Unique glucose oxidation catalysis of Gluconobacter oxydans constitutes an efficient cellulosic gluconic acid fermentation free of inhibitory compounds disturbance.

Toxic inhibitory compounds from lignocellulose pretreatment are the major obstacle to achieve high bioconversion efficiency in biorefinery fermentations. This study shows a unique glucose oxidation catalysis of Gluconobacter oxydans with its gluconic acid productivity free of inhibitor disturbance. The microbial experimentations and the transcriptome analysis revealed that both the activity of the membrane-bound glucose dehydrogenase and the transcription level of the genes in periplasmic glucose oxidation respiratory chain of G. oxydans were essentially not affected in the presence of inhibitory compounds. G. oxydans also rapidly converted furan and phenolic aldehyde inhibitors into the less toxic alcohols or acids. The synergy of the robust periplasmic glucose oxidation and the rapid inhibitor conversion of G. oxydans significantly elevated the efficiency of the oxidative fermentation in lignocellulose hydrolysate. The corresponding genes responsible for the conversion of furan and phenolic aldehyde inhibitors were also mined by DNA microarrays. The synergistic mechanism of G. oxydans provided an important option of metabolic modification for enhancing inhibitor tolerance of general fermentation strains.

Adaptive evolution of Gluconobacter oxydans accelerates the conversion rate of non-glucose sugars derived from lignocellulose biomass.

Gluconobacter oxydans is capable of oxidizing various lignocellulose derived sugars into the corresponding sugar acids including glucose, xylose, arabinose, galactose and mannose, but simultaneous utilization of these sugars is difficult. This study attempted an adaptive evolution of G. oxydans by alternate transfer in inhibitors containing hydrolysate and inhibitors free hydrolysate for intensifying sugars simultaneous utilization. After 420 days' continuous culture, the conversion rate of all non-glucose sugars significantly improved by several folds and achieved complete conversion of lignocellulose-derived sugars to the corresponding sugar acids. The significant up-regulation of mGDH gene in the adapted G. oxydans strain (more than 40-fold greater than the parental) was considered as the decisive factor for the improvement of strain performance. This evolution adaptation strategy also could be used to accelerate robust sugars utilization for other fermented strains in lignocellulose biorefinery.

Complete oxidative conversion of lignocellulose derived non-glucose sugars to sugar acids by Gluconobacter oxydans.

Non-glucose sugars derived from lignocellulose cover approximately 40% of the total carbohydrates of lignocellulose biomass. The conversion of the non-glucose sugars to the target products is an important task of lignocellulose biorefining research. Here we report a fast and complete conversion of the total non-glucose sugars from corn stover into the corresponding sugar acids by whole cell catalysis and aerobic fermentation of Gluconobacter oxydans. The conversions include xylose to xylonate, arabinose to arabonate, mannose to mannonate, and galactose to galactonate, as well as with glucose into gluconate. These cellulosic non-glucose sugar acids showed the excellent cement retard setting property. The mixed cellulosic sugar acids could be used as cement retard additives without separation. The conversion of the non-glucose sugars not only makes full use of lignocellulose derived sugars, but also effectively reduces the wastewater treatment burden by removal of residual sugars.