• Title/Summary/Keyword: Fermentation broth

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Gluconacetobacter persimmonis sp. nov., Isolated from Korean Traditional Persimmon Vinegar

  • Yeo, Soo-Hwan;Lee, Oh-Seuk;Lee, In-Seon;Kim, Hyun-Soo;Yu, Tae-Shick;Jeong, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.276-283
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    • 2004
  • Screening was performed to isolate cellulose-producing microorganisms from the Korean traditional fermented persimmon vinegar. The resulting strain, KJ $145^{T}$, was then taxonomically investigated by phenotypic characterization, particularly chemotaxonomic, and by phylogenetic inference based on a 16S rDNA sequence analysis including other related taxa. Strain KJ $145^{T}$ was found to grow rapidly and form pale white colonies with smooth to rough surfaces on a GYC agar. Strain KJ $145^T$ also produced acetate from ethanol, and was tolerable to 10% ethanol in SM medium. In a static culture, a thick cellulose pellicle was produced, and in GYC broth, the strain grew at temperatures ranging from 28 to $40^\circ{C}$ with an optimum pH of 4.0. The genomic DNA G+C content of strain KJ $145^T$ was 61.9 mol%, and the predominant ubiquinone was Q 10 as the major quinone and Q9 as the minor quinone. The major cellular fatty acids were $C_{16:0}$ and the sum in feature 7 ($C_{18:1}$ w9c, w12t and/or w7c). A 16S rRNA-targeted oligonucleotide probe specific for strain KJ $145^T$was constructed, and the phylogenetic position of the new species was derived from a 16S rDNA-based tree. When comparing the 16S rDNA nucleotide sequences, strain KJ $145^T$ was found to be most closely related to G. hansenii LMG $1527^T$ (99.2%), although KJ $145^T$ was still distinct from G. hansenii LMG $l527^T$ and G. xylinus LMG $1515^T$ in certain phenotypic characteristics. Therefore, on the basis of 16S rDNA sequences and taxonomic characteristics, it is proposed that strain KJ $145^T$ should be placed in the genus Gluconacetobacter as a new species, Gluconacetobacter persimmonis sp. nov., under the type-strain KJ $145^T$ (=KCTC =$10175BP^T$=KCCM=$10354^T$).

Studies on Microbial Penicillin Amidase (Part 5) Application of Reinforced Calcium-Alginate Gel Entrappment Method for Immobilization of Penicillin Amidase from Bacillus megaterium (미생물 페니실린 아미다제에 관한 연구 (제 5보) Bacillus megaterium 페니실린 아미다제의 새로운 고정화 방법)

  • Son, Hyeung-Jin;Seong, Baik-Lin;Mheen, Tae-Ick;Han, Moon-Hi
    • Microbiology and Biotechnology Letters
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    • v.9 no.3
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    • pp.159-164
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    • 1981
  • Reinforced Calcium-alginate gel entrappment method for enzyme immobilization is described with an example of penicillin amidase from Bacillus megaterium KFCC 10029, a partially constitutive mutant of B. megaterium ATCC 14945. Penicillin amidase recovered from the fermentation broth by adsorption on celite is mixed with alginate and gelatin solution, and cast into a pellet or noodle form by coagulation in calcium salt solution followed by crosslinking with glutaraldehyde. Optimum pH and temperature of the immobilized enzyme preparation were 8.0 and 6$0^{\circ}C$, respectively. Kinetic constants such as Km value and the inhibition constant of 6-APA and phenylacetic acid were 2.6 mM, 7.4 mM and 21.2 mM, respectively. The enzyme leakage from the adsorbent during operation was successfully prevented owing to the increase of physical strength of gel coat. The half lives in a column reactor were 6 and 30 days at the respective temperature of 4$0^{\circ}C$ and 3$0^{\circ}C$, which were the 6-8 fold increased values as compared with those of without entrappment. The results highly recommended the use of reinforced Calcium-alginate gel entrappment method for the enhancement of physical strength and the operational stability of alginate gel entrapped enzyme.

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Probiotic Characteristics of Lactobacillus rhamnosus Isolated from Kefir (Kefir로부터 분리한 Lactobacillus rhamnosus의 Probiotic 특성)

  • You, Suk-Jin;Cho, Jin-Kook;Hwang, Seong-Gu;Heo, Kang-Chil
    • Food Science of Animal Resources
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    • v.25 no.3
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    • pp.357-364
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    • 2005
  • To search probiotic microorganisms, we isolated Lactobacillus sp. from kefir, The Lactobacillus sp. strain showed $99.5\%$ of identity to species Lactobacillus rhamnosus by API kit. Lactobacillus rhamnosus showed high resistances to acidic environment, which grew well even at pH 2.0 and $1.0\%$ bile salt Enzyme activity of Lactobacillus rhamnosus was higher in amylase ($0.673\;{\mu}mol/min/mg$) than that in xylanase ($0.288\;{\mu}mol/min/mg$), cellulase($0.117\;{\mu}mol/min/mg$) and phytase($0.269\;{\mu}mol/min/mg$). Especially, the Lactobacillus rhamnosus showed high heat stability which remained $1{\times}10^6\;CFU/ml$ at $60^{\circ}C$. The maximum numbers of Lactobacillus rhamnosus on growth owe was reached at 24 h fermentation and pH was decreased to 4.6. The resistances of Lactobacillus rhamnosus to acidic pH and bile salt were better than that of Lactobacillus acidophilus used as control. When Lactobacillus rhamnosus was cultured with E. coli in MRS broth, E. coli was disappeared after 18 h. These result suggest that the isolated Lactobacillus rhamnosus has a useful probiotics properties.

Esterase Production and Culture Characteristics of Bacteria Isolated from Acid Hydrolysed Soybean Protein (산분해 대두 단백질로부터 분리된 Esterase 생성균의 생육 및 효소생성 특성)

  • Oh, Nam-Soon
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.484-489
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    • 1997
  • The characteristics of growth and esterase activity of bacterial strains isolated from acid hydrolysed soybean protein were examined. All the isolated strains having decomposition activity of p-hydroxybenzoic acid butyl ester and esterase producing activity were identified as Bacillus sp. by morphological and biochemical methods. The specific growth rates, esterase activities and p-hydroxybenzoic acid butyl ester decomposition activities of isolated strains were $0.844{\sim}1.213\;h^{-1}$, $21{\sim}222\;mU/ml$ and $5.4{\sim}8.1\;mU/ml$, respectively. In the fermentation of Bacillus sp. KB8 strain which had the highest esterase producing activity, growth, extracellular excretion and intracellular synthesis of esterase were inhibited by adding NaCl in the culture broth. Esterase producing activity gradually increased after late exponential growth phase, until maximum value of 420 mU/ml reached after 64 hours culture period. Esterase of Bacillus sp. KB8 strain was stable up to $50^{\circ}C$ for 30 minutes, but was inactivated by heating for 30 minutes at $70^{\circ}C$. The enzyme activity exponentially decreased during the incubation time at the temperatures of $60^{\circ}C$ and $65^{\circ}C$.

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Thermostable $\alpha$-Amylase Production by Thermophilic Bacillus sp. TR-25 lsolated from Extreme Enviroment (극한환경에서 분리한 고온성 Bacillus sp. TR-25에 위한 내열성 $\alpha$-amylase의 생산)

  • 노석범;손홍주;이종근
    • Journal of Life Science
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    • v.7 no.1
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    • pp.30-38
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    • 1997
  • For screening thermostable $\alpha$-amylase from thermophiles, various samples from extreme environments such as hot spring and sewage near them, and compoat, wereexamined microbial growth in enrichment culture medium at 55$\circ$C on the assumption that enzymes from thermophiles are inevitable thermostable. One strain showing higher $\alpha$-amylase activity was pure cultured and designated as Bacillus sp. TR-25 from the results of morphological, cultural and physiological characteristics. The most important carbon sourses for the enzyme production were soluble starch, dextrin, potato starch and corn starch. Glucose and fructose had a catabolite repression on the enzyme production. The good nitrogen sources for the enzyme production were yeat extract, nutrient broth, tryptone, corn steep liquor and ammonium sulfate. The enzyme production was accelerated by addition of CaCl$_{2}$. $\cdot $ H$_{2}$O. The optimal medium composition for the enzyme production was soluble starch 2.0%, yeast extract 0.55, CaCl$_{2}$ $\cdot $ 2H$_{2}$O 0.015, Tween 80 0.001%, pH8.0, respectively. In jar fermenter culture, this strain shows a rapid growth and required cheaper carbon and nitrogen source. These properties are very useful to fermentation industry. The $\alpha$-amylase of this strain demonstrated a maximum activity at 80$\circ$C, pH 5.0, respectively. And calcium ion did not improve thermostability of the enzyme. At 10$0^{\circ}C$, this enzyme has 235 of relative activity. Transformation was carried out by thermophilic Bacillus sp. TR-25 genomic DNA. As a result, the transformant has increased thermostable $\alpha$-amylase activity.

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Functional Characteristics of Enterococcus faecium SA5 and Its Potential in Conversion of Ginsenoside Rb1 in Ginseng (Enterococcus faecium SA5의 기능적 특성과 인삼 ginsenoside Rb1의 전환)

  • Kim, Eun-Ah;Renchinkhand, Gereltuya;Urgamal, Magsal;Park, Young W.;Nam, Myoung Soo
    • Journal of Life Science
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    • v.27 no.2
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    • pp.172-179
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    • 2017
  • The fermentation of Panax ginseng can yield many compounds from ginsenosides that have a wide variety of biological functions. Lactic acid bacteria (LAB) strains are capable of converting ginsenosides. The purposes of this study were to: (1) characterize Enterococcus faecium SA5, an isolated LAB from Mongolian mare milk, (2) identify the existence of extracellular ${\beta}$-glucosidase activity in the milk, and (3) ascertain if the ${\beta}$-glucosidase has the capacity of converting ginsenoside in Korean ginseng. The results revealed that E. faecium SA5 was acid-resistant, bile salt-resistant, and has antibiotic activities against 4 pathogenic microorganisms (Salmonella typhimurium KCTC 3216, Listeria monocytogenes KCTC 3710, Bacillus cereus KCTC 1012, Staphylococcus aureus KCTC 1621). In addition, E. faecium SA5 had tolerance against some antibiotics such as colistin, gentamycin and neomycin. It was also found that E. faecium SA5 possessed bile salt hydrolase activity, which could lower blood cholesterol level. When incubated in 10% (w/v) skim milk as a yogurt starter, E. faecium SA5 caused to decrease pH of the medium as well as increase in viable cell counts. Using TLC and HPLC analysis on the samples incubated in MRS broth, our study confirmed that E. faecium SA5 can produce ${\beta}$-glucosidase, which was capable of converting ginsenoside $Rb_1$ into new ginsenosides $Rg_3-s$ and $Rg_3-r$. It was concluded that E. faecium SA5 possessed a potential of probiotic activity, which could be applied to yogurt manufacture as well as ginsenoside conversion in ginseng.

L-Arabinose Production from Diluted Sulfuric Acid Hydrolysis of Corn-fiber (Corn-fiber의 희석된 황산 가수분해에 의한 L-arabinose의 생산)

  • Lee, Hyung-Joo;Lee, Won-Kyu;Ryu, Yeon-Woo
    • KSBB Journal
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    • v.22 no.4
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    • pp.201-206
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    • 2007
  • The demand of L-arabinose has been increased recently because of its advantages including clinical effect. L-arabinose can be produced from dilute acid hydrolysis of agricultural wastes. In this study, optimum conditions of L-arabinose production using dilute acid hydrolysis of agricultural wastes and nutshells were determined. Among the tested various agricultural wastes and nutshells, corn fiber was selected as the best raw material for the production of arabinose. The highest arabinose production was achieved an acid hydrolysis of corn fiber for 1 h at 130$^{\circ}C$ with 0.4% sulfuric acid. Above optimal conditions, it was obtained 20.1 g/L glucose, 10.1 g/L xylose, 7.8 g/L arabinose and 1.8 g/L galactose from 90 g/L of corn fiber. For the purification of arabinose, it was carried out to remove all of sugars except arabinose by the Candida tropicalis cultivation of acid hydrolyzate and an organic contaminants such as pigments by the active carbon treatment of fermentation broth. Moreover, experiments were carried out to eliminate an ions by exchange chromatography. Finally, we obtained 3.1 g of partially purified L-arabinose powder with about 40% yield by evaporation and vacuum drying.

Chemosensitization of Fusarium graminearum to Chemical Fungicides Using Cyclic Lipopeptides Produced by Bacillus amyloliquefaciens Strain JCK-12

  • Kim, K.;Lee, Y.;Ha, A.;Kim, Ji-In;Park, A.R.;Yu, N.H.;Son, H.;Choi, G.J.;Park, H.W.;Lee, C.W.;Lee, T.;Lee, Y.W.;Kim, J.C.
    • 한국균학회소식:학술대회논문집
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    • 2018.05a
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    • pp.44-44
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    • 2018
  • Fusarium head blight (FHB) caused by infection with Fusarium graminearum leads to enormous losses to crop growers, and may contaminate grains with a number of Fusarium mycotoxins that pose serious risks to human and animal health. Antagonistic bacteria that are used to prevent FHB offer attractive alternatives or supplements to synthetic fungicides for controlling FHB without the negative effects of chemical management. Out of 500 bacterial strains isolated from soil, Bacillus amyloliquefaciens JCK-12 showed strong antifungal activity and was considered a potential source for control strategies to reduce FHB. B. amyloliquefaciens JCK-12 produces several cyclic lipopeptides (CLPs) including iturin A, fengycin, and surfactin. Iturin A inhibits spore germination of F. graminearum. Fengycin or surfactin alone did not display any inhibitory activity against spore germination at concentrations less than 30 ug/ml, but a mixture of iturin A, fengycin, and surfactin showed a remarkable synergistic inhibitory effect on F. graminearum spore germination. The fermentation broth and formulation of B. amyloliquefaciens JCK-12 strain reduced the disease incidence of FHB in wheat. Furthermore, co-application of B. amyloliquefaciens JCK-12 and chemical fungicides resulted in synergistic in vitro antifungal effects and significant disease control efficacy against FHB under greenhouse and field conditions, suggesting that B. amyloliquefaciens JCK-12 has a strong chemosensitizing effect. The synergistic antifungal effect of B. amyloliquefaciens JCK-12 and chemical fungicides in combination may result from the cell wall damage and altered cell membrane permeability in the phytopathogenic fungi caused by the CLP mixtures and subsequent increased sensitivity of F. graminearum to fungicides. In addition, B. amyloliquefaciens JCK-12 showed the potential to reduce trichothecenes mycotoxin production. The results of this study indicate that B. amyloliquefaciens JCK-12 could be used as an available biocontrol agent or as a chemosensitizer to chemical fungicides for controlling FHB disease and as a strategy for preventing the contamination of harvested crops with mycotoxins.

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Pervaporation of Butanol from their Aqueous Solution using a PDMS-Zeolite Composite Membrane (PDMS-Zeolite 복합막을 이용한 부탄올 투과증발)

  • Kong, Chang-In;Cho, Moon-Hee;Lee, Yong-Taek
    • Korean Chemical Engineering Research
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    • v.49 no.6
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    • pp.816-822
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    • 2011
  • Pervaporation is known to be a low energy consumption process since it needs only an electric power to maintain the permeate side in vacuum. Also, the pervaporation is an environmentally clean technology because it does not use the third material such as an entrainer for either an azeotropic distillation or an extractive distillation. In this study, Silicalite-1 particles are hydrothermally synthesized and polydimethylsiloxane(PDMS)-zeolite composite membranes are prepared with a mixture of synthesized Silicalite-1 particles and PDMS-polymer. They are used to separate n-butanol from its aqueous solution. Pervaporation characteristics such as a permeation flux and a separation factor are investigated as a function of the feed concentration and the weight % of Silicalite-1 particles in the membrane. A 1,000 $cm^3$ aqueous solution containing butanol of low mole fraction such as order of 0.001 was used as a feed to the membrane cell while the pressure of the permeation side was kept about 0.2~0.3 torr. When the butanol concentration in the feed solution was 0.015 mole fraction, the flux of n-butanol significantly increased from 14.5 g/ $m^2$/hr to 186.3 g/$m^2$/hr as the Silicalite-1 content increased from 0 wt% to 10 wt%, indicating that the Silicalite-1 molecular sieve improved the membrane permselectivity from 4.8 to 11.8 due to its unique crystalline microporous structure and its strong hydrophobicity. Consequently, the concentration of n-butanol in the permeate substantially increased from 0.07 to 0.15 mole fraction. This composite membrane could be potentially appliable for separation of n-butanol from insitu fermentation broth where n-butanol is produced at a fairly low concentration of 0.015 mole fraction.

Purification and Characterization of Phospholipase D from Actinomycetes KF923. (방선균 KE923이 생산하는 Phospholipase D의 정제 및 특성)

  • 곽보연;윤석후;김창진;손동화
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.389-394
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    • 2003
  • In order to screen microorganisms producing phopholipase D (PLD) had high transphosphatidylation activity, about 1,000 Actinomycetes strains were isolated from the 63 soil samples, collected over 6 local area in Korea. When the hydrolytic activity in the supernatant was determined, 131 strains produced PLD more than 0.3 U/ml. Among 131 culture broths tested, 23 ones had transphosphatidylation activity higher than 20% and finally one strain (Actinomycetes KF 923), which had highest hydrolytic and transphophadylation activity, was selected. Actinomycetes KF923 showed the highest hydrolytic activity (13 U/ml) and phosphatidylation activity (95%) after 48 h fermentation using the P medium (yeast extract 1%, peptone 1%, glucose 1.5%, glycerol 1%, $CaCo_3$ 0.4%, pH 7.2). PLD was purified from the culture broth of Actinomycetes KF923 and the specific activity of purified PLD was 567 U/mg. The molecular weight of PLD was about 55 kD and the optimum pH and temperature were 6.0 and $60^{\circ}C$, respectively. The stability of PLD toward pH and temperature were high around pH 8.0 and below $40^{\circ}C$. Special metal ions were not necessary to the PLD activity.