Engineering NOG-pathway in Escherichia coli for poly-(3-hydroxybutyrate) production from low cost carbon sources.

Poly-(3-hydroxybutyrate) (P3HB) is a polyester with biodegradable and biocompatible characteristics suitable for bio-plastics and bio-medical use. In order to reduce the raw material cost, cheaper carbon sources such as xylose and glycerol were evaluated for P3HB production. We first conducted genome-scale metabolic network analysis to find the optimal pathways for P3HB production using xylose or glycerol respectively as the sole carbon sources. The results indicated that the non-oxidative glycolysis (NOG) pathway is important to improve the product yields. We then engineered this pathway into E. coli by introducing foreign phophoketolase enzymes. The results showed that the carbon yield improved from 0.19 to 0.24 for xylose and from 0.30 to 0.43 for glycerol. This further proved that the introduction of NOG pathway can be used as a general strategy to improve P3HB production.

Continuous butanol fermentation from xylose with high cell density by cell recycling system.

A continuous butanol production system with high-density Clostridium saccharoperbutylacetonicum N1-4 generated by cell recycling was established to examine the characteristics of butanol fermentation from xylose. In continuous culture without cell recycling, cell washout was avoided by maintaining pH>5.6 at a dilution rate of 0.26 h(-1), indicating pH control was critical to this experiment. Subsequently, continuous culture with cell recycling increased cell concentration to 17.4 g L(-1), which increased butanol productivity to 1.20 g L(-1) h(-1) at a dilution rate of 0.26 h(-1) from 0.529 g L(-1) h(-1) without cell recycling. The effect of dilution rates on butanol production was also investigated in continuous culture with cell recycling. Maximum butanol productivity (3.32 g L(-1) h(-1)) was observed at a dilution rate of 0.78 h(-1), approximately 6-fold higher than observed in continuous culture without cell recycling (0.529 g L(-1) h(-1)).

A new ß-glucosidase producing yeast for lower-cost cellulosic ethanol production from xylose-extracted corncob residues by simultaneous saccharification and fermentation.

This study reports a new yeast strain of Clavispora NRRL Y-50464 that is able to utilize cellobiose as sole source of carbon and produce sufficient native ß-glucosidase enzyme activity for cellulosic ethanol production using SSF. In addition, this yeast is tolerant to the major inhibitors derived from lignocellulosic biomass pre-treatment such as 2-furaldehyde (furfural) and 5-(hydroxymethyl)-2-furaldehyde (HMF), and converted furfural into furan methanol in less than 12h and HMF into furan-2,5-dimethanol within 24h in the presence of 15 mM each of furfural and HMF. Using xylose-extracted corncob residue as cellulosic feedstock, an ethanol production of 23 g/l was obtained using 25% solids loading at 37 °C by SSF without addition of exogenous ß-glucosidase. Development of this yeast aids renewable biofuels development efforts for economic consolidated SSF bio-processing.