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Serratia marcescens is utilized as a significant enterobacteria in the production of various high‐value secondary metabolites. Acetoin serves as a crucial foundational compound of development and finds application in a broad range of fields. Furthermore, S. marcescens HBQA‐7 is capable of utilizing xylose as its exclusive carbon source for acetoin production. The objective of this study was to utilize a constitutive promoter screening strategy to enhance both xylose utilization and acetoin production in S. marcescens HBQA‐7. By utilizing RNA‐seq, we identified the endogenous constitutive promoter P6 that is the most robust, which facilitated the overexpression of the sugar transporter protein GlfL445I, α‐acetyl lactate synthase, and α‐acetyl lactate decarboxylase, respectively. The resultant recombinant strains exhibited enhanced xylose utilization rates and acetoin yields. Subsequently, a recombinant plasmid, denoted as pBBR1MCS‐P6‐glfL445IalsSalsD, was constructed, simultaneously expressing the aforementioned three genes. The resulting recombinant strain, designated as S3, demonstrated a 1.89‐fold boost in xylose consumption rate compared with the original strain during shake flask fermentation. resulting in the accumulation of 7.14 g/L acetoin in the final fermentation medium. Subsequently, in a 5 L fermenter setup, the acetoin yield reached 48.75 g/L, corresponding to a xylose‐to‐acetoin conversion yield of 0.375 g/g. [ABSTRACT FROM AUTHOR]

In order to obtain high yields of chemicals and fuels from biomass, feedstocks need to be selected that contain high amounts of glucose, xylose, or lignin, depending on the end product. The first objective of this study was to evaluate the biomass composition of 20 samples from 11 broadleaf and grass crop species grown at two distinct sites in Montana. Another objective was to investigate the influence of growing location and seed type to determine which cultivars may be best for renewable chemical production in the northern Great Plains. There was a significant effect due to the growing location as the biomass samples from the site that had higher precipitation exhibited a significantly higher acid-insoluble lignin (23.8%) content compared to the biomass samples from the lower precipitation site (20.0%). Additionally, the 357 napus had a significantly higher amount of glucose (1.8% more) and acid-soluble lignin (0.2% more) than compared to the 940 napus both grown at the Froid site that may be attributed to genetic modifications. In terms of sugar content for potential upgrading, the grasses had significantly more glucose (36.1% total), xylose (24.3%), and arabinose (2.3%) than the Brassicaceae biomasses, but the lignin content was not significantly different. Switchgrass, in particular, had the highest total sugar content of close to 70 wt%. Understanding the impact of growing location on biomass composition is an important consideration for optimizing biomass utilization strategies and developing sustainable bioenergy production systems. [ABSTRACT FROM AUTHOR]

BACKGROUND: Glycation is a green processing technology. Based on our previous studies, glycation with l‐arabinose and xylose was beneficial to enhance the texture properties of silver carp mince (SCM) gels. However, the possible enhancement mechanism remained unclear. Therefore, in this study, SCM gels with different types of reducing sugar (glucose, l‐arabinose, and xylose) were prepared based on our previous study. The possible mechanism of texture enhancement of SCM gels was analyzed by investigating the changes in water distribution, protein structures, and microstructure in the gel system. RESULTS: The glycation of l‐arabinose and xylose enhanced the hardness, cohesiveness, chewiness, and resilience of SCM gels. Hardness increased from 1883.04 (control group) to 3624.54 (l‐arabinose group) and 4348.18 (xylose group). Low‐field nuclear magnetic resonance (LF‐NMR) showed that glycation promoted the tight binding of immobilized water to proteins. Raman spectroscopic analysis showed that glycation increased the surface hydrophobicity and promoted the formation of disulfide bonds. Scanning electron microscopy (SEM) showed that glycation promoted the formation of uniform and dense three‐dimensional network structure in SCM gels. CONCLUSION: In summary, glycation enhanced the binding ability of immobilized water to proteins, improved the surface hydrophobicity, promoted the formation of disulfide bonds, and led to a more uniform and dense gel network structure of proteins, thus enhancing the texture properties of SCM gels. This research provided a theoretical basis for a better understanding of the mechanism of the effect of glycation on the quality of gel products and also provided technical support for the application of l‐arabinose and xylose in new functional gel foods. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Glucose oxidase (GOx) catalyzes the oxidation of D -glucose to D -glucono-1,5-lactone and has a wide range of applications in various industries. However, the strict substrate specificity of GOx hampers its application in the conversion of other abundant sugars such as D -xylose. In this study, the substrate preference of GOx from Aspergillus niger (AnGOx) was engineered using a semi-rational design approach. The mutant T110V/F414L exhibited a 5.7-fold increase in D -xylose oxidation activity compared to that of the wild-type enzyme, which was attributed to its enhanced affinity for the substrate. Molecular dynamics simulations indicated that the T110V and F414L mutations may mitigate the non-productive binding of D -xylose at the entrance of the substrate-binding pocket, and therefore, are beneficial for providing access of its C1 hydroxyl group to the catalytic residues. Moreover, the mutant simultaneously oxidized D -xylose and D -glucose in the corncob hydrolysate to the corresponding aldonic acids when coupled with catalase. These findings provide new insights into substrate recognition by GOx and offer a new method for the utilization of D -xylose from lignocellulosic feedstocks. [ABSTRACT FROM AUTHOR]

This study aims to utilize the microbial resources found within Laphet-so, a traditional fermented tea in Myanmar. A total of 18 isolates of thermotolerant yeasts were obtained from eight samples of Laphet-so collected from southern Shan state, Myanmar. All isolates demonstrated the tannin tolerance, and six isolates were resistant to 5% (w/v) tannin concentration. All 18 isolates were capable of carboxy-methyl cellulose (CMC) degrading, but only the isolate DK showed ethanol production at 45 °C noticed by gas formation. This ethanol producing yeast was identified to be Cyberlindnera rhodanensis based on the sequence analysis of the D1/D2 domain on rRNA gene. C. rhodanensis DK produced 1.70 ± 0.01 U of thermostable extracellular β-glucosidase when cultured at 37 °C for 24 h using 0.5% (w/v) CMC as a carbon source. The best two carbon sources for extracellular β-glucosidase production were found to be either xylose or xylan, with β-glucosidase activity of 3.07–3.08 U/mL when the yeast was cultivated in the yeast malt extract (YM) broth containing either 1% (w/v) xylose or xylan as a sole carbon source at 37 °C for 48 h. The optimal medium compositions for enzyme production predicted by Plackett–Burman design and central composite design (CCD) was composed of yeast extract 5.83 g/L, peptone 10.81 g/L and xylose 20.20 g/L, resulting in a production of 7.96 U/mL, while the medium composed (g/L) of yeast extract 5.79, peptone 13.68 and xylan 20.16 gave 9.45 ± 0.03 U/mL for 48 h cultivation at 37 °C. Crude β-glucosidase exhibited a remarkable stability of 100%, 88% and 75% stable for 3 h at 35, 45 and 55 °C, respectively. [ABSTRACT FROM AUTHOR]

The enzymatic synthesis of xylose fatty acid esters was carried out in two steps: hydrolysis of degummed soybean oil (DSO) and esterification of the produced and purified free fatty acids (FFA) with xylose. Different lipases immobilized on a hydrophobic support were evaluated in the hydrolysis of DSO and an experimental design was used to optimize the reaction. Eversa® Transform 2.0 (EV-Purolite) yielded 93% conversion after 6 h using biocatalyst load of 6.53 wt%, water/oil mass ratio of 5.5 and temperature of 48.4 °C. High operational stability for EV-Purolite was found using a sequential batch strategy and the possibility of concentrating glycerol. In the esterification stage, lipase from Thermomyces lanuginosus (TLL-Purolite) and lipase B from Candida antarctica showed better performance than lipase from porcine pancreas and EV-Purolite. About 17% of FFAs (0.85 molecules of FAA per xylose molecule) was consumed using an enzyme load of 5 wt% of TLL-Purolite and FFA/xylose molar ratio of 5. Sequential esterification batches showed low operational stability of TTL-Purolite due to the desorption of TLL from the support. The final mixture of the reaction medium containing xylose fatty acid esters showed emulsifying properties similar to those of commercial surfactant. [Display omitted] • Xylose fatty acid esters production by degummed soybean oil hydroesterification. • Hydrolysis catalyzed by immobilized Eversa produced 92% of FFAs after 6 h. • Reuse of immobilized Eversa for five consecutive 3 h-batches of hydrolysis. • Modification of 85% xylose as monoester using immobilized TLL. • Xylose esters showed emulsifying properties similar to commercial surfactants. [ABSTRACT FROM AUTHOR]

Heterojunction photocatalysts receive significant interest due to their high performance and easy fabrication. An S-scheme heterojunction is developed on the basis of conventional type II heterojunctions, which can further promote charge separation and migration. In this work, a photocatalyst with a dual S-scheme heterojunction (denoted as C-s-ZIS/CN) is developed by the hybridization of CuInS2 quantum dot-sensitized ZnIn2S4 nanosheets and g-C3N4 nanosheets. CuInS2 quantum dot sensitization efficiently enhanced light absorption. Meanwhile, the unique charge migration pathway in the dual S-scheme heterojunction accelerated the separation and transfer of photogenerated carriers. The photocatalyst was innovatively employed in the simultaneous photocatalytic production of xylonic acid and CO. Under 10 W LED light irradiation (790 mW cm−2), the CO evolution rate reached a high of 228.94 μmol g−1 h−1. When a xenon lamp was employed, the CO evolution rate increased to 964.27 μmol g−1 h−1. Moreover, the optimal xylonic acid yield achieved was up to 58.56%. [ABSTRACT FROM AUTHOR]

In this study, bacterial cellulose (BC) was obtained from Komagataeibacter xylinus S4 and characterized in detail. The effects of a various of carbon sources and media, different pH conditions, incubation temperatures, Surface area/Volume ratios, and incubation durations were determined for BC production. Considering the carbon types, the amount of BC production from high to low was realized as sucrose, fructose, mannitol, xylose, arabinose, and lactose. The highest BC amount (1.303 g/L) was achieved by combining M1A05P5 broth, 30 °C, 1.06 cm-1 Surface area/Volume ratio, pH 3.5 and 21 days. According to scanning electron microscope (SEM) analysis, the cellulose fibril diameters were 34.87-45.97 nm at pH 3.5 and 29.71-102.3 nm at pH 6.5 in M1A05P5. Also, TGA analysis exhibited that the weight loss of BC in the removal of water step initialized between 50 °C and 150 °C and the degradation step initialized between 215 °C and 228 °C. Finally, the electrical conductivity values of the BC samples were determined on the 27-137 °C temperature scale. It was observed that the conductivity was temperature dependent, and the conductivity increased exponentially as the temperature increased. In conclusion, the cellulose from K. xylinus S4 typically showed a semiconducting behavior. [ABSTRACT FROM AUTHOR]

Catalytic hydrogenolysis of xylose to ethylene glycol (EG) and 1,2-propanediol (1,2-PG) is an important way for the high-value valorization of biomass. Herein, Ni-W/CeO2 catalysts prepared by incipient-wetness impregnation method were used for the hydrogenolysis of xylose to EG and 1,2-PG. Several characterizations such as X-ray diffraction (XRD), Transmission electron microscopy (TEM) and X-ray photoelectronic spectroscopy (XPS) were used to analyze the physicochemical properties of the catalysts. The effects of different Ni/W ratios and reaction conditions on the hydrogenolysis performance of the catalyst were investigated. The doping of W promoted the selective cleavage of C–C and C-O bonds of xylose, thus improved the yield of EG 1,2-PG. The doping of W can form interactions with Ni and increase the oxygen vacancy concentration of CeO2, which have a significant impact on the catalyst activity. When the Ni: W ratio is around 1:1, the catalyst has the best activity, and the total yield of diols can reach about 43.2%.. When xylan was used as the substrate, the total yield of diols can reach to 58.7%. The possible reaction mechanism of xylose hydrogenolysis was also discussed. This work developed a new catalyst system for the directional conversion of xylose to EG and 1,2-PG, which provided a new idea for the design of catalyst to converting lignocellulose, especially carbohydrate into high-value chemicals. Ni-W/CeO2 with different Ni/W ratio are efficient catalysts for hydrogenolysis of xylose to EG and 1,2-PG. [ABSTRACT FROM AUTHOR]

The present work models the fermentation process parameters of the newly isolated, Meyerozyma caribbica CP02 for enhanced xylitol production and its fermentability study on rice straw hydrolysate. The study examined the impact of each of the process variables by one variable at a time optimization followed by statistical validation. Temperature of 32 °C, pH of 3.5, agitation of 200 rpm, 1.5% (v/v) inoculum, 80 gL−1 initial xylose was optimized. Subsequently, a sequential two-stage agitation approach was adopted for fermentation. At these optimized conditions, xylitol yield of 0.77 gg−1 and 0.64 gg−1 was achieved using media containing commercial and rice straw derived xylose, respectively. For scale up, in 3L batch bioreactor, the highest xylitol yield (0.63 gg−1) was attained at 72 h with rice straw hydrolysate media containing initial xylose (59.48 ± 0.82 gL−1) along with inhibitors (1.55 ± 0.10 gL−1 aliphatic acids, 0.0.048 ± 0.11 gL−1 furans, 0.64 ± 0.23 gL−1 total phenols). The results imply that even under circumstances characterized by an acidic pH and elevated initial xylose level, M. caribbica CP02, as an isolate, displays robustness and shows favorable fermentability of rice straw hydrolysate. Therefore, isolate CP02 has potential to be used in bio-refineries for high yield xylitol production with minimal hydrolysate processing requirements. [ABSTRACT FROM AUTHOR]

Corn cob is a major waste mass-produced in corn agriculture. Corn cob hydrolysate containing xylose, arabinose, and glucose is the hydrolysis product of corn cob. Herein, a recombinant Escherichia coli strain BT-10 was constructed to transform corn cob hydrolysate into 1,2,4-butanetriol, a platform substance with diversified applications. To eliminate catabolite repression and enhance NADPH supply for alcohol dehydrogenase YqhD catalyzed 1,2,4-butanetriol generation, ptsG encoding glucose transporter EIICBGlc and pgi encoding phosphoglucose isomerase were deleted. With four heterologous enzymes including xylose dehydrogenase, xylonolactonase, xylonate dehydratase, α-ketoacid decarboxylase and endogenous YqhD, E. coli BT-10 can produce 36.63 g/L 1,2,4-butanetriol with a productivity of 1.14 g/[L·h] using xylose as substrate. When corn cob hydrolysate was used as the substrate, 43.4 g/L 1,2,4-butanetriol was generated with a productivity of 1.09 g/[L·h] and a yield of 0.9 mol/mol. With its desirable characteristics, E. coli BT-10 is a promising strain for commercial 1,2,4-butanetriol production. [ABSTRACT FROM AUTHOR]

Corn cob is a major waste mass-produced in corn agriculture. Corn cob hydrolysate containing xylose, arabinose, and glucose is the hydrolysis product of corn cob. Herein, a recombinant Escherichia coli strain BT-10 was constructed to transform corn cob hydrolysate into 1,2,4-butanetriol, a platform substance with diversified applications. To eliminate catabolite repression and enhance NADPH supply for alcohol dehydrogenase YqhD catalyzed 1,2,4-butanetriol generation, ptsG encoding glucose transporter EIICBGlc and pgi encoding phosphoglucose isomerase were deleted. With four heterologous enzymes including xylose dehydrogenase, xylonolactonase, xylonate dehydratase, α-ketoacid decarboxylase and endogenous YqhD, E. coli BT-10 can produce 36.63 g/L 1,2,4-butanetriol with a productivity of 1.14 g/[L·h] using xylose as substrate. When corn cob hydrolysate was used as the substrate, 43.4 g/L 1,2,4-butanetriol was generated with a productivity of 1.09 g/[L·h] and a yield of 0.9 mol/mol. With its desirable characteristics, E. coli BT-10 is a promising strain for commercial 1,2,4-butanetriol production. [ABSTRACT FROM AUTHOR]

Background: Cost-effective production of biofuels from lignocellulose requires the fermentation of d-xylose. Many yeast species within and closely related to the genera Spathaspora and Scheffersomyces (both of the order Serinales) natively assimilate and ferment xylose. Other species consume xylose inefficiently, leading to extracellular accumulation of xylitol. Xylitol excretion is thought to be due to the different cofactor requirements of the first two steps of xylose metabolism. Xylose reductase (XR) generally uses NADPH to reduce xylose to xylitol, while xylitol dehydrogenase (XDH) generally uses NAD+ to oxidize xylitol to xylulose, creating an imbalanced redox pathway. This imbalance is thought to be particularly consequential in hypoxic or anoxic environments. Results: We screened the growth of xylose-fermenting yeast species in high and moderate aeration and identified both ethanol producers and xylitol producers. Selected species were further characterized for their XR and XDH cofactor preferences by enzyme assays and gene expression patterns by RNA-Seq. Our data revealed that xylose metabolism is more redox balanced in some species, but it is strongly affected by oxygen levels. Under high aeration, most species switched from ethanol production to xylitol accumulation, despite the availability of ample oxygen to accept electrons from NADH. This switch was followed by decreases in enzyme activity and the expression of genes related to xylose metabolism, suggesting that bottlenecks in xylose fermentation are not always due to cofactor preferences. Finally, we expressed XYL genes from multiple Scheffersomyces species in a strain of Saccharomyces cerevisiae. Recombinant S. cerevisiae expressing XYL1 from Scheffersomyces xylosifermentans, which encodes an XR without a cofactor preference, showed improved anaerobic growth on xylose as the primary carbon source compared to S. cerevisiae strain expressing XYL genes from Scheffersomyces stipitis. Conclusion: Collectively, our data do not support the hypothesis that xylitol accumulation occurs primarily due to differences in cofactor preferences between xylose reductase and xylitol dehydrogenase; instead, gene expression plays a major role in response to oxygen levels. We have also identified the yeast Sc. xylosifermentans as a potential source for genes that can be engineered into S. cerevisiae to improve xylose fermentation and biofuel production. [ABSTRACT FROM AUTHOR]

As advances are made toward the industrial feasibility of mass-producing biofuels and commodity chemicals with sugar-fermenting microbes, high feedstock costs continue to inhibit commercial application. Hydrolyzed lignocellulosic biomass represents an ideal feedstock for these purposes as it is cheap and prevalent. However, many microbes, including Escherichia coli, struggle to efficiently utilize this mixture of hexose and pentose sugars due to the regulation of the carbon catabolite repression (CCR) system. CCR causes a sequential utilization of sugars, rather than simultaneous utilization, resulting in reduced carbon yield and complex process implications in fed-batch fermentation. A mutant of the gene encoding the cyclic AMP receptor protein, crp*, has been shown to disable CCR and improve the co-utilization of mixed sugar substrates. Here, we present the strain construction and characterization of a site-specific crp* chromosomal mutant in E. coli BL21 star (DE3). The crp* mutant strain demonstrates simultaneous consumption of glucose and xylose, suggesting a deregulated CCR system. The proteomics further showed that glucose was routed to the C5 carbon utilization pathways to support both de novo nucleotide synthesis and energy production in the crp* mutant strain. Metabolite analyses further show that overflow metabolism contributes to the slower growth in the crp* mutant. This highly characterized strain can be particularly beneficial for chemical production by simultaneously utilizing both C5 and C6 substrates from lignocellulosic biomass. [ABSTRACT FROM AUTHOR]

1,2,4-butanetriol (BT) is a polyol with unique chemical properties, which has a stereocenter and can be divided into D-BT (the S-enantiomer) and L-BT (the R-enantiomer). BT can be used for the synthesis of 1,2,4-butanetriol trinitrate, 3-hydroxytetrahydrofuran, polyurethane, and other chemicals. It is widely used in the military industry, medicine, tobacco, polymer. At present, the BT is mainly synthesized by chemical methods, which are accompanied by harsh reaction conditions, poor selectivity, many by-products, and environmental pollution. Therefore, BT biosynthesis methods with the advantages of mild reaction conditions and green sustainability have become a current research hotspot. In this paper, the research status of microbial synthesis of BT was summarized from the following three aspects: (1) the biosynthetic pathway establishment for BT from xylose; (2) metabolic engineering strategies employed for improving BT production from xylose; (3) other substrates for BT production. Finally, the challenges and prospects of biosynthetic BT were discussed for future methods to improve competitiveness for industrial production. [ABSTRACT FROM AUTHOR]

Corn straw is an abundant and renewable alternative for microbial biopolymer production. In this paper, an engineered Sphingomonas sanxanigenens NXG‐P916 capable of co‐utilising glucose and xylose from corn straw total hydrolysate to produce xanthan gum was constructed. This strain was obtained by introducing the xanthan gum synthetic operon gum as a module into the genome of the constructed chassis strain NXdPE that could mass produce activated precursors of polysaccharide, and in which the transcriptional levels of gum genes were optimised by screening for a more appropriate promoter, P916. As a result, strain NXG‐P916 produced 9.48 ± 0.34 g of xanthan gum per kg of fermentation broth (g/kg) when glucose was used as a carbon source, which was 2.1 times improved over the original engineering strain NXdPE::gum. Furthermore, in batch fermentation, 12.72 ± 0.75 g/kg xanthan gum was produced from the corn straw total hydrolysate containing both glucose and xylose, and the producing xanthan gum showed an ultrahigh molecular weight (UHMW) of 6.04 × 107 Da, which was increased by 15.8 times. Therefore, the great potential of producing UHMW xanthan gum by Sphingomonas sanxanigenens was proved, and the chassis NXdPE has the prospect of becoming an attractive platform organism producing polysaccharides derived from biomass hydrolysates. [ABSTRACT FROM AUTHOR]

BACKGROUND: Pistachio shell is a valuable lignocellulosic biomass because almost 90% of its hemicellulose fraction is xylan, which can be converted into high value‐added compounds such as xylooligosaccarides, xylose, xylitol and furfural. The present study represents a green and novel approach to produce xylose from lignocellulosic biomass. Microwave‐assisted high‐pressure CO2/H2O hydrolysis (MW‐HPCO2) comprising a combination never previously used was performed to produce xylose from pistachio shell. RESULTS: Response surface methodology with a Box–Behnken design was implemented to optimize microwave‐assisted high‐pressure CO2/H2O hydrolysis (MW‐HPCO2). The effect of temperature, time and liquid‐to‐solid ratio was studied in the ranges of 180–210 °C, 10–30 min and 5–30 mL g−1, respectively. A maximum xylose yield of 61.39% and minimum degradation compounds (5‐hydroxymethyl furfural and furfural) of 11.07% were attained under reaction conditions of 190 °C, 30 min and 18 mL g−1. CONCLUSION: The results showed that hydrolysis temperature, time and liquid‐to‐solid ratio had a strong influence on the xylose yield, as well as on the formation of degradation compounds. MW‐HPCO2 significantly increased accessibility to cellulose‐derived products in the subsequent enzymatic hydrolysis. The results of the present study reveal that MW‐HPCO2 can be a promising green technique for the hydrolysis of lignocellulosic biomass. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

During the normal production of ethanol from pretreated lignocellulosic biomass, only the solid fraction containing C6 sugars is generally used by unmodified yeasts in the fermentation process – the remaining liquid fraction, rich in xylose, is used almost exclusively by genetically modified organisms (GMOs) or for other purposes. As the liquor can be used in the manufacture of a series of chemical inputs, microorganisms that metabolize C5 sugars from different sources were initially isolated with the aim of converting these sugars into acetic acid (AA) and lactic acid (LA). Experiments were carried out with pretreated elephant grass liquor, evaluating the effect of adding commonly referenced nutrients through experimental design. Thus, the present work involves the isolation, screening, and molecular characterization of bacteria, easily found in nature, with the potential to metabolize xylose and produce AA and LA, with yields similar or superior to those shown in studies employing GMOs. Using wild‐type strains, high yields were obtained with Acetobacter cerevisiae (0.96 g LA and 0.71 g AA per g total sugars) and Levilactobacillus brevis (0.77 g LA and 0.61 g AA per g total sugars) in relation to the organic acids of interest, highlighting the total conversion of the sugar blend (xylose, glucose, and arabinose) in 24 h. Ammonium citrate also significantly influenced the final yields, being responsible for the increase in production. The results are useful for biorefinery platforms and may contribute to the full use of sugars released in the lignocellulosic pretreatment of raw materials. [ABSTRACT FROM AUTHOR]

Anthranilate and its derivatives are important basic chemicals for the synthesis of polyurethanes as well as various dyes and food additives. Today, anthranilate is mainly chemically produced from petroleum‐derived xylene, but this shikimate pathway intermediate could be also obtained biotechnologically. In this study, Corynebacterium glutamicum was engineered for the microbial production of anthranilate from a carbon source mixture of glucose and xylose. First, a feedback‐resistant 3‐deoxy‐arabinoheptulosonate‐7‐phosphate synthase from Escherichia coli, catalysing the first step of the shikimate pathway, was functionally introduced into C. glutamicum to enable anthranilate production. Modulation of the translation efficiency of the genes for the shikimate kinase (aroK) and the anthranilate phosphoribosyltransferase (trpD) improved product formation. Deletion of two genes, one for a putative phosphatase (nagD) and one for a quinate/shikimate dehydrogenase (qsuD), abolished by‐product formation of glycerol and quinate. However, the introduction of an engineered anthranilate synthase (TrpEG) unresponsive to feedback inhibition by tryptophan had the most pronounced effect on anthranilate production. Component I of this enzyme (TrpE) was engineered using a biosensor‐based in vivo screening strategy for identifying variants with increased feedback resistance in a semi‐rational library of TrpE muteins. The final strain accumulated up to 5.9 g/L (43 mM) anthranilate in a defined CGXII medium from a mixture of glucose and xylose in bioreactor cultivations. We believe that the constructed C. glutamicum variants are not only limited to anthranilate production but could also be suitable for the synthesis of other biotechnologically interesting shikimate pathway intermediates or any other aromatic compound derived thereof. [ABSTRACT FROM AUTHOR]

Areca nut husk is the most promising alternative source of low-cost raw materials because it contains a considerable amount of five-carbon monosaccharide sugar in the form of xylose. This polymeric sugar can be isolated and transformed into a value-added chemical using fermentation. To extract sugars from areca nut husk fibers, preliminary pretreatment, such as dilute acid hydrolysis (H2SO4), was performed. The hemicellulosic hydrolysate of areca nut husk can produce xylitol through fermentation, but toxic components inhibit the growth of microorganisms. To overcome this, a series of detoxification treatments, including pH adjustment, activated charcoal, and ion exchange resin, were carried out to reduce the concentration of inhibitors in the hydrolysate. This study reports a remarkable 99% removal of inhibitors in the hemicellulosic hydrolysate. Subsequently, a fermentation process using Candida tropicalis (MTCC6192) was executed with the detoxified hemicellulosic hydrolysate of areca nut husk, yielding an optimum xylitol yield of 0.66 g/g. This study concludes that detoxification techniques like pH adjustment, activated charcoal, and ion exchange resins are the most economical and effective methods for eliminating toxic compounds in hemicellulosic hydrolysates. Therefore, the medium derived after detoxification from areca nut hydrolysate may be considered to have significant potential for xylitol production. [ABSTRACT FROM AUTHOR]

[Display omitted] • Enhanced oxidation capacity of Mn-Cd-S photocatalysts by bandgap engineering. • Cogeneration of high value-added chemicals and clean energy from biomass. • Innovatively selective oxidation of xylose to C3 organic acids by modulating photocatalyst band structure. • Possible pathways and mechanisms for photoreforming xylose conversion. The simultaneous reforming of biomass into high value-added chemicals and H 2 production by water splitting in a green and environmentally clean way is a very challenging task. Herein, we demonstrate the design of bifunctional Mn x Cd 1-x S photocatalyst with a controllable band gap by bandgap engineering. Bandgap engineering effectively regulates the oxidation and reduction capacity of materials. The design of photocatalysts with suitable conduction bands and valence bands makes the targeted conversion of xylose possible. Innovative conversion of xylose to glyceric acid, lactic acid, and propanoic acid. The optimized Mn 0.7 Cd 0.3 S catalyst showed excellent performance in the production of H 2 (14.06 mmol·g cat -1·h−1, 29.9 times more than CdS and 351.5 times more than MnS), xylose conversion (90%), and C3 organic acid yield (59.2%) without cocatalyst and any scavengers under visible light irradiation. This work shows that a rational photocatalyst design can achieve efficient simultaneous production of high value-added chemicals and clean energy. [ABSTRACT FROM AUTHOR]

Cordycepin, a nucleoside analog, is the main antioxidative and antimicrobial substance in Cordyceps militaris. To improve the metabolism of cordycepin, carbon sources, nitrogen sources, trace elements, and precursors were studied by single factor, Plackett–Burman, and central composite designs in C. militaris mycelial fermentation. Under the regulation of the multifactorial interactions of selenite, ferrous chloride, xylose, and glycine, cordycepin production was increased by 5.2-fold compared with the control. The gene expression of hexokinase, ATP phosphoribosyltransferase, adenylosuccinate synthetase, and cns1-3 in the glycolysis, pentose phosphate, and adenosine synthesis pathways were increased by 3.2–7.5 times due to multifactorial interactions, while the gene expression of histidine biosynthesis trifunctional protein and histidinol-phosphate aminotransferase in histidine synthesis pathway were decreased by 23.4%-56.2%. Increasing with cordycepin production, glucose uptake was accelerated, mycelia growth was inhibited, and the cell wall was damaged. Selenomethionine (SeMet), selenocysteine (SeCys), and selenium nanoparticles (SeNPs) were the major Se species in C. militaris mycelia. This study provides a new insight for promoting cordycepin production by regulating glycolysis, pentose phosphate, and histidine metabolism. Key points: • Cordycepin production in the CCDmax group was 5.2-fold than that of the control. • Glucose uptake of the CCDmax group was accelerated and cell wall was damaged. • The metabolic flux was concentrated to the cordycepin synthesis pathway. [ABSTRACT FROM AUTHOR]

Surfactin is an excellent biosurfactant with a wide range of application prospects in many industrial fields. However, its low productivity and high cost have largely limited its commercial applications. In this review, the pathways for surfactin synthesis in Bacillus strains are summarized and discussed. Further, the latest strategies for improving surfactin production, including: medium optimization, genome engineering methods (rational genetic engineering, genome reduction, and genome shuffling), heterologous synthesis, and the use of synthetic biology combined with metabolic engineering approaches to construct high-quality artificial cells for surfactin production using xylose, are described. Finally, the prospects for improving surfactin synthesis are discussed in detail. [ABSTRACT FROM AUTHOR]

Background: Significant genetic diversity exists across Saccharomyces strains. Natural isolates and domesticated brewery and industrial strains are typically more robust than laboratory strains when challenged with inhibitory lignocellulosic hydrolysates. These strains also contain genes that are not present in lab strains and likely contribute to their superior inhibitor tolerance. However, many of these strains have poor sporulation efficiencies and low spore viability making subsequent gene analysis, further metabolic engineering, and genomic analyses of the strains challenging. This work aimed to develop an inhibitor tolerant haploid with stable mating type from S. cerevisiae YB-2625, which was originally isolated from bagasse. Results: Haploid spores isolated from four tetrads from strain YB-2625 were tested for tolerance to furfural and HMF. Due to natural mutations present in the HO-endonuclease, all haploid strains maintained a stable mating type. One of the haploids, YRH1946, did not flocculate and showed enhanced tolerance to furfural and HMF. The tolerant haploid strain was further engineered for xylose fermentation by integration of the genes for xylose metabolism at two separate genomic locations (ho∆ and pho13∆). In fermentations supplemented with inhibitors from acid hydrolyzed corn stover, the engineered haploid strain derived from YB-2625 was able to ferment all of the glucose and 19% of the xylose, whereas the engineered lab strains performed poorly in fermentations. Conclusions: Understanding the molecular mechanisms of inhibitor tolerance will aid in developing strains with improved growth and fermentation performance using biomass-derived sugars. The inhibitor tolerant, xylose fermenting, haploid strain described in this work has potential to serve as a platform strain for identifying pathways required for inhibitor tolerance, and for metabolic engineering to produce fuels and chemicals from undiluted lignocellulosic hydrolysates. [ABSTRACT FROM AUTHOR]

Sustainable and renewable sources of liquid and solid fuels are essential to prevent fossil fuel use from damaging the environment. Secondary agricultural residues, which are already transported to food processing centers, have great potential to be converted into biofuels. The wastes from coffee roasting, sugar production, and rice milling have been investigated using hydrothermal carbonization (HTC) to produce aqueous products containing monosaccharides alongside solid biofuels. These sugar-laden liquid products were characterized after pretreating coffee silverskins, sugarcane bagasse, and rice husks with HTC. They were then concentrated using direct contact membrane distillation (DCMD), a low-energy process that can use waste heat from other biorefinery processes. The higher heating value of the solid products was also characterized by bomb calorimetry. The liquid products from HTC of these wastes from food production were found to contain varying concentrations of glucose, xylose, galactose, and arabinose. DCMD was capable of concentrating the liquid products up to three times their original concentrations. Little difference was found among the higher heating values of the solid products after 180 °C HTC pretreatment compared to 200 °C pretreatment. HTC of waste from food processing can provide solid biofuels and liquid products containing sugars that can be concentrated using DCMD. [ABSTRACT FROM AUTHOR]

In this study, coal fly ash was functionalized, using a simple one-step process (loading with Al3+ and sulfonation), to yield a solid acid catalyst (S/Al-CFA) with strong acid sites. The catalyst was then used to produce furfural from xylose in a biphasic system (H2O(NaCl)/tetrahydrofuran). The furfural yield reached 82% at 180 °C–60 min with catalyst/xylose ratio of 0.2:1.0 (w/w). With the reaction completed, all of the components could be effectively separated, and the furfural was 97.6% pure. The cycle and regeneration of the catalyst were evaluated, and the catalyst deactivation mechanism was investigated. [ABSTRACT FROM AUTHOR]

Despite recent improvements, second-generation (2G) biorefineries still have challenges to overcome, especially regarding cellulose and xylan hydrolysis and xylose fermentation. Hydrolases account for a high percentage of the process' total cost, and industrial yeasts may not assimilate xylose properly. In this sense, prospecting microorganisms that can both metabolize this sugar and hydrolyze cellulose and xylan may lead to significant advances in 2G processes. Since the gut microbiota of herbivorous insects has proven to be the major player in their lignocellulose digestion, yeasts were isolated from the gut of Spodoptera frugiperda larvae, and their cellulolytic, xylanolytic, and xylose-assimilating capacity were analyzed. From a hundred larvae guts, forty-six yeasts were selected for their growth capacity in xylose-containing media. Fourteen strains were able to hydrolyze cellulose, and three showed xylanolytic activity. Two strains, taxonomically identified as Papiliotrema laurentii and Meyerozyma caribbica, stood out with the highest cellular biomass yield and productivity when submitted to a high-throughput microscale growth analysis under different culture conditions (varying pH and sugar concentration). While the P. laurentii strain displayed the highest xylanolytic activity (205.48 U/mL) and an intermediate cellulolytic capacity (31.25 U/mL), the M. caribbica strain showed, besides cellulase activity (37.46 U/mL), high ethanol and xylitol yields (0.36 gethanol/gglucose and 0.35 gxylitol/gxylose, respectively) in media with the most common fermentation inhibitors in 2G bioprocesses. Through a Central Composite Rotatable Design, the optimized condition for the simultaneous production of ethanol and xylitol was found to be in a 55–85 g/L range of sugar concentration and between pH values of 6.0 to 8.5. Also, this experimental design analysis allowed the validation of empiric-coded models. These results indicate the high biotechnological potential of prospecting yeasts in S. frugiperda guts. [ABSTRACT FROM AUTHOR]

Background: Yarrowia lipolytica, one of the most charming chassis cells in synthetic biology, is unable to use xylose and cellodextrins. Results: Herein, we present work to tackle for the first time the engineering of Y. lipolytica to produce lipids from cellodextrins and xylose by employing rational and combinatorial strategies. This includes constructing a cellodextrin-phosphorolytic Y. lipolytica by overexpressing Neurospora crassa cellodextrin transporter, Clostridium thermocellum cellobiose/cellodextrin phosphorylase and Saccharomyces cerevisiae phosphoglucomutase. The effect of glucose repression on xylose consumption was relieved by installing a xylose uptake facilitator combined with enhanced PPP pathway and increased cytoplasmic NADPH supply. Further enhancing lipid production and interrupting its consumption conferred the obese phenotype to the engineered yeast. The strain is able to co-ferment glucose, xylose and cellodextrins efficiently, achieving a similar μmax of 0.19 h−1, a qs of 0.34 g-s/g-DCW/h and a YX/S of 0.54 DCW-g/g-s on these substrates, and an accumulation of up to 40% of lipids on the sugar mixture and on wheat straw hydrolysate. Conclusions: Therefore, engineering Y. lipolytica capable of assimilating xylose and cellodextrins is a vital step towards a simultaneous saccharification and fermentation (SSF) process of LC biomass, allowing improved substrate conversion rate and reduced production cost due to low demand of external glucosidase. [ABSTRACT FROM AUTHOR]

The conventional fractionation of lignocellulose into cellulose, hemicellulose, and lignin was generally carried out at high temperatures and under highly acidic conditions owing to the rigid matrix of lignocellulose and crystallinity of cellulose, causing the degradation of carbohydrates and condensation of lignin, thus hindering their value-added utilization. In this study, a stepwise fractionation method was proposed in which microwave-assisted fractionation of hemicellulose under mild conditions contributed to 81.9% of xylose yield; then, the cellulose part solid was rapidly dissolved in LiBr molten salt hydrate (LiBr-MSH) solution, followed by a filtration process to isolate lignin solid. It was found that the removal of hemicellulose provided more pores for the next dissolution of cellulose, and LiBr-MSH caused the cleavage of the hydrogen bonds of cellulose. Consequently, the cellulose part solid was rapidly dissolved in LiBr-MSH. The results showed that under the optimal fractionation conditions, 78.0% yield of glucan oligomers and 89.7% purity of noncondensed lignin were obtained from cellulose fractionation and filtration, respectively. A high β-O-4 bond content close to 87.3% of the theoretical maximum yield was detected in the obtained lignin, indicating that most β-O-4 bond structure of lignin was well preserved. Additionally, the isolated lignin can be selectively depolymerized into aromatic monomers with a high yield of 20.4%, which is 74.2% of the theoretical maximum yield. [ABSTRACT FROM AUTHOR]

Background: Over the last decades, many strategies to procure and improve xylose consumption in Saccharomyces cerevisiae have been reported. This includes the introduction of efficient xylose-assimilating enzymes, the improvement of xylose transport, or the alteration of the sugar signaling response. However, different strain backgrounds are often used, making it difficult to determine if the findings are transferrable both to other xylose-consuming strains and to other xylose-assimilation pathways. For example, the influence of anomerization rates between α- and β-xylopyranose in pathway optimization and sugar sensing is relatively unexplored. Results: In this study, we tested the effect of expressing a xylose epimerase in S. cerevisiae strains carrying different xylose-consuming routes. First, XIs originating from three different species in isogenic S. cerevisiae strains were tested and the XI from Lachnoclostridium phytofermentans was found to give the best performance. The benefit of increasing the anomerization rate of xylose by adding a xylose epimerase to the XI strains was confirmed, as higher biomass formation and faster xylose consumption were obtained. However, the impact of xylose epimerase was XI-dependent, indicating that anomer preference may differ from enzyme to enzyme. The addition of the xylose epimerase in xylose reductase/xylitol dehydrogenase (XR/XDH)-carrying strains gave no improvement in xylose assimilation, suggesting that the XR from Spathaspora passalidarum had no anomer preference, in contrast to other reported XRs. The reduction in accumulated xylitol that was observed when the xylose epimerase was added may be associated with the upregulation of genes encoding endogenous aldose reductases which could be affected by the anomerization rate. Finally, xylose epimerase addition did not affect the sugar signaling, whereas the type of xylose pathway (XI vs. XR/XDH) did. Conclusions: Although xylose anomer specificity is often overlooked, the addition of xylose epimerase should be considered as a key engineering step, especially when using the best-performing XI enzyme from L. phytofermentans. Additional research into the binding mechanism of xylose is needed to elucidate the enzyme-specific effect and decrease in xylitol accumulation. Finally, the differences in sugar signaling responses between XI and XR/XDH strains indicate that either the redox balance or the growth rate impacts the SNF1/Mig1p sensing pathway. [ABSTRACT FROM AUTHOR]

Starting from 3,4,5-trimethoxyacetophenone and 3,5-dimethoxy-4-hydroxyacetophenone (acetosyringone), 4-(3,4,5-trimethoxyphenyl)- and 4-(3,5-dimethoxy-4-hydroxyphenyl)-1,2,3-thiadiazoles and selenodiazoles were synthesized. Trimethoxy derivatives are stable, whereas dimethoxyhydroxy compounds decompose in slightly alkaline media under the action of oxygen traces. In contrast, acetates of β-glucoside, β-galactoside, and β-xyloside acetosyringone formed carbethoxyhydrazones and semicarbazones. Carboethoxyhydrazones, when treated with thionyl chloride, converted to corresponding 1,2,3-thiadiazoles. In case of oxidation of semicarbazones with selenium dioxide, 1,2,3-selenodiazoles were formed only from the xylose and galactose derivatives. [ABSTRACT FROM AUTHOR]

Lignocellulosic biomass represents a carbon neutral cheap and versatile source of carbon which can be converted to biofuels. A pretreatment step is frequently used to make the lignocellulosic carbon bioavailable for microbial metabolism. Dilute acid pretreatment at high temperature and pressure is commonly utilized to efficiently solubilize the pentose fraction by hydrolyzing the hemicellulose fibers and the process results in formation of furans—furfural and 5-hydroxymethyl furfural—and other inhibitors which are detrimental to metabolism. The presence of inhibitors in the medium reduce productivity of microbial biocatalysts and result in increased production costs. Furfural is the key furan inhibitor which acts synergistically along with other inhibitors present in the hydrolysate. In this review, the mode of furfural toxicity on microbial metabolism and metabolic strategies to increase tolerance is discussed. Shared cellular targets between furfural and acetic acid are compared followed by discussing further strategies to engineer tolerance. Finally, the possibility to use furfural as a model inhibitor of dilute acid pretreated lignocellulosic hydrolysate is discussed. The furfural tolerant strains will harbor an efficient lignocellulosic carbon to pyruvate conversion mechanism in presence of stressors in the medium. The pyruvate can be channeled to any metabolite of interest by appropriate modulation of downstream pathway of interest. The aim of this review is to emphasize the use of hydrolysate as a carbon source for bioproduction of biofuels and other compounds of industrial importance. [ABSTRACT FROM AUTHOR]

Water-extractable arabinoxylan (WEAX) may cause major problems during clarification processes in a brewery owing to its ability to form gel networks. However, high WEAX contents can also enhance the nutritional quality of the final product as they play an important role in the human diet. Therefore, precise quantification of WEAX is required. Current methods are very time- and resource-consuming as well as limited in the number of samples and in some cases provide low accuracy. Thus, a reproducible high-throughput method for the quantification of WEAX optimized for beer was developed, reaching recovery rates (RRs) of almost 100%. The assay is based on Douglas's colorimetric method. Hydrolysis was conducted using glacial acetic acid to induce the formation of red color complexes resulting from the interaction between pentose degradation products and phloroglucinol. The method was successfully transferred to a multi-mode microplate reader to minimize the loss of color intensity over time and to obtain a high throughput. By using 96-well plates, up to 40% of the previous analysis time could be saved, and a larger number of samples could be analyzed in one batch. The collected data determined xylose as an optimal calibration standard due to high accuracy and reproducibility. The respective AX control standards showed RR within the range of 95–105% without exception. To validate and show the ruggedness of the modified method, WEAX concentration in seven commercial German beers (e.g., lager, pilsner, wheat beer, non-alcoholic beer) was quantified. Interfering hexose sugars that lead to measurement errors when analyzing samples with high amounts of fermentable sugars (e.g., non-alcoholic beer produced by limited fermentation) were eliminated by Saccharomyces diastaticus fermentation. Further investigations were carried out by means of LC-MS in order to obtain additional information about the reddish product in the hydrolyzed samples. In this context, C16H12O6 could be identified as one of numerous condensation products, contributing to the coloring. The collected data showed the impact of diverse factors on the measured AX concentration and helped optimize the experimental procedure for a high sample throughput with precise and highly reproducible results. The proposed quantification method should be primarily used in completely fermented finished beer to emphasize the time aspect. Wort samples and non-alcoholic beer produced by limited fermentation can be also analyzed, but only after fermentation with S. diastaticus. [ABSTRACT FROM AUTHOR]

Carbon dots (CDs) are interesting carbon nanomaterials that exhibit great photoluminescent features, low cytotoxicity, and excellent water stability and solubility. For these reasons, many fields are starting to integrate their use for a variety of purposes. The catalytic performance of VOPO4 has been evaluated in the synthesis of nitrogen-doped carbon dots (N-CDs). The synthesis reaction was carried out at 180 °C using VOPO4 as a heterogeneous catalyst for 2 to 4 h of reaction time. After reaction, the N-CDs were purified using a novel method for the protection of the functional groups over the surfaces of the N-CDs. The morphological, superficial, and photoelectronic properties of the N-CDs were thoroughly studied by means of TEM, HRTEM, XPS, and photoluminescence measurements. The conversion of the carbon precursor was followed by HPLC. After three catalytic runs, the catalyst was still active while ensuring the quality of the N-CDs obtained. After the third cycle, the catalyst was regenerated, and it recovered its full activity. The obtained N-CDs showed a great degree of oxidized groups in their surfaces that translated into high photoluminescence when irradiated under different lasers. Due to the observed photoelectronic properties, they were then assayed in the photocatalytic degradation of methyl orange. [ABSTRACT FROM AUTHOR]

D-Xylitol, a biomass-derived sweetener, is increasingly used in cosmetics and pharmaceutical products. The raw material for D-xylitol production, Dxylose, is easily accessible from dissolving pulp production. D-xylitol production involves the heterogeneously catalyzed hydrogenation of Dxylose; this process is energy intensive, as the use of H2 requires high pressure and temperature. This work examined catalytic transfer hydrogenation for xylose conversion into xylitol. Formic acid (FA) was used to replace H2 as the H-donor, as it is easily available, inexpensive, may be obtained from renewable sources, and it avoids the risks associated with the use of high-pressure inflammable gas. A variety of commercially available catalysts were screened to reveal the one enabling the highest yield. The experiments were performed at 40, 80, and 140 °C, with pure xylose as a model compound. Triethylamine (Et3N) was added to ensure sufficient conversion rates. Based on the preliminary studies an experimental design was created (Design Expert®), including the two best performing catalysts Ru/Al2O3 and Ru/C, to investigate the influence of temperature and H-donor and base concentration on xylitol yield. Ru/C resulted in maximum D-xylitol yield of 73.2 % at 100 °C, FA to D-xylose ratio 5:1 and Et3N to FA ratio 0.4. [ABSTRACT FROM AUTHOR]

Cellulose and hemicellulose are major components of woody biomass, pyrolysis of which would inevitably produce anhydrate sugars or sugar-like organics. These sugars might tailor property of biochar via interactions when they pass through inner and outer surface of biochar, which were investigated herein in pyrolysis of poplar sawdust impregnated with various loading of xylose, a model sugar from hemicellulose, at 600 °C. The results suggested that the addition of xylose enhanced the cracking reactions to form more gaseous products but less biochar and bio-oil. Nevertheless, the intermediates from xylose enhanced aromatization of volatiles to form more light phenolics while aided precipitation of heavy organics on surface of biochar. The accelerated carbonization, deoxygenation and dehydrogenation reactions with presence of xylose enhanced aromatization reactions, making the biochar carbon-rich with increased heating value, crystallinity and hydrophobicity. Additionally, xylose addition could substantially enhance thermal stability of the biochar, even with loading of only 5 wt%. The DRIFTS characterization showed that the degradation of xylose produced abundant carbonyls-containing species that react with organics on surface of biochar, forming large aromatics of fused ring structures. [Display omitted] [ABSTRACT FROM AUTHOR]

Banana fiber is a rich lignocellulosic biomass source that has not been widely explored. The hemicellulose components (15 - 20 %) of banana fiber can be a feedstock for producing high-value commodity chemicals. Hemicellulose is extracted by physical, chemical, and biological methods, in which combining hydrothermal treatment with alkaline mode of extraction provides an enhanced recovery percentage. Thus, the present study aimed to optimize the hydrothermal-assisted alkaline method of xylan extraction from the banana fiber biomass. Initially, xylan was extracted with a conventional-based alkali method. A maximum of about 43 and 35 % was recovered from pretreated and raw banana fiber at 12% NaOH concentration when incubated at 55 °C for 24 h. To improve the xylan yield, the hydrothermal assisted alkali method experimented in which 67.1% and 58.3 % of xylan were recovered when treated at 121 °C for 1 h at 12% NaOH. To further enhance the xylan recovery, a two-step alkali process by combining conventional and hydrothermal-assisted alkali methods resulted in the highest xylan (81%) recovery from pretreated banana fiber when incubated with 12 % alkali for 8 h followed by steam treatment. On the other hand, a maximum of 73 % of xylan was recovered when steam treated after incubation for 24 h from raw banana fiber. Thus, the alkali incubation followed by steam treatment significantly showed the highest xylan recovery from the banana fiber biomass. The extracted xylan might be utilized as a source for various xylan-based products, including furfural, xylooligosaccharides, xylose, and xylitol, all of which have significant roles in the pharmaceutical and food industries. [ABSTRACT FROM AUTHOR]

The synthesis of sugar fatty acid esters (SFAEs) from lignocellulosic biomass and oleic acid (C18:1) was catalyzed by immobilized‐stabilized derivatives of Candida antarctica lipase B in a methyl ethyl ketone medium. After steam‐explosion pretreatment of mixed hardwoods and enzymatic hydrolysis at 15%wt solids, xylose and glucose were purified/concentrated to a mass ratio of ~3 to 1. These lignocellulosic sugars were superior to commercial sugars as the carbohydrate source for the esterification reaction in terms of sugar conversions. The highest conversions were obtained using 1.5% w/v of Novozyme 435 (N435, uncoated) as the biocatalyst for the synthesis of SFAEs. Coating the N435 with polyethyleneimine (PEI) prevented enzyme leakage into the reaction medium and produced 35% and 50% higher xylose and glucose conversions to SFAEs, respectively, at the same enzyme loading. After six 24 h reuse cycles with the PEI‐coated N435, xylose conversion decreased by 44%, while a 65% reduction in xylose conversion was observed with the uncoated lipase. Mass spectrometry analysis confirmed the production of xylose and glucose mono‐ and di‐esters. Our purified product presented an emulsion capacity (EC) close to that of a commercial sugar ester and the ECs of the xylose oleate, laurate, and palmitate synthesized in previous studies. © 2023 The Authors. Biofuels, Bioproducts and Biorefining published by Society of Industrial Chemistry and John Wiley & Sons Ltd. [ABSTRACT FROM AUTHOR]

Hydrogen (H 2) is a promising energy vector for mitigating greenhouse gas emissions. Lignocellulosic biomass waste has been introduced as one of the abundant and carbon-neutral H 2 sources. Among those, xylose with its short carbon chain has emerged attractive, where H 2 can be catalytically released in an aqueous reactor. In this study, a composite catalyst system consisting of silica (SiO 2)-supported platinum (Pt)-cobalt (Co) bimetallic nanoparticles was developed for aqueous phase reforming of xylose conducted at 225 °C and 29.3 bar. The PtCo/SiO 2 catalyst showed a significantly higher H 2 production rate and selectivity than that of Pt/SiO 2 , whereas Co/SiO 2 shows no activity in H 2 production. The highest selectivity for useful liquid byproducts was obtained with PtCo/SiO 2. Moreover, CO 2 emissions throughout the reaction were reduced compared to those of monometallic Pt/SiO 2. The PtCo bimetallic nanocatalyst offers an inexpensive, sustainable, and durable solution with high chemical selectivity for scalable reforming of hard-to-ferment pentose sugars. [Display omitted] • Silica (SiO 2)-supported PtCo bimetallic nano-catalyst was tested for APR of xylose. • PtCo/SiO 2 demonstrated higher H 2 selectivity than monometallic Pt/SiO 2 and Co/SiO 2. • Co/SiO 2 showed no catalytic activity for the APR of xylose. • PtCo catalyst produced ethylene glycol, resulting in a higher H 2 production rate. • DFT calculations confirmed that the surface layer of Co atoms enhances the activity. [ABSTRACT FROM AUTHOR]

Background: Poly-β-hydroxybutyrate (PHB), produced by a variety of microbial organisms, is a good substitute for petrochemically derived plastics due to its excellent properties such as biocompatibility and biodegradability. The high cost of PHB production is a huge barrier for application and popularization of such bioplastics. Thus, the reduction of the cost is of great interest. Using low-cost substrates for PHB production is an efficient and feasible means to reduce manufacturing costs, and the construction of microbial cell factories is also a potential way to reduce the cost. Results: In this study, an engineered Sphingomonas sanxanigenens strain to produce PHB by blocking the biosynthetic pathway of exopolysaccharide was constructed, and the resulting strain was named NXdE. NXdE could produce 9.24 ± 0.11 g/L PHB with a content of 84.0% cell dry weight (CDW) using glucose as a sole carbon source, which was significantly increased by 76.3% compared with the original strain NX02. Subsequently, the PHB yield of NXdE under the co-substrate with different proportions of glucose and xylose was also investigated, and results showed that the addition of xylose would reduce the PHB production. Hence, the Dahms pathway, which directly converted D-xylose into pyruvate in four sequential enzymatic steps, was enhanced by overexpressing the genes xylB, xylC, and kdpgA encoding xylose dehydrogenase, gluconolactonase, and aldolase in different combinations. The final strain NX02 (ΔssB, pBTxylBxylCkdpgA) (named NXdE II) could successfully co-utilize glucose and xylose from corn straw total hydrolysate (CSTH) to produce 21.49 ± 0.67 g/L PHB with a content of 91.2% CDW, representing a 4.10-fold increase compared to the original strain NX02. Conclusion: The engineered strain NXdE II could co-utilize glucose and xylose from corn straw hydrolysate, and had a significant increase not only in cell growth but also in PHB yield and content. This work provided a new host strain and strategy for utilization of lignocellulosic biomass such as corn straw to produce intracellular products like PHB. [ABSTRACT FROM AUTHOR]

Background: The non-conventional yeast Rhodotorula toruloides is an emerging host organism in biotechnology by merit of its natural capacity to accumulate high levels of carotenoids and intracellular storage lipids from a variety of carbon sources. While the number of genetic engineering strategies that employ R. toruloides is increasing, the lack of genetic tools available for modification of this yeast is still limiting strain development. For instance, several strong, constitutive R. toruloides promoters have been characterized, but to date, only five inducible promoters have been identified. Although nitrogen-limited cultivation conditions are commonly used to induce lipid accumulation in this yeast, no promoters regulated by nitrogen starvation have been described for R. toruloides. Results: In this study, we used a combination of genomics and transcriptomics methods to identify novel R. toruloides promoter sequences that are either inducible or repressible by nitrogen starvation. RNA sequencing was used to assess gene expression in the recently isolated strain R. toruloides BOT-A2 during exponential growth and during nitrogen starvation, when cultivated with either glucose or xylose as the carbon source. The genome of BOT-A2 was sequenced using a combination of long- and short-read sequencing and annotated with support of the RNAseq data. Differential expression analysis was used to identify genes with a |log2 fold change|≥ 2 when comparing their expression during nitrogen depletion to that during exponential growth. The promoter regions from 16 of these genes were evaluated for their ability to drive the expression of a fluorescent reporter gene. Three promoters that were clearly upregulated under nitrogen starvation and three that were downregulated were selected and further characterized. One promoter, derived from gene RTBOTA2_003877, was found to function like an on–off switch, as it was only upregulated under full nitrogen depletion and downregulated in the presence of the nitrogen source. Conclusions: Six new R. toruloides promoters that were either upregulated or downregulated under nitrogen-starvation were identified. These substantially contribute to the available promoters when engineering this organism and are foreseen to be particularly useful for future engineering strategies requiring specific regulation of target genes in accordance with nitrogen availability. [ABSTRACT FROM AUTHOR]

The catalytic effect of Lewis acid on xylose conversion to furfural has been widely reported, while the underlying reaction network and kinetics are not fully elucidated. This work presents experimental and kinetic modelling studies on xylose conversion to furfural, using AlCl3/HCl as combined Lewis/Brønsted acid catalysts in monophasic water or a biphasic water–methyl isobutyl ketone system in a flow microreactor. The reaction network and kinetics were developed, where Al(OH)2+ was identified as the catalytically active species for isomerization between xylose, lyxose and xylulose, while both Al(OH)2+ and H+ catalyze the sugar dehydration to furfural and side reactions leading to humins (e.g., sugar condensation and furfural degradation). The promoting effect of AlCl3 is attributed to not only the tandem catalysis through xylulose, but also the parallel Al(OH)2+-catalyzed sugar dehydration. Within the studied range (120 to 180 °C, 0.05–0.4 M HCl, 0.04–0.12 M AlCl3), the furfural selectivity relied mainly on the concentration ratio of HCl to AlCl3 rather than their individual concentrations or the temperature. The volcano-like evolution of the maximum furfural yield with increasing HCl/AlCl3 ratio is related to (i) the lower reaction orders in Al(OH)2+ for reactions forming furfural than for side reactions; and (ii) a gradual shift of the dominant reaction pathway from the Al(OH)2+-catalyzed one towards H+-catalyzed one. An optimized furfural yield up to 90% was achieved with 40 mM AlCl3 and 100 mM HCl at 160 °C from 1 M xylose in a continuous slug flow microreactor. The catalytic aqueous phase could be recycled and reused four times without performance loss. [ABSTRACT FROM AUTHOR]

Seven Kluyveromyces marxianus isolates from the elaboration process of pulque and henequen mezcal were characterized. The isolates were identified based on the sequences of the D1/D2 domain of the 26S rRNA gene and the internal transcribed spacer (ITS-5.8S) region. Genetic differences were found between pulque and henequen mezcal isolates and within henequen mezcal isolates, as shown by different branching patterns in the ITS-5.8S phylogenetic tree and (GTG)5 microsatellite profiles, suggesting that the substrate and process selective conditions may give rise to different K. marxianus populations. All the isolates fermented and assimilated inulin and lactose and some henequen isolates could also assimilate xylose and cellobiose. Henequen isolates were more thermotolerant than pulque ones, which, in contrast, presented more tolerance to the cell wall-disturbing agent calcofluor white (CFW), suggesting that they had different cell wall structures. Additionally, depending on their origin, the isolates presented different maximum specific growth rate (µmax) patterns at different temperatures. Concerning tolerance to stress factors relevant for lignocellulosic hydrolysates fermentation, their tolerance limits were lower at 42 than 30 °C, except for glucose and furfural. Pulque isolates were less tolerant to ethanol, NaCl, and Cd. Finally, all the isolates could produce ethanol by simultaneous saccharification and fermentation (SSF) of a corncob hydrolysate under laboratory conditions at 42 °C. [ABSTRACT FROM AUTHOR]

Genetically engineering microorganisms to produce chemicals has changed the industrialized world. The budding yeast Saccharomyces cerevisiae is frequently used in industry due to its genetic tractability and unique metabolic capabilities. S. cerevisiae has been engineered to produce novel compounds from diverse sugars found in lignocellulosic biomass, including pentose sugars, like xylose, not recognized by the organism. Engineering high flux toward novel compounds has proved to be more challenging than anticipated since simply introducing pathway components is often not enough. Several studies show that the rewiring of upstream signaling is required to direct products toward pathways of interest, but doing so can diminish stress tolerance, which is important in industrial conditions. As an example of these challenges, we reviewed S. cerevisiae engineering efforts, enabling anaerobic xylose fermentation as a model system and showcasing the regulatory interplay's controlling growth, metabolism, and stress defense. Enabling xylose fermentation in S. cerevisiae requires the introduction of several key metabolic enzymes but also regulatory rewiring of three signaling pathways at the intersection of the growth and stress defense responses: the RAS/PKA, Snf1, and high osmolarity glycerol (HOG) pathways. The current studies reviewed here suggest the modulation of global signaling pathways should be adopted into biorefinery microbial engineering pipelines to increase efficient product yields. [ABSTRACT FROM AUTHOR]

Domestication is an excellent case study for understanding adaptation and multiple fungal lineages have been domesticated for fermenting food products. Studying domestication in fungi has thus both fundamental and applied interest. Genomic studies have revealed the existence of four populations within the blue‐cheese‐making fungus Penicillium roqueforti. The two cheese populations show footprints of domestication, but the adaptation of the two non‐cheese populations to their ecological niches (i.e., silage/spoiled food and lumber/spoiled food) has not been investigated yet. Here, we reveal the existence of a new P. roqueforti population, specific to French Termignon cheeses, produced using small‐scale traditional practices, with spontaneous blue mould colonisation. This Termignon population is genetically differentiated from the four previously identified populations, providing a novel source of genetic diversity for cheese making. The Termignon population indeed displayed substantial genetic diversity, both mating types, horizontally transferred regions previously detected in the non‐Roquefort population, and intermediate phenotypes between cheese and non‐cheese populations. Phenotypically, the non‐Roquefort cheese population was the most differentiated, with specific traits beneficial for cheese making, in particular higher tolerance to salt, to acidic pH and to lactic acid. Our results support the view that this clonal population, used for many cheese types in multiple countries, is a domesticated lineage on which humans exerted strong selection. The lumber/spoiled food and silage/spoiled food populations were not more tolerant to crop fungicides but showed faster growth in various carbon sources (e.g., dextrose, pectin, sucrose, xylose and/or lactose), which can be beneficial in their ecological niches. Such contrasted phenotypes between P. roqueforti populations, with beneficial traits for cheese‐making in the cheese populations and enhanced ability to metabolise sugars in the lumber/spoiled food population, support the inference of domestication in cheese fungi and more generally of adaptation to anthropized environments. [ABSTRACT FROM AUTHOR]