Scale-up of hydrothermal processing: Liquid hot water and pilot-scale tubular steam explosion batch reactor for bioethanol production using macroalgae Sargassum spp biomass.

Sargassum spp. is a biomass that can potentially use as an alternative for bioethanol production. Hydrothermal processes (liquid hot water and steam explosion pretreatment) were carried out at different operational conditions. Enzymatic hydrolysis performed a preliminary test with different ratios 1:1 and 1:2 (cellulases and hemicellulases) of enzyme loading, once selected 1:2 ratio was obtained conversion yield of 99.91% and therefore carried a scale-up in stirred bioreactor getting 95.92% saccharification yield. Pre-simultaneous saccharification and fermentation strategy was performed in a continuous stirred tank bioreactor (CSTBR), producing ethanol yield of 57.69%, and for simultaneous saccharification and fermentation strategy was performed in a bubble column reactor was 71.37% ethanol yield. The energy efficiency was analyzed in different scenarios; the best data was 30.19 (gsugar/MJ) in the bioreactor enzymatic hydrolysis process. This development allows for establishing the conditions for a third-generation biorefinery on a circular bioeconomy using Sargassum biomass.

Green and simple approach for low-cost bioproducts preparation and CO2 capture.

This study has demonstrated, for the first time, a simple, fast and flexible microwave processing method for the simultaneous preparation of bio-products (bio-oil, bio-gas and biochar) using a methodology that avoids any form of catalyst or chemical activation. The dielectric properties of biomass and physicochemical characterisation such as TGA, elemental and proximate analysis, XRD, SEM/EDX and textural properties, showed that 8 kJ g-1 of microwave energy can produce superior biochars for applications in CO2 capture. The maximum CO2 uptake capacity for biochar produced was 2.5 mmol g-1 and 2.0 mmol g-1 at 0 and 25 °C and 1 bar, which and also exhibited high gas selectivity compared with N2, fast kinetics of adsorption (<10 min) and desirable reusability (>95%) after 20 cycles. GC-MS analysis of generated bio-oil products revealed that higher microwave energies (>8 kJ g-1) significantly enhanced the amount of bio-oil produced (39%) and specifically the formation of levoglucosan, furfural and phenolics compounds, and bio-gas analysis identified trace levels of H2 and CH4. The results from this study confirm a green, inexpensive and efficient approach for biomass valorisation which can easily be embedded within bio-refinery process, and also demonstrates the potential of biochars for post-combustion CO2 uptake.

High-pressure technology for Sargassum spp biomass pretreatment and fractionation in the third generation of bioethanol production.

Sargassum spp is an invasive macroalgae and an alternative feedstock for bioethanol production. Sargassum spp biomass was subjected to high-pressure technology for biomass fractionation under different operating conditions of temperature and residence time to obtain glucan enriched pretreated solids (32.22 g/100 g of raw material). Enzyme hydrolysis process at high pretreated solid loading (13%, w/v) and enzyme loading of 10 FPU/g of glucan was performed, obtaining 43.01 g/L of glucose corresponding to a conversion yield of 92.12%. Finally, a pre-simultaneous saccharification and fermentation strategy (PSSF) was performed to produce bioethanol. This operational strategy produced 45.66 g/L of glucose in the pre-saccharification stage, and 18.14 g/L of bioethanol was produced with a glucose to bioethanol conversion yield of 76.23%. The development of this process highlights the feasibility of bioethanol production from macroalgal biomass in the biorefinery concept.

2,3-Dihydroxyisovalerate production by Klebsiella pneumoniae.

2,3-Dihydroxyisovalerate is an intermediate of valine and leucine biosynthesis pathway; however, no natural microorganism has been found yet that can accumulate this compound. Klebsiella pneumoniae is a useful bacterium that can be used as a workhorse for the production of a range of industrially desirable chemicals. Dihydroxy acid dehydratase, encoded by the ilvD gene, catalyzes the reaction of 2-ketoisovalerate formation from 2,3-dihydroxyisovalerate. In this study, an ilvD disrupted strain was constructed which resulted in the inability to synthesize 2-ketoisovalerate, yet accumulate 2,3-dihydroxyisovalerate in its culture broth. 2,3-Butanediol is the main metabolite of K. pneumoniae and its synthesis pathway and the branched-chain amino acid synthesis pathway share the same step of the α-acetolactate synthesis. By knocking out the budA gene, carbon flow into the branched-chain amino acid synthesis pathway was upregulated, which resulted in a distinct increase in 2,3-dihydroxyisovalerate levels. Lactic acid was identified as a by-product of the process and by blocking the lactic acid synthesis pathway, a further increase in 2,3-dihydroxyisovalerate levels was obtained. The culture parameters of 2,3-dihydroxyisovalerate fermentation were optimized, which include acidic pH and medium level oxygen supplementation to favor 2,3-dihydroxyisovalerate synthesis. At optimal conditions (pH 6.5, 400 rpm), 36.5 g/L of 2,3-dihydroxyisovalerate was produced in fed-batch fermentation over 45 h, with a conversion ratio of 0.49 mol/mol glucose. Thus, a biological route of 2,3-dihydroxyisovalerate production with high conversion ratio and final titer was developed, providing a basis for an industrial process. Key Points • A biological route of 2,3-dihydroxyisovalerate production was setup. • Disruption of budA causes 2,3-dihydroxuisovalerate accumulation in K. pneumoniae. • Disruption of ilvD prevents 2,3-dihydroxyisovalerate reuse by the cell. • 36.5 g/L of 2,3-dihydroxyisovalerate was obtained in fed-batch fermentation.

Bioethanol Production from UK Seaweeds: Investigating Variable Pre-treatment and Enzyme Hydrolysis Parameters.

This study describes the method development for bioethanol production from three species of seaweed. Laminaria digitata, Ulva lactuca and for the first time Dilsea carnosa were used as representatives of brown, green and red species of seaweed, respectively. Acid thermo-chemical and entirely aqueous (water) based pre-treatments were evaluated, using a range of sulphuric acid concentrations (0.125-2.5 M) and solids loading contents (5-25 % [w/v]; biomass: reactant) and different reaction times (5-30 min), with the aim of maximising the release of glucose following enzyme hydrolysis. A pre-treatment step for each of the three seaweeds was required and pre-treatment conditions were found to be specific to each seaweed species. Dilsea carnosa and U. lactuca were more suited with an aqueous (water-based) pre-treatment (yielding 125.0 and 360.0 mg of glucose/g of pre-treated seaweed, respectively), yet interestingly non pre-treated D. carnosa yielded 106.4 g g-1 glucose. Laminaria digitata required a dilute acid thermo-chemical pre-treatment in order to liberate maximal glucose yields (218.9 mg glucose/g pre-treated seaweed). Fermentations with S. cerevisiae NCYC2592 of the generated hydrolysates gave ethanol yields of 5.4 g L-1, 7.8 g L-1 and 3.2 g L-1 from D. carnosa, U. lactuca and L. digitata, respectively. This study highlighted that entirely aqueous based pre-treatments are effective for seaweed biomass, yet bioethanol production alone may not make such bio-processes economically viable at large scale.

Complete Acid-Based Hydrolysis Assay for Carbohydrate Quantification in Seaweed: A Species-Specific Optimized Approach.

Accurate quantification of the carbohydrate content of biomass is crucial for many bio-refining processes. The most commonly followed protocol is typically a modification of the NREL-based assay (specifically designed for carbohydrate analysis from lignocellulosic biomass). However, this NREL protocol was revealed to be excessively thermochemically harsh for seaweed biomass. This can result in erroneously low total sugar quantification as the reaction severity can degrade a proportion of the liberated sugars to decomposition products such as furans. Here we describe an optimization of the total acid hydrolysis protocol for accurate quantification of the carbohydrate content of seaweeds. Different species of seaweed can be accurately evaluated for their carbohydrate contents by following this optimized method.

Selection of yeast strains for bioethanol production from UK seaweeds.

Macroalgae (seaweeds) are a promising feedstock for the production of third generation bioethanol, since they have high carbohydrate contents, contain little or no lignin and are available in abundance. However, seaweeds typically contain a more diverse array of monomeric sugars than are commonly present in feedstocks derived from lignocellulosic material which are currently used for bioethanol production. Hence, identification of a suitable fermentative microorganism that can utilise the principal sugars released from the hydrolysis of macroalgae remains a major objective. The present study used a phenotypic microarray technique to screen 24 different yeast strains for their ability to metabolise individual monosaccharides commonly found in seaweeds, as well as hydrolysates following an acid pre-treatment of five native UK seaweed species (Laminaria digitata, Fucus serratus, Chondrus crispus, Palmaria palmata and Ulva lactuca). Five strains of yeast (three Saccharomyces spp, one Pichia sp and one Candida sp) were selected and subsequently evaluated for bioethanol production during fermentation of the hydrolysates. Four out of the five selected strains converted these monomeric sugars into bioethanol, with the highest ethanol yield (13 g L-1) resulting from a fermentation using C. crispus hydrolysate with Saccharomyces cerevisiae YPS128. This study demonstrated the novel application of a phenotypic microarray technique to screen for yeast capable of metabolising sugars present in seaweed hydrolysates; however, metabolic activity did not always imply fermentative production of ethanol.