재조합 탄산무수화 효소 첨가 생산배지를 이용한 Actinobacillus succinogenes 유래의 숙신산 생산성 향상

Abstract

Succinic acid, a representative biomass-derived platform chemical, is a major fermentation product of Actinobacillus succinogenes. It is well known that carbon dioxide is consumed during the succinate fermentation, but the biochemical mechanism behind this phenomenon is not yet understood well. In this study, it was found that the addition of carbonic anhydrase (CA)s into media significantly enhances the succinic acid production by A. succinogenes during the fermentation supplied with carbon dioxide. It is likely that the (bi) carbonate produced by the CA activity from gaseous carbon dioxide is favoured by A. succinogenes for consumption and utilization. Therefore, the MgCO₃ requirement could be significantly reduced without compromising the succinate productivity. Furthermore, because of too high price of the commercial carbonic anhydrase, it was undertaken to economically overproduce a cyanobacterial carbonic anhydrase by the use of a recombinant Pichia pastoris. An expression vector system was constructed with the carbonic anhydrase gene PCR-cloned from Cyanobacterium Synechocystis sp., and introduced into P. pastoris for fermentation studies. About 95.9 g/L of succinic acid was produced in the production medium with 30 ppm of carbonic anhydrase, approximately 2 fold higher productivity compared to the parallel process with no supplementation of the enzyme. It is expected that this method can provide a valuable way of overcoming inefficiencies inherent in gas supply during CO₂-based bioprocesses like succinic acid fermentation.