연속식 고정화 및 현탁 세포 생물 반응기에 의한 에탄을 생성중 지질 첨가 영향

The Roles of Lipid Supplements in Ethanol Production Using a Continuous Immobilized and Suspended Cell Bioreactor

Saccharomyces cerevisiae의 고정화 및 현탁 세포로 구성된 연속식 생물반응기에서 에탄올 생성시 지질 첨가 영향을 연구하였다. 여러가지 배양조건하에서 에탄을 생산량 및 현탁 세포의 알코올 탈수소효소(alcohol dehydrogenase, ADH)의 비활성도를 측정하였다. 무통기 조건하에서 ergosterol과 oleic acid를 세포 배양액에 첨가하였을때, 에탄을 생산량과 균체 생육이 현저히 증가하였으나, 알코올 탈수소효소의 비활성도는 영향을 받지 않았다. 특히 무통기 조건 및 통기 조건하에서 얻어진 현탁 세포간의 알코올 탈수소효소의 비활성도는 차이가 없었다. 계면활성제 첨가시에도 에탄올 생성, 균체 생육, 알코올 탈수소효소의 비활성도가 크게 증가하였다. 고농도($40\;g/{\ell}$ 이상) 에탄올에 노출된 세포배양액에 ergosterol과 oletic acid 첨가시에도 에탄올 생성량, 균체 생육, 알코올 탈수소효소의 비활성도가 증가하였으나, 계면활성제 첨가시에는 효과가 없었다. 따라서, 지질 첨가효과는 저농도 에탄을 조건에 비해 고농도 에탄을 존재시 크게 작용하였다. 여러가지 매양조건에서 얻어진 현탁 세포의 알코올 탈수소효소의 isozyme patteren을 전기영동법에 의해 조사한 결과 ADH I으로 추정되는 한개의 isozyme만이 확인되었으며, isozyme의 이동거리는 세포의 배양조건에 따라 약간의 차이가 있었다. 에탄올 생성량과 알코올 탈수소효소의 비활성도사이의 상관관계는 성립되지 않았으며, 알코올 탈수소효소의 비활성도보다는 균체량이 에탄올 생성에 더 중요한 인자로 작용하였다.

A one-stage, continuous-flow bioreactor with both immobilized and suspended cells was used to investigate the roles of lipid supplements in ethanol production by Saccharomyces cerevisiae. The reactor performance and the level of alcohol dehydrogenase(ADH) activities of the suspended cells, grown under various conditions, were measured. When ergosterol and/or oleic acid were added with surfactants to the yeast culture grown under non-aerated conditions, remarkable increases in ethanol production and cell growth was achieved, but specific ADH activities were not affected. Especially, no difference of specific ADH activities of the suspended cells grown under aerated and non-aerated condition was observed. The addition of the surfactant as a supplement also resulted in significant increases in ethanol production, cell growth, and specific ADH activity. When ergosterol and oleic acid were added to the yeast culture exposed to higher ethanol concentration($>40\;g/{\ell}$) level, ethanol production, cell growth, and specific ADH activity were increased, but the addition of surfactant was as effective as at lower ethanol concentration level. The results indicated that lipid supplements were more effective at higher ethanol concentration level than at lower ethanol concentration level during ethanol production. ADH isozyme patterns of the yeast cultures grown under various conditions on starch gel electrophoresis showed only one major band, probably ADH I. The migrating distance of the major isozyme, however, varied slightly according to the culture conditions of the cells. No apparent correlation was found between specific ADH activity and amount of ethanol produced. Cell mass was more important factor for ethanol production than specific ADH activity of the cells.