• Korean Journal of Soil Science and Fertilizer
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• 제25권4호
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• pp.401-406
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• 1992

Antifungal substances against three plant pathogenic fungi, Phytophthora capsici, Phytophthora nicotianae var. parasitica and Rhizoctionaia solani were fractionated cultures of Streptomyces sp. A-2 strain isolated in Korean soils. Characterizations of active substances related with antagonistic effects were follows : 1. The excellent media which showed the transfer efficiency of antagonistic substances from Streptomyces sp. A-2 strain G.Y.B and B.H.I. among four that are glucose yeast broth (G.Y.B), $M\ddot{u}eller$, brain heart infusion(B.H.I.) and Czapek media. Active substances which were transfered into ethylacetate or left in residual aqueous phase did not lose antagonistic activity in spite of autoclavation. This indicated that bonds of these compounds were rigid enough to keep activity under such conditions. 2. Antagonistic substances were extracted according to adjustment of pH 3 or pH12 to 5 day-old B.H.I. broth cultures of Streptomyces sp. A-2 strain. Comparative analysis fluorescent bands on HPTLC to antagonsitic spectra against three phytopathogenic fungi indicated that major substances with antagonistic activity were extracted regardless of different pH adjustment to broth cultures. Since UV spectrum of these fractions scanned from 500nm to 200nm was similiar to that of polyene macrolide, major substances related with antagonistic activities were assumed to be polyene derivatives antibiotics.

• Journal of Microbiology and Biotechnology
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• 제14권1호
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• pp.90-96
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• 2004

A synthetic defined medium, fortified with amino acids, was developed for the stable production of the kringle fragments of human apolipoprotein(a) (apo(a)), rhLK68. Using a complex rich medium containing yeast extract and a high-cell-density fed-batch culture, the expression level of rhLK68 reached 17% of the total cellular protein, which corresponded to $5\;g\;l^{-1}$ of the culture. To replace the complex media with chemically defined media, several amino acids that positively affect cell growth and gene expression were chosen by a statistical method. The various combinations of the selected amino acids were tested for its fortifying effect on a minimal defined medium. When glutamine only was added, the overall expression level of rhLK68 reached 93% of the complex rich medium increasing the specific expression level by 22.4% and decreasing the cell growth by 24%. Moreover, the addition of glutamine resulted in a 2-fold increase in the concentration of rhLK68 in the culture broth, compared with the minimal defined medium. The synthetic defined media developed in this study could be generally applied to high-cell-density cultures of the recombinant Escherichia coli BL21(DE3), especially for the production of therapeutic proteins that require a strict quality control of the culture media and fermentation processes.

• Journal of Life Science
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• 제22권6호
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• pp.823-827
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• 2012

A bacterial strain producing a fibrinolytic enzyme, subtilisin CP-1, was isolated from Doen-Jang, a Korean traditional fermentation food. Based on the analysis of gene sequence of 16S rRNA and biochemical analysis, the strain was identified as Bacillus sp. and named as Bacillus sp. CP-1. To investigate the effect of the medium on the production of fibrinolytic enzyme from Bacillus sp. CP-1, two commercial bacterial culture media, tryptic soy broth (TSB) and Luria-Bertani (LB), were applied to the cultivation of Bacillus sp. CP-1. The strain secreted only one proteolytic enzyme (subtilisin CP-1) in the culture broth. The molecular weight of subtilisin CP-1 was estimated to be 28 kDa. Subtilisin CP-1 was optimally active at pH 9.0 and $45^{\circ}C$, and exhibited high specificity for Meo-Suc-Arg-Pro-Tyr-pNA (S-2586), a synthetic chromogenic substrate for chymotrypsin. The first eight amino acid residues of the N-terminal sequence of the enzyme are AQSVPYGI; this sequence is identical to that of subtilisin NAT and E.

• Applied Biological Chemistry
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• 제46권4호
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• pp.291-298
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• 2003

A bacterial strain producing a high level of an extracellular phytase was isolated from livestock waste water, identified as a strain of Pseudomonas fragi and designated as Pseudomonas fragi Y9451. Under the phytase production medium, the activity of phytase reached the highest level after 120 hours of incubation. On the effect of carbon sources on the phytase production, the most favorable carbon source for phytase production was fructose. As for the effect of nitrogen sources, high levels of phytase activity were detected in the medium containing nutrient broth as the nitrogen source. Free $PO_4^{3-}$ inhibited phytase production with increasing concentration of $KE_2PO_4$ and phytate in the media. The addition of $CaCl_2$ and $MgSO_4$ also resulted in the inhibition of phytase production. To investigate the effect of aeration on the phytase production, different volumes of culture broth in Erlenmeyer flasks were incubated in rotary shaker at the speed of 200 rpm. As a result, a high level of phytase activity was detected at small volume of culture broth as compared to larger volume because of its more aerobic condition.

• The Korean Journal of Pesticide Science
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• 제20권2호
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• pp.152-158
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• 2016

The cause of death was investigated with several dead cabbage moth larvae in breeding box. Bacterial strains were isolated and selected from the dead larvae by bioassay. One of them was identified as Serratia marcescens used by morphological characteristics and gene sequencing. S. marcescens were cultured by Luria Bertani (LB) media broth for bioassay. When 100-fold dilution of culture broth to third larvae of diamondback moth, Plutella xylostella, it was showed a 100% mortality at 2 days after treatment, and only 10-fold dilution of supernatant liquid was showed 86.6% mortality. When the culture broth of S. marcescens was applied to the larvae of beet armyworm, Spodoptera exigua, contact and feeding toxicity were 20 and 8% of mortality, respectively. Otherwise, when the culture broth of S. marcescens was applied to 5 major plant pathogens, antibacterial activities against Fusarium oxysporum, Rhizoctonia solani, Colletotrichum gloeosporioides, Phytophthora capsici and Sclerotinias clerotiorum were 4.7, 11.3, 20, 15.7 and 42.6%, respectively. Also, degradation ability of S. marcescens against protein and chitin were examined.

• Korean Journal of Food Science and Technology
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• 제26권5호
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• pp.644-648
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• 1994

Lactic acid bacteria(LAB) and its metabolites were tested for their inhibitory effect on the growth of Staphylococcus aureus in vitro. When S. aureus and LAB were incubated simultaneously in MRS broth, the growth of S. aureus began to be suppressed after 12 hour of incubation, completely inactivated within 24 hour of incubation by L. lactis and 48 hour by L. casei, L. acidophilus, and Sc. thermophilus. The pH values of media incubated by S. aureus and LAB were about 4.5 at 12 hour of incubation and 3.5 at 48 hour. The metabolites of the four LAB all exerted antibacterial activity on the growth of S. aureus in TS broth, but is got lost the antibacterial activity by heating ($100^{\circ}C,\;20\;min$). On treating metabolites with catalase, only L. lactis were lost its antibacterial activity. Organic acids (acetate, lactate) showed more active inhibition than inorganic acid (HCl) at pH 3.5, 4.5 but there is no significant differance at pH 5.5, 6.5.

• Korean Journal of Microbiology
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• 제40권2호
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• pp.100-103
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• 2004

The culture condition and medium composition for the enhanced mycelial growth of Inonotus mikadoi IMSNU 32058 were investigated. The optimal temperature and pH for the mycelial growth were $27^{\circ}C$ and pH 4.5, respectively. Among the complex media tested, the malt extract medium and Phellinus igniarius medium were very good for mycelial growth of I. mikadoi. When Czapek-Dox medium was used as a minimal medium for cultivation of mycelia, xylose, raffinose and carboxymethyl cellulose were very excellent as a carbon and energy source. With respect to carbohydrate, sucrose and glucose were very good carbon sources. In general, organic complex nitrogen sources were better than other inorganic ones. As the organic complex nitrogen sources tested, yeast extract, soytone, proteose peptone and bacto peptone were the best as a source of organic nitrogen. When ammonium sulfate as an inorganic source of nitrogen was used, the enhanced mycelial growth was shown. p-Aminobenzoic acid was proved to be most appropriate source of vitamin. After the mycelia of I. mikadoi was cultivated at $27^{\circ}C$ for 5 days in MEM broth (pH 4.5), the activities of both exomycelial and endo-mycelial enzymes were determined. Among endomycelial enzymes assayed, the specific activity of laccase was much higher than those of other enzymes. When the fungus was grown in MEM broth, exomycelial specific enzyme activity of laccase was comparatively high. However, little or no enzyme activities of protease, chitinase and lipase were found.

• Journal of dental hygiene science
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• 제12권3호
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• pp.209-215
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• 2012

Somok, the heart wood of Caesalpinia sappan is used in traditional Chinese medicine. This study was performed to investigate the effect of growth and culture conditions of brazilin from C. sappan against S. mutans ATCC 25175. The bacteria were cultured in brain heart infusion (BHI) broth, and then incubated under 5% $CO_2$ at $37^{\circ}C$ for 18-24 hours. The effect of brazilin against S. mutans was confirmed under the changes of the culture conditions, such as growth curve and the change of pH, protein, and total carbohydrate. The growth of S. mutans in control medium was the highest at 24 hr, while brazilin-added medium (0.3 mg/ml) showed maximum growth at 32 hr. The pH values of the control medium was 5.25 at 16 hr, but the media supplemented with brazilin (0.3 mg/ml) was 7.0 at 16 hr. The amounts of total carbohydrate of the control medium was 11 mg/ml at 8 hr, but the brazilin-added media (0.3 mg/ml) was 18 mg/ml at 8 hr. In the protein change of the culture medium, the control culture broth and the brazilin supplemented-cultures was 2.4 mg/ml and 2.54 mg/ml at 24 hr, respectively. Polysaccharide contents of the control medium and test media were 3 mg/ml and 2 mg/ml at 8 hr, respectively. Thus, the application of C. sappan can be considered a useful and practical material for the prevention of dental caries.

• The Korean Journal of Pesticide Science
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• 제12권2호
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• pp.184-191
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• 2008

Entomopathogenic bacteria (Xenorhabdus nematophila, X. sp. and Photorhabdus temperata subsp. temperata) isolated from entomopathogenic nematodes express potent insecticidal activity in insect hemocoel. They are also known to suppress insect immune mediation by inhibiting phospholipase $A_2$, leading to host immunosuppression. This study analyzed effects of their cultured broths on inhibiting insect immunosuppression. For this, we removed all bacterial cells using $0.2\;{\mu}m$ pore sized membrane from the bacteria-cultured broth. All three sterilized cultured media, in dose-dependent manners, significantly inhibited hemocyte-spreading behavior of 5th instar larvae of Spodoptera exigua. However, they showed differential inhibitory activities among different bacterial species, in which X. nematophila showed the most potent inhibitory activity. This immunosuppressive effect was applied to increase the pathogenicity of Bacillus thuringiensis (Bt). All three bacterial cultured broths including bacterial cells significantly potentiated Bt pathogenicity against young S. exigua larvae when each of them was orally administered in a mixture of low dose of Bt. Finally, we tested the effect of oral administration of the cultured media containing the immunosuppressive compound(s) secreted by the bacteria. The membrane-sterilized cultured broths were mixed with the low dose of Bt and then orally administered to the young S. exigua. Only the cultured medium of X. nematophila showed increase of Bt pathogenicity. These results indicated that the; cultured media of the three bacteria possessed immunosuppressive factor(s), which may act to potentiate Bt toxicity to young S. exigua larvae.

• Journal of Fisheries and Marine Sciences Education
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• 제26권6호
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• pp.1366-1372
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• 2014

This study was conducted to select the media for the formalin killed vaccine (FKC) production of Vibrio harveyi and its application for olive flounder, Paralichthys olivaceus. For this, we have investigated the immune effects of Vibrio harveyi FKC vaccines grown in 3 different media i.e. Tryptic Soy Broth (TSB), TSB containing 2-2'-dipyridyl (TSB-D), Brain Heart Infusion Broth (BHIB) on the production of agglutinating antibody and protection against Vibrio harveyi in olive flounder. Additionally, a dual vaccine was prepared by combining Streptococcus parauberis vaccine to V. harveyi vaccine and its efficacy was also analyzed with the determination of optimal administration dosage. Consequently, olive flounder immunized with FKC grown in TSB-D showed the same protection with the vaccine grown in BHIB and the optimal dose of the vaccine was 10mg/kg of body weight. Indeed the dual vaccine showed higher agglutination titer and protection than control fish. The optimal dose for dual vaccine was 10mg/kg body weight for each of two vaccines.