Fermentative valorisation of xylose-rich hemicellulosic hydrolysates from agricultural waste residues for lactic acid production under non-sterile conditions.

Lactic acid (LA) is a platform chemical with diverse industrial applications. Presently, commercial production of LA is dominated by microbial fermentation using sugary or starch-based feedstocks. Research pursuits emphasizing towards sustainable production of LA using non-edible and renewable feedstocks have accelerated the use of lignocellulosic biomass (LCB). The present study focuses on the valorisation of xylose derived from sugarcane bagasse (SCB) and olive pits (OP) through hydrothermal and dilute acid pretreatment, respectively. The xylose-rich hydrolysate obtained was used for LA production by homo-fermentative and thermophilic Bacillus coagulans DSM2314 strain under non-sterile conditions. The fed-batch mode of fermentation resulted in maximum LA titers of 97.8, 52.4 and 61.3 g/L with a yield of 0.77, 0.66 and 0.71 g/g using pure xylose, xylose-rich SCB and OP hydrolysates, respectively. Further, a two-step aqueous two-phase system (ATPS) extraction technique was employed for the separation and recovery of LA accumulated on pure and crude xylose. The LA recovery was 45 - 65% in the first step and enhanced to 80-90% in the second step.The study demonstrated an efficient integrated biorefinery approach to valorising the xylose-rich stream for cost-effective LA production and recovery.

Revisiting the Effect of U-Bends, Flow Parameters, and Feasibility for Scale-Up on Residence Time Distribution Curves for a Continuous Bioprocessing Oscillatory Baffled Flow Reactor.

An oscillatory baffled flow reactor (OBR) has been designed with 60 interbaffled cells. The baffled columns of 40 mm internal diameter together result in a reactor length of 5740 mm. The oscillatory amplitude and frequency were in the range of 2-12 mm and 0.3-2 Hz, respectively. The report investigates the impact of U-bends and the number of reactor sections on axial dispersion for scale-up feasibility. A prediction model using operating parameters has been developed to maximize plug flow conditions using the tanks-in-series (TiS) model. The maximum TiS value was 13.38 in a single column compared to 43.68 in the full reactor at a velocity ratio of 2.27 using oscillatory parameters 8 mm and 0.3 Hz. The mixing efficiency along the reactor was found to decrease after each column at amplitudes <6 mm compared to amplitudes up to 12 mm, where a negligible impact was observed. U-bend geometry had a significant role in the decrease of TiS values.

Development of Hypertolerant Strain of Yarrowia lipolytica Accumulating Succinic Acid Using High Levels of Acetate.

Acetate is emerging as a promising feedstock for biorefineries as it can serve as an alternate carbon source for microbial cell factories. In this study, we expressed acetyl-CoA synthase in Yarrowia lipolytica PSA02004PP, and the recombinant strain grew on acetate as the sole carbon source and accumulated succinic acid or succinate (SA). Unlike traditional feedstocks, acetate is a toxic substrate for microorganisms; therefore, the recombinant strain was further subjected to adaptive laboratory evolution to alleviate toxicity and improve tolerance against acetate. At high acetate concentrations, the adapted strain Y. lipolytica ACS 5.0 grew rapidly and accumulated lipids and SA. Bioreactor cultivation of ACS 5.0 with 22.5 g/L acetate in a batch mode resulted in a maximum cell OD600 of 9.2, with lipid and SA accumulation being 0.84 and 5.1 g/L, respectively. However, its fed-batch cultivation yielded a cell OD600 of 23.5, SA titer of 6.5 g/L, and lipid production of 1.5 g/L with an acetate uptake rate of 0.2 g/L h, about 2.86 times higher than the parent strain. Cofermentation of acetate and glucose significantly enhanced the SA titer and lipid accumulation to 12.2 and 1.8 g/L, respectively, with marginal increment in cell growth (OD600: 26.7). Furthermore, metabolic flux analysis has drawn insights into utilizing acetate for the production of metabolites that are downstream to acetyl-CoA. To the best of our knowledge, this is the first report on SA production from acetate by Y. lipolytica and demonstrates a path for direct valorization of sugar-rich biomass hydrolysates with elevated acetate levels to SA.

Recycling bread waste into chemical building blocks using a circular biorefining approach.

Food waste is a global problem, causing significant environmental harm and resulting in substantial economic losses globally. Bread is the commonly wasted food item in the developed world and presents a severe problem for the majority of European nations. It is the second most wasted food item in the UK after potatoes, with an equivalent of 20 million slices of bread thrown away daily. Bread is a starchy material and a rich and clean source of easily extractable fermentable sugars - this is in direct contrast to lignocellulosic feedstocks where harsh physical, chemical and/or enzymatic pretreatment processes are required for release of fermentable sugars. Furthermore, these necessary lignocellulosic pretreatment methods often produce sugars contaminated with fermentation inhibitors. Therefore, bread waste presents a clear opportunity as a potential carbon source for novel commercial processes and, to this end, several alternative routes have been developed to utilize bread waste. Possibilities for direct recycling of bread waste within the food industry are limited due to the relatively short material lifetime, stringent process and hygiene requirements. Anaerobic digestion (AD) and incineration are commonly employed methods for the valorisation of bread waste, generating limited amounts of green energy but with little other environmental or economic benefits. Most food wastes and by-products in the UK including bakery waste are treated through AD processes that fail to harness the full potential of the these wastes. This short communication reviews the challenges of handling bread waste, with a focus on a specific UK scenario. The review will consider how bread waste is generated across the supply chain, current practices to deal with the waste and logistics challenges in waste collection. The presence of clean and high-quality fermentable sugars, proteins and other nutrients in bread make it an ideal substrate for generating chemicals, fuels, bioplastics, pharmaceuticals and other renewable products through microbial fermentations. We suggest potential applications for recycling bread waste into its chemical building blocks through a fermentative route where a circular biorefining approach could maximize resource recovery and environmental savings and eliminate waste to as close to zero as possible.

Valorisation of xylose to renewable fuels and chemicals, an essential step in augmenting the commercial viability of lignocellulosic biorefineries.

Biologists and engineers are making tremendous efforts in contributing to a sustainable and green society. To that end, there is growing interest in waste management and valorisation. Lignocellulosic biomass (LCB) is the most abundant material on the earth and an inevitable waste predominantly originating from agricultural residues, forest biomass and municipal solid waste streams. LCB serves as the renewable feedstock for clean and sustainable processes and products with low carbon emission. Cellulose and hemicellulose constitute the polymeric structure of LCB, which on depolymerisation liberates oligomeric or monomeric glucose and xylose, respectively. The preferential utilization of glucose and/or absence of the xylose metabolic pathway in microbial systems cause xylose valorization to be alienated and abandoned, a major bottleneck in the commercial viability of LCB-based biorefineries. Xylose is the second most abundant sugar in LCB, but a non-conventional industrial substrate unlike glucose. The current review seeks to summarize the recent developments in the biological conversion of xylose into a myriad of sustainable products and associated challenges. The review discusses the microbiology, genetics, and biochemistry of xylose metabolism with hurdles requiring debottlenecking for efficient xylose assimilation. It further describes the product formation by microbial cell factories which can assimilate xylose naturally and rewiring of metabolic networks to ameliorate xylose-based bioproduction in native as well as non-native strains. The review also includes a case study that provides an argument on a suitable pathway for optimal cell growth and succinic acid (SA) production from xylose through elementary flux mode analysis. Finally, a product portfolio from xylose bioconversion has been evaluated along with significant developments made through enzyme, metabolic and process engineering approaches, to maximize the product titers and yield, eventually empowering LCB-based biorefineries. Towards the end, the review is wrapped up with current challenges, concluding remarks, and prospects with an argument for intense future research into xylose-based biorefineries.