• Title/Summary/Keyword: stationary phase

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Characteristics and Partial Purification of a Bacteriocin Produced by Pediococcus damnosus JNU 534 (Pediococcus damnosus JNU 534가 생산하는 박테리오신의 특성 및 정제)

  • Lee, Jae-Won;Han, Su-Min;Yun, Bo-Hyun;Oh, Se-Jong
    • Food Science of Animal Resources
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    • v.31 no.6
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    • pp.952-959
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    • 2011
  • A new bacteriocin-producing lactic acid bacteria (LAB) which has been isolated from kimchi was identified as Pediococcus damnosus by use of API kit and 16S rDNA sequencing, and designated as P. damnosus JNU 534. The bacteriocin produced by P. damnosus JNU 534 markedly inhibited the growth of some of LAB and Listeria monocytogenes, whereas other pathogens including Gram negative bacteria were not susceptible. The production of bacteriocin started at the beginning of exponential phase and reached maximum activity at the early stationary phase. The bacteriocin was stable on the wide pH range of 2-9 and heat treatment up to $100^{\circ}C$ for 15 min. The antimicrobial compound was inactivated by treatments of proteolytic enzymes indicating its proteinaceous in nature. The bacteriocin was purified by 30% ammonium sulfate precipitation followed by hydrophobic interaction column and $C_{18}$ column chromatography. The estimated molecular weight of the bacteriocin using tricine SDS-PAGE was approximately 3.4 kDa and the identified N-terminal amino acid sequence was $NH_2$-ILLEELNV.

Cultural Conditions of Lactobacillus sp. GM7311 for the Production of Bacteriocin (Lactobacillus sp. GM7311에 의한 박테리오신의 생산 조건)

  • LEE Myung Suk;CHANG Dong Suck;KANG Ji-Hee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.5
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    • pp.834-841
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    • 1997
  • A lactic acid bacteria which showed antimicrobial activity was isolated from dairy products and identified as Lactobacillus sp. according to the morphological, physiological and biochemical properties, which was named Lactobacillus sp. SH 7311. The bacteriocin of Lactobacillus sp. GM 7311 showed a broad range of inhibitory spectrum against some gram positive and negative bacteria. Especially, Proteus mirabilis was highly sensitive to bacteriocin and used as indicator strain for further investigation. The optimal condition for the production of bacteriocin was showed on MRS broth at $37^{\circ}C$ and pM 6.0. Bacteriocin production of this strain cultured under optimal condition was increased late logarithmic phase to early stationary phase. This bacteriocin was fully active at the pH range $2.0\~5.0$, also was stable at $100^{\circ}C$ for 60 min. at pH 5.0, But about $40\%$ of bacteriocin activity was diminished by the treatment of acetone, ethanol, iso-butanol and ethyl ether during 2 hours at $4^{\circ}C$.

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A study on the comparison of operation for long & short time in the Dranco process (건식단상혐기성소화조 장.단기 운영의 비교연구)

  • Hong, Jong-Soon;Kim, Jae-Woo;Shin, Dae-Yewn
    • Journal of environmental and Sanitary engineering
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    • v.23 no.4
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    • pp.73-82
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    • 2008
  • In this study, a dry single-phase anaerobic digestion process (Dranco system) was investigated to evaluate the optimum operational conditions. Several factors such as injection rate of organic waste, biogas production, $CH_4$ content in the biogas, pH of the sludge, $NH_3$-N and VFA concentration were investigated based on the operation of the digestion process for 2 months (short term) and 8 months (long-term). The operation results showed that a small quantity of food waste should be injected every week and that a 10% increase of the microorganism injection rate should be needed. However, normal operation was conducted after 11 weeks based on the designed quantity. The $CH_4$ content in the biogas was high at the beginning and the end of the food injection. However, it was low during week days. When the biogas production was high, the $CH_4$ concentration was low. The biogas production increased with an increase of the injection rate. $100m^3$/ton of biogas was produced from normal operation of the digestion process based on the designed quantity. The pH values of the digestion tank based on short-term operation ranged from 8 to 8.5. However, the pH values ranged from 7.45 to 8.15 after 4 weeks of long-term operation. The $NH_3$-N concentration of short-term operation ranged from 4,500 to 5,500 ppm and it gradually decreased to 2,000ppm after normal operation was commenced. For long-term operation, it was 5,000ppm initially and 3,800ppm after normal operation was commenced. The VFA concentration of sludge was less than 900ppm and 2,500ppm for short and long-term operations, respectively, after normal operation. Overall, the differences between sludge pH, $NH_3$-N and VFA concentrations may be due to the different types of microorganisms and the digestion ability of the microorganisms which exist in the accumulation of non digested organics. Moreover, it may be also caused by the type of food waste. Further investigation is needed to confirm these relationships.

Effects of Cell-free Culture Fluids for the Expression of Putative Acyltransferase in Corynebacterium glutamicum (코리네형 균주의 Acyltransferase 발현에 미치는 세균배양액의 효과)

  • Kim, Yong-Jae;Lee, Heung-Shick;Ha, Un-Hwan
    • Korean Journal of Microbiology
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    • v.48 no.3
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    • pp.207-211
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    • 2012
  • Autoinduction is mediated by signaling molecules known as autoinducers (AIs) that are produced, released and detected by bacterium itself. We recently reported that Corynebacterium glutamicum possesses an autoinduction system which secretes autoinducers during the stationary-phase of growth, triggering the expression of acyltransferase gene. However, it is still not clear what may act as autoinducers for the autoinduction in C. glutamicum. In this study, we compared the inducing effects of cell-free culture fluids obtained from a number of microbes including Agrobacterium tumefaciens, Vibrio harveyi, and Escherichia coli. Fluids from A. tumefaciens did not increase the expression of acyltransferase, whereas fluids from V. harveyi BB120 ($AI-1^+$, $AI-2^+$) did. Interestingly, the expression was increased by the fluids obtained from the early exponential-phase culture of BB120. Furthermore, this induction was not observed by the fluids from autoinducer mutants of V. harveyi MM77 ($AI-1^-$, $AI-2^-$) and BB152 ($AI-1^-$, $AI-2^+$). Unlike the effect shown by BB152, fluids from E. coli ($AI-1^-$, $AI-2^+$) still induced the acyltransferase expression. Taken together, these results suggest that C. glutamicum autoinducers seem to be unidentified molecules which do not belong to AI-1 or AI-2.

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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Characterization of Bacteriocin, lacticin YH-10, Produced by Lactococcus lactis subsp. lactis YH-10 Isolated from Kimchi (김치유산균인 Lactococcus lactis subsp. lactis YH-10가 생산하는 박테리오신의 특성)

  • Park, Eun-Min;Kim, Young-Hwa;Park, So-Jin;Kim, Yun-Im;Ha, Yu-Mi;Kim, Sung-Koo
    • Journal of Life Science
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    • v.14 no.4
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    • pp.683-688
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    • 2004
  • A bacteriocin-producing lactic acid bacteria was isolated from Kimchi on MRS selective media with the use of Lactobacillus delbrueckii subsp. delbrueckii as an indicator strain. The strain YH-10 was identified as Lactococcus lactis subsp. lactis through the API test. The crude bacteriocin (freeze-dried 50% ammonium sulfate precipitate of culture supernatant) produced by the strain was named as lacticin YH-10. Lacticin YH-10 showed the growth inhibitory activity against Gram positive pathogenic bacteria and other lactic acid bacteria. The bacteriocin was inactivated by proteases such as protamex and aroase AP-10 and partially inactivated by amylase, proteinase K, trypsin, and papain. The lacticin YH-10 remained its activity with the treatment of heat at 10$0^{\circ}C$ for 60 min or the changes of pH 2 to 11. However, the activity was lost at high pH combined with the exposure to 10$0^{\circ}C$. The bacteriocin production of the strain was started in the exponential phase and stopped in the stationary phase. The approximate molecular mass of the bacteriocin produced by the strain was approximate 14 kDa in the analysis on SDS-PAGE.

Characteristics of yeast with low temperature adaptation for Yakju brewed (약주 제조를 위한 저온 적응성 효모의 특성)

  • Seo, Dong-Jun;Yeo, Soo-Hwan;Mun, Ji-Young;Jung, Woo-Jin;Cho, Yong Sik;Baek, Seong Yeol
    • Food Science and Preservation
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    • v.22 no.6
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    • pp.908-914
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    • 2015
  • The objectives of this study were to isolate and characterize low temperature adaptation yeast and to obtain suitable yeasts strains for manufacturing Yakju. In this study, we isolated 482 wild yeasts from fermented foods. Out of these, 5 yeast strains were selected based on increased growth at low temperature ($15^{\circ}C$) and high ${\beta}$-glucosidase activity. To screen the aromatic level of isolates, media containing cerulenin and 5,5,5-trifluor-DL-leucine (TFL) were used. Y297 strain demonstrated tolerance against TFL and produced more than 13% alcohol. Y297 strain was identified a Saccharomyces cerevisiae based on the 26S rDNA gene sequences. Maximum cell growth was observed after 19 hr and 38 hr of incubation at $25^{\circ}C$ and $15^{\circ}C$, respectively. The exponential phase was followed by a lengthy stationary phase, at $15^{\circ}C$, when the cells remained high viable. Y297 strain demonstrated tolerance against alcohol (10%), glucose (60%) and salt(NaCl, 8%). ${\beta}$-glucosidase and esterase activity in Y297 were higher than those of controls at $15^{\circ}C$. Overall, these results indicated that using wild yeast strain, isolated from fermented food, affects the chemical characteristics of the brewed Yakju.

Characteristics of Chungkookjang Produced by Bacillus subtillus MC31 (B. subtilis MC 31를 이용한 청국장의 품질특성)

  • Mann, So-Yon;Kim, Eun-Ah;Lee, Ga-Young;Kim, Ro-Ui;Hwang, Dae-Youn;Son, Hong-Joo;Lee, Byong-Won;Lee, Chung-Yeol;Kim, Dong-Seob
    • Journal of Life Science
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    • v.23 no.4
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    • pp.560-568
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    • 2013
  • Chungkookjang was fermented by B. subtilis MC31, a ${\gamma}$-amino butyric acid (GABA) producing microorganism. The characteristics of Chungkookjang were investigated while fermenting. Twenty four amino acids were detected in Chungkookjang, leucine was the highest of them all. Total cell populations of B. subtilis MC31 phase were between log $9.52{\pm}0.5$ ~ log $9.049{\pm}0.5$ CFU/g at stationary phase. Contents of moisture, crude ash, crude protein, crude lipid and crude fiber are $61.07{\pm}0.01%$, $1.52{\pm}0.01%$, $17.66{\pm}0.04%$, $8.96{\pm}0.03%$ and 2.61%, respectively. Contents of ammonia type nitrogen, amino type nitrogen and reducing sugar were increased during fermentation at $40^{\circ}C$ for 72 hr, however those of titratable acidity and total sugar were decreased. pH was slowly alkalized during fermentation. Viscous substance and protease contents in Chungkookjang were $4.7{\pm}0.05%$ and $0.519{\pm}7.36$ g/l, apiece. When the fibrin plate and Robbin method for fibrinolytic activity were applied, B. subtilis MC31 showed high activity. These results suggested that B. subtilis MC31 is suitable to be used as a starter to enhance the quality of Chungkookjang.

Characteristics of the Extracellular Proteolytic Enzyme Produced by Candida lipolytica FM5 Isolated from Mackerel (Scomber japonicus) (Candida lipolytica FM5가 생산하는 균체외 단백분해효소의 특성)

  • PARK Mi-Yeon;OH Eun-Gyong;CHANG Dong-Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.5
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    • pp.631-636
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    • 1998
  • Candida lipolytica(C. lipolytica) FM5 was selected as one of the strong saprophytic yeasts isolated from mackerel (Scomber japonicus). The selected strain could produce extracellular proteolytic enzyme. The effective medium for production of proteolytic enzyme by C. lipolytica FM5 was TPPY broth containing Bacto-tryptone $0.5\%$, proteose peptone $0.5\%$, yeast extract $0.25\%$, NaCl $0.5\%$ and $CaCl_2\;0.2\%$. The pattern of proteolytic enzyme production by C. lipolytica FM5 was the almost same as that of growth curve of the strain. Namely, the enzyme production was begun from the early stage of exponential phase and it was reached the highest at the begining of the stationary phase of the yeast growth. The optimum toeperature of the produced proteolytic enzyme was $35^{\circ}C$ and its activity was not significantly changed by pH between 6.5$\~$9.0 and also it was not significantly affected by several kinds of cations such as $Ca^{2+},\;Cu^{2+},\;Fe^{2+}$ and $Mg^{2+}$ but it was affected negatively by some cations such as $Zn^{2+},\;Mn^{2+}$ and $K^+$.

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Development and Validation of an Analytical Method for Glucuronolactone in Energy Drinks by Hydrophilic Interaction Liquid Chromatography-electrospray Tandem Mass Spectrometry

  • Oh, Mi Hyune;Lim, Moo Song;Chai, Jeung Young;Kim, Eun Jung;Cho, Joong Hoon;Lim, Chul Joo;Choi, Sun Ok
    • Journal of Food Hygiene and Safety
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    • v.32 no.2
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    • pp.89-95
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    • 2017
  • A rapid, sensitive analytical method for glucuronolactone in beverages was developed and validated using hydrophilic interaction liquid chromatography coupled to electrospray ionization tandem mass spectrometry (HILIC-ESI-MS/MS). To determine the optimum analytical conditions for glucuronolactone, three different kinds of HILIC columns and two mobile phases with different pH values were examined. An amide-bonded stationary phase with a pH 9 acetonitrile-rich mobile phase was the best condition in terms of column retention, ESI-MS/MS response area, and signal-to-noise ratio. After extraction, glucuronolactone was separated through the HILIC amide column and detected by negative ESI-MS/MS in selected reaction monitoring (SRM) mode. Nine energy drinks sold in Korea were spiked with glucuronolactone at a concentration of 5 ng/mL; the Monster $Energy^{TM}$ sample showed the smallest peak area and its signal-to-noise ratio was used for method validation. Good linearity was obtained in the concentration range from 20 to 1500 ng/mL with a correlation coefficient > 0.998. The developed method had a limit of detection (LOD) of 6 ng/mL and a limit of quantitation (LOQ) of 20 ng/mL. The recovery of this method at concentration of 20, 100, 500, and 1000 ng/mL was 96.3%-99.2% with relative standard deviations (RSD) of 1.6%-14.0%. A reproducibility precision assessment at concentration of 100 and 500 ng/mL was carried out among three laboratories. The recovery of that evaluation was 95.1%-102.3% with RSD of 2.7%-7.0%. An analysis of variance indicated that there was no difference between the recovery results of the three laboratories at the 5% significance level. The validated method is applicable to inspecting beverages adulterated with glucuronolactone in Korea.