Candida parapsilosis 돌연변이주에 의한 Xylitol 생산조건의 최적화

Optimization of Culture Conditions for Xylitol Production by A Mutant of Candida parapsilosis

Candida parapsilosis ATCC 22019 돌연변이주를 이용하여 xylitol 생산에 영향을 주는 배양 조건인 pH와 온도 그리고 교반속도 및 산소전달속도등의 환경인자가 Xylitol의 생성에 미치는 영향을 살펴보았다. 발효조에서 pH가 증가 할수록 균체농도와 기질소비속도가 증가하여 발효시간이 단축되었다. 그러나, Xylitol생산은 pH 4.5와 5.5에서 큰 차이 없이 50g/l의 xylose로 부터 약 34g/l로 최대농도를 보여주었다. 온도가 증가 할수록 최대 비증식속도가 증가하였지만 최종 균체농도는 감소하였고, xylitol 생산성은 $30^{\circ}C$에서 최대값을 보여주었다. 산소전달속도의 영향을 조사하기 위하여 발효조의 교반속도를 변화시키면서 배양한 결과 균체농도는 산소 전달속도가 높을수록 증가하였지만, xylitol 생산은 크게 감소하였다. 교반속도를 150rpm(산소전달속도 $30\;hr^{-1}$에 해당)으로 배양할때 발효시간 62시간에서 50g/l의 xylose로 부터 xylitol 농도가 35.8g/l로 최대값을 나타내었다. Xylitol 생산성을 증가시키기 위하여 1차 발효가 끝난 발효조에서 균체를 회수하여 20g/l로 농축하여 최적조건인 pH 4.5, $30^{\circ}C$, 산소전달속도 $30\;hr^{-1}$에서 재배양을 하였을 때 50g/l의 xylose가 배양시간 약 18시간만에 모두 이용되었고 전환수율 80%에 해당하는 40g/l의 Xylitol이 생성되었다. 이때 Xylitol의 생산성은 2.22g/l-hr으로 일반 발효때 얻은 $0.5{\sim}g/l-hr$ 보다 약 $3{\sim}4$배 증가되었다.

Effect of culture conditions such as pH, temperature, agitation speed and oxygen transfer rate on xylitol production from xylose by Candide parapsilosis ATCC 21019 mutant was investigated in a jar fermentor. The initial concentration of xylosr was fixed at 50 g/l in this experiment. When pH was increased, cell growth and xylose consumption rate were increased, but maximum xylitol production was shown in the range of pH 4.5 and 5.5 with a yield of 0.68 g/g-xylose. The optimal temperature for xylitol production was determined to be $30^{\circ}C$. Considering the importance of dissolved oxygen tension, for xylitol production, the effect of oxygen transfer rate coefficient $(k_La)$ on fermentation parameters was carefully evaluated in the range of $20{\sim}85\;hr{-1}\;of\;k_La$ (corresponding to $100{\sim}300$rpm of agitation speed). The xylitol production was maximized at $30\;hr^{-1}\;of\;k_La$(150 rpm). A higher oxygen transfer rate supported better cell growth with lower xylitol yield. It was determined that maximum xylitol concentration, xylitol yield and productivity was 35.8 g/l, 71.6% and $0.58\;g/l{\sim}hr^{-1}$, respectively, at $30\;hr^{-1}\;of\;k_La$ In order to further increase xylitol productivity, ferementation using the concentrated biomass(20 g/l) was carried out at the conditions of pH 4.5, $30^{\circ}C$ and $30\;hr\;1$ of oxygen transfer rate. The final xylitol concentration of 40 g/l was obtained at 18 hours of culture time. From this result, it was calculated that xylitol yield was 80ft on the basis of xylose consumption and volumetric productivity was $2.22\;g/l{\sim}hr$ which was increased by $3{\sim}4$ fold compared with $0.5{\sim}0.7\;g/l-hr$ obtained in a normal fermentation condition.