• KSBB Journal
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• v.15 no.3
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• pp.318-322
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• 2000

An oxygen-sensitive fumarate reductase has been purified from the cytosol fraction of the Enterococcus faecalis RKY1 grown anaerobically on a defined medium containing glycerol and fumarate. A major portion of the purification was performed with employing Triton X-100 and reducing agents by Phenyl-sepharose CL-4B DEAE-sepharose and Dephadex G-150 The final activity was 0.42 unit/mg. The deduced molecular mass of active band was 66 kDa. The optimal pH and temperature for the activity were 7.0 and 38$^{\circ}C$ respectively. The enzyme activity was not affected by 1mM metal ions such as bacl2 $.$2H2O HgCl2 MnCl2$.$4H2O ZnCl2 CuCl2$.$2H2O Mgcl2$.$6H2O FeSo4$.$7H2O and by EDTA. Partially purified enzyme ws yellow in color ; spectroscopic study indicated the presence of flavins as a cofactor.

• KSBB Journal
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• v.19 no.5
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• pp.410-414
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• 2004

Fed-batch cultures of Enterococcus faecalis RKY1 were performed to maximize the L(+)-Iactic acid concentration in the bioreactor. The highest lactic acid concentration was obtained at around 225 g/L by intermittent feeding the concentrated glucose media containing 500 g/L of glucose and 15 g/L (or 75 g/L) of yeast extract. However, in all fed-batch cultures, volumetric productivities of lactic acid gradually decreased due to the inhibitory effect of lactic acid produced during the fermentation. The highest value of lactic acid concentration obtained in this work corresponded to around 1.5-fold increase compared with conventional batch fermentation.

• KSBB Journal
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• v.17 no.2
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• pp.182-188
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• 2002

Enterococcus faecalis RKY1 cells were immobilized in an asymmetric hollow fiber bioreactor for application to the continuous production of succinic acid. The media was fed into shell-side of the module using a peristaltic pump, and the products were collected through lumen-side outlet. The number of hollow fibers within the module did not affect the bioreactor efficiency in the transverse operated hollow fiber bioreactor. The steady state at the outlet of hollow fiber bioreactor was reached after 24 hr cultivation at flow rate of 0.25 mL/mim, 12 hr at 0.5 L/min, and 9 hr at 1.0 mL/mm, respectively. The succinate and fumarate concentrations within the hollow fiber bioreactor, however, were as changeful as increasing the flow rate. During continuous operation with the flow rates between 0.5 and 2.0 mL, the productivity of succinate was 8.0-10.9 g/L $.$ hr at 30 g/L fumarate, 4.9-14.9 g/L hr at 50 g/L fumarate, and 7.2-17.1 g/L hr at 80 galL fumarate, respectively.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.218-221
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• 2003

Lactic acid fermentation from sucrose as a carbon source was experimented. E. faecalis RKY1 metabolized sucrose efficiently into lactic acid through homolactic fermentation pathway. The optimal sucrose concentration for lactic acid production was found to be 150 g/l with the yield and productivity of 0.97 g/g and $3.7\;g/l\;{\cdot}\;h$, respectively. Lactic acid produced from sucrose was almost L(+)-lactic acid up to 98% based on total lactic acid produced. Therefore, sucrose was thought to be potential carbon source for L(+)-lactic acid production using E. faecalis RKY1.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.185-188
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• 2003

The possible inhibitory effect of wood hydrolyzate in lactic acid fermentation was partially decreased by adaptation of Enterococcus faecalis RKY1 to wood hydrolyzate-based medium. The strain RKY1 grown on wood hydrolyzate-based medium was tolerant to wood hydrolyzate medium with fermentation efficiency being improved to approximately 20% higher than that obtained with the strain RKY1 grown on glucose-based medium.

• Microbiology and Biotechnology Letters
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• v.30 no.3
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• pp.258-263
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• 2002

In this study the effects of temperature, pH, and addition of some minerals were investigated in lactic acid fermentation using Enterococcus faecalis RKYI . As a result, strain of RKYl had retained biological activity at the wide range of temperature($34-46^{\circ}C$) and pH(6.0-9.0), and the optimum temperature and pH were $42^{\circ}C$ and 7.0, respectively. When the effect of various phosphate sources added on lactic acid fermentation was studied, di-basic forms of phos-phate(especially, ammonium phosphate and potassium phosphate) had more stimulating effect rather than mono-basic phosphate sources. And there were no perceivable effect of manganese and magnecium salts addition on lactic acid fermentation.

• Journal of Microbiology and Biotechnology
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• v.10 no.1
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• pp.1-7
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• 2000

Bioconversion of fumarate to succinate was anaerobically conduced in a synthetic medium containing glycerol as a hydrogen donor and fumarate as a hydrogen acceptor. We investigated the effects of pH, carbon and nitrogen sources, conversion substrate, and other culture conditions on the production of succinate using a nwely isoloated Enterococcus facalis PKY1. Addition of a variety of carbonates to the medium significantly increasd the rates of production of succinate. The production of succinate and cell growth were relatively satisfactory in the pH range of 7.0-7.6. By using glycerol as a hydrogen donor, high purity succinate was produced with few byproducts. Yeast extract as a sole nitrogen source was the most effective for producing succinalte. As a result, the optimum condition of biconversion was obtained at a medium containing 20g/I glycerol, 50 g/l fumarate, 15 g/l yeast extract, 10 g/l $K_2HPO_4$, 1 g/I NaCl, 50ppm $MgCl_2{\cdot}6H_2O$, 10ppm $FeSo_4{\cdot}7H_2O$, and 5 g/I $Na_2CO_3$ at pH 7.0-7.6. Under the optimum condition, a succinate concentration of 153 g/I was produced in 36 h. The total volumetric production rate and the molar yield of succinate were 4.3 g/l/h and 85%, respectively.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.201-203
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• 2003

In this study, succinic acid production using fumaric acid fermentation broth was investigated. We tried to produce fumaric aicd from glucose by Rhizopus oryzae and then convert fumaric acid fermentation broth into succinic acid by Enterococcus faecalis RKY1. Conversion ratio of succinic acid was more than 0.90 g/g-fumaric acid. Furthermore, we optimized conditions of conversion from fermentation broth. As a result, when fumaric acid fermentation broth for succinic acid production was employed, we could decrease the amount used of glycerol and yeast extract.

• Journal of Microbiology and Biotechnology
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• v.24 no.5
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• pp.661-666
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• 2014

Both lactic acid productivity and cell growth were linearly correlated with yeast extract supplementation in batch fermentation. During conventional continuous operation, although fresh feed was introduced into the bioreactor with a significantly low dilution rate (0.04 $h^{-1}$), the amount of yeast extract employed was not enough to maintain the growth of microorganism. However, when the fresh feed contained 100 g/l glucose and 2 g/l yeast extract during cell-recycle continuous operation at a dilution rate of 0.04 $h^{-1}$, more than 90 g/l lactic acid was continuously produced, with the average productivity of 3.72 $g/l{\cdot}h$. In this experiment, 82 g of yeast extract (77% of reduction yield) could be reduced for the production of 1 kg of lactic acid compared with batch fermentation of a similar volumetric productivity.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.297-300
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• 2000

Succinic acid was produced by Enterococcus faecalis RKY1 cells immobilized in hollow fiber bioreactor as an alternatively immobilized culture in bioconversion of fumaric acid to succinic acid. The feed was pumped through the shell side. As the flow rate of the feed was increased, the steady state was obtained more quickly. The steady state was reached after 24 hr cultivation in 0.25 ml/min, 12 hr in 0.5 ml/min, and 9 hr in 1.0 ml/min, respectively. The effect of medium pH on succinate production was also investigated. By changing the medium pH of 8.0, the succinic acid produced was increased about 16% than that of pH 7.0.