• Title/Summary/Keyword: cell-free culture broth

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Regulation of extracellular alkaline protease biosynthesis in a strain of streptomyces sp. (Streptomyces sp. 일주에서 균체외 호염기성 단백질분해 효소의 생합성 조절)

  • 신현승;이계준
    • Korean Journal of Microbiology
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    • v.24 no.1
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    • pp.32-37
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    • 1986
  • In fermentation studies it revealed that Streptomyces sp. SMF 3001 started to synthesize extracellular alkaline protease from early exponential phase of cell growth. The biosynthesis of the alkaline protease was greatly induced by skim milk as a sola nitrogen source and further stimulation was observed under inorganic sulphur limited culture. However, it was found that the biosynthesis was apparently repressed by $NH_4^+$ and free amino acids, specially by cysteine. It was considered that the strain SMF 301 of Streptomyces sp. would produce the alkaline protease for the uptake of sulphur compounds from protein contained in the culture broth.

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Biodegradation and Removal of PAHs by Bacillus velezensis Isolated from Fermented Food

  • Sultana, Omme Fatema;Lee, Saebim;Seo, Hoonhee;Al Mahmud, Hafij;Kim, Sukyung;Seo, Ahyoung;Kim, Mijung;Song, Ho-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.999-1010
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    • 2021
  • Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in the environment. They are highly toxigenic and carcinogenic. Probiotic bacteria isolated from fermented foods were tested to check their ability to degrade and/or detoxify PAHs. Five probiotic bacteria with distinct morphologies were isolated from a mixture of 26 fermented foods co-cultured with benzo(a)pyrene (BaP) containing Bushnell Haas minimal broth. Among them, B. velezensis (PMC10) significantly reduced the abundance of BaP in the broth. PMC10 completely degraded BaP presented at a lower concentration in broth culture. B. velezensis also showed a clear zone of degradation on a BaP-coated Bushnell Haas agar plate. Gene expression profiling showed significant increases of PAH ring-hydroxylating dioxygenases and 4-hydroxybenzoate 3-monooxygenase genes in B. velezensis in response to BaP treatment. In addtion, both live and heat-killed B. velezensis removed BaP and naphthalene (Nap) from phosphate buffer solution. Live B. velezensis did not show any cytotoxicity to macrophage or human dermal fibroblast cells. Live-cell and cell-free supernatant of B. velezensis showed potential anti-inflammatory effects. Cell-free supernatant and extract of B. velezensis also showed free radical scavenging effects. These results highlight the prospective ability of B. velezensis to biodegrade and remove toxic PAHs from the human body and suggest that the biodegradation of BaP might be regulated by ring-hydroxylating dioxygenase-initiated metabolic pathway.

Purification and Characterization of the Regulatory Substance of Furfural Biodegradation in Pseudomonas fluorescens (Pseudomonas fluorescens에 의한 Furfural의 분해대사 조절물질에 관하여)

  • 이병웅;유병설;이계준;하영칠
    • Korean Journal of Microbiology
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    • v.23 no.4
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    • pp.241-247
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    • 1985
  • The objectives of this study were to isolate and identify ninhydrin positive substande(s) produced in the culture broth of Pseudomonas fluorescens. It was found that the NPS could stimulate bioconversion of furfural into furoic acid. In order to isolate the NPS from the culture broth, cell free filtrate was subjected to ion-exchange chromatography, gel-permeation and finally to cellulose column chromatography. The purified NPS was white amorphous power and very soluble in water, slightly soluble in methanol and very insoluble in organic solvents. UV, and IR absorption spectra. $^H$ and $^{13}C-NMR$ were measured in order to identify the chemical structure of the NPS.

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Isolation and Identification of a Symbiotic Bacterium from Steinernema carpocapsae

  • Park, Sun-Ho;Yu, Yeon-Su
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.4 no.1
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    • pp.12-16
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    • 1999
  • Xenorhabdus nematophilus sp., an insect-pathogenic bacterium, was newly isolated from Korean entomopathogenic nematode of Steinernema carpocapsae, which can be used as a useful bioinsecticide. Primary and secondary form variants of Xenorhabdus nematophilus were observed when cultured in vitro. Primary form variants adsorbed bromothymol blue, while secondary form did not. However, many other characters of two variants were very similar. The variants were all rod-shaped and cell size was highly variable ranging from 0.5 by 2.0 ${\mu}$m to 1.0 by 5.0 ${\mu}$m. Both produced highly toxic substances and killed the insect larva within 20∼38 hr, indicating that insect pathogenicity of Xenorhabdus is not directly associated with its phase variation. In addition, cell-free culture supernatant of Xenorhabdus was sufficient to kill the insect larva by injecting it ito insect hemolymph; however, cell-harboring culture broth was more effective for killing the insect. The use of Xenorhabdus nematophilus may provide a potential alternative to Bacillus thuringiensis (Bt) toxins.

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Hydrolysis of the Nitrile group in $\alpha$-Aminophenylacetonitrile by Nitrilase;Development of a New Biotechnology for Stereospecific Production of S-$\alpha$-Phenylglycine

  • Choi, Soo-Young;Goo, Yang-M
    • Archives of Pharmacal Research
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    • v.9 no.1
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    • pp.45-47
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    • 1986
  • Phenylglycine was obtained as the sole metabolite when .alpha.-aminophenylacetonitrile was ted to the culture broth of Aspergillus furmigatus furmigatus. The isolated phenylglycine showed L-configuration with 80% optical purity. Examination of the hydrolysis of the substrate to phenylglycine with cell free extracts, and the supernatant fraction and the particulate fraction both of which were obtained after ultracentrifugation of the cell free extract at 100,000g, indicated that the nitrile group hydrolyzing enzymes, nitrilase existed not only in cytoplasm, but in microsome fractions.

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고정상세포분리기의 개발 및 Cyclosporin A 생산을 위한 고정화 연속배양공정에의 적용

  • Lee, Tae-Ho;Park, Sung-Kwan;Chang, Yong-Keun;Chun, Gie-Taek
    • Microbiology and Biotechnology Letters
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    • v.24 no.6
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    • pp.717-725
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    • 1996
  • We have developed an efficient immobilized cell separator for continuous operation of immobilized fungal cell cultures, and applied this separator to actual fermentation process for the production of cyclosporin A (CyA), a powerful immunosuppressant. In the experiments employing highly viscous polymer (carboxymethyl cellulose) solution, the decantor showed good separating performances at high solution viscosites and fast dilution rates. Air duct and cylindrical separator installed inside the decantor turned out to play key roles for the efficient separation of the immobilized cells. By installing the decantor in an immobilized perfusion reactor system (IPRS), continuous immobilized culture was stably carried out even at high dilution rate for a long period, leading to high productivities of free cells and CyA. Almost no immobilized biomass existed in effuluent stream of the IPRS, demonstrating the effectiveness of the decan- tor system for a long-term continuous fermentation. It was noteworthy that we could obtain these results despite of the unfavorable fermentation conditions, i.e., reduced density of the biosupports caused by overgrowth of cells inside the bead particles and existence of high density of suspended fungal cells (10g/l) in the fermentation broth.

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Characteristics of the Algal Growth inhibition Substances Produced by Alteromonas sp. SR-14 (Alteromonas sp. SR-14가 생산하는 조류증식 저해 물질의 특성)

  • 김지회;이희정;이태식;김형락;이명숙;장독석
    • Journal of Food Hygiene and Safety
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    • v.14 no.3
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    • pp.270-276
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    • 1999
  • In previous reports, the authors isolated the algicidal marine bacterium, Alteromonas sp. SR-14 and demonstrated its growth inhibition of diatom, Chaetoceros calcitrans (C. calcitrans). In this paper, we studied the effects of cell free culture filtrate of Alteromonas sp. SR-14 on the growth of C. calcitrans, and the characteristics of the algal growth inhibition substance. The culture filtrate of Alteromonas sp. SR-14 grown in peptone broth showed growth inhibition activity against C. calcitrans. The reasonable culture conditions of the bacterium for producing of algal growth inhibition substances were $15~20^{\circ}$ in temperature, 7.0-9.0 in pH and $23~30{\textperthousand}$ in salinity, respectively. The algal growth inhibition activity of culture filtrate was increased from stationary phase in growth curve of Alteromonas sp. SR-14. The molecular weights of algal growth inhibition substances produced by Alteromonas sp. SR-14 were ranged about from 3 KDa to 12 KDa. Among the substances, less than 10 KDa fraction were stable by heating at $100^{\circ}$ for 10 minutes, while more than 10 KDa fraction were heat labile. According to the experimental results, the algal growth inhibition substance produced by the bacterium was not a single compound.

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Charaterization of Nisin Production and Resistance of Lactococcus lactis ssp. lactis $ML_8$ (Lactococcus lactis ssp. lactis $ML_8$의 Nisin 생산 및 저항 특성)

  • 김등양;이형주
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.619-623
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    • 1991
  • To investigate nisin production and resistance of Lactococcus lactis ssp. tactis ML (L. lactis $ML_8$, effects of medium, pH of culture broth, and cell growth on the nisin activity, and effect of nisin with or without $Ca^[2+}$ ion on the growth of L. lactzs were analyzed. In the bio-assay of nisin by the agar diffusion method, inhibition-zone diameter of Micrococcus Javus was propotional to the logarithm of nisin concentration ranged 0.5~20 unitlml (12.5~500 ng/mf). Nisin activity of the pasteurized culture filtrates of L. lactis MLs was high at pH 2!3 but was inactivated completely at pH over 6.0. Nisin production of the L. lactis $ML_8$ cultured on LTB broth increased at late logarithmic phase and reached 10.5 unitlml after 16 hr. The cell growth of L. lactis LM 0230, a plasmid free and nisin sensitive strain, was inhibited on agar medium containing 7 unitlrnl of nisin, while L. lactis $ML_8$ showed high survival ability at 20 unitld of nisin. When 40 mM $Ca^[2+}$ ion was added to Elliker broth with 8 unitlml of nisin, the growth pattern of L. lactis $ML_8$ was similiar to that on control medium which did not contain nisin and $Ca^[2+}$ ion, and this suggested that $Ca^[2+}$ increased the nisin resistance of the L. lactis.

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Two Entomopathogenic Bacteria, Xenorhabdus nematophila K1 and Photorhabdus temperata subsp. temperata ANU101 Secrete Factors Enhancing Bt Pathogenicity against the Diamondback Moth, Plutella xylostella (배추좀나방(Plutella xylostella)에 대한 두 곤충병원세균(Xenorhabdus nematophila K1과 Photorhabdus temperata subsp. temperata ANU101) 배양물질의 Bt 병원성 제고 효과)

  • Seo, Sam-Yeol;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.48 no.3
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    • pp.385-392
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    • 2009
  • Two entomopathogenic bacteria, Xenorhabdus nematophila and Photorhabdus temperata subsp. temperata, are known to be potent against the diamondback moth, Plutella xylostella, when the bacteria are injected into the hemocoel. This study investigated any pathogenic effect of their culture broth on P. xylostella by oral administration. Only culture broth of both bacterial species did not give enough pathogenic effects by the oral administration. However, when the culture broth was orally treated together with Bacillus thuringiensis (Bt), both cell-free culture broth significantly enhanced Bt pathogenicity against the 3rd instar larvae of P. xylostella. The culture broth was then fractionated into hexane, ethyl acetate, and aqueous extracts. Most synergistic effect on Bt pathogenicity was found in ethyl acetate extracts of both bacterial species. Thin layer chromatography of these extracts clearly showed that ethyl acetate extracts of both bacterial culture broths possessed metabolites that were different to those of hexane and aqueous extracts. These results suggest that the both entomopathogenic bacteria produce and secrete different factors to give significant synergistic effect on Bt pathogenicity.

Optimal conditions and effects of prebiotics for growth and antimicrobial substances production of Lactobacillus brevis BK11 (Lactobacillus brevis BK11의 증식과 항균물질 생산을 위한 최적 배양조건 및 prebiotics의 영향)

  • Lim, Eun-Seo
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.288-299
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    • 2015
  • Lactobacillus brevis BK11 obtained from Baikkimchi was selected to study the effects of culture medium, initial pH, atmosphere composition, incubation temperature and time, and prebiotics on growth and production of antimicrobial substances. Growth and antimicrobial substances production of L. brevis BK11 were significantly higher in MRS broth than in BHI or M17 broth. The production of cell mass, lactic acid, and bacteriocin by BK11 strain was at maximum in MRS broth adjusted to pH 6.0. Aerobic and microaerobic conditions were favored cell growth and antimicrobial substances production than anaerobic condition. Biomass and lactic acid production and antimicrobial substances activity of BK 11 were significantly better at 30 and $37^{\circ}C$ than at $25^{\circ}C$. Growth of the strain BK11 entered the stationary growth stage at 24 h after inoculation, and decreased after 36 h. Antimicrobial activities of cell-free culture supernatant and bacteriocin solution were highest when cultured in MRS broth with an initial pH 6.0 for 24-30 h at $37^{\circ}C$. In addition, the highest cell number and lactic acid and bacteriocin production were recorded in the presence of 1 and 2% (w/v) fructooligosaccharide (FOS), however, inulin and raffinose did not affect biological and physicochemical characteristics and antimicrobial activities of L. brevis BK11 cultures. According to these results, production of antimicrobial substances by L. brevis KB11 was closely associated with cell density. Under optimal conditions for antimicrobial substances production, L. brevis BK11 effectively inhibited the growth of Helicobacter pylori ATCC 43504.