• Title/Summary/Keyword: Stationary phase

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Transcriptional and Mycolic Acid Profiling in Mycobacterium bovis BCG In Vitro Show an Effect for c-di-GMP and Overlap between Dormancy and Biofilms

  • Cruz, Miguel A. De la;Ares, Miguel A.;Rodriguez-Valverde, Diana;Vallejo-Cardona, Alba Adriana;Flores-Valdez, Mario Alberto;Nunez, Iris Denisse Cota;Aceves-Sanchez, Michel de Jesus;Lira-Chavez, Jonahtan;Rodriguez-Campos, Jacobo;Bravo-Madrigal, Jorge
    • Journal of Microbiology and Biotechnology
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    • v.30 no.6
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    • pp.811-821
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    • 2020
  • Mycobacterium tuberculosis produces mycolic acids which are relevant for persistence, recalcitrance to antibiotics and defiance to host immunity. c-di-GMP is a second messenger involved in transition from planktonic cells to biofilms, whose levels are controlled by diguanylate cyclases (DGC) and phosphodiesterases (PDE). The transcriptional regulator dosR, is involved in response to low oxygen, a condition likely happening to a subset of cells within biofilms. Here, we found that in M. bovis BCG, expression of both BCG1416c and BCG1419c genes, which code for a DGC and a PDE, respectively, decreased in both stationary phase and during biofilm production. The kasA, kasB, and fas genes, which are involved in mycolic acid biosynthesis, were induced in biofilm cultures, as was dosR, therefore suggesting an inverse correlation in their expression compared with that of genes involved in c-di-GMP metabolism. The relative abundance within trehalose dimycolate (TDM) of α-mycolates decreased during biofilm maturation, with methoxy mycolates increasing over time, and keto species remaining practically stable. Moreover, addition of synthetic c-di-GMP to mid-log phase BCG cultures reduced methoxy mycolates, increased keto species and practically did not affect α-mycolates, showing a differential effect of c-di-GMP on keto- and methoxy-mycolic acid metabolism.

Effect of Gamma Irradiation on the Expression of Gene Endoding Metalloprotease in Vibrio vulnificus (감마선 조사가 vibrio vulnificus의 Metalloprotease 유전자 발현에 미치는 영향)

  • Jung, Jin-Woo;Lim, Sang-Yong;Joe, Min-Ho;Yun, Hye-Jeong;Hur, Jung-Mu;Kim, Dong-Ho
    • Microbiology and Biotechnology Letters
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    • v.36 no.1
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    • pp.6-11
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    • 2008
  • To check the microbiological safety with respect to increased virulence of surviving pathogens after irradiation, in this study, the transcriptional change of vvp gene encoding metalloprotease, which is one of the typical virulence factors of Vibrio mulnificus, was monitored by real-time PCR during the course of growth cycle after reinoculation of irradiated Vibrio. When V. vulnificus was exposed to a dose of 0.5 and 1 kGy, the lag period before growth resumption of sub-cultures became longer than non-irradiated counterpart as increase of irradiation dose. In the case of non-irradiated culture, the transcription of vvp was significantly activated at 15 h after inoculation, when bacterial growth reached the stationary phase, and the highest level of pretense activity (686 U/mL) was measured at the same time. Interestingly, vvp expression of irradiated Vibrio was turned up earlier than non-irradiated Vibrio during the mid log phase of growth, whereas these rapid induction of vvp expression from irradiated cells didn't result in an increase of metalloprotease production. When Vibrio was irradiated at 0.5 and 1 kGy, the protease activities peaked at 18 h after inoculation and the levels of activities were lower 1.2- and 1.4-fold, respectively, compared to the non-irradiated counterpart. Results from this study indicate that gamma radiation is not likely to activate the virulence ability of surviving Vibrio.

Growth and fatty Acid Composition with Growth Conditions for Spirulina platensis platensis (배양 조건에 따른 Spirulina platensis의 성장 및 지방산 조성)

  • Joo Dong-Sik;CHO Man-Gi;Buchholz Rainer;LEE Eung-H
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.3
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    • pp.409-416
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    • 1998
  • Owing to their high growth rate, marine microalgae such as Chlorella, Spirulina, Porphpidium and Dunaliella have been believed to be potentially useful for the production of foods, drugs and energy from light, $CO_2$ and minerals. In this study, we investigated the relationship between the growth and fatty acid composition in the blue green alga Spirulina platensis when the temperature and light intensity of culture conditions were changed. The optimal growth conditions for Spirulina platensis from the biomass and lipid contents were $30^{\circ}C$ on 6391 $\mu$E/$m^3$/sec and $35^{\circ}C$ on 4235 $\mu$E/$m^3$/sec. The difference of lipid contents between exponential phase and stationary phase were very large according to growth conditions. According to growth conditions the fatty acid compositions of Spirulina platensis differed, but regardless of growth conditions the main fatty acids were C16 : 0 and C20 : 0 in saturated fatty acid, C16 : 1, C18 : 1 and C 18 : 2 in unsaturated fatty acid.

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Polyamine, Cytochrome c and Enzymes Related to the Utilization of Methanol in Methylobacterium extorquens AMI Growing at Different pHs (상이한 수소이온농도에서 성장하는 Methylobacterium extorquens AM1의 메탄올 이용 관련효소와 Cytochrome c 및 폴리아민)

  • 박기정;이순희;김영민
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.533-538
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    • 1992
  • The generation time of Methylobacterium extorquens AMI growing on methanol at pH 5.5 and 7.0 was found to be 23 hand 8.3 h. respectively. The bacterium grown at pH 7.0 were found to contain more amounts of spermidine and putrescine than the cell grown at pH 5.5. Cells grown at both conditions exhibited strong methanol dehydrogenase (MDH) activity at the mid-exponential growth phase. The amounts of MDH. however. were found to be almost equal through all gro~1h phases. Cells growing at the stationary phase contained large amounts of cytochrome c. The cytochrome c content was higher in cells growing at pH 7.0 than the cells growing at pH 5.5. Cells growing at pH 5.5 in the presence of putrescine or spermidine contained increased amounts of putrescine. The level of spermine, however. was decreased and that of spermidine was not changed. Spermine added into the medium was found to have no effect on the level of cellular polyamines. Putrescine or spermidine added into the medium stimulated MDH and hydroxypyruvate reductase activities. but did not affect the contents of MDH and cytochrome c. It was found that preincubation of cell-free extracts with polyamines does not stimulate MDH and hydroxypyruvate reductase activities.

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Effects of Inducible Substrates on the Co-production of Glucoamylase and Exopolygalacturonase from Cryptococcus laurentii Y-23 (Cryptococcus laurentii Y-23의 glucoamylase와 exopolygalacturonase의 동시발효에 미치는 유도기질의 영향)

  • Kim, Chang-Hwa;Paik, Sang-Kyoo;Yun, Hye-Sun;Jin, Ing-Nyol;Yu, Choon-Bal
    • Korean Journal of Food Science and Technology
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    • v.32 no.4
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    • pp.875-880
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    • 2000
  • The production of glucoamylase and exopolygalacturonase from Cryptococcus laurentii Y-23 were investigated with the inducible substrates and mineral salts. Soluble starch induced only glucoamylase wherease pectin induced exopolygalacturonase as well as glucoamylase, and glucose did not induce glucoamylase whereas pectic acid induced a little amount of exopolygalacturonase. At the productions of two enzymes by inducible substrates for the 5 day-cultivation, the yeasts started log phase around 12 hours and mostly reached stationary phase around 36 hours. The best productivity of glucoamylase was observed with addition of soluble starch in the cultivation for 72 to 86 hours, and the high productivity of exopolygalacturonase was done by addition of both pectin and soluble starch in the cultivation for more than 72 hours. Without ammonium sulfate in the medium, however, cultural pH was so increased gradually that production of both enzymes were decreased and delayed as well. $Mn^{2+}$ increased both productivities of glucoamylase and exopolygalacturonase with 21% and 18%, respectively.

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Isolation and Characterization of Bacillus subtilis CA105 from Spent Mushroom (Pleurotus ostreatus) Substrates (느타리버섯 수확후배지로부터 분리한 Bacillus subtilis CA105의 특성)

  • Kim, Hye Soo;Kim, Chul Hwan;Kwon, Hyun Sook;Lee, Chan-Jung;Kong, Won-Sik;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.13 no.4
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    • pp.305-309
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    • 2015
  • In order to isolate compost-promoting bacteria with high activity of cellulase and xylanase, spent mushroom substrates with sawdust were collected from mushroom cultivation farm, Jinju, Gyeongnam in Korea. Among of the isolates, one strain, designated CA105 was selected by agar diffusion method. The strain CA105 was identified as members of the Bacillus subtilis by biochemical characteristics using VITEK 2 system. Comparative 16S rRNA gene sequence analysis showed that isolate CA105 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rRNA gene sequence similarity of 98.9%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, isolate CA105 was classified within the genus Bacillus subtilis, for which the name Bacillus subtilis CA105 is proposed. The cellulase and xylanase activity of B. subtilis CA105 was slightly increased according to bacterial population from exponential phase to stationary phase in growth curve for Bacillus sp. CA105.

Study on the Nicotine-Degrading Bacteria(2) -The Optimal Growth Condition of Nicotinophiles- (니코틴 분해 세균에 관한 연구(2) -니코틴 분해 세균의 최적 생장조건 연구-)

  • 강은희
    • Journal of the Korean Society of Tobacco Science
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    • v.2 no.2
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    • pp.20-37
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    • 1980
  • Among the 34 strains of Nicotinophiles selected in the previous experiments, strain NCT27 identified with Pseudomonas putida and strain NCT30 identified with Arthrobacter oxydans biotype nan thus were Investigated for optimization of growth conditions for nicotine degradation and other cultural characteristics. The compositions of optimized medium were to be following: $KH_2PO_4$ 2.Ogr, KCI 5.Ogr, $MgSO_4$.$7H_2O$ 20mg, $MnSO_4$.$6H_2O$ 0.2mg, $FeSO_4$.$7H_2O$ 1.Omg, Col$^{++}$ (Cobalt Acetate),2.O$\gamma$, N1$^{++}$ (NiSO4,6H2O) 0.5$\gamma$, and yeast extract 80mg per liter. The optimum initial concentrations of nicotine for growth were 0.4% for Pseudomonas and 0.1% for Arthrobacter, respectively. The optimum temperature and pH were 3$0^{\circ}C$ and 7.0 for both of strains. The pH of culture medium of Pseudomonas was changed from acidic condition to basic one in going from the logarithmic growth phase to the stationary growth phase. In contrast with Pseudomonas, it remained constant in case of Arthrobacter. The growth of Arthrobacter was completely inhibited in the nicotine concentration of 0.7&. However, Pseudomonas could grow even in the nicotine concentration of 1.0%. Moreover, it could grow successfully in the tobacco extract media as well as media containing carbon and nitrogen sources other than nicotine. The maximum rates of nicotine degradation were to be 1.22 gr./hr./liter for Pseudomonas and 0.186 gr./hr./liter for Arthrobacter, respectively.

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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.