Biphasic pretreatment excels over conventional sulfuric acid in pinewood biorefinery: An environmental analysis.

This study assesses the environmental impact of pine chip-based biorefinery processes, focusing on bioethanol, xylonic acid, and lignin production. A cradle-to-gate Life Cycle Assessment (LCA) is employed, comparing a novel biphasic pretreatment method (p-toluenesulfonic acid (TsOH)/pentanol, Sc-1) with conventional sulfuric acid pretreatment (H2SO4, Sc-2). The analysis spans biomass handling, pretreatment, enzymatic hydrolysis, yeast fermentation, and distillation. Sc-1 yielded an environmental impact of 1.45E+01 kPt, predominantly affecting human health (96.55%), followed by ecosystems (3.07%) and resources (0.38%). Bioethanol, xylonic acid, and lignin contributed 32.61%, 29.28%, and 38.11% to the total environmental burdens, respectively. Sc-2 resulted in an environmental burden of 1.64E+01 kPt, with a primary impact on human health (96.56%) and smaller roles for ecosystems (3.07%) and resources (0.38%). Bioethanol, xylonic acid, and lignin contributed differently at 22.59%, 12.5%, and 64.91%, respectively. Electricity generation was predominant in both scenarios, accounting for 99.05% of the environmental impact, primarily driven by its extensive usage in biomass handling and pretreatment processes. Sc-1 demonstrated a 13.05% lower environmental impact than Sc-2 due to decreased electricity consumption and increased bioethanol and xylonic acid outputs. This study highlights the pivotal role of pretreatment methods in wood-based biorefineries and underscores the urgency of sustainable alternatives like TsOH/pentanol. Additionally, adopting greener electricity generation, advanced technologies, and process optimization are crucial for reducing the environmental footprint of waste-based biorefineries while preserving valuable bioproduct production.

The Mechanical Properties and Water-Reducing and Retarding Mechanism of a Xylonic Cement Admixture.

This study explores the mechanical properties, as well as the water-reducing and setting delay mechanism, of a novel xylonic acid-based water reducer applied to cementitious materials. Four xylonic acid water reducers were synthesized in this study: XACa (PX) from pure xylose, XACa (HS) from hemicellulose hydrolysate, XANa (PX) from pure xylose, and XANa (HS) from hemicellulose hydrolysate. These were generated through the whole-cell catalysis of Gluconobacter oxydans bacteria, using pure xylose and hemicellulose hydrolysate as substrates. The findings indicate that the xylonic acid-based water reducer can attain a water-reducing capability between 14% and 16% when the dosage (expressed as a mass fraction of cement) is roughly 0.2%. In initial and final setting tests, XACa (PX) demonstrated a pronounced retarding influence at admixture levels below 0.15%, reaching its apex at 0.10%. This delayed the initial setting time by 76% and the final setting time by 136% relative to the control group. However, a slight pro-setting effect was noted beyond a 0.2% dosage. In the compressive and flexural tests of concrete, under the same slump, the XA group improved its mechanical properties by 5% to 10% compared to the SodiuM lignosulfonate (SL) group. In the air content and chloride ion migration resistance tests, the XA group reduced the air content by 38% compared to the SL group, but also increased the data of rapid chloride migration (DRCM) by 16%. Characterization studies revealed that the carboxyl and hydroxyl groups in xylonic acid undergo chemisorption with the Si-O bonds on the surface of cement particles. These groups interact with the Si-O bonds on cement particles, contributing to water-reducing effects and delaying the setting process by impeding Ca2+ ion aggregation in the calcium-silicate-hydrate gel. Its significant water-reducing effect, adjustable setting time, and excellent mechanical and durability properties suggest its viability as an alternative to lignosulfonate series water-reducing agents.

Kinetic modeling of xylonic acid production by Gluconobacter oxydans: effects of hydrodynamic conditions.

In this study, the synthesis of xylonic acid from xylose by Gluconobacter oxydans NL71 has been investigated. According to the relationship between oxygen transfer rate and oxygen uptake rate, three different kinetic models of product formation were established and the nonlinear fitting was carried out. The results showed that G. oxydans has critical dissolved oxygen under different strain concentrations, and the relationship between respiration intensity and dissolved oxygen conformed to the Monod equation [Formula: see text]. The maximum reaction rate per unit cell mass and the theoretical maximum specific productivity of G. oxydans obtained by the kinetic model are 0.042 mol/L/h and 6.97 g/gx/h, respectively. These results will assist in determining the best balance between the airflow rate and cell concentration in the reaction and improve the production efficiency of xylonic acid.

Modified acid pretreatment to alter physicochemical properties of biomass for full cellulose/hemicellulose utilization.

Acid pretreatment of biomass decomposed hemicelluloses but could not effectively remove lignin, which hindered biomass saccharification and carbohydrates utilization. In this work, 2-naphthol-7-sulfonate (NS) and sodium bisulfite (SUL) were simultaneously added to acid pretreatment, which was found to synergistically increase hydrolysis yield of cellulose from 47.9 % to 90.6 %. Based on in-depth investigations, strong linear correlations were observed between cellulose accessibility and lignin removal, fiber swelling, CrI/cellulose ratio, cellulose crystallite size, respectively, indicating that some physicochemical characteristics of cellulose played significant roles in improving cellulose hydrolysis yield. After enzymatic hydrolysis, 84 % carbohydrates could be liberated and recovered as fermentable sugars for subsequent utilization. Mass balance illustrated that for 100 kg raw biomass, 15.1 kg xylonic acid and 20.5 kg ethanol could be co-produced, indicating the efficient utilization of biomass carbohydrates.

Screening of Gluconobacter oxydans in xylonic acid fermentation for tolerance of the inhibitors formed dilute acid pretreatment.

Pre-hydrolysate liquor, as an inevitable by-product, contains a large amount of xylose, and is therefore an inexpensive feedstock that can be upgraded to value-added chemical xylonic acid. However, inhibitors, simultaneously formed in lignocellulose pretreatment process, are regarded as the major obstacle for effectively bio-converting xylose in pre-hydrolysate into xylonic acid. In this study, Gluconobacter oxydans, with highly selective and efficient, was employed for xylonic acid production; the impacts of five typical toxic inhibitory compounds on xylonic acid productivity and the activity of the membrane-bound dehydrogenase were evaluated. The results revealed that the inhibitors showed different degrees of influence toward xylonic acid production, and the order of inhibitory effect for acidic inhibitors was formic acid > acetic acid > levulinic acid; the inhibitory effect of aldehyde inhibitors was furfural > 5-hydroxymethyl-furfural. This study provides an important basis of metabolic modification and detoxification process for enhancing inhibitor tolerance and xylonic acid productivity.

Oxygen mass transfer enhancement by activated carbon particles in xylose fermentation media.

In this work, the effect of activated carbon particles on the production of xylonic acid from xylose by Gluconobacter oxydans in a stirred tank bioreactor was investigated. The enhancement of the oxygen transfer coefficient by activated carbon particles was experimentally evaluated under different solids volume fractions, agitation and aeration rates conditions. The experimental conditions optimized by response surface methodology (agitation speed 800 rpm, aeration rate 7 L min-1, and activated carbon 0.002%) showed a maximum oxygen transfer coefficient of 520.7 h-1, 40.4% higher than the control runs without activated carbon particles. Under the maximum oxygen transfer coefficient condition, the xylonic acid titer reached 108.2 g/L with a volumetric productivity of 13.53 g L-1 h-1 and a specific productivity of 6.52 g/gx/h. In conclusion, the addition of activated carbon particles effectively enhanced the oxygen mass transfer rate. These results demonstrate that activated carbon particles enhanced cultivation for xylonic acid production an inexpensive and attractive alternative.

An integrated process for co-producing fermentable sugars and xylonate from sugarcane bagasse based on xylonic acid assisted pretreatment.

In this study, a renewable organic acid (xylonic acid), which can be prepared by the biooxidation of xylose, is used for pretreating sugarcane bagasse. The effects of reaction temperature and time on the release of fermentable xylose and glucose were investigated. On the basis of guaranteeing the good enzymatic hydrolysis efficiency and minimizing the effects of inhibitors, the pretreatment with 1 % xylnoic acid at 190 °C for 30 min was selected after optimization. In this case, 70 % xylose was released, while enzymatic hydrolysis yield was also up to 86.5 %. Meanwhile, the pretreated hydrolysate liquor was proved that it could be used for producing xylonate by biooxidation of Gluconobacter oxydans. Finally, the sequential process of the xylonic acid pretreatment and saccharification also clear the path for recycling the lignin as value-added bioproducts. Overall, this study presents a green-like strategy for fully exploiting sugarcane bagasse.

A smart self-balancing biosystem with reversible competitive adsorption of in-situ anion exchange resin for whole-cell catalysis preparation of lignocellulosic xylonic acid.

Xylonic acid (XA) bioproduction via whole-cell catalysis of Gluconobacter oxydans is a promising strategy for xylose bioconversion, which is hindered by inhibitor formation during lignocellulosic hydrolysates. Therefore, it is important to develop a catalytic system that can directly utilize hydrolysate and efficiently produce XA. Determination of the dynamic adsorption characteristics of 335 anion exchange resin resulted in a unique and interesting reversible competitive adsorption between acetic acid-like bioinhibitor, fermentable sugar and XA. Xylose in crude lignocellulosic hydrolysates was completely oxidized to 52.52 g/L XA in unprecedented self-balancing biological system through reversible competition. The obtained results showed that in-situ resin adsorption significantly affected the direct utilization of crude lignocellulosic hydrolysate for XA bioproduction (p ≤ 0.05). In addition, the resin adsorbed ca. 90 % of XA during bioconversion. The study achieved a multiple functions and integrated system, "detoxification, neutralization and product separation" for one-pot bioreaction of lignocellulosic hydrolysate.

Chemometrics and genome mining reveal an unprecedented family of sugar acid-containing fungal nonribosomal cyclodepsipeptides.

Xylomyrocins, a unique group of nonribosomal peptide secondary metabolites, were discovered in Paramyrothecium and Colletotrichum spp. fungi by employing a combination of high-resolution tandem mass spectrometry (HRMS/MS)-based chemometrics, comparative genome mining, gene disruption, stable isotope feeding, and chemical complementation techniques. These polyol cyclodepsipeptides all feature an unprecedented d-xylonic acid moiety as part of their macrocyclic scaffold. This biosynthon is derived from d-xylose supplied by xylooligosaccharide catabolic enzymes encoded in the xylomyrocin biosynthetic gene cluster, revealing a novel link between carbohydrate catabolism and nonribosomal peptide biosynthesis. Xylomyrocins from different fungal isolates differ in the number and nature of their amino acid building blocks that are nevertheless incorporated by orthologous nonribosomal peptide synthetase (NRPS) enzymes. Another source of structural diversity is the variable choice of the nucleophile for intramolecular macrocyclic ester formation during xylomyrocin chain termination. This nucleophile is selected from the multiple available alcohol functionalities of the polyol moiety, revealing a surprising polyspecificity for the NRPS terminal condensation domain. Some xylomyrocin congeners also feature N-methylated amino acid residues in positions where the corresponding NRPS modules lack N-methyltransferase (M) domains, providing a rare example of promiscuous methylation in the context of an NRPS with an otherwise canonical, collinear biosynthetic program.

Recent progress in the microbial production of xylonic acid.

Interest in the production of renewable chemicals from biomass has increased in the past years. Among these chemicals, carboxylic acids represent a significant part of the most desirable bio-based products. Xylonic acid is a five-carbon sugar-acid obtained from xylose oxidation that can be used in several industrial applications, including food, pharmaceutical, and construction industries. So far, the production of xylonic acid has not yet been available at an industrial scale; however, several microbial bio-based production processes are under development. This review summarizes the recent advances in pathway characterization, genetic engineering, and fermentative strategies to improve xylonic acid production by microorganisms from xylose or lignocellulosic hydrolysates. In addition, the strengths of the available microbial strains and processes and the major requirements for achieving biotechnological production of xylonic acid at a commercial scale are discussed. Efficient native and engineered microbial strains have been reported. Xylonic acid titers as high as 586 and 171 g L-1 were obtained from bacterial and yeast strains, respectively, in a laboratory medium. Furthermore, relevant academic and industrial players associated with xylonic acid production will be presented.

Enzymatic membrane reactor in xylose bioconversion with simultaneous cofactor regeneration.

In this study, we present the concept of co-immobilization of xylose dehydrogenase and alcohol dehydrogenase from Saccharomyces cerevisiae on an XN45 nanofiltration membrane for application in the process of xylose bioconversion to xylonic acid with simultaneous cofactor regeneration and membrane separation of reaction products. During the research, the effectiveness of the co-immobilization of enzymes was confirmed, and changes in the properties of the membrane after the processes were determined. Using the obtained biocatalytic system it was possible to obtain 99% xylonic acid production efficiency under optimal conditions, which were 5 mM xylose, 5 mM formaldehyde, ratio of NAD+:NADH 1:1, and 60 min of reaction. Additionally, the co-immobilization of enzymes made it possible to improve stability of the co-immobilized enzymes and to carry out xylose conversion in six consecutive cycles and after 7 days of storage at 4 °C with over 90% efficiency. The presented data confirm the effectiveness of the co-immobilization, improvement of the stability and reusability of the biocatalysts, and show that the obtained enzymatic system is promising for use in xylose bioconversion and simultaneous regeneration of nicotinamide cofactor.

Overexpression of mGDH in Gluconobacter oxydans to improve D-xylonic acid production from corn stover hydrolysate.

BACKGROUND: D-Xylonic acid is a versatile platform chemical with broad potential applications as a water reducer and disperser for cement and as a precursor for 1,4-butanediol and 1,2,4-tributantriol. Microbial production of D-xylonic acid with bacteria such as Gluconobacter oxydans from inexpensive lignocellulosic feedstock is generally regarded as one of the most promising and cost-effective methods for industrial production. However, high substrate concentrations and hydrolysate inhibitors reduce xylonic acid productivity. RESULTS: The D-xylonic acid productivity of G. oxydans DSM2003 was improved by overexpressing the mGDH gene, which encodes membrane-bound glucose dehydrogenase. Using the mutated plasmids based on pBBR1MCS-5 in our previous work, the recombinant strain G. oxydans/pBBR-R3510-mGDH was obtained with a significant improvement in D-xylonic acid production and a strengthened tolerance to hydrolysate inhibitors. The fed-batch biotransformation of D-xylose by this recombinant strain reached a high titer (588.7 g/L), yield (99.4%), and volumetric productivity (8.66 g/L/h). Moreover, up to 246.4 g/L D-xylonic acid was produced directly from corn stover hydrolysate without detoxification at a yield of 98.9% and volumetric productivity of 11.2 g/L/h. In addition, G. oxydans/pBBR-R3510-mGDH exhibited a strong tolerance to typical inhibitors, i.e., formic acid, furfural, and 5-hydroxymethylfurfural. CONCLUSION: Through overexpressing mgdh in G. oxydans, we obtained the recombinant strain G. oxydans/pBBR-R3510-mGDH, and it was capable of efficiently producing xylonic acid from corn stover hydrolysate under high inhibitor concentrations. The high D-xylonic acid productivity of G. oxydans/pBBR-R3510-mGDH made it an attractive choice for biotechnological production.

An overview of the metabolically engineered strains and innovative processes used for the value addition of biomass derived xylose to xylitol and xylonic acid.

Xylose, the most abundant pentose sugar of the hemicellulosic fraction of lignocellulosic biomass, has to be utilized rationally for the commercial viability of biorefineries. An effective pre-treatment strategy for the release of xylose from the biomass and an appropriate microbe of the status of an Industrial strain for the utilization of this pentose sugar are key challenges which need special attention for the economic success of the biomass value addition to chemicals. Xylitol and xylonic acid, the alcohol and acid derivatives of xylose are highly demanded commodity chemicals globally with plenty of applications in the food and pharma industries. This review emphasis on the natural and metabolically engineered strains utilizing xylose and the progressive and innovative fermentation strategies for the production and subsequent recovery of the above said chemicals from pre-treated biomass medium.

Optimization of Specific Productivity for Xylonic Acid Production by Gluconobacter oxydans Using Response Surface Methodology.

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.

Engineering Zymomonas mobilis for the Production of Xylonic Acid from Sugarcane Bagasse Hydrolysate.

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.

Understanding D-xylonic acid accumulation: a cornerstone for better metabolic engineering approaches.

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.

A cost-practical cell-recycling process for xylonic acid bioproduction from acidic lignocellulosic hydrolysate with whole-cell catalysis of Gluconobacter oxydans.

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.

Ethylene glycol and glycolic acid production by wild-type Escherichia coli.

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-type Escherichia coli W3110 within 10 H of cultivation with a substrate conversion ratio of 0.5 mol/mol. Furthermore, E. coli strains that produce solely ethylene glycol or glycolic acid were constructed. 10.3 g/L of glycolic acid was produced by E. coli ΔyqhD+aldA, and the achieved conversion ratio was 0.56 mol/mol. Similarly, the E. coli ΔaldA+yqhD produced 8.0 g/L of ethylene glycol with a conversion ratio of 0.71 mol/mol. Ethylene glycol and glycolic acid production by E. coli on xylonic acid as a carbon source provides new information on the biosynthesis pathway of these products and opens a novel way of biomass utilization.

Optimization of carbon source efficiency for lipid production with the oleaginous yeast Saitozyma podzolica DSM 27192 applying automated continuous feeding.

BACKGROUND: Biotechnologically produced microbial lipids are of interest as potential alternatives for crude and plant oils. Their lipid profile is similar to plant oils and can therefore be a substitute for the production of biofuels, additives for food and cosmetics industry as well as building blocks for oleochemicals. Commercial microbial lipids production, however, is still not profitable and research on process optimization and cost reduction is required. This study reports on the process optimization using glucose or xylose with the unconventional oleaginous yeast Saitozyma podzolica DSM 27192 aiming to reduce the applied carbon source amount without sacrificing lipid productivity. RESULTS: By optimizing the process parameters temperature and pH, lipid productivity was enhanced by 40%. Thereupon, by establishing a two-phase strategy with an initial batch phase and a subsequent fed-batch phase for lipid production in which a constant sugar concentration of about 10 g/L was maintained, resulted in saving of ~ 41% of total glucose and ~ 26% of total xylose. By performing the automated continuous sugar feed the total sugar uptake was improved to ~ 91% for glucose and ~ 92% for xylose and thus, prevented waste of unused carbon source in the cultivation medium. In addition, reduced glucose cultivation resulted in to 28% higher cell growth and 19% increase of lipid titer. By using xylose, the by-product xylonic acid was identified for the first time as by-product of S. podzolica. CONCLUSIONS: These findings provide a broad view of different cultivation process strategies with subsequent comparison and evaluation for lipid production with S. podzolica. Additionally, new biotechnological characteristics of this yeast were highlighted regarding the ability to produce valuable organic acids from sustainable and renewable sugars.

Preparation of highly flexible and sustainable lignin-rich nanocellulose film containing xylonic acid (XA), and its application as an antibacterial agent.

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.