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Enhanced Production of Astaxanthin by Metabolically Engineered Non-mevalonate Pathway in Escherichia coli

  • Jeong, Tae Hyug;Cho, Youn Su;Choi, Seong-Seok;Kim, Gun-Do;Lim, Han Kyu
    • Microbiology and Biotechnology Letters
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    • v.46 no.2
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    • pp.114-119
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    • 2018
  • Astaxanthin is one of the major carotenoids used in pigment has a great economical value in pharmaceutical markets, feeding, nutraceutical and food industries. This study was to increase the production of astaxanthin by co-expression with transformed Escherichia coli using six genes involved in the non-mevalonate pathway. Involved in the non-mevalonate biosynthetic pathway of the strain Kocuria gwangalliensis were cloned dxs, ispC, ispD, ispE, ispF, ispG, ispH and idi genes in order to increase astaxanthin production from the transformed E. coli. And co-expression with the genes to compared the amount of astaxanthin production. This engineered E. coli, containing both the non-mevalonate pathway gene and the astaxanthin biosynthesis gene cluster, produced astaxanthin at $1,100{\mu}g/g$ DCW (dry cell weight), resulting in approximately three times the production of astaxanthin.

Enantioselective Bioconversion Using Escherichia coli Cells Expressing Saccharomyces cerevisiae Reductase and Bacillus subtilis Glucose Dehydrogenase

  • Park, Hyun-Joo;Jung, Ji-Hye;Choi, Hye-Jeong;Uhm, Ki-Nam;Kim, Hyung-Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.20 no.9
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    • pp.1300-1306
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    • 2010
  • Ethyl (R, S)-4-chloro-3-hydroxybutanoate (ECHB) is a useful chiral building block for the synthesis of L-carnitine and hypercholesterolemia drugs. The yeast reductase, YOL151W (GenBank locus tag), exhibits an enantioselective reduction activity, converting ethyl-4-chlorooxobutanoate (ECOB) exclusively into (R)-ECHB. YOL151W was generated in Escherichia coli cells and purified via Ni-NTA and desalting column chromatography. It evidenced an optimum temperature of $45^{\circ}C$ and an optimum pH of 6.5-7.5. Bacillus subtilis glucose dehydrogenase (GDH) was also expressed in Escherichia coli, and was used for the recycling of NADPH, required for the reduction reaction. Thereafter, Escherichia coli cells co-expressing YOL151W and GDH were constructed. After permeablization treatment, the Escherichia coli whole cells were utilized for ECHB synthesis. Through the use of this system, the 30 mM ECOB substrate could be converted to (R)-ECHB.

Cloning and Experssion of a Human tau Gene cDNA in Escherichia coli (인체 tau 유전자의 cDNA 클로닝 및 Escherichia coli에서의 발현)

  • Chung, Sang-Ho;Maeda, Tadakazu;Yanagawa, Hiroshi
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.28-33
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    • 1994
  • In normal cells tau protein is associated with axonal microtubules, whereas in Alxheimer's disease it is immobilized in the somatodendritic compartment of certain nerve cells as a major component of the paired helical filament. As a part of the study to analyze the nature of the paired helical filament (PHF) deposits and some related factors in brain, we have cloned and expressed a human tau gene cDNA in Escherichia coli to obtain the recombinant human tau protein in abundance.

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In vitro Investigation of the Harmful Effects of Smoke Plume Produced by Pulsed Nd: YAG Laser Treatment (Pulsed Nd : YAG 레이저조사시 발생되는 연기의 유해효과에 관한 실험실적 연구)

  • Kyung-Hun Kim;Keum-Back Shin;Bok Choi
    • Journal of Oral Medicine and Pain
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    • v.20 no.2
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    • pp.283-289
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    • 1995
  • 레이저가 의학영역에서 사용된 후 구강내 연조직과 경조직 병소에 대한 임상적 적용에 관한 관심이 커져왔다. 레이저가 조사되는 부위에서는 대부분 연기가 발생한다. 이 경우 병원체가 존재하는 병소에 대한 레이저치료시 발생되는 연기속에 병원체가 포함되어 이Tekas 환자와 술자는 이의 흡입가능성이 크므로 균혈증 및 상기도나 호흡계의 의원성 감염이 유발될 수 있다. 저자는 레이저치료시 발생되는 연기속에 병원체가 포함되어 있는지의 여부를 규명하기위해 각각 10개의 실험군에 통기성 미생물인 Escherichia coli(E.coli)가 배양된 brain geart infusion (BHI) 배지와 E.coli 가 존재하는 치근단 병소를 실험적으로 만든 후 각각 pulsed Nd:YAG 레이저를 조사해 연기를 채취, 배양, 분석하여 다음과 같은 결과를 얻었다. 1. Pulsed Nd:YAG 레이저를 E. colir 배양된 agar substrate 와 실험적으로 만들어진 치근단부위의 E.coli 현탁액에 조사시 발생된 연기에서 처음 접종된 것과 같은 E.coli가 검출되었다. 2. 감염병소에 pulsed Nd:YAG 레이저를 조사시 발생되는 연기의 유해효과로부터 환자와 술자 및 보조자의 보호를 위해 효과적인 흡입기를 사용이 권장된다.

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Transcriptional Regulation of Escherichia coli serC-aroA Operon : Further Support for cAMP-Dependent Expression

  • Sa, Jae-Hoon;Park, Soo-Sun;Lim, Chang-Jin
    • BMB Reports
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    • v.28 no.1
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    • pp.21-26
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    • 1995
  • The Escherichia coli mixed-function serC-aroA operon encodes biosynthethic enzymes for unrelated pathways leading to the syntheses of serine and aromatic amino acids. It has been proposed that the operon is expressed in a cAMP-dependent manner. In this work experiments were performed to investigate the cAMP-dependent expression of the operon. Exogenous cAMP increased ${\beta}$-galactosidase synthesis in the $cya^+$ and cya strains harboring the serC-aroA-lac fusion plasmid. This enhancement was more dramatic in the $cya^-$ strain grown in a minimal medium. In a dot blot assay the serC-aroA mRNA content increased in a concentration-dependent pattern after the addition of exogenous cAMP. The activity of phosphoserine aminotransferase, encoded by the serC gene, apparently increased in E. coli cells after the addition of cAMP. All results obtained confirmed that the expression of the E. coli serC-aroA operon is positively regulated by cAMP at the level of transcription.

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A Bacteriological Assessment for Salmonella and Escherichia coli in Some Selected Fresh Water Prawn (Macrobrachium rosenbergii) Farms and Depots

  • Haider, M.N.;Faridullah, M.;Kamal, M.;Islam, M.N.;Khan, M.N.A.
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.1
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    • pp.40-47
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    • 2007
  • Golda farms and depots of selected areas of the different districts of Bangladesh viz. Khulna, Bagerhat, Jessore and Norial area were sampled for the detection of Salmonella sp. and Escherichia coli. Incidence of Salmonella positive samples was 39%, 25%, 50% and 42% in the farms and 30%, 20%, 20% and 30% in the depots of Dumuria under Khulna, Bagerhat Sadar under Bagerhat, Avoynagar under Jessore and Kalia under Norail district respectively. On the other hand, E. coli positive samples was 23%, 42%, 25% and 17% in the farms and 70%, 30%, 50% and 30% in the depots of Dumuria (Khulna), Bagerhat Sadar (Bagerhat), Avoynagar (Jessore) and Kalia (Norail) region respectively. The overall results indicate that the trend of Samonella and E. coli contamination in farms and depots of all the regions is more or less similar although some variations were observed among the farms and depots of different location and region.

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Determination of the Intracellular Concentrations of Metabolites in Escherichia coli Collected during the Exponential and Stationary Growth Phases using Liquid Chromatography-Mass Spectrometry

  • Park, Chang-Hun;Park, Chang-Hun;Lee, Youn-Jin;Lee, Sang-Yup;Oh, Han-Bin;Lee, Jin-Won
    • Bulletin of the Korean Chemical Society
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    • v.32 no.2
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    • pp.524-530
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    • 2011
  • In the present study, we demonstrate that SRM LC-MS/MS method developed by Luo et al. (ref. 10) can be successfully applied to the quantitative analysis of intracellular metabolites in E. coli that are collected at the exponential and stationary growth phases. A focus is given on measuring the changes in the concentrations of intracellular metabolites in batch cultures, which were induced during both the dynamically changing exponential and stationary growth phases. The following intracellular metabolites are quantified in the exponential and stationary phases of E. coli growth, using the SRM mode of a triple quadrupole mass spectrometer: glucose-1-phosphate, fructose-1,6-bisphosphate, phosphoenolpyruvate, pyruvate, acetyl-coenzyme A, 6-phosphogluconate, ribulose-5-phosphate, xylulose-5-phosphate, erythrose-4-phosphate. The determined intracellular metabolite concentration profiles are shown to be in a good agreement with the growth profiles of E. coli, which clearly indicates that SRM LC-MS/MS can be successfully used for following the metabolite changes induced at different growth stages.

Antibacterial and therapeutic effects of a combination of Coptidis rhizoma and Galla rhois extracts in piglets challenged with Campylobacter coli

  • Lee, Soo-Mi;Cho, Byung-Wook;Yoo, Chang-Yeol;Kim, Suk;Son, Song-Ee;Lee, Hu-Jang
    • Korean Journal of Veterinary Research
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    • v.55 no.4
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    • pp.259-262
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    • 2015
  • The antibacterial effects of a combination of Coptidis rhizoma and Galla rhois extracts (CGE) were evaluated in piglets. The minimum bactericidal concentration of CGE was 2.0 mg/mL. Thirty 5-week-old piglets were challenged with Campylobacter (C.) coli after allocation to three different groups, a control and two treatment groups fed with CGE at 2.0 or 4.0 g/kg feed for 7 days. On day 7, C. coli in the feces of the CGE-treated groups were significantly lower than in the control (p < 0.01). These results suggest that CGE can be used to control C. coli in piglets.

Characteristics and antimicrobial susceptibility of Escherichia coli isolated from horse (말에서 분리한 Escherichia coli의 특성 및 항생제 감수성)

  • Yun, Sung-Wook;Kwon, Do-Yeon;Choi, Seong-kyoon;Lee, Hee-Soo;Cho, Gil-Jae
    • Korean Journal of Veterinary Research
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    • v.50 no.3
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    • pp.231-237
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    • 2010
  • This study was conducted to investigate characteristics and antimicrobial susceptibility of Escherichia (E.) coli isolates isolated from vaginal mucosa and clitorial fossa of 105 Thoroughbred mares suspicious of the genital disease in Korea during the period from March 2006 to July 2007. Ninety six E. coli isolates were identified as standard biochemical properties and using BIOLOG system. Fifty three isolates (55.2%) could be classified into a total of 21 O serotypes and forty three isolates (44.8%) were non-typeable with 51 O antisera used in this study. The verotoxin 1 (VT 1) and verotoxin 2 genes were analyzed by multiplex PCR. Among them, one isolate was detected VT 1 gene (130 bp). Most of isolates showed a high susceptibility in ciprofloxacin (100%), enrofloxacin (100%), norfloxacin (100%), cefoxitin (96.9%), gentamicin (96.9%), sulphamethoxazole (96.9%), nitrofurantoin (94.8%), amikacin (93.8%), nalidixic acid (92.7%) and tetracycline (90.6%). These results may provide the basic information to establish strategies for the treatment and prevention of reproductive disease in Thoroughbred mares in Korea.

Studies on the Biological Function and Antibacterial Effect of Lactoperoxidase System in Milk (우유내의 LP system의 생리기능 및 항균성에 관한여구 1. Escherichia coli O157:H7에 대한 항균효과)

  • 정충일;남은숙;김대원;전형일
    • Journal of Food Hygiene and Safety
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    • v.13 no.1
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    • pp.1-5
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    • 1998
  • This study was carried out to measure the antibacterial effect of lactoperoxidase/ thiocyanate/hydrogen peroxide system (LP system) against E. coli 0157:H7. When the initial inoculum levels (($10^{2},\;10^{4},\;10^{7}cfu/ml$), concentration of LP (10 ppm, 20 ppm, 30 ppm), culture media (TSB-YE, UHT milk, raw milk) and storage temperatures ($5^{\circ}C,\;10^{\circ}C,$15$^{\circ}C$) were set up differently for the experiment and the antibacterial effect was compared, the highest antibacterial effect of LP system was shown at $10^{2}cfu/ml$ of initial inoculum level, 10 ppm of LP concentration and $5^{\circ}C$ of incubation temperature.

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