Reconstruction of a Genome-Scale Metabolic Model to Simulate Uncoupled Growth of Zymomonas mobilis

Zymomonas mobilis is known as an example of the uncoupled growth phenomenon. This microorganism also has a unique metabolism that degrades glucose by the Entner–Doudoroff (ED) pathway. In this paper, a genome-scale metabolic model including 434 genes, 757 reactions and 691 metabolites was reconstructed to simulate uncoupled growth and study its effect on flux distribution in the central metabolism. The model properly predicted that ATPase was activated in experimental growth yields of Z. mobilis. Flux distribution obtained from model indicates that the major carbon flux passed through ED pathway that resulted in the production of ethanol. Small amounts of carbon source were entered into pentose phosphate pathway and TCA cycle to produce biomass precursors. Predicted flux distribution was in good agreement with experimental data. The model results also indicated that Z. mobilis metabolism is able to produce biomass with maximum growth yield of 123.7 g (mol glucose)-1 if ATP synthase is coupled with growth and produces 82 mmol ATP gDCW-1h-1. Coupling the growth and energy reduced ethanol secretion and changed the flux distribution to produce biomass precursors.

Production of High-Content Fructo-Oligosaccharides

Fructo-oligosaccharides (FOS) are produced from sucrose by Aureobasidium pullulans in yields between 40-60% (w/w). To increase the amount of FOS it is necessary to remove the small, non-prebiotic sugars, present. Two methods for producing high-purity FOS have been developed: the use of microorganisms able to consume small saccharides; and the use of continuous chromatography to separate sugars: simulated moving bed (SMB). It is herein proposed the combination of both methods. The aim of this study is to optimize the composition of the fermentative broth (in terms of salts and sugars) that will be further purified by SMB. A yield of 0.63 gFOS.gSucrose^-1 was obtained with A. pullulans using low amounts of salts in the initial fermentative broth. By removing the small sugars, Saccharomyces cerevisiae and Zymomonas mobilis increased the percentage of FOS from around 56.0% to 83% (w/w) in average, losing only 10% (w/w) of FOS during the recovery process.

The Composition of Rice Bran Hydrolysate and Its Possibility to Use in the Ethanol Production by Zymomonas mobilis Biofilm

Rice bran has been abandoned as agricultural waste for million tonnes per year in Thailand, therefore they have been proposed to be utilized as a rich carbon source in the production of bioethanol. Many toxic compounds are possibly released during the pretreatment of rice bran prior the fermentation process. This study aims to analyze on the availability of toxic compounds and the amount of glucose obtained from 2 different pretreatments using sulfuric acid and mixed cellulase enzymes (without and with delignification/ activated charcoal). The concentration of furfural, 5- hydroxymethyl furfural (5-HMF), levulinic acid, vanillin, syringaldehyde and4-hydroxybenzaldehyde (4-HB) and the percent acetic acid were found to be 0.0517 ± 0.049 mg/L, 0.032 ± 0.06 mg/L, 21074 ± 1685.62 mg/L, 126.265 ± 6.005 mg/L, 2.89 ± 0.30 mg/L, 0.37 ± 0.031mg/L and 0.72% under the pretreatment process without delignification/ activated charcoal treatment and 384.47 ± 99.02 g/L, 0.068 mg/L, 142107.62 ± 8664.6 mg/L, 0.19 mg/L, 5.43 ± 3.29 mg/L, 4.80 ± 0.76 mg/L and 0.254% under the pretreatment process with delignification/ activated charcoal treatment respectively. The presence of high concentration of acetic acid was found to impede the growth of Zymomonas mobilis strain TISTR 551 despite the present of high concentration of levulinic acid. Z. mobilis strain TISTR 551 was found to produce 8.96 ± 4.06 g/L of ethanol under 4 days fementation period in biofilm stage in which represented 40% theoretical yield.