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Rational design and facile preparation of low-cost and efficient catalysts for the selective converting of biomass-derived monosaccharides into high value-added chemicals is highly demanded, yet challenging. Herein, we first demonstrate a N doped defect-rich carbon (NC-800-5) as metal-free catalyst for the selective oxidation of d -xylose into d -xylonic acid in alkaline aqueous solution at 100 °C for 30 min, with 57.4% yield. The doped graphitic N is found to be the active site and hydroxyl ion participating in the oxidation of d -xylose. Hydroxyl ion and d -xylose first adsorb on NC-800-5 surface, and the aldehyde group of d -xylose is catalyzed to form germinal diols ion. Then, C–H bond break to yield carboxylic group. Furthermore, NC-800-5 catalyst shows high stability in recycled test.

The preparation of inexpensive, high-performance non-noble metal catalysts for selective oxidation of biomass-derived monosaccharides to high value-added chemicals is challenging but in high demand. Herein, we successfully synthesized a nickel-embedded carbon nanotube (Ni-NCNT) catalyst to selectively convert the monosaccharides into sugar acids under mild condition. The Ni-NCNT catalyst was highly reactive towards glucose and xylose oxidation, reaching 90.8% yield of gluconic acid and 88.4% yield of xylonic acid under the optimal condition. Theoretical calculations revealed that the intrinsic reaction mechanism of xylose oxidation involves the activation of the oxidant, adsorption of xylose, dissociation of the formyl C–H bond in xylose, formation of chemisorbed xylonic acid, and desorption of xylonic acid. The dissociation of the formyl C–H bond and the formation of adsorbed xylonic acid are key steps in the oxidation of xylose. The OH−, instead of O2, acted as a direct oxidant and was crucial for the xylose oxidation. The highly active and cost-effective Ni-NCNT developed in this study provided a promising route to generate valuable chemicals from biomass.

The xylose oxidative pathway (XOP) has been engineered in microorganisms for the production of a wide range of industrially relevant compounds. However, the performance of metabolically engineered XOP-utilizing microorganisms is typically hindered by D-xylonic acid accumulation. It acidifies the media and perturbs cell growth due to toxicity, thus curtailing enzymatic activity and target product formation. Fortunately, from the growing portfolio of genetic tools, several strategies that can be adapted for the generation of efficient microbial cell factories have been implemented to address D-xylonic acid accumulation. This review centers its discussion on the causes of D-xylonic acid accumulation and how to address it through different engineering and synthetic biology techniques with emphasis given on bacterial strains. In the first part of this review, the ability of certain microorganisms to produce and tolerate D-xylonic acid is also tackled as an important aspect in developing efficient microbial cell factories. Overall, this review could shed some insights and clarity to those working on XOP in bacteria and its engineering for the development of industrially applicable product-specialist strains. KEY POINTS: D-Xylonic acid accumulation is attributed to the overexpression of xylose dehydrogenase concomitant with basal or inefficient expression of enzymes involved in D-xylonic acid assimilation. Redox imbalance and insufficient cofactors contribute to D-xylonic acid accumulation. Overcoming D-xylonic acid accumulation can increase product formation among engineered strains. Engineering strategies involving enzyme engineering, evolutionary engineering, coutilization of different sugar substrates, and synergy of different pathways could potentially address D-xylonic acid accumulation.

The selective oxidation of monosaccharides into sugar acids is an important reaction for the production of biomass-based chemicals, but the design of an efficient catalyst remains a big challenge. Herein we report a series of efficient and stable surface functionalized carbon nanotube supported palladium catalysts for the selective oxidation of monosaccharides into sugar acids at room temperature. The palladium nanoparticles supported on alkaline carbon nanotubes (Pd/CNT-N) showed the highest reaction activity. The maximum gluconic acid yield of 98.3% and xylonic acid yield of 95.8% were obtained at 30 °C within 180 min. DFT calculations revealed that the mechanism of xylose oxidation to xylonic acid involved the activation of OH−, adsorption of xylose, dissociation of the formyl C–H bond in xylose, formation of chemisorbed xylonic acid, and desorption of xylonic acid. The activation energy barrier for xylose oxidation over Pd/CNT-N is lower than that of Pd/CNT-O, explaining that the basic groups on CNT are more beneficial for accelerating monosaccharide oxidation and enhancing the sugar acid selectivity. This work not only presents a new facile route to produce sugar acid at room temperature, but also provides a basis to design and develop efficient catalysts with surface functionalized supports for various catalytic reactions.

The photocatalytic selective oxidation of biomass-derived feedstocks to high-value organic acids is promising and challenging, especially for the production of xylonic acid. Herein, novel phosphorus-doped carbon nitride with grafted sulfonic acid groups (P@CN-SO3H) was prepared and successfully used in the photocatalytic-reforming of xylose to xylonic acid. The successful doping of phosphorus and grafting of sulfonic acid groups significantly extend the visible light adsorption range and decrease the photoluminescence intensity of the resulting carbon nitride, thereby enhancing its photocatalytic activity. P@CN-SO3H can efficiently catalyze the synthesis of xylonic acid (88.1%) from xylose with excellent stability and reusability. Electron spin-resonance results indicate that the contents of h+, ˙OH, ·O2−, and 1O2 increase with the irradiation time, facilitating the synthesis of xylonic acid, in which ˙O2− plays a primary role. The economic benefits and results of the present xylonic acid production system indicate its huge potential for industrial applications.

Peanut is frequently constrained by extreme environmental conditions such as drought. To reveal the involvement of metabolites, TAG 24 (drought-tolerant) and JL 24 (drought-sensitive) peanut genotypes were investigated under control and 20% PEG 6000-mediated water scarcity conditions at the seedling stage. Samples were analyzed by gas chromatography-mass spectrometry (GC-MS) to identify untargeted metabolites and targeted metabolites, i.e., polyamines and polyphenols by high-performance liquid chromatography (HPLC) and ultrahigh-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), respectively. The principal component analysis (PCA), partial least-squares discriminant analysis (PLS-DA), heat map, and cluster analysis were applied to the metabolomics data obtained by the GC-MS technique to determine the important metabolites for drought tolerance. Among 46 resulting metabolites, pentitol, phytol, xylonic acid, d-xylopyranose, stearic acid, and d-ribose were important drought-responsive metabolites. Agmatine and cadaverine were present in TAG 24 leaves and roots, respectively, during water-deficit conditions and believed to be the potential polyamines for drought tolerance. Polyphenols such as syringic acid and vanillic acid were produced more in the leaves of TAG 24, while catechin production was high in JL 24 during stress conditions. Seven metabolic pathways, namely, galactose metabolism, starch and sucrose metabolism, fructose and mannose metabolism, pentose and glucuronate interconversion, propanoate metabolism, amino sugar and nucleotide sugar metabolism, and biosynthesis of unsaturated fatty acids were significantly affected by water-deficit conditions. This study provides valuable information about the metabolic response of peanut to drought stress and metabolites identified, which encourages further study by transcriptome and proteomics to improve drought tolerance in peanut.

For high value utilization of depectinized celery, in this work. Sulfuric acid (1%, 160 °C, 60 min) treatments, followed by high pressure homogenization, were used to isolate lignin-rich nanocellulose (LRNC) from depectinized celery. LRNC yield from celery was 43.9%. LRNC solutions containing up to 20% xylonic acid (XA) were cast into films, which exhibited significantly improved flexibility, transparency, and hydrophilic properties. Moreover, the antibacterial property of the hybrid films was determined by the content of XA, and better antibacterial property were gained with higher amounts of XA. In total, > 61.6% depectinized celery was used as the storage of food yield. This study provided a value-added utilization technology for celery and other vegetables.

As natural metabolites, organic acids have been widely applied in food, pharmaceutical, and bio-based materials industries. Particularly, the short-chain organic acids, including C2, C3, C4, C5, and C6 organic acids, are necessary intermediate metabolites in cells and are also alternatives to some commercial chemical products. As the necessary metabolites in cells, most major short-chain organic acids can be produced through microbial fermentation. Specifically, with the development of synthetic biology, metabolic engineering could endow cells with the ability to produce more short-chain organic acid products including propionic acid, pyruvate, lactic acid, 3-hydroxypropionic acid, malic acid, succinic acid, fumaric acid, butyric acid, itaconic acid, α-ketoglutaric acid, glutaric acid, citric acid, gluconic acid, muconic acid, adipic acid, xylonic acid, and so on. The recent advances in the biological production of short-chain organic acids, as well as the challenges and perspectives, are summarized in this review to promote the generation of microbial cell factories for the production of short-chain organic acids.Key points• Outlines the production strategy of short-chain organic acids• Provide guidance for efficient synthesis of short-chain organic acids• Impacts the necessary factor of acid resistance on the successful production of host cells.

Ethylene glycol and glycolic acid are bulk chemicals with a broad range of applications. The ethylene glycol and glycolic acid biosynthesis pathways have been produced by microorganisms and used as a biological route for their production. Unlike the methods that use xylose or glucose as carbon sources, xylonic acid was used as a carbon source to produce ethylene glycol and glycolic acid in this study. Amounts of 4.2 g/L of ethylene glycol and 0.7 g/L of glycolic acid were produced by a wild-typeEscherichia coliW3110 within 10 H of cultivation with a substrate conversion ratio of 0.5 mol/mol. Furthermore,E. colistrains that produce solely ethylene glycol or glycolic acid were constructed. 10.3 g/L of glycolic acid was produced byE. coli Delta yqhD+aldA, and the achieved conversion ratio was 0.56 mol/mol. Similarly, theE. coli Delta aldA+yqhDproduced 8.0 g/L of ethylene glycol with a conversion ratio of 0.71 mol/mol. Ethylene glycol and glycolic acid production byE. colion xylonic acid as a carbon source provides new information on the biosynthesis pathway of these products and opens a novel way of biomass utilization.

Xylonic acid is a promising compound for the substitution of gluconic acid. Gluconobacter oxydans DSM 2343 has proven to be a highly potent biocatalyst for the conversion of xylose to xylonic acid. In the present study, different nitrogen sources for the growth of G. oxydans and subsequent xylonic acid production were investigated for the first time with minimal medium. Application of 0.32 g/L glutamate supplemented with 0.15 g/L ammonium sulfate as a cheap nitrogen source enabled a xylonic acid productivity of 2.92 g/(Lh) which is similar to findings involving a complex medium (3.20 g/(Lh)). The study further investigated the impact of the xylose source on the growth and production of G. oxydans. Dose-response curves confirmed that G. oxydans is mainly insensitive towards the main inhibitory compounds, acetate and hydroxymethylfurfural, up to a concentration of 5 g/L and 2.5 g/L, respectively. However, batch investigations indicated that substitution of 25 % of the pure xylose with hemicellulosic xylose resulted in a xylonic acid yield of 90 % compared to the control approach without hemicellulosic xylose. The feeding of hemicellulosic xylose in a pulsed fed-batch mode even enabled the use of 50 g/L demonstrating that the proper selection of a feeding strategy for the hemicellulosic xylose greatly improves the production of xylonic acid.

Co-production of two or more desirable compounds from low-cost substrates by a single microbial catalyst could greatly improve the economic competitiveness of many biotechnological processes. However, reports demonstrating the adoption of such co-production strategy are still scarce. In this study, the ability of genome-edited strain Psudomonas putida EM42 to simultaneously valorise D-xylose and D-cellobiose - two important lignocellulosic carbohydrates - by converting them into the platform chemical D-xylonic acid and medium chain length polyhydroxyalkanoates, respectively, was investigated. Biotransformation experiments performed with P. putida resting cells showed that promiscuous periplasmic glucose oxidation route can efficiently generate extracellular xylonate with high yield reaching 0.97 g per g of supplied xylose. Xylose oxidation was subsequently coupled to the growth of P. putida with cytoplasmic beta-glucosidase BglC from Thermobifida fusca on D-cellobiose. This disaccharide turned out to be a better co-substrate for xylose-to-xylonate biotransformation than monomeric glucose. This was because unlike glucose, cellobiose did not block oxidation of the pentose by periplasmic glucose dehydrogenase Gcd, but, similarly to glucose, it was a suitable substrate for polyhydroxyalkanoate formation in P. putida. Co-production of extracellular xylose-born xylonate and intracellular cellobiose-born medium chain length polyhydroxyalkanoates was established in proof-of-concept experiments with P. putida grown on the disaccharide. This study highlights the potential of P. putida EM42 as a microbial platform for the production of xylonic acid, identifies cellobiose as a new substrate for mcl-PHA production, and proposes a fresh strategy for the simultaneous valorisation of xylose and cellobiose.

The xylose oxidative pathway (XOP) is continuously gaining prominence as an alternative for the traditional pentose assimilative pathways in prokaryotes. It begins with the oxidation of D-xylose to D-xylonic acid, which is further converted to α-ketoglutarate or pyruvate + glycolaldehyde through a series of enzyme reactions. The persistent drawback of XOP is the accumulation of D-xylonic acid intermediate that causes culture media acidification. This study addresses this issue through the development of a novel pH-responsive synthetic genetic controller that uses a modified transmembrane transcription factor called CadCΔ. This genetic circuit was tested for its ability to detect extracellular pH and to control the buildup of D-xylonic acid in the culture media. Results showed that the pH-responsive genetic sensor confers dynamic regulation of D-xylonic acid accumulation, which adjusts with the perturbation of culture media pH. This is the first report demonstrating the use of a pH-responsive transmembrane transcription factor as a transducer in a synthetic genetic circuit that was designed for XOP. This may serve as a benchmark for the development of other genetic controllers for similar pathways that involve acidic intermediates.

Background The critical issue in the competitiveness between bioengineering and chemical engineering is the products titer and the volume productivity. The most direct and effective approach usually employs high-density biocatalyst, while the weakened mass transfer and evoked foam problem accompany ultrahigh-density biocatalyst loading and substrate/product titer. In high-density obligate aerobic bioconversion, oxygen as electron acceptor is a speed-limiting step in bioprocesses, but sufficient oxygen supply will lead to the foaming which results in a significant reduction in oxygen utilization and the use of additional defoamers. In this study, we designed a novel sealed-oxygen supply (SOS) biotechnology to resolve the formidable barrier of oxygen transferring rate (OTR), for bio-based fuels and chemical production process. Results Based on systemic analysis of whole-cell catalysis in Gluconobacter oxydans, a novel sealed-oxygen supply technology was smartly designed and experimentally performed for biocatalytic oxidation of alcohols, sugars and so on. By a simple operation skill of automatic online supply of oxygen in a sealed stirring tank bioreactor of SOS, OTR barrier and foaming problem was resolved with great ease. We finally obtained ultrahigh-titer products of xylonic acid (XA), 3-hydroxypropionic acid (3-HPA), and erythrulose at 588.4 g/L, 69.4 g/L, and 364.7 g/L, respectively. Moreover, the volume productivity of three chemical products was improved by 150–250% compared with normal biotechnology. This SOS technology provides a promising approach to promote bioengineering competitiveness and advantages over chemical engineering. Conclusion SOS technology was demonstrated as an economic and universally applicable approach to bio-based fuels and chemicals production by whole-cell catalysis. The novel technology greatly promotes the competitiveness of bioengineering for chemical engineering, and provides a promising platform for the green and environmental use of biofuels.

Candida glycerinogenes, an industrial yeast with excellent multi-stress tolerance, has been applied to glycerol production for decades. However, its genetic manipulation was limited by the absence of meiosis, the diploid genome, and the lack of molecular tools. We described here the implementation of a transient CRISPR-Cas9 system for efficient genome editing in C. glycerinogenes. By targeting the counterselectable marker genes (TRP1, URA3), single and double gene knock-outs were achieved and the auxotroph obtained can be used as a background for targeting other gene (HOG1) at a mutation efficiency of 80%. Further, a xylonic acid producing C. glycerinogenes strain was constructed by knock-in of the xylose dehydrogenase gene, which produced up to 28 g/L ethanol and 9 g/L xylonic acid simultaneously from simulated lignocellulosic hydrolysate (contained 70 g/L glucose and 24 g/L xylose). These results indicated that the CRSIPR-Cas9 system developed here can facilitate the study of gene functions and metabolic pathways in C. glycerinogenes.

Cellulosic sugar acids from lignocellulose biomass are composed of gluconic acid, xylonic acid, arabonic acid, galactonic acid and mannonic acid. The major application of cellulosic sugar acids is cement retarder additive. This study experimentally investigated the performance of cellulosic sugar acids on cement hydration and compared with the commercial sugar acid from crop starch feedstock. The excellent retarding efficiency of cellulosic sugar acids was demonstrated while cement strength and soundness maintained similar to that of starch based sugar acid product. X-ray diffractograms, thermograms, and microstructure analyses showed that the hydration of cement reactions and the formation of cement products were significantly delayed by the addition of cellulosic sugar acids. This study indicates that cellulosic product from low-cost agriculture residue is a promising alternative of commercial product from crop starch feedstock as cement additive.

Large amounts of xylose cannot be efficiently metabolized and fermented due to strain limitations in lignocellulosic biorefinery. The conversion of xylose into high value chemicals can help to reduce the cost of commercialization. Therefore, xylonic acid with potential value in the construction industry offers a valuable alternative for xylose biorefinery. However, low productivity is the main challenge for xylonic acid fermentation. This study investigated the effect of three reaction parameters (agitation, aeration, and biomass concentration) on xylose acid production and optimized the key process parameters using response surface methodology The second order polynomial model was able to fit the experimental data by using multiple regression analysis. The maximum specific productivity was achieved with a value of 6.64 ± 0.20 g gx−1 h−1 at the optimal process parameters (agitation speed 728 rpm, aeration rate 7 L min−1, and biomass concentration 1.11 g L−1). These results may help to improve the production efficiency during xylose acid biotransformation from xylose.

Background: D-xylonic acid is a versatile platform chemical with wide potential applications as the water reducer and disperser for cement and the precursor for 1,4-butanediol and 1,2,4-tributantriol. Microbial production of D-xylonic acid with bacteria like Gluconobacter oxydans from cheap lignocellulosic feedstock is generally regarded as one of the most promising and cost-effective methods to industrial production, but xylonic acid productivity is reduced by high substrate inhibition and hydrolysate inhibitors. Results: D-xylonic acid productivity of G. oxydans DSM2003 was improved by overexpressing the mGDH gene, which encoded the membrane-bound glucose dehydrogenase. Using the mutated plasmids based on pBBR1MCS-5 in our previous work, the recombinant strain G. oxydans/pBBR-R3510-mGDH with the significant improvement in D-xylonic acid production and the strengthened tolerance to hydrolysate inhibitors was obtained. The fed-batch biotransformation of D-xylose by this recombinant strain reached the record high titer (588.7 g/L), yield (99.4%), and space-time yield (8.72 g/L/h). Moreover, up to 246.4 g/L D-xylonic acid was produced directly from corn stover hydroxylates without detoxification at a yield of 98.9% and a space-time yield of 11.2 g/L/h. In addition, G. oxydans/pBBR-R3510-mGDH performed strong tolerance to typical inhibitors, i.e., formic acid, furfural, and 5-hydroxymethylfurfural. Conclusion: Through overexpression of mgdh in G. oxydans, we obtained a recombinant strain G. oxydans/pBBR-R3510-mGDH. It was capable to efficiently produce xylonic acid from the corn stover hydrolysate under high concentrations of inhibitors. The high D-xylonic acid productivity made G. oxydans/pBBR-R3510-mGDH an attractive choice for biotechnical production.

Sugarcane bagasse is an agricultural residue rich in xylose, which may be used as a feedstock for the production of high-value-added chemicals, such as xylonic acid, an organic acid listed as one of the top 30 value-added chemicals on a NREL report. Here, Zymomonas mobilis was engineered for the first time to produce xylonic acid from sugarcane bagasse hydrolysate. Seven coding genes for xylose dehydrogenase (XDH) were tested. The expression of XDH gene from Paraburkholderia xenovorans allowed the highest production of xylonic acid (26.17 ± 0.58 g L−1) from 50 g L−1 xylose in shake flasks, with a productivity of 1.85 ± 0.06 g L−1 h−1 and a yield of 1.04 ± 0.04 gAX/gX. Deletion of the xylose reductase gene further increased the production of xylonic acid to 56.44 ± 1.93 g L−1 from 54.27 ± 0.26 g L−1 xylose in a bioreactor. Strain performance was also evaluated in sugarcane bagasse hydrolysate as a cheap feedstock, which resulted in the production of 11.13 g L−1 xylonic acid from 10 g L−1 xylose. The results show that Z. mobilis may be regarded as a potential platform for the production of organic acids from cheap lignocellulosic biomass in the context of biorefineries.

Xylooligosaccharides (XOS) have gained considerable attention worldwide as prebiotics due to their immune-strengthening activity and beneficial gut bacteria development and can be produced from xylan-rich resources by acid hydrolysis. The present study proved the organic acid hydrolysis to be beneficial for XOS yield. In this study, a recyclable and separable organic acid, i.e., furoic acid, was used for hydrolyzing xylan to produce XOS, and the response surface methodology design was applied to maximize the XOS yield; the results indicated that the quadratic model terms of the interaction between reaction temperature and hydrolysis time showed the most significant impact on XOS yields (P < 0.05). The predicted maximum yield of XOS was 49.0% with 1.2% furoic acid at 167°C for 33 min, being close to the experimental value (49.2%), indicating that the fitted models were in good agreement with the experimental results. Meanwhile, the primary byproducts, including xylose and furfural, were concurrently bio-oxidized into xylonic acid and furoic acid by Gluconobacter oxydans and separated by electrodialysis. Subsequently, the furoic acid with low solubility (

Background Methods to produce XOS have been intensively investigated, including enzymatic hydrolysis, steam explosion, and acid hydrolysis. Acid hydrolysis is currently the most widely used method to produce XOS due to its advantages of fewer processing steps, stronger raw material adaptability, higher yield, and better reproducibility. Especially, organic acids such as acetic acid, formic acid and xylonic acid work better as compared with mineral acids. However, the catalytic mechanism of different organic acids has been little studied. In this paper, four different organic acids, including formic acid, glycolic acid, lactic acid, and acetic acid were selected to compare their hydrolytic effects. Results Using pKa values as the benchmark, the yield of xylo-oligosaccharide (XOS) increased with the increasing value of pKa. The yield of XOS was 37% when hydrolyzed by 5% acetic acid (pKa = 4.75) at 170 ℃ for 20 min. Combined severity (CS), a parameter associated with temperature and reaction time was proposed, was proposed to evaluate the hydrolysis effect. The results of CS were consistent with that of pKa values on both the yield of XOS and the inhibitor. Conclusion The results based on pKa values and combined severity, a parameter associated with temperature and reaction time, concluded that acetic acid is a preferred catalyst. Combining the techno-economic analysis and environmental benefits, acetic acid hydrolysis process has lower factory production costs, and it is also an important metabolite and a carbon source for wastewater anaerobic biological treatment. In conclusion, production of xylo-oligosaccharides by acetic acid is an inexpensive, environment-friendly, and sustainable processing technique.

The effects of cell voltage and of concentration of sugars (glucose and xylose) on the performances of their electro-reforming have been evaluated at a Pd3Au7/C anode in 0.10 mol L&minus, 1 NaOH solution. The catalyst synthesized by a wet chemistry route is first comprehensively characterized by physicochemical and electrochemical techniques. The supported catalyst consists in alloyed Pd3Au7 nanoparticles of circa 6 nm mean diameter deposited on a Vulcan XC72 carbon support, with a metal loading close to 40 wt%. Six-hour chronoamperometry measurements are performed at 293 K in a 25 cm2 electrolysis cell for the electro-conversion of 0.10 mol L&minus, 1 and 0.50 mol L&minus, 1 glucose and xylose at cell voltages of +0.4 V, +0.6 V and +0.8 V. Reaction products are analyzed every hour by high performance liquid chromatography. The main products are gluconate and xylonate for glucose and xylose electro-reforming, respectively, but the faradaic yield, the selectivity and the formation rate of gluconate/xylonate decrease with the increase of aldose concentration, whereas lower faradaic yields and higher formation rates of gluconate/xylonate are observed at +0.8 V than at +0.4 V (higher chemical yields).

Xylonic acid is one of the top 30 highly valuable chemicals that can be obtained by microbial conversion. One of the recently used microorganisms to produce xylonic acid from xylose is the recombinant Escherichia coli. In this study, the effect of medium and nitrogen source concentration on the growth of the recombinant E. coli BL21 (DE3) and xylonic acid production were investigated. The fermentation was carried out in shake flasks for 24 h at 37 °C and 200 rpm. Hydroxamate analysis was performed to determine the concentration of xylonic acid. The recombinant E. coli BL21 (DE3) produced up to 8.69 g/L xylonic acid with yield of 0.86 g/g from 10 g/L xylose in SOB medium with 20 g/L tryptone and 5 g/L yeast extract.

ᴅ-xylonic acid is a five-carbon organic acid that has gain increasing interest due to its broadly flexible application in fields such as foods, pharmaceutical and agriculture. Microbial production of ᴅ-xylonic acid can be done by using many species of Pseudomonas, Acetobacter, Gluconobacter and the latest finding was by engineered microorganism. In this study, recombinant E. coli BL21 (DE3) is used to produce ᴅ-xylonic acid from ᴅ-xylose and the effect of temperature, initial pH of medium and agitation rate on ᴅ-xylonic acid production is determined. Fermentation was carried out using super optimal broth medium in shake flasks for 24 hours. Results indicate that ᴅ-xylonic acid concentration was as high as 9.95 g L-1 with yield of 0.91 g/g ᴅ-xylose from 10 g L-1 when governed under temperature of 37°C, initial pH of medium pH 7 and agitation rate 200 rpm.

In bacterial system, direct conversion of xylose to xylonic acid is mediated through NAD-dependent xylose dehydrogenase (xylB) and xylonolactonase (xylC) genes. Heterologous expression of these genes from Caulobacter crescentus into recombinant Corynebacterium glutamicum ATCC 13032 and C. glutamicum ATCC 31831 (with an innate pentose transporter, araE) resulted in an efficient bioconversion process to produce xylonic acid from xylose. Process parameters including the design of production medium was optimized using a statistical tool, Response Surface Methodology (RSM). Maximum xylonic acid of 56.32 g/L from 60 g/L xylose, i.e. about 76.67% of the maximum theoretical yield was obtained after 120 h fermentation from pure xylose with recombinant C. glutamicum ATCC 31831 containing the plasmid pVWEx1 xylB. Under the same condition, the production with recombinant C. glutamicum ATCC 13032 (with pVWEx1 xylB) was 50.66 g/L, i.e. 69% of the theoretical yield. There was no significant improvement in production with the simultaneous expression of xylB and xylC genes together indicating xylose dehydrogenase activity as one of the rate limiting factor in the bioconversion. Finally, proof of concept experiment in utilizing biomass derived pentose sugar, xylose, for xylonic acid production was also carried out and obtained 42.94 g/L xylonic acid from 60 g/L xylose. These results promise a significant value addition for the future bio refinery programs.

A sustainable and efficient two-stage fermentation was developed to produce gluconic acid (GA) and xylonic acid (XA) from dilute acid pretreated corn stover (DA-CS) using Gluconobacter oxydans. Cells (6.2 g/L) were obtained after four cycles of fermentation in DA-CS enzyme hydrolysate at 50 ml scale in a shake flask. With each cycle showing complete utilization of glucose with GA productivity rate at 5.3 g/L/h. The enriched cells were then collected and used to ferment DA-CS liquid stream (generated during pretreatment containing mostly xylose) at 50 ml scale in a shake flask to produce XA at a rate of 1.9 g/L/h. Subsequently, the process was scaled up to 1 L. Both GA and XA was produced at the rate of 8.7 g/L/h and 3.7 g/L/h respectively in 36 h. From 1 kg of corn stover we were able to produce 296.2 g GA and 167.4 g XA by two stage fermentation.

One of the major obstacles in process of lignocellulosic biorefinery is the utilization of pre-hydrolysate from pre-treatment. Although lignocellulosic pre-hydrolysate can serve as an economic starting material for xylonic acid production, the advancement of xylonic acid or xylonate is still limited by further commercial value or applications. In the present study, xylose in the high concentration wheat straw pre-hydrolysate was first in-situ biooxidized to xylonate by Gluconobacter oxydans. To meet the needs of commercialization, crude powdered calcium xylonate was prepared by drying process and calcium xylonate content in the prepared crude product was more than 70%. Then, the calcium xylonate product was evaluated as concrete admixture without any complex purification steps and the results demonstrated that xylonate could improve the performance of concrete. Overall, the crude xylonate product directly produced from low-cost wheat straw pre-hydrolysate can potentially be developed as retarding reducer, which could subsequently benefit lignocellulosic biorefinery.

In this work, the influence of oxygen transfer and uptake rates on xylonic acid production from xylose by Gluconobacter oxydans in a stirred tank bioreactor is studied. The effect of oxygen transfer coefficient is experimentally evaluated under different cell concentrations, agitation and aeration rates. Under non-oxygen limitation conditions, the maximum specific oxygen uptake rate and xylonic acid production rate, respectively, are about 18.0 mmolO2/gx/h and 6.0 g/gx/h. For a cell concentration of 1 and 1.5 g/L, respectively, a minimum kla value of 105.5 and 134.2 h−1 is found to prevent the oxygen transfer limitation and guarantee the highest cell activity. These results will assist in determining optimal hydrodynamic operating conditions for maximizing xylonic acid production, and encourage future utilization of C5 sugars from lignocellulosic biomass.

Microbial production of xylonic acid can be achieved via xylose oxidation catalyzed by the xylose dehydrogenase enzyme. In the present study, 11 putative xylose dehydrogenases (XDH) from bacterial and fungi were identified through in silico analysis using the sequences of the two XDH previously described. Among them, 6 genes were successfully cloned and expressed in Komagataella phaffii (Pichia pastoris) as demonstrated by the ability of the strains to produce xylonic acid. The best strain was capable of producing up to 37.1 ± 1.9 and 11.7 ± 1.6 g/L of xylonic acid with yields of 0.96 ± 0.02 and 0.40 ± 0.06 g/g in mineral medium and sugarcane bagasse hydrolysate, respectively. The results presented here demonstrated the functionality of 6 newly identified xylose dehydrogenases, increasing the number of XDH described. In addition, the efficiency of K. phaffii as a xylonic acid producer is showed for the first time.

A novel bio-refinery sequence yielding varieties of co-products was developed using straw pulping solid residue. This process utilizes neutral sulfite pretreatment which under optimal conditions (160 °C and 3% (w/v) sulfite charge) provides 64.3% delignification while retaining 90% of cellulose and 67.3% of xylan. The pretreated solids exhibited excellent enzymatic digestibility, with saccharification yields of 86.9% and 81.1% for cellulose and xylan, respectively. After pretreatment, the process of semi-simultaneous saccharification and fermentation (S-SSF) and bio-catalysis was investigated. The results revealed that decreased ethanol yields were achieved when solid loading increased from 5% to 30%. An acceptable ethanol yield of 76.8% was obtained at 20% solid loading. After fermentation, bio-catalysis of xylose remaining in fermentation broth resulted in near 100% xylonic acid (XA) yield at varied solid loadings. To complete the co-product portfolio, oxidation ammoniation of the dissolved lignin successfully transformed it into biodegradable slow-release nitrogen fertilizer with excellent agricultural properties.

Selective oxidation of biomass-based monosaccharides into value-added sugar acids is highly desired, but limited success of producing D-xylonic acid has been achieved. Herein, we report an efficient catalyst system, viz., Au nanoparticles anchored on the inner walls of hollow Al2O3 nanospheres (Au@h-Al2O3), which could catalyze the selective oxidation of D-xylose into D-xylonic acid under base-free conditions. The mesoporous Al2O3 shell as the adsorbent first adsorbed D-xylose. Then, the interface of Au nanoparticles and Al2O3 as active sites spontaneously dissociated O2, and the exposed Au nanoparticle surface as the catalytic site drove the transformation. With this catalyst system, the valuable D-xylonic acid was produced with excellent yields in the aerobic oxidation of D-xylose. Extensive investigation showed that Au@h-Al2O3 is an efficient catalyst with high stability and recyclability.

This is the first review presenting and discussing Burkholderia sacchari as a bacterial chassis. B. sacchari is a distinguished polyhydroxyalkanoates producer strain, with low biological risk, reaching high biopolymer yields from sucrose (0.29 g/g), and xylose (0.38 g/g). It has great potential for integration into a biorefinery using residues from biomass, achieving 146 g/L cell dry weight containing 72% polyhydroxyalkanoates. Xylitol (about 70 g/L) and xylonic acid [about 390 g/L, productivity 7.7 g/(L.h)] are produced by the wild-type B. sacchari. Recombinants were constructed to allow the production and monomer composition control of diverse tailor-made polyhydroxyalkanoates, and some applications have been tested. 3-hydroxyvalerate and 3-hydroxyhexanoate yields from substrate reached 80% and 50%, respectively. The genome-scale reconstruction of its metabolic network, associated with the improvement of tools for genetic modification, and metabolic fluxes understanding by future research, will consolidate its potential as a bioproduction chassis.

Herein, a new hydrothermal method has been reported for one-step controllable synthesis of ZnS@Bi2S3 nanosheets, which were used for the highly selective oxidation of biomass-derived monosaccharides to produce value-added sugar acids. The results of the photocatalytic oxidation by ZnS@Bi2S3 were compared with that by pure Bi2S3, under the same reaction conditions. Compared to pure Bi2S3, ZnS@Bi2S3 nanosheets has showed broadened band gap and improved stability. The relative band alignment indicated that the ZnS@Bi2S3 nanosheets could suppress the recombination of photogenerated charge carriers to improve the photocatalytic activity. Density functional theory (DFT) calculations were performed to investigate the photocatalytic reaction mechanism where xylose and oxygen are adsorbed on Bi2S3 (113) and ZnS (111). The xylose adsorbed on the catalytic surfaces reacted with the active free radicals generated from the photocatalytic process, and then was selectively oxidized to xylonic acid.

Oxygen supply for microbial cultures is often identified as a limiting factor for aerobic fermentation. Through implementation of an integrated oxygen control strategy, the high oxygen mass transfer rate satisfied cellular metabolic demands. Gluconobacter oxydans NL71 fermentation of xylose to xylonic acid was improved remarkably. Finally, the productivity of xylonic acid from xylose by biooxidation was markedly increased to 32.5±3.1g/L/h compared to production levels using conventional laboratory-scale bioreactors. By improving microbial fermentative vitality, we successfully bio-converted 1800g xylose to 1813±36g xylonic acid by combination of a fed-batch addition of xylose substrate as well as a cell-recycling strategy. Bioconversion results demonstrated a highly efficient fermentation model that performs continuous bioreaction, assisting the effort to industrialize microbial xylonic acid production.

The thermodynamic dissociation constants of xylonic acid and gluconic acid were studied via potentiometric methods, and the results were verified using lactic acid, which has a known pKa value, as a model compound. Solutions of xylonic acid and gluconic acid were titrated with a standard solution of sodium hydroxide. The determined pKa data were processed via the method of derivative plots using computer software, and the accuracy was validated using the Gran method. The dissociation constants associated with the carboxylic acid group of xylonic and gluconic acids were determined to be pKa1 = 3.56 ± 0.07 and pKa1 = 3.74 ± 0.06, respectively. Further, the experimental data showed that the second deprotonation constants associated with a hydroxyl group of each of the two acids were pKa2 = 8.58 ± 0.12 and pKa2 = 7.06 ± 0.08, respectively. The deprotonation behavior of polyhydroxy carboxylic acids was altered using various ratios with Cu(II) to form complexes in solution, and this led to proposing a hypothesis for further study.

An economically-prudent pretreatment is a crucial first step towards realization of the industrial lignocellulosic biorefinery. The aim of this study was to utilize lignocellulosic biomass to co-produce xylo-oligosaccharides (XOS) and glucose starting from a novel self-providing xylonic acid (XA) acidolysis method. Based on the optimization results of main acidolysis pretreatment parameters by uniform design experiments, we found that among various lignocellulosic materials, the highest yield of XOS from xylan was 54.16% with corncob, followed by 39.19% with wheat straw, 29.01% with corn straw and 30.23% with poplar sawdust. By effective degradation and removal of xylan constituents with XA acidolysis, enzymatic hydrolysabilities of inert cellulose constituents of corn cob, corn straw, wheat straw and poplar sawdust were achieved to 100%, 72.94%, 75.35% and 38.97%. Comparative mass balance diagrams of xylan and cellulose reveal that XA acidolysis pretreatment is environmental-friendly and effective for three agricultural residues, apart from woody poplar.

Background Biological routes for ethylene glycol production have been developed in recent years by constructing the synthesis pathways in different microorganisms. However, no microorganisms have been reported yet to produce ethylene glycol naturally. Results Xylonic acid utilizing microorganisms were screened from natural environments, and an Enterobacter cloacae strain was isolated. The major metabolites of this strain were ethylene glycol and glycolic acid. However, the metabolites were switched to 2,3-butanediol, acetoin or acetic acid when this strain was cultured with other carbon sources. The metabolic pathway of ethylene glycol synthesis from xylonic acid in this bacterium was identified. Xylonic acid was converted to 2-dehydro-3-deoxy-d-pentonate catalyzed by d-xylonic acid dehydratase. 2-Dehydro-3-deoxy-d-pentonate was converted to form pyruvate and glycolaldehyde, and this reaction was catalyzed by an aldolase. d-Xylonic acid dehydratase and 2-dehydro-3-deoxy-d-pentonate aldolase were encoded by yjhG and yjhH, respectively. The two genes are part of the same operon and are located adjacent on the chromosome. Besides yjhG and yjhH, this operon contains four other genes. However, individually inactivation of these four genes had no effect on either ethylene glycol or glycolic acid production; both formed from glycolaldehyde. YqhD exhibits ethylene glycol dehydrogenase activity in vitro. However, a low level of ethylene glycol was still synthesized by E. cloacae ΔyqhD. Fermentation parameters for ethylene glycol and glycolic acid production by the E. cloacae strain were optimized, and aerobic cultivation at neutral pH were found to be optimal. In fed batch culture, 34 g/L of ethylene glycol and 13 g/L of glycolic acid were produced in 46 h, with a total conversion ratio of 0.99 mol/mol xylonic acid. Conclusions A novel route of xylose biorefinery via xylonic acid as an intermediate has been established.

Background Obtaining high-value products from lignocellulosic biomass is central for the realization of industrial biorefinery. Acid pretreatment has been reported to yield xylooligosaccharides (XOS) and improve enzymatic hydrolysis. Moreover, xylose, an inevitable byproduct, can be upgraded to xylonic acid (XA). The aim of this study was to valorize sugarcane bagasse (SB) by starting with XA pretreatment for XOS and glucose production within a multi-product biorefinery framework. Results SB was primarily subjected to XA pretreatment to maximize the XOS yield by the response surface method (RSM). A maximum XOS yield of 44.5% was achieved by acid pretreatment using 0.64 M XA for 42 min at 154 °C. Furthermore, XA pretreatment can efficiently improve enzymatic digestibility, and achieved a 90.8% cellulose conversion. In addition, xylose, the inevitable byproduct of the acid-hydrolysis of xylan, can be completely converted to XA via bio-oxidation of Gluconobacter oxydans (G. oxydans). Subsequently, XA and XOS can be simultaneously separated by electrodialysis. Conclusions XA pretreatment was explored and exhibited a promising ability to depolymerize xylan into XOS. Mass balance analysis showed that the maximum XOS and fermentable sugars yields reached 10.5 g and 30.9 g per 100 g raw SB, respectively. In summary, by concurrently producing XOS and fermentable sugars with high yields, SB was thus valorized as a promising feedstock of lignocellulosic biorefinery for value-added products.

Utilization of both C5 and C6 sugars to produce biofuels and bioproducts is a key goal for the development of integrated lignocellulosic biorefineries. Previously we found that although engineered Zymomonas mobilis 2032 was able to ferment glucose to ethanol when fermenting highly concentrated hydrolyzates such as 9% glucan-loading AFEX-pretreated corn stover hydrolyzate (9% ACSH), xylose conversion after glucose depletion was greatly impaired. We hypothesized that impaired xylose conversion was caused by lignocellulose-derived inhibitors (LDIs) in hydrolyzates. To investigate the effects of LDIs on the cellular physiology of Z. mobilis during fermentation of hydrolyzates, including impacts on xylose utilization, we generated synthetic hydrolyzates (SynHs) that contained nutrients and LDIs at concentrations found in 9% ACSH. Comparative fermentations of Z. mobilis 2032 using SynH with or without LDIs were performed, and samples were collected for end product, transcriptomic, metabolomic, and proteomic analyses. Several LDI-specific effects were observed at various timepoints during fermentation including upregulation of sulfur assimilation and cysteine biosynthesis, upregulation of RND family efflux pump systems (ZMO0282-0285) and ZMO1429-1432, downregulation of a Type I secretion system (ZMO0252-0255), depletion of reduced glutathione, and intracellular accumulation of mannose-1P and mannose-6P. Furthermore, when grown in SynH containing LDIs, Z. mobilis 2032 only metabolized ∼50% of xylose, compared to ∼80% in SynH without LDIs, recapitulating the poor xylose utilization observed in 9% ACSH. Our metabolomic data suggest that the overall flux of xylose metabolism is reduced in the presence of LDIs. However, the expression of most genes involved in glucose and xylose assimilation was not affected by LDIs, nor did we observe blocks in glucose and xylose metabolic pathways. Accumulations of intracellular xylitol and xylonic acid was observed in both SynH with and without LDIs, which decreased overall xylose-to-ethanol conversion efficiency. Our results suggest that xylose metabolism in Z. mobilis 2032 may not be able to support the cellular demands of LDI mitigation and detoxification during fermentation of highly concentrated lignocellulosic hydrolyzates with elevated levels of LDIs. Together, our findings identify several cellular responses to LDIs and possible causes of impaired xylose conversion that will enable future strain engineering of Z. mobilis.

Background Poplars are considered suitable dedicated energy crops, with abundant cellulose and hemicellulose, and huge surplus biomass potential in China. Xylan, the major hemicellulosic component, contributes to the structural stability of wood and represents a tremendous quantity of biobased chemicals for fuel production. Monomeric xylose conversion to value-added chemicals such as furfural, xylitol, and xylonic acid could greatly improve the economics of pulp-paper industry and biorefinery. Acetic acid (HAc) is used as a friendly and recyclable selective catalyst amenable to xylan degradation and xylooligosaccharides production from lignocellulosic materials. However, HAc catalyst usually works much feebly at inert woods than agricultural straws. In this study, effects of different iron species in HAc media on poplar xylan degradation were systematically compared, and a preferable Fe3+-assisted HAc hydrolysis process was proposed for comparable xylose-hydrolysate recovery (XHR) and enzymatic saccharification of cellulose. Results In presence of 6.5% HAc with 0.17–0.25 wt% Fe3+, xylose yield ranged between 72.5 and 73.9%. Additionally, pretreatment was effective in poplar delignification, with a lignin yield falling between 38.6 and 42.5%. Under similar conditions, saccharification efficiency varied between 60.3 and 65.9%. Starting with 100 g poplar biomass, a total amount of 12.7–12.8 g of xylose and 18.8–22.8 g of glucose were harvested from liquid streams during the whole process of Fe3+-HAc hydrolysis coupled with enzymatic saccharification. Furthermore, the enhancement mechanism of Fe3+ coupled with HAc was investigated after proof-of-concept experiments. Beechwood xylan and xylose were treated under the same condition as poplar sawdust fractionation, giving understanding of the effect of catalysts on the hydrolysis pathway from wood xylan to xylose and furfural by Fe3+-HAc. Conclusions The Fe3+-assisted HAc hydrolysis process was demonstrated as an effective approach to the wood xylose and other monosaccharides production. Synergistic effect of Lewis acid site and aqueous acetic acid provided a promising strategy for catalytic valorization of poplar biomass.

Efficient utilization of lignocellulose is an economically relevant practice for improving the financial prospects of biorefineries. Lignocellulose contains significant levels of xylose that can be converted into valuable xylonic acid. However, some inhibitors of bioconversion processes are produced after pretreatment. Xylonic acid production in bacteria, such as Gluconobacter oxydans, is hindered by poor bacterial tolerance to contaminants. Therefore, in order to enhance bacterial resistance to inhibitors, a recombinant strain of G. oxydans was created by the introduction of the thioredoxin gene. Thioredoxin is a key protein responsible for maintaining cellular redox potential and is critical to the conversion of xylose to xylonate. Overexpression of thioredoxin was confirmed at the enzymatic level, while the recombinant strain showed increased catalytic activity when inhibitors, such as formic acid or p-hydroxybenzaldehyde (PHBA), were added to the synthetic xylose medium (17% and 7% improvement in xylonic acid yield, respectively). To probe the molecular mechanism behind the recombinant strain response to inhibitors, the expression levels of various genes were analyzed by qRT-PCR, which revealed five differentially expressed genes (DEGs) upon exposure to formic acid or PHBA.

The conversion of biomass components catalyzed via immobilized enzymes is a promising way of obtaining valuable compounds with high efficiency under mild conditions. However, simultaneous transformation of glucose and xylose into gluconic acid and xylonic acid, respectively, is an overlooked research area. Therefore, in this work we have undertaken a study focused on the co-immobilization of glucose dehydrogenase (GDH, EC 1.1.1.118) and xylose dehydrogenase (XDH, EC 1.1.1.175) using mesoporous Santa Barbara Amorphous silica (SBA 15) for the simultaneous production of gluconic acid and xylonic acid. The effective co-immobilization of enzymes onto the surface and into the pores of the silica support was confirmed. A GDH:XDH ratio equal to 1:5 was the most suitable for the conversion of xylose and glucose, as the reaction yield reached over 90% for both monosaccharides after 45 min of the process. Upon co-immobilization, reaction yields exceeding 80% were noticed over wide pH (7&ndash, 9) and temperature (40&ndash, 60 &deg, C) ranges. Additionally, the co-immobilized GDH and XDH exhibited a significant enhancement of their thermal, chemical and storage stability. Furthermore, the co-immobilized enzymes are characterized by good reusability, as they facilitated the reaction yields by over 80%, even after 5 consecutive reaction steps.

SummaryCo-production of two or more desirable compounds from low-cost substrates by a single microbial catalyst could greatly improve the economic competitiveness of many biotechnological processes. However, reports demonstrating the adoption of such co-production strategy are still scarce. In this study, the ability of genome-edited strainPsudomonas putidaEM42 to simultaneously valorise D-xylose and D-cellobiose -two important lignocellulosic carbohydrates -by converting them into the platform chemical D-xylonic acid and medium chain length polyhydroxyalkanoates, respectively, was investigated. Biotransformation experiments performed withP. putidaresting cells showed that promiscuous periplasmic glucose oxidation route can efficiently generate extracellular xylonate with high yield reaching 0.97 g per g of supplied xylose. Xylose oxidation was subsequently coupled to the growth ofP. putidawith cytoplasmic β-glucosidase BglC fromThermobifida fuscaon D-cellobiose. This disaccharide turned out to be a better co-substrate for xylose-to-xylonate biotransformation than monomeric glucose. This was because unlike glucose, cellobiose did not block oxidation of the pentose by periplasmic glucose dehydrogenase Gcd, but, similarly to glucose, it was a suitable substrate for polyhydroxyalkanoate formation inP. putida. Co-production of extracellular xylose-born xylonate and intracellular cellobiose-born medium chain length polyhydroxyalkanoates was established in proof-of-concept experiments withP. putidagrown on the disaccharide. This study highlights the potential ofP. putidaEM42 as a microbial platform for the production of xylonic acid, identifies cellobiose as a new substrate for mcl-PHA production, and proposes a fresh strategy for the simultaneous valorisation of xylose and cellobiose.

The aim of this study was to determine hydrothermally induced changes in the physico-chemical properties of a fine microfibrillated cellulose (MFC). The MFC was prepared from a monocomponent endoglucanase-treated never-dried birch kraft pulp which had been run several times through a microfluidizer in order to obtain a sample substantially free from fibers or fiber fragments. The fine MFC was subjected to hydrothermal treatment at $$150\,^{\circ }\hbox {C}$$ for 0.5, 2.5, 4.5, 8.5 and 20.5 h, after which the viscosity, water-retention capacity, surface charge, discoloration and formation of hydrolysis products were determined. The viscosity, surface charge and water-retention capacity of MFC decreased as a result of the treatment but in oscillatory measurements the storage and loss moduli increased. Hydrothermal treatment also caused discoloration of the sample which could not be fully washed away with water. Surface analysis with XPS revealed no significant changes in the surface structure of the dried MFC cakes but the filtrates after the hydrothermal treatment were complex mixtures of sugars, organic acids and furans. The main degradation products detected were xylose and xylobiose, but isomerization products of sugars (xylulose, fructose) were also found in the filtrates. In addition numerous organic acids, including formic, glycolic, lactic and xylonic acid, were identified in the filtrates, showing that both acid and alkaline catalyzed reactions occur due to dissociation of water into $$\hbox {H}^{+}$$ and $$\hbox {OH}^{-}$$ .

In a sustainable society based on circular economy, the use of waste lignocellulosic biomass (LB) as feedstock for biorefineries is a promising solution, since LB is the world’s most abundant renewable and non-edible raw material. LB is available as a by-product from agricultural and forestry processes, and its main components are cellulose, hemicellulose, and lignin. Following suitable physical, enzymatic, and chemical steps, the different fractions can be processed and/or converted to value-added products such as fuels and biochemicals used in several branches of industry through the implementation of the biorefinery concept. Upon hydrolysis, the carbohydrate-rich fraction may comprise several simple sugars (e.g., glucose, xylose, arabinose, and mannose) that can then be fed to fermentation units. Unlike pentoses, glucose and other hexoses are readily processed by microorganisms. Some wild-type and genetically modified bacteria can metabolize xylose through three different main pathways of metabolism: xylose isomerase pathway, oxidoreductase pathway, and non-phosphorylative pathway (including Weimberg and Dahms pathways). Two of the commercially interesting intermediates of these pathways are xylitol and xylonic acid, which can accumulate in the medium either through manipulation of the culture conditions or through genetic modification of the bacteria. This paper provides a state-of-the art perspective regarding the current knowledge on xylose transport and metabolism in bacteria as well as envisaged strategies to further increase xylose conversion into valuable products.

Xylonic acid (XA), as a bio-based platform chemical, is of considerable interest for xylose bioconversion. The whole-cell catalysis of Gluconobacter oxydans presents a promising application potential, while the hard works of cell culture still severely hinder XA business from the crude toxics-containing lignocellulosic hydrolysate. Hence, the bacterial cells should be recycled to reduce commercial production cost. The implementation of diatomite detoxification not only absorbs most of the degraded inhibitors in hydrolysate, but also confines the sugar contents loss with 10% and allows the bacterial cells to maintain 90% bioconversion performance during cell-recycling operation. Additionally, a scale-up of XA bioproduction was achieved in a sealed oxygen supply fermenter. Finally, 210 g XA was produced from 1000 g corncob originated hydrolysate within 24 h of whole-cell catalysis. Diatomite treatment provides an efficient and cost-practical approach for the commercial bioproduction of biochemicals like XA from lignocellulosic biomass.

Paraburkholderia sacchari revealed an outstanding ability to metabolize D-xylose to xylonic acid. The work described here aimed at finding the operational conditions to promote the production of xylonic acid in bioreactors. Fed-batch assays using xylose as the sole substrate for growth and production attained a xylonic acid concentration of 150 g L−1, corresponding to a productivity of 1.5 g L−1 h−1 and a yield of xylonic acid over xylose (YP/S) of 0.63 g XylAc/gXyl. When glucose and xylose were used sequentially for growth and production respectively, xylonic acid concentrations in the range 370−390 g L−1 were observed, resulting in a volumetric productivity of 6.4–7.7 g L−1 h−1 and a yield of 1.1 gXylAc/gXyl, matching the theoretical yield. These very competitive results were achieved using a simple process set-up.

Lignocellulosic biomass represents a potentially large resource to supply the world's fuel and chemical feedstocks. Enzymatic bioconversion of this substrate offers a reliable strategy for accessing this material under mild reaction conditions. Owing to the complex nature of lignocellulose, many different enzymatic activities are required to function in concert to perform efficient transformation. In nature, large multienzyme complexes are known to effectively hydrolyze lignocellulose into constituent monomeric sugars. We created artificial complexes of enzymes, called rosettazymes, in order to hydrolyze glucuronoxylan, a common lignocellulose component, into its cognate sugar D-xylose and then further convert the D-xylose into D-xylonic acid, a Department of Energy top-30 platform chemical. Four different types of enzymes (endoxylanase, α-glucuronidase, β-xylosidase, and xylose dehydrogenase) were incorporated into the artificial complexes. We demonstrated that tethering our enzymes in a complex resulted in significantly more activity (up to 71%) than the same amount of enzymes free in solution. We also determined that varying the enzyme composition affected the level of complex-related activity enhancement as well as overall yield.

An oxidative production process of xylonic acid using xylose in distillation stillage of cellulosic ethanol fermentation broth was designed, experimentally investigated, and evaluated. Dry dilute acid pretreated and biodetoxified corn stover was simultaneously saccharified and fermented into 59.80g/L of ethanol (no xylose utilization). 65.39g/L of xylose was obtained in the distillation stillage without any concentrating step after ethanol was distillated. Then the xylose was completely converted into 66.42g/L of xylonic acid by Gluconobacter oxydans. The rigorous Aspen Plus modeling shows that the wastewater generation and energy consumption was significantly reduced comparing to the previous xylonic acid production process using xylose in pretreatment liquid. This study provided a practical process option for xylonic acid production from lignocellulose feedstock with significant reduction of wastewater and energy consumption.

"Efficient production of poly-3-hydroxybutyrate (P(3HB)) based on glucose-xylose mixtures simulating different types of lignocellulosic hydrolysate (LCH) was addressed using Burkholderia sacchari, a wild strain capable of metabolizing both sugars and producing P(3HB). Carbon catabolite repression was avoided by maintaining glucose concentration below 10g/L. Xylose concentrations above 30g/L were inhibitory for growth and production. In fed-batch cultivations, pulse size and feed addition rate were controlled in order to reach high productivities and efficient sugar consumptions. High xylose uptake and P(3HB) productivity were attained with glucose-rich mixtures (glucose/xylose ratio in the feed=1.5w/w) using high feeding rates, while with xylose-richer feeds (glucose/xylose=0.8w/w), a lower feeding rate is a robust strategy to avoid xylose build-up in the medium. Xylitol production was observed with xylose concentrations in the medium above 30-40g/L. With sugar mixtures featuring even lower glucose/xylose ratios, i.e. xylose-richer feeds (glucose/xylose=0.5), xylonic acid (a second byproduct) was produced. This is the first report of the ability of Burkholderia sacchari to produce both xylitol and xylonic acid." info:eu-repo/semantics/publishedVersion

High titer gluconic acid and xylonic acid were simultaneously fermented by Gluconobacter oxydans DSM 2003 using corn stover feedstock after dry dilute sulfuric acid pretreatment, biodetoxification and high solids content hydrolysis. Maximum sodium gluconate and xylonate were produced at the titer of 132.46g/L and 38.86g/L with the overall yield of 97.12% from glucose and 90.02% from xylose, respectively. The drawbacks of filamentous fungus Aspergillus niger including weak inhibitor tolerance, large pellet formation and no xylose utilization were solved by using the bacterium strain G. oxydans. The obtained sodium gluconate/xylonate product was highly competitive as cement retarder additive to the commercial product from corn feedstock. The techno-economic analysis (TEA) based on the Aspen Plus modeling was performed and the minimum sodium gluconate/xylonate product selling price (MGSP) was calculated as $0.404/kg. This study provided a practical and economic competitive process of lignocellulose utilization for production of value-added biobased chemicals.