A novel fermentation strategy for removing the key inhibitor acetic acid and efficiently utilizing the mixed sugars from lignocellulosic hydrolysates

[摘要] As part of preliminary research efforts, we have completed several experiments which demonstrate 'proof of concept.' These experiments addressed the following three questions: (1) Can a synthetic mixed sugar solution of glucose and xylose be efficiently consumed using the multi-organism approach? (2) Can this approach be used to accumulate a model product? (3) Can this approach be applied to the removal of an inhibitor, acetate, selectively from mixtures of xylose and glucose? To answer the question of whether this multi-organism approach can effectively consume synthetic mixed sugar solutions, we first tested substrate-selective uptake using two strains, one unable to consume glucose and one unable to consume xylose. The xylose-selective strain ALS998 has mutations in the three genes involved in glucose uptake, rendering it unable to consume glucose: ptsG codes for the Enzyme IICB{sup Glc} of the phosphotransferase system (PTS) for carbohydrate transport (Postma et al., 1993), manZ codes for the IID{sup Man} domain of the mannose PTS permease (Huber, 1996), glk codes for glucokinase (Curtis and Epstein 1975) We also constructed strain ALS1008 which has a knockout in the xylA gene encoding for xylose isomerase, rendering ALS1008 unable to consume xylose. Two batch experiments and one continuous bioprocess were completed. In the first experiment, each strain was grown separately in a defined medium of 8 g/L xylose and 15 g/L glucose which represented xylose and glucose concentrations that can be generated by actual biomass. In the second experiment, the two strains were grown together in batch in the same defined, mixed-sugar medium. In a third experiment, we grew the strains continuously in a 'chemostat', except that we shifted the concentrations of glucose and xylose periodically to observe how the system would respond. (For example, we shifted the glucose concentration suddenly from 15 g/L to 30 g/L in the feed).

[发布日期] 2009-02-11

[学科分类] 燃料技术

[关键词] ACETIC ACID;BIOMASS;CARBOHYDRATES;ENZYMES;FERMENTATION;GENES;GLUCOSE;MANNOSE;MIXTURES;MUTATIONS;PHOSPHOTRANSFERASES;REMOVAL;SACCHARIDES;SACCHAROSE;STRAINS;TRANSPORT;XYLOSE