• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.1
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• pp.11-17
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• 2001

To enhance the productivity and activity of nitrile hydratase in Rhodococcus rhodochrous M33, a glucose-limited fed-batch culture was performed. In a fed-batch culture where the glucose was controlled at a limited level and cobalt was supplemented during the fermentation period, the cell mass and total activity of nitrile hydratase both increased 3.3-fold compared to that in the batch fermentation. The productivity of nitrile hydratase also increased 1.9-fold compared to that in the batch fermentation. The specific activity of nitrile hydratase in the whole cell preparation when using a fed-batch culture was 120 units/mg-DCW, which was similar to that in the batch culture.

• Journal of Microbiology and Biotechnology
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• v.16 no.5
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• pp.778-785
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• 2006

Catechol 1,2-dioxygenase was purified from cells of R. rhodochrous N75 grown at the expense of benzoate and p-toluate as the sole sources of carbon. A single catechol 1,2-dioxygenase was found to be induced with either growth substrate. The enzyme has an estimated $M_r$ of 71,000 consisting of two identical subunits. Catechol 1,2-dioxygenase from R. rhodochrous N75 exhibits some unusual properties including: broad substrate specificity, extradiol cleavage activity with 4-methylcatechol and low $K_m$ values for halocatechols, suggesting that this enzyme is distinct from other known catechol and chlorocatechol 1,2-dioxygenases.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.6
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• pp.465-468
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• 2000

Methyl mercaptan oxidase was successfully induced from Rhodococcus rhodochrous IGTS8 using methyl mercaptan gas and purified to homogeneity for the detection of mercaptans. The purification procedure involved DEAE-Sephacel and Superose 12 column chromatography with recovery yields of 85.8 and 83.3%, and a specific activity of 92.7 and 303.4 units/mg-protein, respectively. The molecular weight of purified methyl mercaptan oxidase was determined to be 64.5 kDa by SDS-PAGE. The extract from gel filtration chromatography oxidizes methyl mercaptan to produce formaldehyde, which can be easily detected by the purpald-coloring method. Optimum temperature for activity was achieved at 60$^{\circ}C$. This enzyme was inhibited by both K$_2$SO$_4$and NaCl at concentration of less than 100mM and recovered to original activity at concentration of 200mM. In the presence of methanol, the activity decreased by 33%.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.540-543
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• 2000

Microbial desulfurization using a biocatalyst which is capable of selectively liberating sulfur from HDS-refractory organic sulfur compounds is an alternative biotechnology to the current technology of hydrodesulfurization. The system used in the experiments is a two phase system consisting of 0.1%(w/v) dibenzothiophene in hexadecane as model oil and a mineral medium for cell growth. Rhodococcus rhodochrous IGTS8, a desulfurization strain, was grown in flask culture at different oil phase ratio with 10% and 30%. Most of the dibenzothiophene was converted to 2-hydroxybiphenyl when the oil ratio was 10%, but wasn't when the oil ratio was 30%. However, the total degraded DBT amounts were similar. In experiments of adjusting pH to improve the efficiency of degradation, the amounts of degraded DBT were increased by 50%. When the modified medium which has two-fold nutrients than those of minimal salt medium was used, the amounts of degraded DBT were increased by 32%. When both of the methods were used, the efficiency was increased by 136%.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.549-551
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• 2001

The heavy use of petroleum products in modern livings has brought ubiquitous environmental contaminants of aromatic compounds, which persist in aquatic and geo-environment without the substantial degradation. The persistence and accumulation of the aromatic compounds, which include xylene, phenol, toluene, phthalate, and so on are known to cause serious problems in our environments. Some of soil and aquatic microorganisms facilitate their growth by degrading aromatic compounds and utilizing degrading products as growth substrates, the biodegradation helps the reentry of carbons of aromatic compounds, preventing their accumulation in our environments. The metabolic studies on the degradation of aromatic compounds by microoganisms were extensively carried out along with their genetic studies. A Rhodococcus sp. EL-GT isolated in activated sludges has shown the excellent ability to grow on phenol as a sole carbon source. In the present study investigated a gene encoding phenol-degrading enzymes from a Rhodococcus sp. EL-GT.

• Proceedings of the Korean Aquaculture Society Conference
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• 2006.05a
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• pp.375-376
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• 2006

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.4
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• pp.194-200
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• 2002

The cells of Rhodococcus rhodochrous M33, which produce a nitrile hydratase enzyme, were immobilized in acrylamide-based polymer gels. The optimum pH and temperature for the activity of nitrile hydratase in both the free and Immobilized cells were 7.4 and 45$\^{C}$, respectively, yet the optimum temperature for acrylamide production by the immobilized cells was 20$\^{C}$. The nitrile hydratase of the immobilized cells was more stable with acrylamide than that of the free cells. Under optimal conditions, the final acrylamide concentration reached about 400 g/L with a conversion yield of almost 100% after 8 h of reaction when using 150 g/L of immobilized cells corresponding to a 1.91 g-dry cell weight/L. The enzyme activity of the immobilized cells rapidly de-creased with repeated use. However, the quality of the acrylamide produced by the immobilized cells was much better than that produced by the free cells in terms of color, salt content, turbidity, and foam formation. The quality of the aqueous acrylamide solution obtained was found to be of commercial use without further purification.

• Journal of Applied Biological Chemistry
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• v.51 no.6
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• pp.307-314
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• 2008

To explore the optimal conditions for the removal of $H_{2}S$ gas by biofiltration, various conditions, including inlet $H_{2}S$ concentration, flow rate, moisture, and cell number, were examined. Heterotrophic bacteria were isolated from the compost of the animal excreta. A strain that effectively removed $H_{2}S$ was selected and identified as Rhodococcus rhodochrous B261 by analysis of its 16S rDNA sequence. A cell number of $10^{7}\;cfu/g^{-}compost$ was sufficient to dominate the microbiota, and an effective removal was observed at $H_{2}S$ gas concentrations below 220 mg/L. The moisture content of 33-38% was suitable for activation of the microbial activity and delaying the desiccation. Higher flow rates resulted in lower removal rates of the $H_{2}S$ gas. Under the conditions of $10^7\;cfu/g^{-}compost$, $H_{2}S$ gas concentrations of 220 mg/L, and moisture content of 33-38%, the inlet $H_{2}S$ gas concentrations of 120 and 400 mg/L were completely removed for 34 and 12 days, respectively. The amount of sulfur removed was $2.99{\times}10^{-9}H_{2}S-S/cell$, which was suggested as the amount of sulfur removed by a single cell. The biofilter consisting of the compost and R. rhodochrous B261 could be suitable for a long-term biofilteration for the removal of $H_{2}S$ and other malodorous compounds.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.121-125
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• 2002

Rhodococcus rhodochrous IGTSS (ATCC 5396S) can break organo sulfur compounds such as dibenzothiophene. Since the environment for biodesulfurization process is invariably hydrophobic, parameters in hydrophobic systems should be examined. For the model oil, hexadecane-containing 5.43mM dibenzothiophene, the volumetric desulfurization rate was decreased with the oil-to-aqueous phase ratio up to 50%. The rate declined sharply after 48h because the cell activity, which is refreshed by medium exchange, was lost. To supply the exhausted nutrients, medium exchange was performed. At 30% oil phase, most of DBT was removed by medium exchange on 48h, and the rate was 2.03mg $DBT_{removed}/L_{dispersion}-hr.$ At 50% oil phase, medium exchange on 60h was performed and the rate was 1.79mg $DBT_{removed}/L_{dispersion}-hr.$ The 300mL flask system was scaled up to a 5-L bioreactor system. On 60 h, a medium exchange was performed and the rate was 5.28mg $DBT_{removed}/L_{dispersion}-hr.$ and all of DBT was removed. It means that we can use the biodesulfurization process even 10 the high oil-to-water phase by some appropriate methods such as controlled feeding of key nutrients and the dilution or removal of some toxic metabolites by continuous reactor.

• Proceedings of the Korean Environmental Sciences Society Conference
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• 2002.05b
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• pp.209-212
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• 2002

The heavy use of petroleum products in modern livings has brought ubiquitous environmental contaminants of aromatic compounds, which persist in aquatic and geo-environment without the substantial degradation. The persistence and accumulation of the aromatic compounds, which include xylene, phenol, toluene, phthalate, and so on are known to cause serious problems in our environments. Some of soil and aquatic microorganisms facilitate their growth by degrading aromatic compounds and utilizing degrading products as growth substrates, the biodegradation helps the reentry of carbons of aromatic compounds, preventing their accumulation in our environments. The metabolic studies on the degradation of aromatic compounds by microoganlsms were extensively carried out along with their genetic studies. A Rhodococcus sp. isolated in activated sludges has shown the excellent ability to grow on phenol as a sole carbon source. In the present study investigated a gene encoding phenol-degrading enzymes from a Rhodococcus sp.