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Developing Escherichia coli strains that are tolerant to acetate toxicity is important in light of an increased interest in the efficient utilization of lignocellulosic biomass feedstocks for the biosynthesis of value-added products. In this study, four strains known to produce polyhydroxyalkanoates (PHAs) from the typical hemicellulosic sugar xylose were tested for their tolerance to acetate. E. coli RSC10 was found to be tolerant of acetate, both in growth and fermentation studies. In the presence of acetate the strain showed a >2-fold increase in overall yields compared to using xylose alone as the feedstock. More importantly, the strain was found to be able to utilize acetate as a feedstock for biosynthesis of PHAs, with complete depletion of acetate (25 mM) at 9 h when acetate was the sole feedstock. Higher concentrations of acetate showed greater inhibition of fermentation than growth with a reduction of 90% in PHA yields at 100 mM. Additionally, the present work provides data to support the potential of acetate as a modulator for the control of composition of PHAs that incorporate lactate (LA) monomers into the copolymer from hemicellulose derived sugars., (Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Consolidated bioprocessing of lignocellulose is an attractive strategy for the sustainable production of petroleum-based alternatives. One of the underutilized sources of carbon in lignocellulose is the hemicellulosic fraction which largely consists of the polysaccharide xylan. In this study, Escherichia coli JW0885 (pyruvate formate lyase activator protein mutant, pflA(-)) was engineered to express recombinant xylanases and polyhydroxyalkanoate (PHA)-producing enzymes for the biosynthesis of poly(lactate-co-3-hydroxybutyrate) [P(LA-co-3HB)] from xylan as a consolidated bioprocess. The results show that E. coli JW0885 was capable of producing P(LA-co-3HB) when xylan was the only feedstock and different feeding and growth parameters were examined in order to improve upon initial yields. The highest yields of P(LA-co-3HB) copolymer obtained in this study occurred when xylan was added during mid-exponential growth after cells had been grown at high shaking-speeds (290 rpm). The results showed an inverse relationship between total PHA production and LA-monomer incorporation into the copolymer. Proton nuclear magnetic resonance ((1)H NMR), gel permeation chromatography (GPC), and differential scanning calorimetry (DSC) analyses corroborate that the polymers produced maintain physical properties characteristic of LA-incorporating PHB-based copolymers. The present study achieves the first ever engineering of a consolidated bioprocessing bacterial system for the production of a bioplastic from a hemicelluosic feedstock., (Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Conversion of lignocellulosic biomass to value added products such as ethanol and other platform chemicals is enhanced by pretreatment, which reduces the crystallinity and molecular weight of cell wall polymers, thus increasing the available reaction sites. In this study, switchgrass (Panicum virgatum L.) was pretreated with high energy electron beam (EB) irradiation to reduce its recalcitrance and achieve higher sugar conversion rates during treatment with cellulases and β-glucosidase. Conversion rates to sugars were compared before and after hot water (HW) extraction of EB-treated and control samples of switchgrass. Thermogravimetric analysis (TGA) was employed to determine peak degradation temperature of these EB-treated biomass samples before and after HW extraction, and near infrared spectroscopy (NIR) was used as a rapid technique to determine cellulose, hemicellulose, and lignin contents in the samples. TGA data confirm previously reported results that EB pretreatment reduces the molecular weight and crystallinity of cellulose and hemicellulose. This leaves hemicellulose more amenable to HW extraction and creates more cellulase-accessible sites, as shown by NIR and glucose yield data, respectively. Hemicellulose content was reduced from 30.2 to 16.9% after HW extraction and 1000 kGy EB treatment, and ultimate glucose yield after cellulase hydrolysis increased more than 4-fold. This study provides evidence that when EB pretreatment is utilized in combination with HW extraction, higher conversion rates and yields of glucose can be obtained from the cellulosic fraction of switchgrass., (Copyright © 2013 Elsevier Ltd. All rights reserved.)