• KSBB Journal
• /
• 제15권3호
• /
• pp.318-322
• /
• 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
• /
• 제19권5호
• /
• pp.410-414
• /
• 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
• /
• 제17권2호
• /
• pp.182-188
• /
• 2002

푸마르산으로부터 숙신산 생물전환을 위하여 E. faecalis RKY1을 HFBR에 친정화하여 연속생산 공정에 대한 가능성을 모색하였다. E. faecalis RKY1은 실관의 spongy 부분에 효과적으로 고정화되었으며, transverse mode로 HFBR 조업시 실관가닥이 50개 및 200개일 경우 거의 비슷한 경향을 나타냈다. 또한 배지의 공급속도를 0.25, 0.5, 1.0 mL/min로 증가시키면서 조업한 결과, 정상상태에 도달하는 시간이 각각 24, 12, 9시간으로서 유속이 증가할수록 정상상태에 도달하는 시간은 단축되었지만, 반응기 내의 기질 및 생성물 분포는 그 변화가 심하였다. 실관 생물반응기를 이용하여 숙신산 생물 전환시 배지의 공금 유속이 증가할수록 숙신산 생산성은 증가하였지만 전환수율이 감소하여 미반응 푸마르산은 증가하였다. 최대 숙신산 생산성 및 이 때의 수율은 푸마르산염 농도 80 g/L 및 배지 공금 유속 2.0 mL/min일 때 17.1 g/L ·hr 및 0.54 g/g이었으며, 최적 숙신산 생산성 및 이 때의 수율은 푸마르산염 농도 50 g/L 및 배지 공급 유속 1.0 mL/min일때 9.0 g/L · 및 0.90 g/g으로서 회분식 생물전환의 경우보다 더 우수하였다.

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2003년도 생물공학의 동향(XII)
• /
• pp.218-221
• /
• 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
• /
• 한국생물공학회 2003년도 생물공학의 동향(XII)
• /
• pp.185-188
• /
• 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.

• 한국미생물·생명공학회지
• /
• 제30권3호
• /
• pp.258-263
• /
• 2002

E. faecalis RKYl을 이용한 젖산발효에서 세포성장과 젖산 생산에 대한 배양온도 및 pH의 영향을 조사한 결과온도가 낮을수록 세포성장은 향상되었으나 발효시간이 지연되어 젖산생산이 저하되었으며$ 50^{\circ}C$에서는 세포성장이 거의 없었다. pH 7.0-9.0에서 거의 비슷한 세포성장 및 젖산생산을 보였으나 pH 6.0에서는 세포성장이 저하되었고 발효시간이 지연되었으며, pH 10.0에서는 세포성장이 거의 없었다. E. faecalis RKY에 의한 젖산발효는 비교적 넓은 범위의 배양온도($34- 46^{\circ}C$)및 pH(7.0-9.0)에서 안정하였으며 최적 배양온도 및 pH 는 각각 $42^{\circ}C$, 7.0이었다. 또한 일반적으로 일인산염에 비해 이인산염을 첨가해 주었을 때 젖산생산성이 향상되었으며, 효모추출물 10g·$L^{-1}$을 질소원으로 사용하여 배양하였을때 $Mg^{2+}$ 과 $Mn^{2+}$의 첨가효과는 없는 것으로 나타났다.

• Journal of Microbiology and Biotechnology
• /
• 제10권1호
• /
• pp.1-7
• /
• 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
• /
• 한국생물공학회 2003년도 생물공학의 동향(XII)
• /
• pp.201-203
• /
• 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
• /
• 제24권5호
• /
• pp.661-666
• /
• 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
• /
• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
• /
• pp.297-300
• /
• 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.