity of refined clays which a are important properties required for porcelain clays were improved as increasing sucrose concentration. The degree of s shrinking, however, did not changed. the redness among the chromaticity of refined clays is favorably reduced through the r ripening by the iron reducing bacteria. Considering iron removal efficiency and the change of physical properties, the optimal c concentration of sucrose was 4%(w/w) in the clay.

• Journal of Microbiology and Biotechnology
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• v.20 no.3
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• pp.615-621
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• 2010

The viability of low-temperature sulfate reduction with hydrogen as electron donor was studied with a bench-scale gas-lift bioreactor (GLB) operated at $9^{\circ}C$. Prior to the GLB experiment, the temperature range of sulfate reduction of the inoculum was assayed. The results of the temperature gradient assay indicated that the inoculum was a psychrotolerant mesophilic enrichment culture that had an optimal temperature for sulfate reduction of $31^{\circ}C$, and minimum and maximum temperatures of $7^{\circ}C$ and $41^{\circ}C$, respectively. In the GLB experiment at $9^{\circ}C$, a sulfate reduction rate of 500-600 mg $l^{-1}d^{-1}$, corresponding to a specific activity of 173 mg ${SO_4}^{2-}g\;VSS^{-1}d^{-1}$, was obtained. The electron flow from the consumed $H_2$-gas to sulfate reduction varied between 27% and 52%, whereas the electron flow to acetate production decreased steadily from 15% to 5%. No methane was produced. Acetate was produced from $CO_2$ and $H_2$ by homoacetogenic bacteria. Acetate supported the growth of some heterotrophic sulfate-reducing bacteria. The sulfate reduction rate in the GLB was limited by the slow biomass growth rate at $9^{\circ}C$ and low biomass retention in the reactor. Nevertheless, this study demonstrated the potential sulfate reduction rate of psychrotolerant sulfate-reducing mesophiles at suboptimal temperature.