• Title/Summary/Keyword: microbial cell growth

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Isolation and Characterization of the Eicosapentaenoic Acid Biosynthesis Gene Cluster from Shewanella sp. BR-2

  • Lee, Su-Jin;Seo, Pil-Soo;Kim, Chul-Ho;Kwon, Oh-Suk;Hur, Byung-Ki;Seo, Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.881-887
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    • 2009
  • Forty-four eicosapentaenoic acid (EPA)-producing microbial strains were isolated from the intestines of marine fishes. Among them, one strain showing a maximum level of EPA (4.78% of total fatty acids) was identified as Shewanella sp. BR-2 on the basis of its 168 rRNA sequence. The EPA content reached a maximum level during the mid-exponential phase of cell growth, and gradually decreased with further growth of the cells. A cosmid DNA including the EPA biosynthesis gene cluster consisting of pfaA-E was isolated from a cosmid library of genomic DNA of Shewanella sp. BR-2, named pCosEPA-BR2. An E. coli clone harboring pCosEPA-BR2 produced EPA at a maximum level of 7.5% of total fatty acids, confirming the EPA biosynthesis activity of the cloned gene cluster.

Mcl-PHAs Produced by Pseudomonas sp. Gl01 Using Fed-Batch Cultivation with Waste Rapeseed Oil as Carbon Source

  • Mozejko, Justyna;Wilke, Andreas;Przybylek, Grzegorz;Ciesielski, Slawomir
    • Journal of Microbiology and Biotechnology
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    • v.22 no.3
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    • pp.371-377
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    • 2012
  • The present study describes medium-chain-length polyhydroxyalkanoates (mcl-PHAs) production by the Pseudomonas Gl01 strain isolated from mixed microbial communities utilized for PHAs synthesis. A two-step fed-batch fermentation was conducted with glucose and waste rapeseed oil as the main carbon source for obtaining cell growth and mcl-PHAs accumulation, respectively. The results show that the Pseudomonas Gl01 strain is capable of growing and accumulating mcl-PHAs using a waste oily carbon source. The biomass value reached 3.0 g/l of CDW with 20% of PHAs content within 48 h of cultivation. The polymer was purified from lyophilized cells and analyzed by gas chromatography (GC). The results revealed that the monomeric composition of the obtained polyesters depended on the available substrate. When glucose was used in the growth phase, 3-hydroxyundecanoate and 3-hydroxydodecanoate were found in the polymer composition, whereas in the PHAs-accumulating stage, the Pseudomonas Gl01 strain synthesized mcl-PHAs consisting mainly of 3-hydroxyoctanoate and 3-hydroxydecanoate. The transcriptional analysis using reverse-transcription real-time PCR reaction revealed that the phaC1 gene could be transcribed simultaneously to the phaZ gene.

The Study on the Grazing Rate of Protozooplankton tin the Microbial Food Web of Inchon Coastal Waters (仁川 沿岸 미세 생물 먹이망 내에서 浮游 原생動物의 포식율에 관한 연구)

  • 최중기;김시균
    • 한국해양학회지
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    • v.30 no.5
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    • pp.458-466
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    • 1995
  • To clarify the role of protozooplankton in the microbial food web of Inchon coastal ecosystem, the grazing capacities and growth rates of heterotrophic flagellated (HFL) and ciliates were investigated. We measured the growth rates of bacteria, HFL, autotrophic flagellated (AFL) and ciliates using reverse flow (RF) filter set and obtained the clearance rates and ingestion rates of HFL and dilates by fluorescent labelled bacteria (FLB) method and isolation culture method from natural seawater of Inchon coastal area. The standing crops of bacteria, HFL, AFL and ciliates in this study area were 0.7∼1.5${\times}$10/SUP 6/ cells$.$ml/SUP -1/ 1,680∼2,180 cells$.$ml/SUP -1/, 1,090∼2,180 cells$.$ml, and 3,600∼5,700 cells$.$l/SUP -1/, respectively. The mean growth rates of bacteria, HFL, AFL and ciliates were 0.039 h/SUP -1/, 0.032∼0.033h/SUP -1/, 0.015∼ 0.025h/SUP -1/and 0.021h/SUP -1/, respectively. The growth rates of protozooplankton of Inchon coastal waters were relatively low. Mean grazing rate of HFL, AFL and ciliates were0.039 h/SUP -1/, 0.032∼0.033h/SUP -1/,0.015∼ 0.025h/SUP -1/and 0.021h/SUP -1/, respectively. The growth rates of protozooplankton of Inchon coastal waters were relatively low. Mean grazing rate of HFL on bacteria was 0.055h/SUP -1/. Mean grazing rates of ciliates on HFL was relatively high of 0.057 h/SUP -1/, while the grazing rate of ciliates on AFL was low of 0.019h/SUP -1/. The clearance rates and ingestion rates of HFL on bacteria were 3.6∼61.8nl$.$HFL$.$h/SUP -1/ and 6∼52 BAC. HFL$.$h/SUP -1/ respectively. FLB grazing experiment showed low values, while natural bacteria experiment showed high value meaning high grazing pressure on total bacteria. The clearance rates and ingestion rates of Tintinnopsis sp., dominant ciliates in Inchon coastal waters, were 3.2∼4.6 ul$.$CIL/SUP -1/$.$h/SUP -1/ and 5∼6 cells$.$CIL/SUP -1/$.$h/SUP -1/ respectively, showing higher values in the high tide than low tide. The ingestion rates of ciliates implies that ciliate took up 17∼20% of total nanoflagellates of Inchon coastal waters.

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Isolation and Characterization of Endophytic Bacteria from Rice Root Cultivated in Korea (한반도 중부지방의 벼 뿌리로부터 내생 세균의 분리와 특성 분석)

  • Park, Soo-Young;Yang, Sung-Hyun;Choi, Soo-Keun;Kim, Ji-Hyeon;Kim, Jong-Guk;Park, Seung-Hwan
    • Microbiology and Biotechnology Letters
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    • v.35 no.1
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    • pp.1-10
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    • 2007
  • The 44 endophytic bacterial strains were isolated from surface-sterilized root of rice cultivated in seven different locations of Chungcheong province, Korea. Each isolate was introduced into rice seedlings grown gnotobiotically by inoculating scissor-cut first true leaf with cell suspensions, and the colonization capacity of each isolate in root tissue was analyzed at 7 days after inoculation. Sixteen out of 44 isolates were re-isolated from root successfully with the frequency of $10^{3-5}$ CFU/g tissue. Interestingly, seven out of 16 isolates were identified as Burkholderia species. The identity between inoculated strains and re-isolates was confirmed by genomic finger-printing and 16S rDNA sequence analysis. By a confocal laser scanning microscopic observation it was revealed that KJ001 strain, one of the sixteen isolates tagged with gfp colonized in root tissue especially around xylem. Six out of seven Burkholderia strains obtained in this study showed antagonizing activities against seven different fungal pathogens, contain nifH gene, and five of them enhanced growth of cucumber over 30%. The isolates showed no hypersensitive response on tobacco leaves and no pathogenecity in rice. From these results it was found that the endophytic Burkholderia strains will be useful in agriculture to develop a biocontrol agent or a bio-fertilizer.

Studies on Protoplast Formation and Regeneration of Lyophyllum decastes (Lyophyllum decastes의 원형질체 분리와 재생에 관한 연구)

  • Bok, Jin-Woo;Kim, Jong-Pil;Jin, Mi-Rim;Choi, Eung-Chil;Kim, Byong-Kak
    • The Korean Journal of Mycology
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    • v.22 no.2
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    • pp.130-137
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    • 1994
  • This experiment was carried out to investigate proper conditions for protoplast isolation and regeneration from mycelia of Lyophyllum decastes. Novozym 234(10 mg/ml) with 0.6 M $MgSO_4$ in phosphate buffer(pH 4.0) was proper for protoplast isolation. The optimal reaction time of the mycelium with the lytic enzyme was four hours in shaking condition at 120 strokes per min. When the mycelium of L. decastes was cultured at $24^{\circ}C$ for 5 days, the formation of protoplasts was effective. The liquid medium was more effective for protoplast isolation than the solid medium. In the liquid medium, high yields of protoplasts were obtained from 0.6 M $MgSO_4$ osmotic stabilizer. Protoplasts of L. decastes were regenerated to normal hyphal growth and the regeneration frequency of the protoplasts in the complete agar medium containing Triton X-100(0.0025%) was $5.94{\sim}8.32%$. The regeneration medium stabilized with 0.6 M sucrose was the best for regeneration of the protoplasts. In contrast to protoplast formation, regeneration was inhibited by the inorganic salts used as osmotic stabilizer.

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Studies on Microbial Utilization of Agricultural Wastes (Part IV) Cellulosic Waste Materials as Substrate on the Production of Cellulosic Single Cell Protein. (농산폐자원의 미생물학적 이용에 관한 연구 (제육보) 섬유소단세포단백 생산에서의 천연기질의 이용성)

  • Bae, Moo;Ko, Young-Hee
    • Microbiology and Biotechnology Letters
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    • v.5 no.1
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    • pp.18-23
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    • 1977
  • Experiments were carried out to pursue the availability and the feasibility of utilizable cellulosic materials as substrate for the production of cellulosic single cell protein. The resuluts were obtained as fellows. 1. Effects of carbolydrates as a sole carton source on the growth of Cellulomonas flavigena KIST 321 were examined. The result showed that cellulose and xylose would be most utlizable for cell mass production. 2. Alkaline treated waste papers and clothes resulted in good growth of the organism than intact ones did. However the waste papers as substrate of cellulosic fermentation were not digestible, even if the meterial was treated with alkalies. 3. Rice straw, rape straw and panic grass appeared to be good substrates for the cell mass production. 4. Leaves were proved to be a good substrate for the cell mass production, but wood sawdust was hardly digested by merely alkaline treatment. 5. When cellulosic wastes as the substrate were examined into the concentration of alkaline solution, the result suggested that the best productivity of cell mass from cellulosic materials was obtained on treatment with 0.8∼1.0% NaOH solution. 6. The productivity of cell mass was increased by washing out with water after alkaline treatment of newspaper, pine sawdust, lime sawdust and pine leaf.

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Isolation, Root Colonization and Evaluation of Some Plant Growth-promoting Rhizobacteria in Paddy Rice

  • Kang, Ui-Gum;Park, Hyang-Mi;Ko, Jee-Yeon;Lee, Jae-Saeng;Jeon, Weon-Tai;Park, Chang-Young;Park, Ki-Do;Chebotar, Vladimir K.
    • Korean Journal of Soil Science and Fertilizer
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    • v.50 no.3
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    • pp.135-149
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    • 2017
  • In order to obtain promising rice growth-promoting microbial strains that can be used as substitutes for chemical fertilizers, 172 bacterial strains were isolated from rice roots grown in Korean and Russian soils. Out of them, the strains KR076, KR083, KR181 and RRj228 showed plant growth-promoting activities on maize seedlings. Bacillus megaterium KR076 and Bacillus sp. KR083 showed both nitrogen-fixing and plant growth-promoting activities, while Rhizobium sp. KR181 and Pseudomonas sp. RRj228 appeared to support only plant growth-promotion, but not $N_2$ fixation. Especially, RRj228 showed high growth promoting activity at low concentrations. Inoculation studies with KR083 and RRj228 revealed a high affinity to the Japonica rice variety such as Junambyeo than the Korean Tongil type variety such as Arumbyeo. Both KR083 and RRj228 strains showed rhizoplane and/or endophytic colonization in Japonica and Tongil types rice when soaked with the bacterial suspension of $1.1{\times}10^5cfu\;ml^{-1}$ for six and twelve hours. However, the total bacterial cell numbers were higher in the roots of Japonica variety than in the Tongil type. In inoculation trials with Daesanbyeo rice variety, the seedlings inoculated with KR181 and RRj228 at the rate of $2.0{\times}10^6cfu\;ml^{-1}$ showed yield increment of 35% and 33% (p < 0.01), respectively, so that they contributed to the replacement of chemical fertilizer at half doses of N, $P_2O_5$, and $K_2O$ in pots. In Junambyeo rice seedlings, the strain RRj228, when inoculated with a cell suspension of $1.8{\times}10^6cfu\;ml^{-1}$, promoted 3.4% higher yield at 70% dose than at a full dose level of N $110kg\;ha^{-1}$ in field. These results suggest that the rhizobacteria KR181 and RRj228 are prospective strains for enhancing rice performance.

Degradation of Polyvinyl Alcohol in Dye-Processing Wastewater by Agar-Acrylamide Microbial Immobilization Method (한천-아크릴아마이드 미생물 고정화법에 의한 폐수 중 폴리비닐알콜의 분해)

  • 김재훈;김정목조무환
    • KSBB Journal
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    • v.10 no.3
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    • pp.241-248
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    • 1995
  • For the treatment of poorly biodegradable polyvinyl alcohol(PVA) in dye-processing wastewater, immobilized microbial beads were prepared by uslng agar-acrylamide method. PVA removal efficiency for the synthetic wastewater was 85% at the PVA volume loading rate of $3.1g/\ell$.day. In case of real desizing wastewater, PVA removal efficiency was 81.3% at the PVA volume loading rate of $3.25g/\ell$.day. In observation of cross section of immobilized bead passed 5 months with diameter of 2.4mm, the growth of cell was limited by the resistance of substrate and oxygen transfer for the inners region of more than 48% of bead radius from the surface. It was estimated that 70% of total removed PVA was degraded by the immobilized cells in the continuous immobilized reactor. Substrate utilization rate in the suspended reactor was decreased with increasing dilution rates above 0.083 hr-1, but that in the immobilized reactor was increased with increasing dilution rates up to 0.125hr-1. The substrate removal efficiency of immobilized reactor was much superior to that of suspended reactor with increasing dilution rates. Saturation constant of substrate utilization rate equation, Ks was $6.6 g PVA/\ell$, and maximum specific substrate utilization. k was 0.175g PVA/g cell.hr

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Lactobacillus plantarum APsulloc 331261 Fermented Products as Potential Skin Microbial Modulation Cosmetic Ingredients (Lactobacillus plantarum APsulloc 331261 발효 용해물의 피부 미생물 조절 효과)

  • Kim, Hanbyul;Myoung, Kilsun;Lee, Hyun Gee;Choi, Eun-Jeong;Park, Taehun;An, Susun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.1
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    • pp.23-29
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    • 2020
  • The skin is colonized by a large number of microorganisms with a stable composition of species. However, disease states of skin such as acne vulgaris, psoriasis, and atopic dermatitis have specific microbiome compositions that are different from those of healthy skin. The target modulation of the skin microbiome can be a potential treatment for these skin diseases. Quorum sensing (QS), a bacterial cell-cell communication system, can control the survival of bacteria and increase cell density. Also, QS affects the pathogenicity of bacteria such as biofilm formation and protease production. In this study, we confirmed anti-QS activity of Amorepacific patented ingredients, which are Lactobacillus ferment lysate (using Lactobacillus plantarum APsulloc 331261, KCCM 11179P) through bio-reporter bacterial strain Chromobacterium violaceum. The purple pigment production of C. violaceum controlled by QS was reduced 27.3% by adding 10 ㎍/mL of Lactobacillus ferment lysate (freeze dried). In addition, the Lactobacillus ferment lysate increased growth of Staphylococcus epidermidis 12% and decreased growth of Pseudomonas aeruginosa 38.5% and its biofilm formation 17.7% at a concentration of 10 ㎍/mL compared to the untreated control group. Moreover, S. epidermidis was co-cultured with the representative dermatological bacterium Staphylococcus aureus in the same genus, the growth of S. epidermidis was increased 134 % and the growth of S. aureus was decreased 13%. These results suggest that fermented lysate using Lactobacillus plantarum APsulloc 331261 may be useful as a cosmetic ingredient that can control the balance of skin microbiome.

Antimicrobial Activity of Rhus javanica Extracts Against Animal Husbandry Disease-Related Bacteria (가축질병 균주에 대한 오배자 추출물의 항균활성)

  • Choi, Il
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.8
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    • pp.1214-1220
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    • 2003
  • Antimicrobial activity of Rhus javanica (RJ) extract against animal husbandry disease-related bacteria was determined by a paper disc method. The RJ extracts showed a significant antimicrobial activity against Gram positive (+) bacteria and especially the activity was most potent against L. monocytogenes and S. epidermidis. Minimum inhibitory concentrations (MIC) of the MeOH and EtOH extracts of RJ were in the range of 0.8 ∼ 16 mg/mL and 0.8 ∼ 10 mg/mL, respectively. Furthermore, among five solvent fractions (n-hexane, CHC1$_3$, EtOAc, n-BuOH and $H_2O$ frs.) from MeOH extract of RJ, the EtOAc fr. exhibited the most significant antimicrobial activity The antimicrobial activities of RJ extracts against most microbial strains were unstable by either heat treatment or acid treatment. The inhibitory effect of RJ extracts on microbial cell growth was further examined by the addition of 0, 100, 300, and 500 ppm of RJ extracts into growth medium. The growth of gram positive (+) bacteria, S. aureus, S. epidermidis and L. monocytogenes was inhibited for 72 hours when at least 300 ppm of RJ extracts added, but the growth of gram negative (-) bacteria was only inhibited when at least 500 ppm of RJ extracts were added. Taken together, tile antimicrobial activities of RJ extracts were more effective against gram positive (+) bacteria compared to those against gram negative (-) bacteria.