• Title/Summary/Keyword: microbial strain

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A Strategy to Increase Microbial Hydrogen Production, Facilitating Intracellular Energy Reserves

  • Lee, Hyo Jung;Park, Jihoon;Lee, Joo-Young;Kim, Pil
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1452-1456
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    • 2016
  • Overexpression of the genes encoding phosphoeneolpyruvate carboxykinase (pckA) and NAD-dependent malic enzyme (maeA) facilitates higher intracellular ATP and NAD(P)H concentrations, respectively, under aerobic conditions in Escherichia coli. To verify a hypothesis that higher intracellular energy reserves might contribute to H2 fermentation, wild-type E. coli strains overexpressing pckA and maeA were cultured under anaerobic conditions in a glucose minimal medium. Overexpression of pckA and maeA enabled E. coli to produce 3-times and 4-times greater H2 (193 and 284 nmol, respectively) than the wild type (66 nmol H2). The pckA and maeA genes were further overexpressed in a hydrogenase-3-enhanced E. coli strain. The hydrogenase-3-enhanced strain (W3110+fhlA) produced 322 nmol H2, whereas the ATP-enhanced strain (W3110+fhlA+pckA) produced 50% increased H2 (443 nmol). Total H2 in the NAD(P)H-enhanced strain (W3110+fhlA+maeA) was similar to that in the control strain at 319 nmol H2. Possible explanations for the contribution of the increased cellular energy reserves to the enhanced hydrogen fermentation observed are discussed based on the viewpoint of metabolic engineering strategy.

Changes in the ginsenoside content during the fermentation process using microbial strains

  • Lee, So Jin;Kim, Yunjeong;Kim, Min-Gul
    • Journal of Ginseng Research
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    • v.39 no.4
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    • pp.392-397
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    • 2015
  • Background: Red ginseng (RG) is processed from Panax ginseng via several methods including heat treatment, mild acid hydrolysis, and microbial conversion to transform the major ginsenosides into minor ginsenosides, which have greater pharmaceutical activities. During the fermentation process using microbial strains in a machine for making red ginseng, a change of composition occurs after heating. Therefore, we confirmed that fermentation had occurred using only microbial strains and evaluated the changes in the ginsenosides and their chemical composition. Methods: To confirm the fermentation by microbial strains, the fermented red ginseng was made with microbial strains (w-FRG) or without microbial strains (n-FRG), and the fermentation process was performed to tertiary fermentation. The changes in the ginsenoside composition of the self-manufactured FRG using the machine were evaluated using HPLC, and the 20 ginsenosides were analyzed. Additionally, we investigated changes of the reducing sugar and polyphenol contents during fermentation process. Results: In the fermentation process, ginsenosides Re, Rg1, and Rb1 decreased but ginsenosides Rh1, F2, Rg3, and Compound Y (C.Y) increased in primary FRG more than in the raw ginseng and RG. The content of phenolic compounds was high in FRG and the highest in the tertiary w-FRG. Moreover, the reducing sugar content was approximately three times higher in the tertiary w-FRG than in the other n-FRG. Conclusion: As the results indicate, we confirmed the changes in the ginsenoside content and the role of microbial strains in the fermentation process.

Genomic epidemiology for microbial evolutionary studies and the use of Oxford Nanopore sequencing technology (미생물 진화 연구를 위한 유전체 역학과 옥스포드 나노포어 염기서열분석 기술의 활용)

  • Choi, Sang Chul
    • Korean Journal of Microbiology
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    • v.54 no.3
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    • pp.188-199
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    • 2018
  • Genomic epidemiology exploits various basic microbial research areas. High-throughput sequencing technologies dramatically have been expanding the number of microbial genome sequences available. Abundant genomic data provide an opportunity to perform strain typing more effectively, helping identify microbial species and strains at a higher resolution than ever before. Genomic epidemiology needs to find antimicrobial resistance genes in addition to standard genome annotations. Strain typing and antimicrobial resistance gene finding are static aspects of genomic epidemiology. Finding which hosts infected which other hosts requires the inference of transient transmission routes among infected hosts. The strain typing, antimicrobial resistance gene finding, and transmission tree inference would allow for better surveillance of microbial infectious diseases, which is one of the ultimate goals of genomic epidemiology. Among several high-throughput sequencing technologies, genomic epidemiology will benefit from the more portability and shorter sequencing time of the Oxford Nanopore Technologies's MinION, the third-generation sequencing technology. Here, this study reviewed computational methods for quantifying antimicrobial resistance genes and inferring disease transmission trees. In addition, the MinION's applications to genomic epidemiology were discussed.

Mass Cultivation of Rhodococcus sp. 3-2, a Carbendazim-Degrading Microorganism, and Development of Microbial Agents (카벤다짐 분해 미생물인 Rhodococcus sp. 3-2의 대량 배양 및 미생물 제제 개발)

  • Jun-Kyung Park;Seonghun Im;Jeong Won Kim;Jung-Hwan Ji;Kong-Min Kim;Haeseong Park;Yeong-Seok Yoon;Hang-Yeon Weon;Gui Hwan Han
    • Korean Journal of Environmental Agriculture
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    • v.42 no.4
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    • pp.259-268
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    • 2023
  • Rhodococcus sp. 3-2 strain has been reported to degrade benzimidazole-based pesticides, such as benomyl and carbendazim. Therefore, this study aimed to optimize culture medium composition and culture conditions to achieve cost-effective and efficient large-scale production of the Rhodococcus sp. 3-2 strain. The study identified that the optimal media composition for mass culture comprised 0.5% glucose, 0.5% yeast extract, 0.15% NaCl, 0.5% K2HPO4, 0.5% sodium succinate, and 0.1% MgSO4. Additionally, a microbial agent was developed using a 1.5-ton fermenter, with skim milk (20%), monosodium glutamate (15%), and vitamin C (2%) as key components. The storage stability of the microbial agent has been confirmed, with advantages of low temperature conservation, which helps to sustain efficacy for at least six months. We also assessed the benomyl degradation activity of the microbial agent within field soil. The results revealed an over 90% degradation rate when the concentration of viable cells exceeded 2.65 × 106 CFU/g after a minimum of five weeks had elapsed. Based on these findings, Rhodococcus sp. 3-2 strain can be considered a cost-effective microbial agent with diverse agricultural applications.

Phylogenetic Analysis of Culturable Arctic Bacteria

  • Lee, Yoo-Kyung;Kim, Hyo-Won;Cho, Kyeung-Hee;Kang, Sung-Ho;Lee, Hong-Kum;Kim, Yea-Dong
    • Ocean and Polar Research
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    • v.26 no.1
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    • pp.51-58
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    • 2004
  • We isolated and identified culturable Arctic bacteria that had inhabited soils around the Korean Arctic Research Station Dasan located at Ny-Alsund, Svalbard, Norway $(79^{\circ}N,\;12^{\circ}E)$. The collected soils were diluted in distilled water; the diluted soil-water was spread on 3M petri-films at Dasan Station. The petri-films were transported to the laboratory at KORDI, and cultured at $4^{\circ}C$. Colonies grown on the petri-films were subsequently cultured on nutrient agar plates at $4^{\circ}C$ every 7 days. The pure colonies were inoculated into nutrient liquid media, genomic DNA was extracted, and phylogenetic analysis was performed on the basis of 165 rDNA sequences. A total of 227 strains of bacteria were isolated. Among them, 16S rDNA sequences of 185 strains were identical with those of known strains isolated in this study, and 42 strains were finally identified. Phylogenetic analysis using 16S rDNA indicated that the 30 strains belonged to Pseudomonas, 7 strains to Arthrobacter, two strains to Flavobacterium, and the remaining to Achromobacter, Pedobacter, and Psychrobacter. Among the 42 strains, 14 bacteria produced protease: they were 6 strains of Pseudomonax, 4 strains of Arthrobater, an Achromobacter strain, 2 strains of Flavobacterium, and a Pedohacter strain. We expect these Arctic bacteria can be used for screening to develop new industrial enzymes that are active at low temperatures.

Protein Expression Analysis of Halobacillus dabanensis $D-8^T$ Subjected to Salt Shock

  • Feng De Qin;Zhang Bo;Lu Wei Dong;Yang Su Sheng
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.369-374
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    • 2006
  • To investigate the mechanism of salt tolerance of gram-positive moderately halophilic bacteria, two-dimensional gel electrophoresis (2-D PAGE) was employed to achieve high resolution maps of proteins of Halobacillus dabanensis $D-8^T$. Approximately 700 spots of proteins were identified from these 2-D PAGE maps. The majority of these proteins had molecular weights between 17.5 and 66 kDa, and most of them were distributed between the isoelectric points (pI) 4.0 and 5.9. Some protein spots were distributed in the more acidic region of the 2-D gel (pI <4.0). This pattern indicated that a number of proteins in the strain $D-8^T$ are acidic. To understand the adaptation mechanisms of moderately halophilic bacteria in response to sudden environmental changes, differential protein profiles of this strain were investigated by 2-D PAGE and $Imagemaster^{TM}$ 2D Platinum software after the cells were subjected to salt shock of 1 to 25% salinity for 5 and 50 min. Analysis showed 59 proteins with an altered level of expression as the result of the exposure to salt shock. Eighteen proteins had increased expression, S proteins were induced, and the expression of 33 proteins was down-regulated. Eight of the up-regulated proteins were identified using MALDI-TOF/MS and MASCOT, and were similar to proteins involved in signal transduction, proteins participating in energy metabolism pathways and proteins involved in stress.

Screening of Antagonistic Bacteria having Antifungal Activity against Various Phytopathogens (다양한 식물병원성 곰팡이에 항진균 활성을 갖는 길항미생물의 탐색)

  • Yang, Hee-Jong;Jeong, Su-Ji;Jeong, Seong-Yeop;Jeong, Do-Youn
    • The Korean Journal of Mycology
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    • v.42 no.4
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    • pp.333-340
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    • 2014
  • The aim of this study was to isolate a potential multifunctional biocontrol agent from bacteria for control of multiple plant diseases as an alternative to fungicides. A total of 201 strains were isolated from soil undamaged by repeated cultivation in Sunchang and their ability to produce antibiotics, siderophores and extracellular enzymes such as protease, cellulase and amylase was investigated. Selected strain SCS3 produced cellulose, protease and amylase. This strain also produced siderophores and showed excellent antifungal activity against various phytopathogens. SCS3 was identified as Bacillus subtilis using 16S rRNA sequencing, and named Bacillus subtilis SCS3. Finally, physiological and biochemical characteristics of B. subtilis SCS3 were examined. From the results, B. subtilis SCS3 was found to be a useful multifunctional biocontrol agent against various phytopathogens.

A Study on the Driving Characteristics of Microbial Fuel Cell Using Mixed Strains in Domestic Wastewater (생활폐수 내 혼합균주를 이용한 미생물 연료전지의 구동 특성에 관한 연구)

  • KIM, SANG KYU;YOO, DONG JIN
    • Transactions of the Korean hydrogen and new energy society
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    • v.32 no.6
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    • pp.506-513
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    • 2021
  • The use of fossil fuels is a major contributor to the increase atmospheric greenhouse gas emissions. As such problems arise, interest in new and renewable energy devices, particularly fuel cells, is greatly increasing. In this study, various characteristics of mixed strains were observed in wastewater collected by the Jeonju Environment Office to investigate the effects of microorganisms on voltage generation and voltage generation of substrates, electrode materials, electrons, electron transport media, and ash microbial fuel cells. As a result of separately measuring the voltage generated during inoculation, the inoculation voltage of Escherichia coli K12 (E. coli K12) was 0.45 V, and the maximum inoculation voltage of the mixed strain was 1.2 V. Thereafter, voltage values were collected using a digital multimeter and the amount of voltage generated over time was measured. In the case of E. coli K12, the maximum voltage reached 0.45 V, and the cell voltage was maintained above 0.23 V for 140 hours. In contrast, for the mixed strain, the maximum voltage reached 1.2 V and the voltage was slowly decreased to 0.97 V. In addition, the degree of microbial adsorption to the electrod surface after the inoculation test was confirmed using a scanning electron microscope. Therefore, these results showed the possibility of purifying pollutants at the same time as power generation through the production of hydrogen ions using microorganisms and wastewater.

Effect of Different Pretreatment Methods on the Bioconversion of Rice Bran into Ethanol

  • Eyini, M.;Rajapandy, V.;Parani, K.;Lee, Min-Woong
    • Mycobiology
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    • v.32 no.4
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    • pp.170-172
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    • 2004
  • The efficiency of acid, enzyme and microbial pretreatment of rice bran was compared based on the content of cellulose, hemicellulose, reducing sugars and xylose in the substrate. An isolate of Aspergillus niger or a strain of Trichoderma viride(MTCC 800) was employed for microbial pretreatment of rice bran in solid state. Acid pretreatment resulted in the highest amount of reducing sugars followed by enzyme and microbial pretreatment. A. niger showed a higher rate of hydrolysis than T. viride. The rice bran hydrolysate obtained from the different methods was subsequently fermented to ethanol either by Zymomonas mobilis(NCIM 806) or by Pichia stipitis(NCIM 3497). P. stipitis fermentation resulted in higher ethanol(37% higher) and biomass production($76{\sim}83%$ higher) than those of Z. mobilis. Maximum ethanol production resulted at 12h in Zymomonas fermentation, while in Pichia fermentation, it was observed at 60h. Microbial pretreatment of rice bran by A. niger followed by fermentation employing P. stipitis was more efficient but slower than the other microbial pretreatment and fermentation.

Strain Improvement of Leuconostoc mesenteroides as a Acid-Resistant Mutant and Effect on Kimchi fermentation as a Starter (Leuconostoc mesenteroides의 내산성 변이주의 김치발효에 미치는 효과)

  • Kim Young-Hwan;Kim Hee-Zoong;Kim Ji-Young;Choi Tae-Bu;Kang Sang-Mo
    • Microbiology and Biotechnology Letters
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    • v.33 no.1
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    • pp.41-50
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    • 2005
  • An organic acid tolerance mutant (M-200) was obtained from Leuconostoc mesenteroides KCCM 35471, followed by the screening procedure using a specific organic acid medium (lactic acid: acetic acid, 2:1). The characteristics of the acid tolerance M-200 and the wild type LM-W were examined at various temperature and pH ranges $(l0-30^{\circ}C$ of temp, 3.5-4.5 of pH). The growth of strain M-200 at HCl adjusted medium $(10^{\circ}C\;and\;pH 3.5)$ was observed. In the case of organic acid adjusted medium, the strain showed its growth at the pH range of 3.8. When the strain M-200 was used as a starter for Kimchi fermentation, a constant acid level (0.55) was observed during the whole fermentation period. This result indicates that the strain produces a proper level of acid content for the Kimchi fermentation. This result also indicates that the edible period of Kimchi can be extended to 3.5 fold compare to the result obtained from the LM-W used Kimchi fermentation. However the excess use of the strain M-200 showed the inhibition of growth of Lactobacillus plantarum, low lactic acid level content and low level of organoleptic test. In the case of organic acid content during the Kimchi fermentation, the strain M-200 showed relatively low production rate compare to the wild type (M-200: 3.5 mg/L at 21 days of fermentation, LM-W: 7 mg/L at 21 days of fermentation). Therefore a mixed Kimchi starter containing M-200 and other strains probably maintain a good Kimchi quality during the fermentation.