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Soil stabilization is a very important method of science and engineering for improving the properties of soil. This paper aims to stabilize expansive black cotton soil through a biological approach involving plant extracts, plant waste materials, and microorganisms. While chemical methods exist, i.e., lime stabilization, geotextiles, etc., they are not economically feasible for large-scale applications. The primary issue with black cotton soil is due to the presence of montmorillonite clay mineral, which makes it unsuitable for the construction of roads and airfields. The cation exchange capacity (CEC) can be defined as the ability of soil to absorb and exchange positively charged ions; thus, if free positively charged ions are not available, the soil will not exchange them with others. The CEC of the soil is diminished, and ultimately, the soil is stabilized to some extent. This paper explores the preparation of plant extract, which contains a high number of anions, and directly inoculates it with soil, which nullifies the positive charge of the soil and diminishes the CEC. The use of cellulose and lignin-degrading microorganisms as an energy source and other minerals that are needed for their growth will be utilized from the soil to reduce CEC, i.e., Mg required for DNA replication and Ca required for their growth and maintenance. Another approach to diminishing the CEC is to use the microorganisms that produce EPS, which require Ca and Mg as adhesions for the formation of biofilm, i.e., Pseudomonas aeruginosa, Bacillus subtilis, and Escherichia coli. The use of microorganisms that have specific enzymes is also used in the diminishing soil CEC, i.e., by using ureolytic enzyme-producing bacteria like Sporosarcina pasteurii, Bacillus paramycoides, Citrobacter sedlakii, and Enterobacter bugadensis. [ABSTRACT FROM AUTHOR]

For the targeted selection of microbial communities that provide cellulose degradation, soil samples containing cellulolytic microorganisms and specific plant residues as a substrate can be used. The details of this process have not been studied: in particular, whether the use of different soils determines the varying efficiency of communities; whether these established cellulolytic communities will have substrate specificity, and other factors. To answer these questions, four soil microbial communities with different cellulolytic activity (Podzol and the soil of Chernevaya taiga) and substrates (oat straw and hemp shives) with different levels of cellulose availability were used, followed by trained communities that were tested on botrooth substrates (in all possible combinations). Based on the analysis of the taxonomic structure of all communities and their efficiency across all substrates (decomposition level, carbon, and nitrogen content), it was shown that the most important taxa of all trained microbial cellulolytic communities are recruited from secondary soil taxa. The original soil does not affect the efficiency of cellulose decomposition: both soils produce equally active communities. Unexpectedly, the resulting communities trained on oats were more effective on hemp than the communities trained on hemp. In general, the usage of pre-trained microbial communities increases the efficiency of decomposition. [ABSTRACT FROM AUTHOR]

Context: The production of bioethanol from renewable raw materials is a decisive factor in the economic development of many countries. However, the complexity of the processes and the numerous experimental variables involved require a deeper understanding of the chemical reactions that take place during bioethanol production to define optimal parameters. Here, we have employed density functional theory–based calculations to investigate the local reactivity of oligomeric systems by considering crystalline and amorphous cellulose models to better understand some details regarding pulp pretreatment processes. Our results evidence a higher chemical susceptibility of amorphous portions of cellulose to chemicals typically employed in acid hydrolysis. Additionally, we observed that glucose monomers coming from cellulose hydrolysis may undergo oxidation, leading to the formation of byproducts such as hydroxymethylfurfural (HMF), acetic acid, formic acid, and levulinic acid. The analysis of local chemical softness indexes indicated that cellulose hydrolysis may be associated with intermediate chemical steps. Finally, we investigated the influence of distinct solvents (dielectric constants) on the local reactivity of the systems, evidencing a relevant role of the solvent dielectric constant for cellulose degradation in glucose. Methods: Initial three-dimensional structures were constructed. Pre-optimizations were performed in a Hartree–Fock (HF) approach employing the PM7 semi-empirical hamiltonian. The structures were then re-optimized via density functional theory (DFT). The local reactivity study of the systems was conducted through the condensed-to-atoms Fukui indexes (CAFI). Systematic changes of the dielectric constants were also considered in geometry optimization and CAFI calculations to estimate the influence of solvents on the reactivity of the systems. [ABSTRACT FROM AUTHOR]

The red palm weevil (RPW), Rhynchophorus ferrugineus, poses a significant threat to date palms globally, heavily relying on symbiotic microbes for various physiological and behavioral functions. This comprehensive study delves into the intricate dynamics of RPW gut microbiota, revealing a diverse microbial community consisting of seven genera and eight species from Proteobacteria, Firmicutes, and Actinobacteria. The stability of gut bacteria across different life stages was observed, with notable impacts on larval metabolism attributed to shifts in bacterial composition. Bacillus subtilis emerged as a key player, producing a spectrum of metabolic enzymes. Furthermore, the gut bacteria exhibited remarkable pesticide degradation capabilities, suggesting a potential role in the host’s resistance to pesticides. The Arthrobacter sp. was identified as a promising candidate for eco-friendly pest biocontrol and biodegradation strategies. Investigating the influence of thermal stress on two groups of RPW larvae (conventional-fed and antibiotic-fed) at varying temperatures (15, 27, and 35 °C) unveiled potential survival implications. This study highlights the pivotal role of bacterial symbionts in enabling larvae adaptation and thermal stress tolerance. In essence, this research contributes crucial insights into the diversity and functions of RPW gut bacteria, emphasizing their prospective applications in pest control strategies.

The red palm weevil (RPW), Rhynchophorus ferrugineus, poses a significant threat to date palms globally, heavily relying on symbiotic microbes for various physiological and behavioral functions. This comprehensive study delves into the intricate dynamics of RPW gut microbiota, revealing a diverse microbial community consisting of seven genera and eight species from Proteobacteria, Firmicutes, and Actinobacteria. The stability of gut bacteria across different life stages was observed, with notable impacts on larval metabolism attributed to shifts in bacterial composition. Bacillus subtilis emerged as a key player, producing a spectrum of metabolic enzymes. Furthermore, the gut bacteria exhibited remarkable pesticide degradation capabilities, suggesting a potential role in the host's resistance to pesticides. The Arthrobacter sp. was identified as a promising candidate for eco-friendly pest biocontrol and biodegradation strategies. Investigating the influence of thermal stress on two groups of RPW larvae (conventional-fed and antibiotic-fed) at varying temperatures (15, 27, and 35 °C) unveiled potential survival implications. This study highlights the pivotal role of bacterial symbionts in enabling larvae adaptation and thermal stress tolerance. In essence, this research contributes crucial insights into the diversity and functions of RPW gut bacteria, emphasizing their prospective applications in pest control strategies. [ABSTRACT FROM AUTHOR]

β-glucosidase is a key component of cellulase for its function in hydrolyzing cellobiose to glucose in the final step of cellulose degradation. The high-level expression of β-glucosidase is essential for cellulose conversion. Aspergillus niger ATCC 20611 has the potential for efficient protein expression because of its ability to secret enzymes for the industrial production of fructooligosaccharides, but it lacks robust promoters for high-level protein expression. Here, the development of A. niger 20611 as a powerful protein expression system exploited the conserved constitutive promoter Pgpd1 of the glyceraldehyde-3-phosphate dehydrogenase-encoding gene from Trichoerma reesei to drive the expression of the enhanced green fluorescent protein in A. niger ATCC 20611. The mycelium of the transformant AGE9 exhibited intense fluorescence. Then, the promotor Pgpd1 was used to drive the expression of β-glucosidase and the enzyme activity of transformants AGB1 and AGB33 were 1.02 and 0.51 U/mL, respectively. These results demonstrate that the promotor Pgpd1 from T. reesei was applicable for A. niger ATCC 20611. Furthermore, the T. reesei-specific robust promoter Pcdna1 was used to drive the expression of β-glucosidase. The β-glucosidase exhibited a high-level expression with a yield of 15.2 U/mL, which was over 13.9 times higher than that driven by the promoter Pgpd1. The β-glucosidase was thermally stable and accounted for 85% of the total extracellular proteins. Subsequently, the fermentation broth including β-glucosidase was directly added to the cellulase mixture of T. reesei for saccharification of the acid-treated corncob residues and the delignified corncob residues, which increased the saccharification efficiency by 26.21% and 29.51%, respectively. Thus, β-glucosidase exhibited a high level of expression in A. niger ATCC 20611 and enhanced cellulose degradation by addition in vitro. In addition, the robust promoter Pcdna1 of T. reesei could drive the high-level expression of protein in A. niger ATCC 20611. These results demonstrate that the promoters in filamentous fungi could be employed across species in A. niger ATCC 20611 and further facilitated the efficient expression of β-glucosidase to optimize cellulases for efficient cellulose transformation. [ABSTRACT FROM AUTHOR]

Simple Summary: Rodents, as the most abundant group of mammals, offer a unique opportunity to study the relationship between diet and host gut microecology. In this study, we focused on two species of the genus Myospalax in the Eurasian steppes in China: M. psilurus and M. aspalax. These specialized herbivorous rodents have adapted to a subterranean lifestyle and exhibit distinct dietary choices. Our study aimed to investigate how their diet composition is related to their gut microbial communities and whether there are unique indicator species for their gut microecology. Using 16S amplicon technology and macro-barcoding technology, we analyzed the gut bacterial communities and diet composition of the two zokor species. We found that M. psilurus had a higher diversity of gut microbial bacterial communities compared to M. aspalax. The two species also possessed different gut bacterial indicator species, with unique relationships between their diet and gut microbes. Our results provide new insights into the adaptation of zokors to long-term subterranean life and shed light on their cellulose degradation abilities and ecological niches. As the most abundant group of mammals, rodents possess a very rich ecotype, which makes them ideal for studying the relationship between diet and host gut microecology. Zokors are specialized herbivorous rodents adapted to living underground. Unlike more generalized herbivorous rodents, they feed on the underground parts of grassland plants. There are two species of the genus Myospalax in the Eurasian steppes in China: one is Myospalax psilurus, which inhabits meadow grasslands and forest edge areas, and the other is M. aspalax, which inhabits typical grassland areas. How are the dietary choices of the two species adapted to long-term subterranean life, and what is the relationship of this diet with gut microbes? Are there unique indicator genera for their gut microbial communities? Relevant factors, such as the ability of both species to degrade cellulose, are not yet clear. In this study, we analyzed the gut bacterial communities and diet compositions of two species of zokors using 16S amplicon technology combined with macro-barcoding technology. We found that the diversity of gut microbial bacterial communities in M. psilurus was significantly higher than that in M. aspalax, and that the two species of zokors possessed different gut bacterial indicator genera. Differences in the feeding habits of the two species of zokors stem from food composition rather than diversity. Based on the results of Mantel analyses, the gut bacterial community of M. aspalax showed a significant positive correlation with the creeping-rooted type food, and there was a complementary relationship between the axis root-type-food- and the rhizome-type-food-dominated (containing bulb types and tuberous root types) food groups. Functional prediction based on KEGG found that M. psilurus possessed a stronger degradation ability in the same cellulose degradation pathway. Neutral modeling results show that the gut flora of the M. psilurus has a wider ecological niche compared to that of the M. aspalax. This provides a new perspective for understanding how rodents living underground in grassland areas respond to changes in food conditions. [ABSTRACT FROM AUTHOR]

In this investigation, cellulose-degrading fungi and bacteria were isolated from different partially decomposed cellulose-rich substrates, such as groundnut residues, rice straw, and rotten wood, following dilution plating techniques on carboxymethyl cellulose agar media and screening for potential cellulose degradation ability. The development of a clear halo zone surrounding the microbial colonies during the initial screening process using the Congo red test (20 isolates) suggested cellulose hydrolysis, and the highest cellulase production activity was implied by the isolates with the largest clear zone ratio (9 isolates). Using both macroscopic and microscopic examinations, as well as standard biochemical tests outlined in Bergey’s Manual of Determinative Bacteriology, the genus-level identification of fungi and bacteria was accomplished. In order to molecularly identify the 4 isolated fungal and bacterial strains at the species level after being ultimately selected for cellulase production potential under in vitro studies, fungal and bacterial DNA was extracted and amplified by PCR using the universal primers ITS1 and ITS4 for fungi (ITS rRNA, 5.8S rRNA) and 8F and 1492R for bacterial isolates (16S rRNA). After sequencing, the PCR results were compared to other comparable sequences in GenBank (NCBI). Based on the available NCBI data, phylogenetic analysis of their ribosomal gene partial sequences revealed that DAJ2 (PP086700) shares 100% homology with Aspergillus foetidus, DTJ4 (PP086699) shares 99.74% similarity with Trichoderma atrobrunnium, DBJ6 (PP082584) shares 100% identity with Priestia megaterium, and DMB9 (PP082585) shares 99.88% homology with Micrococcus yunnanensis. The cellulolytic potential of Phanerochaete chrysosporium is well established. Therefore, it was considered a standard culture for comparison and was collected from the MTCC, Chandigarh, India. Overall, all 4 selected isolates and the check organism were mutually compatible or synergistic with each other, and their consortium is useful for the accelerated decomposition of organic constituents during rapid composting. [ABSTRACT FROM AUTHOR]

• For the first time a hydrotalcite was tested for deacidification of cellulosic objects. • A hydrotalcite dispersion with crystallites about 3 nm in isopropanol was prepared and charactarised. • Application by spraying proved to be a successful method for the deacidification. • Anion exchange capability resulted in increase of pH of acid paper samples. Preservation of cellulose cultural objects, particularly books, is a field of research with about 100 years of history. The most dangerous in terms of their lifespan is the use of acidic wood-based paper which represents a problem in both quality and durability. Acid hydrolysis of polymeric cellulose chains is a key factor responsible for loss of visual appearance quality and mechanical strength. Sulphate and aluminium ions together with generated formic, acetic, and other higher carboxylic acids accelerate the rate of degradation catalytically as well as autocatalytically. Alkaline species (alkali and alkali earth hydrogen carbonates, oxides, organocompounds, and amines) suppress the degradation processes. This paper refers to the utilisation of hydrotalcites (HT, Mg x Al 2 (OH) 2x+4 CO 3 , x > 6) as deacidifying agents in a non-halogenated low toxic carrier – isopropanol. Preparation of an HT dispersion, its characterisation, and application to (acidic) test paper are described. Due to its ion exchange and neutralization capabilities, HT increased pH of test papers from about 4 to at least 6. Similarly to the action of HT as antacids, it is possible to suppose the neutralizing capability in deacidifying of aged-acid papers, even at pH lower than 7. A higher increase of pH was achieved only with extremely high loadings of HT; 16 g / m2 allowed to reach a surface pH value equal to 7.8. Both the optical properties and mechanical strength of treated samples were improved at loadings up to 3 g / m2. [Display omitted] [ABSTRACT FROM AUTHOR]

For the targeted selection of microbial communities that provide cellulose degradation, soil samples containing cellulolytic microorganisms and specific plant residues as a substrate can be used. The details of this process have not been studied: in particular, whether the use of different soils determines the varying efficiency of communities; whether these established cellulolytic communities will have substrate specificity, and other factors. To answer these questions, four soil microbial communities with different cellulolytic activity (Podzol and the soil of Chernevaya taiga) and substrates (oat straw and hemp shives) with different levels of cellulose availability were used, followed by trained communities that were tested on botrooth substrates (in all possible combinations). Based on the analysis of the taxonomic structure of all communities and their efficiency across all substrates (decomposition level, carbon, and nitrogen content), it was shown that the most important taxa of all trained microbial cellulolytic communities are recruited from secondary soil taxa. The original soil does not affect the efficiency of cellulose decomposition: both soils produce equally active communities. Unexpectedly, the resulting communities trained on oats were more effective on hemp than the communities trained on hemp. In general, the usage of pre-trained microbial communities increases the efficiency of decomposition.

Cellulolytic microorganisms play a crucial role in agricultural waste disposal. Strain QXD-8T was isolated from soil in northern China. Similarity analyses of the 16S rRNA gene, as well as the 120 conserved genes in the whole-genome sequence, indicate that it represents a novel species within the genus Microbacterium. The Microbacterium sp. QXD-8T was able to grow on the CAM plate with sodium carboxymethyl cellulose as a carbon source at 15 °C, forming a transparent hydrolysis circle after Congo red staining, even though the optimal temperature for the growth and cellulose degradation of strain QXD-8T was 28 °C. In the liquid medium, it effectively degraded cellulose and produced reducing sugars. Functional annotation revealed the presence of encoding genes for the GH5, GH6, and GH10 enzyme families with endoglucanase activity, as well as the GH1, GH3, GH39, and GH116 enzyme families with β-glucosidase activity. Additionally, two proteins in the GH6 family, one in the GH10, and two of nine proteins in the GH3 were predicted to contain a signal peptide and transmembrane region, suggesting their potential for extracellularly degrade cellulose. Based on the physiological features of the type strain QXD-8T, we propose the name Microbacterium psychrotolerans for this novel species. This study expands the diversity of psychrotolerant cellulolytic bacteria and provides a potential microbial resource for straw returning in high-latitude areas at low temperatures. [ABSTRACT FROM AUTHOR]

Abstract The presence of stone cells in pears is recognized as a problem for the pear processing industry. Bacillus and mold can degrade stone cells because of their potential to digest cellulose and lignin, but they cannot be used for the degradation test of pear stone cells. In this study Lactoplantibacillus plantarum JYLP‐326 was used in single culture or in the mixed culture with Saccharomyces cerevisiae Y2 to study their potential to degrade stone cells during fermentation and the mechanism of degradation was further explored. Synergy in cellulase activity was observed in the mixed‐culture where the maximum activity was observed at 96th hour of fermentation. Activities of endoglucanase, exoglucanase and β‐glucosidase were 1.75, 4.58, and 2.31. The degradation rate of stone cells in the mixed‐culture was 37.67%, which was significantly higher than that the results obtained for single cultures. The results of scanning electron microscopy (SEM) showed that the surface of the cultured stone cells became rough. Metabolomics studies confirmed the presence of specific metabolites related to the degradation of stone cells after the fermentation. It was concluded that the mixed‐culture fermentation using the above‐mentioned strains could be exploited by the pear processing industry to degrade stone cells.

β-glucosidase is a key component of cellulase for its function in hydrolyzing cellobiose to glucose in the final step of cellulose degradation. The high-level expression of β-glucosidase is essential for cellulose conversion. Aspergillus niger ATCC 20611 has the potential for efficient protein expression because of its ability to secret enzymes for the industrial production of fructooligosaccharides, but it lacks robust promoters for high-level protein expression. Here, the development of A. niger 20611 as a powerful protein expression system exploited the conserved constitutive promoter Pgpd1 of the glyceraldehyde-3-phosphate dehydrogenase-encoding gene from Trichoerma reesei to drive the expression of the enhanced green fluorescent protein in A. niger ATCC 20611. The mycelium of the transformant AGE9 exhibited intense fluorescence. Then, the promotor Pgpd1 was used to drive the expression of β-glucosidase and the enzyme activity of transformants AGB1 and AGB33 were 1.02 and 0.51 U/mL, respectively. These results demonstrate that the promotor Pgpd1 from T. reesei was applicable for A. niger ATCC 20611. Furthermore, the T. reesei-specific robust promoter Pcdna1 was used to drive the expression of β-glucosidase. The β-glucosidase exhibited a high-level expression with a yield of 15.2 U/mL, which was over 13.9 times higher than that driven by the promoter Pgpd1. The β-glucosidase was thermally stable and accounted for 85% of the total extracellular proteins. Subsequently, the fermentation broth including β-glucosidase was directly added to the cellulase mixture of T. reesei for saccharification of the acid-treated corncob residues and the delignified corncob residues, which increased the saccharification efficiency by 26.21% and 29.51%, respectively. Thus, β-glucosidase exhibited a high level of expression in A. niger ATCC 20611 and enhanced cellulose degradation by addition in vitro. In addition, the robust promoter Pcdna1 of T. reesei could drive the high-level expression of protein in A. niger ATCC 20611. These results demonstrate that the promoters in filamentous fungi could be employed across species in A. niger ATCC 20611 and further facilitated the efficient expression of β-glucosidase to optimize cellulases for efficient cellulose transformation.

As the most abundant group of mammals, rodents possess a very rich ecotype, which makes them ideal for studying the relationship between diet and host gut microecology. Zokors are specialized herbivorous rodents adapted to living underground. Unlike more generalized herbivorous rodents, they feed on the underground parts of grassland plants. There are two species of the genus Myospalax in the Eurasian steppes in China: one is Myospalax psilurus, which inhabits meadow grasslands and forest edge areas, and the other is M. aspalax, which inhabits typical grassland areas. How are the dietary choices of the two species adapted to long-term subterranean life, and what is the relationship of this diet with gut microbes? Are there unique indicator genera for their gut microbial communities? Relevant factors, such as the ability of both species to degrade cellulose, are not yet clear. In this study, we analyzed the gut bacterial communities and diet compositions of two species of zokors using 16S amplicon technology combined with macro-barcoding technology. We found that the diversity of gut microbial bacterial communities in M. psilurus was significantly higher than that in M. aspalax, and that the two species of zokors possessed different gut bacterial indicator genera. Differences in the feeding habits of the two species of zokors stem from food composition rather than diversity. Based on the results of Mantel analyses, the gut bacterial community of M. aspalax showed a significant positive correlation with the creeping-rooted type food, and there was a complementary relationship between the axis root-type-food- and the rhizome-type-food-dominated (containing bulb types and tuberous root types) food groups. Functional prediction based on KEGG found that M. psilurus possessed a stronger degradation ability in the same cellulose degradation pathway. Neutral modeling results show that the gut flora of the M. psilurus has a wider ecological niche compared to that of the M. aspalax. This provides a new perspective for understanding how rodents living underground in grassland areas respond to changes in food conditions.

Cellulose degradation is a critical process in soil ecosystems, playing a vital role in nutrient cycling and organic matter decomposition. Enzymatic degradation of cellulosic biomass is the most sustainable and green method of producing liquid biofuel. It has gained intensive research interest with future perspective as the majority of terrestrial lignocellulose biomass has a great potential to be used as a source of bioenergy. However, the recalcitrant nature of lignocellulose limits its use as a source of energy. Noteworthy enough, enzymatic conversion of cellulose biomass could be a leading future technology. Fungal enzymes play a central role in cellulose degradation. Our understanding of fungal cellulases has substantially redirected in the past few years with the discovery of a new class of enzymes and Cellulosome. Efforts have been made from time to time to develop an economically viable method of cellulose degradation. This review provides insights into the current state of knowledge regarding cellulose degradation in soil and identifies areas where further research is needed.

One of the main challenges in the preservation of paper materials is the development of compatible and eco-friendly cleaning systems suitable for the wet removal of hydrophobic, aged compounds, such as greasy materials or synthetic adhesives, whose presence enhances paper degradation and worsens its optical features. To avoid the use of organic solvents that can be potentially harmful to operators, in this work we propose a new water-based gel obtained by chemical modification of gellan gum gel, a well-known paper cleaning hydrogel. With the aim of obtaining a cleaning system that is able to interact and adsorb not only hydrophilic cellulose degradation byproducts, but also greasy materials and synthetic adhesives, methacrylation has been applied on gellan gum. First, the chemical modification process of gellan gum has been monitored by means of NMR and ATR-FTIR spectroscopy. Then, the mechanical and physical–chemical properties of the resulting gel have been evaluated by using rheological, AFM, and wettability measurements. Finally, the compatibility and cleaning efficiency of the newly synthetized gel have been tested on paper with different age (from eighteenth and twentieth centuries) and composition by means of a multi-technique framework consisting of spectroscopic, chromatographic, SEM, colorimetric and pH measurements. This work shows a first interesting step towards the development of single component paper cleaning system, easy to use, for the removal of both cellulose hydrophilic degradation products and hydrophobic compounds from different paper typologies. [ABSTRACT FROM AUTHOR]

We investigated the cell functions of the Ca2+ signaling genes phospholipase C-1 (plc-1), Ca2+/H+exchanger (cpe-1), and secretory phospholipase A2 (splA2) for stress responses and cellulose utilization in Neurospora crassa. The Δplc-1, Δcpe-1, and ΔsplA2 mutants displayed increased sensitivity to the alkaline pH and reduced survival during induced thermotolerance. The ΔsplA2 mutant also exhibited hypersensitivity to the DTT-induced endoplasmic reticulum (ER) stress, increased microcrystalline cellulose utilization, increased protein secretion, and glucose accumulation in the culture supernatants. Moreover, the ΔsplA2 mutant could not grow on microcrystalline cellulose during ER stress. Furthermore, plc-1, cpe-1, and splA2 synthetically regulate the acquisition of thermotolerance induced by heat shock, responses to alkaline pH and ER stress, and utilization of cellulose and other alternate carbon sources in N. crassa. In addition, expression of the alkaline pH regulator, pac-3, and heat shock proteins, hsp60, and hsp80 was reduced in the Δplc-1, Δcpe-1, and ΔsplA2 single and double mutants. The expression of the unfolded protein response (UPR) markers grp-78 and pdi-1 was also significantly reduced in the mutants showing growth defect during ER stress. The increased cellulolytic activities of the ΔsplA2 and Δcpe-1; ΔsplA2 mutants were due to increased cbh-1, cbh-2, and endo-2 expression in N. crassa. Therefore, plc-1, cpe-1, and splA2 are involved in stress responses and cellulose utilization in N. crassa. [ABSTRACT FROM AUTHOR]

Abstract Background Cellulose degradation can determine mycelial growth rate and affect yield during the growth of Flammulina filiformis. The degradation of cellulose requires the joint action of a variety of cellulases, and some cellulase-related genes have been detected in mushrooms. However, little is known about the transcriptional regulatory mechanisms of cellulose degradation. Results In this study, FfMYB15 that may regulate the expression of cellulase gene FfCEL6B in F. filiformis was identified. RNA interference (RNAi) showed that FfCEL6B positively regulated mycelial growth. Gene expression analyses indicated that the expression patterns of FfCEL6B and FfMYB15 in mycelia cultured on the 0.9% cellulose medium for different times were similar with a correlation coefficient of 0.953. Subcellular localization and transcriptional activity analyses implied that FfMYB15 was located in the nucleus and was a transcriptional activator. Electrophoretic mobility shift assay (EMSA) and dual-luciferase assays demonstrated that FfMYB15 could bind and activate FfCEL6B promoter by recognizing MYB cis-acting element. Conclusions This study indicated that FfCEL6B played an active role in mycelial growth of F. filiformis and was regulated by FfMYB15.

In this study, aqueous extracts of cotton linter paper and "Hanji", which is a Korean traditional paper, were analyzed to evaluate the deterioration of paper cultural heritage. We quantified cello-oligosaccharides, which are water-soluble low-molecular-weight carbohydrates, in extracts from the two types of paper. The degree of polymerization (DP) of aged paper was also evaluated based on the absolute molecular weight of cellulose. The content of water-soluble cello-oligosaccharides (COS) varied according to the degree of cellulose degradation. Therefore, the cellulose hydrolysis rate could be predicted from the linear relationship between the cellulose hydrolysis rate and the water-soluble COS content. The DP calculated using the predicted cellulose hydrolysis rate was accurate if the amount of cello-oligosaccharides extracted exceeded the limit of quantification of COS. Consequently, among the many minimally invasive methods available, this methodology may provide an indirect method to evaluate cellulose degradation by quantifying the carbohydrates dissolved in extracts of cotton linter paper and Hanji. [ABSTRACT FROM AUTHOR]

Purpose: Farmers need alternative approaches to manage nitrogen (N) that meet crop needs while reducing loss to the environment. Identifying crop genotypes that promote decomposition of organic materials and understanding the potential mechanisms responsible could help address this challenge. Consequently, we aimed to 1) determine whether carrot genotypes differ in their potential to facilitate organic matter decomposition in soil, and 2) identify bacterial taxa that are stimulated by carrot roots, and thus could play a role in these processes. Methods: We grew five genotypes expected to differ in N use efficiency in a nutrient-poor soil amended with 15 N-enriched corn residue, tracked changes in carbon (C) and N pools, quantified microbial activity and bacterial community composition, and predicted the potential expression of microbial genes involved in soil C and N cycles. Results: Experimental genotype 8503 had the greatest capacity to promote decomposition of corn residues. This genotype had the highest % of N from the corn residue in its taproots and on average, promoted higher b-glucosidase activity in soils. Distinct bacterial communities from the families Micromonosporaceae, Chromatiaceae, and Rhodospirillaceae were also enriched in the soils of genotypes like 8503 that were most effective in obtaining N from the corn residue, and this was correlated with greater potential expression of genes responsible for β-glucosidase and nitrification activity. Conclusions: Carrot genotypes do differ in their potential to alter soil bacterial communities and stimulate microbially-mediated decomposition of organic materials indicating that it may be possible to begin selecting for this important trait. [ABSTRACT FROM AUTHOR]

Cellulolytic microorganisms play a crucial role in agricultural waste disposal. Strain QXD-8T was isolated from soil in northern China. Similarity analyses of the 16S rRNA gene, as well as the 120 conserved genes in the whole-genome sequence, indicate that it represents a novel species within the genus Microbacterium. The Microbacterium sp. QXD-8T was able to grow on the CAM plate with sodium carboxymethyl cellulose as a carbon source at 15 °C, forming a transparent hydrolysis circle after Congo red staining, even though the optimal temperature for the growth and cellulose degradation of strain QXD-8T was 28 °C. In the liquid medium, it effectively degraded cellulose and produced reducing sugars. Functional annotation revealed the presence of encoding genes for the GH5, GH6, and GH10 enzyme families with endoglucanase activity, as well as the GH1, GH3, GH39, and GH116 enzyme families with β-glucosidase activity. Additionally, two proteins in the GH6 family, one in the GH10, and two of nine proteins in the GH3 were predicted to contain a signal peptide and transmembrane region, suggesting their potential for extracellularly degrade cellulose. Based on the physiological features of the type strain QXD-8T, we propose the name Microbacterium psychrotolerans for this novel species. This study expands the diversity of psychrotolerant cellulolytic bacteria and provides a potential microbial resource for straw returning in high-latitude areas at low temperatures.

This research project examines the effects of X-rays on cellulose and some moulds frequently found on paper. The aim was to identify applications for X-ray irradiation that can be used as a disinfection method for archival material. The question was if X-rays are suitable as an alternative to gamma radiation and if they are less harmful. For this purpose, the minimum X-ray dose required to reduce the microbial count to a harmless level was determined. The material-altering effect was examined on samples treated with X-rays and gamma radiation. Spectrophotometric measurements showed that there is no noticeable colour change with either type of radiation. The determination of the molecular weight distribution, in turn, showed that the molar mass of the cellulose is considerably reduced with both treatment methods. Using mechanical tests, however, it could be demonstrated that this has no significant influence on the tensile strength. The examination of the oxidation behaviour also showed no significant difference between the differently treated samples. The studies demonstrated that both methods have an almost identical effect on cellulose. Thus, X-ray treatment is primarily a supplement to the known disinfection methods and is particularly suitable for objects that would not withstand treatment with alcohol. [ABSTRACT FROM AUTHOR]

Background: The Horned Screamer (Anhima cornuta) is an herbivorous bird that inhabits wetlands of the South American tropical region. We hypothesize that due to its herbivorous niche, its digestive tract compartments may have bacteria specialized in fermenting complex plant carbohydrates. To test this hypothesis, we compared the bacterial communities along the gastrointestinal tract (GIT) of a Horned Screamer captured in Venezuela. Methods: Samples were taken from tissues and content of the proventriculus and the small intestine (considered for this study as upper GIT), and the large intestine and cecum (lower GIT). The bacterial community was characterized by sequencing the V4 region of the 16S rRNA gene. Bioinformatic analysis was performed using QIIME, QIITA and Microbiome Analyst. The association between microbial taxonomy and function was analyzed using their Greengenes OTU IDs and a custom KEGG BRITE hierarchical tree and visualized with BURRITO. Results: The Screamer's gastrointestinal microbiota was composed by seven phyla being Firmicutes and Bacteroidetes the most predominant. The dominant taxa in the upper GIT were Helicobacter, Vibrio, Enterobacter, Acinetobacter and Staphylococcus. The dominant taxa in the lower GIT were Oribacterium, Blautia, Roseburia, Ruminococcus, Desulfovibrio, Intestinimonas, Marvinbryantia and Parabacteroides. Complete degradation of cellulose to the end-products acetate, propanoate, butanoate and acetoacetate was found in the upper and lower GIT without significant differences. Conclusion: Our study confirmed changes in bacterial community composition throughout the GIT of the Horned Screamer primarily associated with the production of metabolic end-products of carbohydrate digestion essential for the fermentation of the herbivorous diet. [ABSTRACT FROM AUTHOR]

On a sample of the Turin Shroud (TS), we applied a new method for dating ancient linen threads by inspecting their structural degradation by means of Wide-Angle X-ray Scattering (WAXS). The X-ray dating method was applied to a sample of the TS consisting of a thread taken in proximity of the 1988/radiocarbon area (corner of the TS corresponding to the feet area of the frontal image, near the so-called Raes sample). The size of the linen sample was about 0.5 mm × 1 mm. We obtained one-dimensional integrated WAXS data profiles for the TS sample, which were fully compatible with the analogous measurements obtained on a linen sample whose dating, according to historical records, is 55–74 AD, Siege of Masada (Israel). The degree of natural aging of the cellulose that constitutes the linen of the investigated sample, obtained by X-ray analysis, showed that the TS fabric is much older than the seven centuries proposed by the 1988 radiocarbon dating. The experimental results are compatible with the hypothesis that the TS is a 2000-year-old relic, as supposed by Christian tradition, under the condition that it was kept at suitable levels of average secular temperature—20.0–22.5 °C—and correlated relative humidity—75–55%—for 13 centuries of unknown history, in addition to the seven centuries of known history in Europe. To make the present result compatible with that of the 1988 radiocarbon test, the TS should have been conserved during its hypothetical seven centuries of life at a secular room temperature very close to the maximum values registered on the earth.

Cellobiohydrolase, a workhorse enzyme for cellulose degradation, has the potential for application in biofuels, biochemicals, biomaterials, etc. However, highly efficient cellobiohydrolase is desirable to improve the economic feasibility of industrial applications. In this work, two variants Tr Cel7AT97A and Tr Cel7AK166A were designed to increase the flexibility of the entrance of the catalytic tunnel. We performed the molecular dynamics simulations using NAMD for 100 ns at 300 K in the NPT (isothermal–isobaric) ensemble and then we found that the region flexibility at the catalytic tunnel entrances of Tr Cel7AT97A and Tr Cel7AK166A was improved, compared to that of Tr Cel7A, based on the calculation of the average root mean square fluctuation. The specific activities of Tr Cel7AT97A and Tr Cel7AK166A were increased by 37 % and 68 %, respectively, compared with that of Tr Cel7A at 45 °C for 12 h. An effective approach named revive amino acid flexibility for enzyme activity based on amino acid flexibility was proposed for protein engineering of cellobiohydrolase Tr Cel7A. [Display omitted] • Tr Cel7AK166A exhibited outstanding cellulose-degrading activity at 45 °C. • Flexibility of catalytic tunnel entrance of Tr Cel7A was improved by engineering. • Cellulose-degrading activity of Tr Cel7A is affected by the amino acid flexibility. [ABSTRACT FROM AUTHOR]

Managing carbon inputs from straw can pave the way towards carbon neutrality and climate change mitigation. Straw decomposition by cooperative microbial actions is an important process of carbon cycling in nature, and in this process, microbial communities are constantly in succession. Soil is rich in microorganisms and can be a source of microbial for straw degradation. In this study, corn straw was mixed with different soil types and incubated in conical flasks for 70 days. Bacterial diversity and community structure were determined using 16S rRNA sequencing. Then, the effects of physicochemical parameters and enzyme activities on the composition of bacterial communities at different stages were evaluated. The results showed that bacterial diversity decreased during co-cultivation. The differences in bacterial communities between all treatments were greater in the later stages, with Pseudomonadota, Actinomycetota, and Bacillota as the major phyla. Among them, the biomarkers at different times for different treatments included Sphingomonas , Mycobacterium , Oceanobacillus , Streptomyces , Pseudomonas , Flavobacterium , and Saccharomonospora. All of them showed cellulose degradation capacity; thus, the organic matter gradually decreased during the co-cultivation. Canonical correspondence analysis (CCA) showed that pH, organic matter (OM), electrical conductivity (EC), cellulase, β-glucosidase, and filter paper (FPase) activities had a significant effect on bacterial communities at different stages. Our findings suggested that soil microbial communities can be an effective source of cellulose-degrading microorganisms, and corn straw co-cultivation with different soil types increased the abundance of cellulose-degrading bacteria, which provides the theoretical basis for efficient cellulose-degrading agent screening. [ABSTRACT FROM AUTHOR]

Cellulose degradation results from the synergistic effect of different enzymes, but which enzyme is involved in the initial stage of cellulose degradation is still not well understood. Cellobiohydrolase 2 (CBH2) attached to the conidial surface is possibly associated with the initial stage. However, its specific mechanism is still incompletely known. This study explored the potential role of CBH2 in initiating cellulose degradation using a constitutive overexpression strategy. First, the CBH2-overexpression Trichoderma reesei strains Qgc2–5 and Qrc2–40 were constructed using the constitutive promoters Pgpd1 and PrpS30, respectively. It was found that cbh2 was expressed at a high level under the glucose conditions and was significantly higher than that of the parental strain QM9414 at the early stage of 29 h when cellulose was used as the carbon source. Particularly, the constitutive overexpression of cbh2 caused the strong expression of major cellulase-encoding genes (cbh1, eg1, and eg2) and the rapid decomposition of cellulosic material. Meanwhile, the scanning electron microscope showed that the groove-like structure of the cellulose surface was eroded seriously owing to CBH2 overexpression, which caused the cellulose surface to be smooth. These results showed that the overexpression of CHB2 caused the major cellulase enzymes to be expressed and contributed to cellulose degradation, showing the potential role of CBH2 in the initial stage of the cellulose hydrolytic process.

Background The Horned Screamer (Anhima cornuta) is an herbivorous bird that inhabits wetlands of the South American tropical region. We hypothesize that due to its herbivorous niche, its digestive tract compartments may have bacteria specialized in fermenting complex plant carbohydrates. To test this hypothesis, we compared the bacterial communities along the gastrointestinal tract (GIT) of a Horned Screamer captured in Venezuela. Methods Samples were taken from tissues and content of the proventriculus and the small intestine (considered for this study as upper GIT), and the large intestine and cecum (lower GIT). The bacterial community was characterized by sequencing the V4 region of the 16S rRNA gene. Bioinformatic analysis was performed using QIIME, QIITA and Microbiome Analyst. The association between microbial taxonomy and function was analyzed using their Greengenes OTU IDs and a custom KEGG BRITE hierarchical tree and visualized with BURRITO. Results The Screamer’s gastrointestinal microbiota was composed by seven phyla being Firmicutes and Bacteroidetes the most predominant. The dominant taxa in the upper GIT were Helicobacter, Vibrio, Enterobacter, Acinetobacter and Staphylococcus. The dominant taxa in the lower GIT were Oribacterium, Blautia, Roseburia, Ruminococcus, Desulfovibrio, Intestinimonas, Marvinbryantia and Parabacteroides. Complete degradation of cellulose to the end-products acetate, propanoate, butanoate and acetoacetate was found in the upper and lower GIT without significant differences. Conclusion Our study confirmed changes in bacterial community composition throughout the GIT of the Horned Screamer primarily associated with the production of metabolic end-products of carbohydrate digestion essential for the fermentation of the herbivorous diet.

Actinomycetes are an important group of bioactive hydrolytic bacteria in any ecosystem. However, the actinomycete biodiversity in tropical ecosystems, particularly in Vietnam, is still underexplored. The aim of this article is to analyze the abundance, taxonomic structure and ecophysiological features of actinomycete complexes of soils and litter in Vietnam's protected areas. A total of 41 samples of soils, plant litter and suspended soils were collected from six of Vietnam's national parks and nature reserves. The direct inoculation technique showed that the total abundance of actinomycetes varied from 2.0 × 104 to 1.0 × 108 CFU/g. According to the luminescent microscopy with acridine orange dye, the length of the actinomycete mycelium was as long as 1000 m/g in the litter of Xuan Son National Park. A total of 80 strains were isolated and tested for antagonistic activity against Bacillus subtilis, Aspergillus niger and Candida albicans. Inoculation on Getchinson's medium showed high cellulolytic activity. The most active strains were isolated from alluvial brown soil, plant litter and suspended soil of the Pu Hoat Nature Reserve. In these samples, actinomycetes adapted to high temperatures and low pH were found to be predominant. High-throughput sequencing of the V3–V4 region of the 16S rRNA gene and bioinformatic analysis confirmed the high taxonomic diversity and high hydrolytic activity of actinomycete complexes of the Pu Hoat Nature Reserve samples. [ABSTRACT FROM AUTHOR]

Background: Cellulose degradation can determine mycelial growth rate and affect yield during the growth of Flammulina filiformis. The degradation of cellulose requires the joint action of a variety of cellulases, and some cellulase-related genes have been detected in mushrooms. However, little is known about the transcriptional regulatory mechanisms of cellulose degradation. Results: In this study, FfMYB15 that may regulate the expression of cellulase gene FfCEL6B in F. filiformis was identified. RNA interference (RNAi) showed that FfCEL6B positively regulated mycelial growth. Gene expression analyses indicated that the expression patterns of FfCEL6B and FfMYB15 in mycelia cultured on the 0.9% cellulose medium for different times were similar with a correlation coefficient of 0.953. Subcellular localization and transcriptional activity analyses implied that FfMYB15 was located in the nucleus and was a transcriptional activator. Electrophoretic mobility shift assay (EMSA) and dual-luciferase assays demonstrated that FfMYB15 could bind and activate FfCEL6B promoter by recognizing MYB cis-acting element. Conclusions: This study indicated that FfCEL6B played an active role in mycelial growth of F. filiformis and was regulated by FfMYB15. [ABSTRACT FROM AUTHOR]

The Gram-negative bacterium Cytophaga hutchinsonii digests cellulose through a novel cellulose degradation mechanism. It possesses the lately characterized type IX secretion system (T9SS). We recently discovered that N-glycosylation of the C-terminal domain (CTD) of a hypothetical T9SS substrate protein in the periplasmic space of C. hutchinsonii affects protein secretion and localization. In this study, green fluorescent protein (GFP)-CTDCel9A recombinant protein was found with increased molecular weight in the periplasm of C. hutchinsonii. Site-directed mutagenesis studies on the CTD of cellulase Cel9A demonstrated that asparagine residue 900 in the D-X-N-X-S motif is important for the processing of the recombinant protein. We found that the glycosyltransferase-related protein GtrA (CHU_0012) located in the cytoplasm of C. hutchinsonii is essential for outer membrane localization of the recombinant protein. The deletion of gtrA decreased the abundance of the outer membrane proteins and affected cellulose degradation by C. hutchinsonii. This study provided a link between the glycosylation system and cellulose degradation in C. hutchinsonii. [ABSTRACT FROM AUTHOR]

In 2021, Wide-angle X-ray Scattering (WAXS) was applied to a sample of the Turin Shroud as a new method for dating ancient linen threads by inspecting their structural degradation. The major result of the research was to estimate the natural aging of the linen of the Shroud, through which it was found to be compatible with the hypothesis of a 2000-year-old relic, and not a medieval fabric. The present work regards the theoretical natural aging of the linen versus different alleged historical–geographical paths of the Shroud through cities where the presence of icons of Jesus Christ is attested. The theoretical results of the natural aging of the Shroud, calculated for the year 2021, agree extremely well with the experimental results for at least one alleged path. The results also show that if the Shroud were medieval and kept in Europe for 7 centuries, natural aging that is one order of magnitude less than that measured by WAXS would have resulted. Today, the Shroud is kept in a reliquary with a controlled atmosphere, at 19~20 °C temperature, and 50% relative humidity. These values are shown to be unsuitable for maintaining the depolymerization of the cellulose at a level that is sufficiently low enough to preserve the image visible on the Shroud for a long time. [ABSTRACT FROM AUTHOR]

Abstract Paper degradation menaces the useful lifetime of books, manuscripts, and works on paper during storage, circulation, and display in libraries, archives, and museums. Severe damages such as embrittlement, decay, and mold often occur to the paper that might threaten to lose cultural heritage. However, the shelf life of papers stored in suitable conditions can be extended by hundreds of years. The most important external factors affecting the deterioration of paper-based materials include, in particular, changes in temperature and air humidity. In this study, the effects of accelerated aging under different conditions, including substantially different relative humidity, were considered relative to the strength properties of the paper sheets. These include the mechanical strength, such as breaking length, tear resistance, and bursting strength of the paper samples before and after dry heat aging and hydrothermal aging. Samples with various content of lignin produced in neutral pH were examined to exclude the adverse influence of acidity on paper properties. The results indicate that impact of moisture on tensile properties and pH-value of paper is much greater than the effect of increased temperature. The results of this work are intended to consolidate and expand the theoretical foundation and provide technical support for the conservators and library staff on the storage of paper cultural relics.

Biofilm commonly forms on the surfaces of cellulosic biomass but its roles in cellulose degradation remain largely unexplored. We used Bacillus subtilis to study possible mechanisms and the contributions of two major biofilm components, extracellular polysaccharides (EPS) and TasA protein, to submerged biofilm formation on cellulose and its degradation. We found that biofilm produced by B. subtilis is able to absorb exogenous cellulase added to the culture medium and also retain self-produced cellulase within the biofilm matrix. The bacteria that produced more biofilm degraded more cellulose compared to strains that produced less biofilm. Knockout strains that lacked both EPS and TasA formed a smaller amount of submerged biofilm on cellulose than the wild-type strain and also degraded less cellulose. Imaging of biofilm on cellulose suggests that bacteria, cellulose, and cellulases form cellulolytic biofilm complexes that facilitate synergistic cellulose degradation. This study brings additional insight into the important functions of biofilm in cellulose degradation and could potentiate the development of biofilm-based technology to enhance biomass degradation for biofuel production. [ABSTRACT FROM AUTHOR]

Brown rot fungi cause a type of wood decay characterized by carbohydrate degradation and lignin modification. The chemical and physical changes caused by brown rot are usually studied using bulk analytical methods, but these methods fail to consider local variations within the wood material. In this study we applied hyperspectral near infrared imaging to Scots pine sapwood samples exposed to the brown rot fungi Coniophora puteana and Rhodonia placenta to obtain position-resolved chemical information on the fungal degradative process. A stacked-sample decay test was used to create a succession of decay stages within the samples. The results showed that the key chemical changes associated with decay were the degradation of amorphous and crystalline carbohydrates and an increase in aromatic and carbonyl functionality in lignin. The position-resolved spectral data revealed that the fungi initiated degradation in earlywood, and that earlywood remained more extensively degraded than latewood even in advanced decay stages. Apart from differences in mass losses, the two fungi produced similar spectral changes in a similar spatial pattern. The results show that near infrared imaging is a useful tool for analyzing brown rot decayed wood and may be used to advance our understanding of fungal degradative processes. [ABSTRACT FROM AUTHOR]

The recently discovered type IX secretion system (T9SS) is limited to the Bacteroidetes phylum. Cytophaga hutchinsonii, a member of the Bacteroidetes phylum widely spread in soil, has complete orthologs of T9SS components and many T9SS substrates. C. hutchinsonii can efficiently degrade crystalline cellulose using a novel strategy, in which bacterial cells must be in direct contact with cellulose. It can rapidly glide over surfaces via unclear mechanisms. Studies have shown that T9SS plays an important role in cellulose degradation, gliding motility, and ion assimilation in C. hutchinsonii. As reported recently, T9SS substrates are N- or O-glycosylated at their C-terminal domains (CTDs), with N-glycosylation being related to the translocation and outer membrane anchoring of these proteins. These findings have deepened our understanding of T9SS in C. hutchinsonii. In this review, we focused on the research progress on diverse substrates and functions of T9SS in C. hutchinsonii and the glycosylation of its substrates. A model of T9SS functions and the glycosylation of its substrates was proposed.

Brown rot fungi cause a type of wood decay characterized by carbohydrate degradation and lignin modification. The chemical and physical changes caused by brown rot are usually studied using bulk analytical methods, but these methods fail to consider local variations within the wood material. In this study we applied hyperspectral near infrared imaging to Scots pine sapwood samples exposed to the brown rot fungi Coniophora puteana and Rhodonia placenta to obtain position-resolved chemical information on the fungal degradative process. A stacked-sample decay test was used to create a succession of decay stages within the samples. The results showed that the key chemical changes associated with decay were the degradation of amorphous and crystalline carbohydrates and an increase in aromatic and carbonyl functionality in lignin. The position-resolved spectral data revealed that the fungi initiated degradation in earlywood, and that earlywood remained more extensively degraded than latewood even in advanced decay stages. Apart from differences in mass losses, the two fungi produced similar spectral changes in a similar spatial pattern. The results show that near infrared imaging is a useful tool for analyzing brown rot decayed wood and may be used to advance our understanding of fungal degradative processes.

Cellulose degrading bacterial diversity of Bhitarkanika mangrove ecosystem, India, was uncovered and the cellulose degradation mechanism in Bacillus haynesii DS7010 under the modifiers such as pH (pCO 2), salinity and lead (Pb) was elucidated in the present study. The abundance of cellulose degrading heterotrophic bacteria was found to be higher in mangrove sediment than in water. The most potential strain, B. haynesii DS7010 showed the presence of endoglucanase, exoglucanase and β-glucosidase with the maximum degradation recorded at 48 h of incubation, with 1% substrate concentration at 41 °C incubation temperature. Two glycoside hydrolase genes, celA and celB were confirmed in this bacterium. 3D structure prediction of the translated CelA and CelB proteins showed maximum similarities with glycoside hydrolase 48 (GH48) and glycoside hydrolase 5 (GH5) respectively. Native PAGE followed by zymogram assay unveiled the presence of eight isoforms of cellulase ranged from 78 kDa to 245 kDa. Among the stressors, most adverse effect was observed under Pb stress at 1400 ppm concentration, followed by pH at pH 4. This was indicated by prolonged lag phase growth, higher reactive oxygen species (ROS) production, lower enzyme activity and downregulation of celA and celB gene expressions. Salinity augmented bacterial metabolism up to 3% NaCl concentration. Mangrove leaf litter degradation by B. haynesii DS7010 indicated a substantial reduction in cellulolytic potential of the bacterium in response to the synergistic effect of the stressors. Microcosm set up with the stressors exhibited 0.97% decrease in total carbon (C%) and 0.02% increase in total nitrogen (N%) after 35 d of degradation while under natural conditions, the reduction in C and the increase in N were 4.05% and 0.2%, respectively. The findings of the study suggest the cellulose degradation mechanism of a mangrove bacterium and its resilience to the future consequences of environmental pollution and climate change. [Display omitted] • GH48 and GH5 were confirmed within the cellulolytic system of B. haynesii DS7010. • Native PAGE with zymogram assay revealed the presence of eight cellulase isoforms. • Variable pH/pCO 2 and lead (Pb) negatively affected bacterial metabolism. • Salinity stress exhibited a positive effect on the bacterium up to 3% concentration. • Bacterial leaf litter degradation was severely affected by cumulative effect of stressors. [ABSTRACT FROM AUTHOR]

On a sample of the Turin Shroud (TS), we applied a new method for dating ancient linen threads by inspecting their structural degradation by means of Wide-Angle X-ray Scattering (WAXS). The X-ray dating method was applied to a sample of the TS consisting of a thread taken in proximity of the 1988/radiocarbon area (corner of the TS corresponding to the feet area of the frontal image, near the so-called Raes sample). The size of the linen sample was about 0.5 mm × 1 mm. We obtained one-dimensional integrated WAXS data profiles for the TS sample, which were fully compatible with the analogous measurements obtained on a linen sample whose dating, according to historical records, is 55–74 AD, Siege of Masada (Israel). The degree of natural aging of the cellulose that constitutes the linen of the investigated sample, obtained by X-ray analysis, showed that the TS fabric is much older than the seven centuries proposed by the 1988 radiocarbon dating. The experimental results are compatible with the hypothesis that the TS is a 2000-year-old relic, as supposed by Christian tradition, under the condition that it was kept at suitable levels of average secular temperature—20.0–22.5 °C—and correlated relative humidity—75–55%—for 13 centuries of unknown history, in addition to the seven centuries of known history in Europe. To make the present result compatible with that of the 1988 radiocarbon test, the TS should have been conserved during its hypothetical seven centuries of life at a secular room temperature very close to the maximum values registered on the earth. [ABSTRACT FROM AUTHOR]

• Diagnostic of ancient paper by completely non-invasive diagnostic approach. • UV–Vis-NIR imaging, FORS and fluorescence spectroscopies performed in-situ. • HPLC and FTIR analyses on extracts obtained by a smart sampling method. • A complete characterization of two letters of St. Francis de Sales (1567–1622) was obtained. We report a non-invasive and multi-analytical physico-chemical method for the characterization of paper artworks, able to identify sizing, inks, and glues and to quantify oxidative degradation by-products. The proposed methodology is mostly carried out in situ by using easy-to-use and cheap portable instrumentation for the acquisition of multispectral images, punctual ultraviolet-visible-near infrared fiber-optics reflectance spectroscopy (FORS) and punctual fiber optics fluorescence spectroscopy. Further analytical information is provided by non-invasive in-situ sampling of paper surface by using soft latex sponges, making possible laboratory chromatographic and infrared spectroscopic analyses on the aqueous sponge extracts. The proposed diagnostics method was applied to two 17th century letters written by St Francis of Sales (1567–1622), collected at the Chigi Palace in the town of Ariccia (Italy). Results show an intense oxidative degradation of the letters, also localized in water spots, and the presence of carboxylic acids by-products. Analysis of FORS spectra provided the concentration of chromophores in the paper substrate. The diagnostic method allowed the identification of gelatin sizing, the presence of starch glue in specific areas of the letters and the type of ink used in the text. Our diagnostic approach aims to offer to conservator-restorer a characterization of a paper artwork, that can be applied to other, for a correct planning of conservation interventions. [ABSTRACT FROM AUTHOR]

As part of free cellulases or scaffolding proteins in cellulosomes, the hydrophilic non-catalytic X2 module is widely distributed in cellulolytic Clostridia or other Firmicutes bacteria. Previous biochemical studies suggest that X2 modules might increase the solubility and substrate binding affinity of X2-bearing proteins. However, their in vivo biological functions remain elusive. Here we employed CRISPR-Cas9 editing to genetically modify X2 modules by deleting the conserved motif (NGNT) from the CipC scaffoldin. Both single and double X2 mutants (X2-N: near the N terminus of CipC; X2-C: near the C terminus of CipC) presented similar stoichiometric compositions in isolated cellulosomes as the wildtype strain (WT). These X2 mutants had an elongated adaptation stage during growth on cellulose compared to cellobiose. Compared to WT, the double mutant ΔX2-NC reduced cellulose degradation by 15% and the amount of released soluble sugars by 63%. Since single X2 mutants did not present such obvious physiological changes as ΔX2-NC, there seems to be a functional redundancy between X2 modules in CipC. The in vivo adhesion assay revealed that ΔX2-NC decreased cell attachment to cellulose by 70% but a weaker effect was also overserved in single X2 mutants. These results highlight the in vivo biological role of X2 in increasing cellulose degradation efficiency by enhancing the binding affinity between cells and cellulose, which provides new perspectives for microbial engineering. [ABSTRACT FROM AUTHOR]

Cytophaga hutchinsonii can efficiently degrade crystalline cellulose, in which the cell surface cellulases secreted by the type IX secretion system (T9SS) play important roles, but the degradation mechanism remains unclear, and the anchor mechanism of cellulases on the outer membrane in C. hutchinsonii has not been studied. Here, chu_2177 was identified by transposon mutagenesis and was proved to be indispensable for cellulose utilization in C. hutchinsonii. Disruption of chu_2177 resulted in O-antigen deficiency and chu_177 could confer O-antigen ligase activity upon an Escherichia coli waal mutant, indicating that chu_2177 encoded the O-ntigen ligase. Moreover, deletion of chu_2177 caused defects in cellulose utilization, cell motility, biofilm formation, and stress resistance. Further study showed that the endoglucanase activity was markedly decreased in the outer membrane but was increased in the culture fluid without chu_2177. Western blot proved that endoglucanase CHU_1336 was not located on the outer membrane but was released in the culture fluid of the Δ2177 mutant. Further proteomics analysis showed that many cargo proteins of T9SS were missing in the outer membrane of the Δ2177 mutant. Our study revealed that the deletion of chu_2177 affected the localization of many T9SS cargo proteins including cellulases on the outer membrane of C. hutchinsonii. [ABSTRACT FROM AUTHOR]

ABSTRACT The complexity of microbial communities hinders our understanding of how microbial diversity and microbe-microbe interactions impact community functions. Here, using six independent communities originating from the refuse dumps of leaf-cutter ants and enriched using the plant polymer cellulose as the sole source of carbon, we examine how changes in bacterial diversity and interactions impact plant biomass decomposition. Over up to 60 serial transfers (∼8 months) using Whatman cellulose filter paper, cellulolytic ability increased and then stabilized in four enrichment lines and was variable in two lines. Bacterial community characterization using 16S rRNA gene amplicon sequencing showed community succession differed between the highly cellulolytic enrichment lines and those that had slower and more variable cellulose degradation rates. Metagenomic and metatranscriptomic analyses revealed that Cellvibrio and/or Cellulomonas dominated each enrichment line and produced the majority of cellulase enzymes, while diverse taxa were retained within these communities over the duration of transfers. Interestingly, the less cellulolytic communities had a higher diversity of organisms competing for the cellulose breakdown product cellobiose, suggesting that cheating slowed cellulose degradation. In addition, we found competitive exclusion as an important factor shaping all of the communities, with a negative correlation of Cellvibrio and Cellulomonas abundance within individual enrichment lines and the expression of genes associated with the production of secondary metabolites, toxins, and other antagonistic compounds. Our results provide insights into how microbial diversity and competition affect the stability and function of cellulose-degrading communities. IMPORTANCE Microbial communities are a key driver of the carbon cycle through the breakdown of complex polysaccharides in diverse environments including soil, marine systems, and the mammalian gut. However, due to the complexity of these communities, the species-species interactions that impact community structure and ultimately shape the rate of decomposition are difficult to define. Here, we performed serial enrichment on cellulose using communities inoculated from leaf-cutter ant refuse dumps, a cellulose-rich environment. By concurrently tracking cellulolytic ability and community composition and through metagenomic and metatranscriptomic sequencing, we analyzed the ecological dynamics of the enrichment lines. Our data suggest that antagonism is prevalent in these communities and that competition for soluble sugars may slow degradation and lead to community instability. Together, these results help reveal the relationships between competition and polysaccharide decomposition, with implications in diverse areas ranging from microbial community ecology to cellulosic biofuels production.

As part of free cellulases or scaffolding proteins in cellulosomes, the hydrophilic non-catalytic X2 module is widely distributed in cellulolytic Clostridia or other Firmicutes bacteria. Previous biochemical studies suggest that X2 modules might increase the solubility and substrate binding affinity of X2-bearing proteins. However, their in vivo biological functions remain elusive. Here we employed CRISPR-Cas9 editing to genetically modify X2 modules by deleting the conserved motif (NGNT) from the CipC scaffoldin. Both single and double X2 mutants (X2-N: near the N terminus of CipC; X2-C: near the C terminus of CipC) presented similar stoichiometric compositions in isolated cellulosomes as the wildtype strain (WT). These X2 mutants had an elongated adaptation stage during growth on cellulose compared to cellobiose. Compared to WT, the double mutant ΔX2-NC reduced cellulose degradation by 15% and the amount of released soluble sugars by 63%. Since single X2 mutants did not present such obvious physiological changes as ΔX2-NC, there seems to be a functional redundancy between X2 modules in CipC. The in vivo adhesion assay revealed that ΔX2-NC decreased cell attachment to cellulose by 70% but a weaker effect was also overserved in single X2 mutants. These results highlight the in vivo biological role of X2 in increasing cellulose degradation efficiency by enhancing the binding affinity between cells and cellulose, which provides new perspectives for microbial engineering.

β-Glucosidases (Bgls) convert cellobiose and other soluble cello-oligomers into glucose and play important roles in fundamental biological processes, providing energy sources in living organisms. Bgls are essential terminal enzymes of cellulose degradation systems and attractive targets for lignocellulose-based biotechnological applications. Characterization of novel Bgls is important for broadening our knowledge of this enzyme class and can provide insights into its further applications. In this study, we report the biochemical and structural analysis of a Bgl from the hemicellulosedegrading thermophilic anaerobe Thermoanaerobacterium saccharolyticum (TsaBgl). TsaBgl exhibited its maximum hydrolase activity on p-nitrophenyl-β-D-glucopyranoside at pH 6.0 and 55 ◦C. The crystal structure of TsaBgl showed a single (β/α)8 TIM-barrel fold, and a β8-α14 loop, which is located around the substrate-binding pocket entrance, showing a unique conformation compared with other structurally known Bgls. A Tris molecule inhibited enzyme activity and was bound to the active site of TsaBgl coordinated by the catalytic residues Glu163 (proton donor) and Glu351 (nucleophile). Titration experiments showed that TsaBgl belongs to the glucose-tolerant Bgl family. The gatekeeper site of TsaBgl is similar to those of other glucose-tolerant Bgls, whereas Trp323 and Leu170, which are involved in glucose tolerance, show a unique configuration. Our results therefore improve our knowledge about the Tris-mediated inhibition and glucose tolerance of Bgl family members, which is essential for their industrial application. [ABSTRACT FROM AUTHOR]