• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.713-713
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• 2003

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1542-1546
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• 2009

Zeaxanthin glucosyltransferase (CrtX) mediates the formation of zeaxanthin to zeaxanthin diglucoside. Here, we report cloning of the crtX gene responsible for zeaxanthin diglucoside biosynthesis from Paracoccus haeundaensis and the production of the corresponding carotenoids in transformed cells carrying this gene. An expression plasmid containing the crtX gene (pSTCRT-X) was constructed, and Escherichia coli cells containing this plasmid produced the recombinant protein of approximately 46 kDa. Biosynthesis of zeaxanthin diglucoside was obtained when the plasmid pSTCRT-X was co-transformed into E. coli containing the pET-44a(+)-CrtEBIYZ carrying crtE, crtB, crtI, crtY, and crtZ genes required for zeaxanthin $\beta$-D-diglucoside biosynthesis.

• Journal of Microbiology and Biotechnology
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• v.27 no.6
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• pp.1106-1111
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• 2017

Escherichia coli was engineered to sense methanol by employing a chimeric two-component system (TCS) strategy. A chimeric MxaY/EnvZ (MxaYZ) TCS was constructed by fusing the Paracoccus denitrificans MxaY with the E. coli EnvZ. Real-time quantitative PCR analysis and GFP-based fluorescence analysis showed maximum transcription of ompC and the fluorescence at 0.01% of methanol, respectively. These results suggested that E. coli was successfully engineered to sense methanol by the introduction of chimeric MxaYZ. By using this strategy, various chimeric TCS-based bacterial biosensors can be constructed and used for the development of biochemical-producing recombinant microorganisms.

• Korean journal of applied entomology
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• v.55 no.3
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• pp.189-196
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• 2016

Previously, only 25 species of Pseudococcidae (mealybugs) were known to occur in Cambodia, and only 18 species in Laos. In 2015, we collected 14 mealybug species in these two countries, including six species not previously known to occur in these areas: Coccidohystrix insolita (Green, 1908) in Cambodia; Antonina graminis (Maskell, 1897) in both countries; and Nipaecoccus viridis (Newstead, 1894), Paracoccus marginatus Williams & Granara de Willink, 1992, Phenacoccus solani (Ferris, 1918), and Rastrococcus iceryoides (Green, 1908) in Laos.

• Journal of Life Science
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• v.11 no.3
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• pp.273-278
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• 2001

Removal of nitrogen compound from waste water is essential and often accomplished by biological process. Deni-trification bacterium. Paracoccus denitrificans(KCTC 2350) is employed to estimate the ability and the characteristics of denitrification. In the immobilized biological reactor system, the measurement of absolute amount of active strain in the reactor is comparatively difficult or impossible. In this study, strain immobilized denitrification reactor was designed with the unwoven texture wrapped peeped hole plastic tube to calculated the absolute amount of active strain by comparing the activity of the immobilized reactor adn the free cell reactor. The reactor system was continuous stirred tank reactor and the rate of substrate consumption was assumed to be Michaelis-Menten equation. As a result, we found that the amount of immobilized active strain was the half of the total active strain in the reactor and the time required to reach in the equilibrium state in the immobilized reactor system was shorter than that of the free cell reactor system.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2004.06a
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• pp.292-296
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• 2004

• Microbiology and Biotechnology Letters
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• v.46 no.2
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• pp.145-153
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• 2018

Among the carotenoid biosynthesis genes, crtI gene encodes the phytoene desaturase (CrtI) enzyme, and phytoene desaturase convert phytoene to lycopene. Phytoene desaturase is involved in the dehydrogenation reaction, in which four single bonds in the phytoene are introduced into a double bond, eliminating eight hydrogen atoms in the process. Phytoene desaturase is one of the key regulating enzyme in carotenoid biosynthetic pathway of various carotenoid biosynthetic organisms. The crtI gene in genomic DNA of Paracoccus haeundaensis was amplified and cloned into a T-vector to analyze the nucleotide sequence. As a result, the crtI gene coding for phytoene desaturase from P. haeundaensis consists of 1,503 base pairs encoding 501 amino acids residues. An expression plasmid containing the crtI gene was constructed, and Escherichia coli cells containing this plasmid produced the recombinant protein of approximately 55 kDa, equivalent to the molecular weight of phytoene desaturase. The expressed protein in cell lysate showed enzymatic activity similar to phytoene desaturase. Phytoene and lycopene were analyzed by HPLC and measured at wavelength of 280 nm and 470 nm, respectively. The $K_m$ values for phytoene and NADPH were $11.1{\mu}M$ and $129.3{\mu}M$, respectively.

• Korean Journal of Food Science and Technology
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• v.30 no.4
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• pp.902-907
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• 1998

In this study the anaerobic degradation of nitrate by in GFM (gel and foam matrix) and bead gel immobilized Paracoccus denitrificans DSM 65 in continous culture was conducted. A novel GFM immobilization system was developed in order to improve conventional system (bead). With increasing nitrate concentration in water, the nitrate reduction rate was increased. The observed maximum denitrification rate by in GFM immobilized cells was 177 mg/L h in buffered water, while that was 33 mg/L h in tap water. In comparison with bead system the reduction activity by GFM system showed $1.2{\sim}2.1$ times better. The denitrification activity was not changed after 16 days storage at $5^{\circ}C$ and also showed better activity than that of free cells or even bead immobilized cells.

• Journal of Microbiology and Biotechnology
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• v.34 no.9
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• pp.1826-1835
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• 2024

Paracoccus denitrificans has been identified as a representative strain with heterotrophic nitrificationaerobic denitrification capabilities (HN-AD), and demonstrates strong denitrification proficiency. Previously, we isolated the DYTN-1 strain from activated sludge, and it has showcased remarkable nitrogen removal abilities and genetic editability, which positions P. denitrificans DYTN-1 as a promising chassis cell for synthetic biology engineering, with versatile pollutant degradation capabilities. However, the strain's low stability in plasmid conjugation transfer efficiency (PCTE) hampers gene editing efficacy, and is attributed to its restriction modification system (R-M system). To overcome this limitation, we characterized the R-M system in P. denitrificans DYTN-1 and identified a DNA endonuclease and 13 DNA methylases, with the DNA endonuclease identified as HNH endonuclease. Subsequently, we developed a plasmid artificial modification approach to enhance conjugation transfer efficiency, which resulted in a remarkable 44-fold improvement in single colony production. This was accompanied by an increase in the frequency of positive colonies from 33.3% to 100%. Simultaneously, we cloned, expressed, and characterized the speculative HNH endonuclease capable of degrading unmethylated DNA at 30℃ without specific cutting site preference. Notably, the impact of DNA methylase M9 modification on the plasmid was discovered, significantly impeding the cutting efficiency of the HNH endonuclease. This revelation unveils a novel R-M system in P. denitrificans and sheds light on protective mechanisms employed against exogenous DNA invasion. These findings pave the way for future engineering endeavors aimed at enhancing the DNA editability of P. denitrificans.

• Journal of Life Science
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• v.27 no.3
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• pp.339-345
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• 2017

Carotenoids are natural lipid-soluble pigments, which are produced primarily by bacteria, algae, and plants. Many studies have focused on the identification, production, and utilization of natural sources of astaxanthin from algae, yeast, and crustacean byproducts as an alternative to the synthetic pigment, which is mostly used today. The aim of the present study was to identify a mutant of Paracoccus haeundaensis by exposure to UV and ethyl methanesulfonate (EMS). The mutant was then exposed to nutrient stress conditions to isolate an astaxanthin-hyperproducing strain, followed by characterization of the mutant. The survival rate decreased in accordance with an increase in the UV exposure time and an increase in the EMS concentration. A mutant of the original P. haeundaensis strain was identified that showed hyperproduction of astaxanthin following exposure to UV irradiation (20 min) and EMS treatment (0.4 M concentration). The optimal culture conditions for the PUE mutant were $25^{\circ}C$, pH 7-8, and 3% NaCl. The effects of various carbon and nitrogen sources on the growth and astaxanthin production of PUE were examined. The addition of 1% raffinose and 3% potassium nitrate influenced cell growth and astaxanthin production. The selected mutant exhibited an increase of 1.58 folds in astaxanthin content compared to initial wild type strain. A genetically stable mutant strain obtained using mutagen (UV irradiation and EMS treatment) may be a suitable candidate for further industrial scale production of astaxanthin.