Plant Cell Wall Components Research Articles

Cellulolytic and Xylanolytic Enzymes from Yeasts: Properties and Industrial Applications.

Lignocellulose, the main component of plant cell walls, comprises polyaromatic lignin and fermentable materials, cellulose and hemicellulose. It is a plentiful and renewable feedstock for chemicals and energy. It can serve as a raw material for the production of various value-added products, including cellulase and xylanase. Cellulase is essentially required in lignocellulose-based biorefineries and is applied in many commercial processes. Likewise, xylanases are industrially important enzymes applied in papermaking and in the manufacture of prebiotics and pharmaceuticals. Owing to the widespread application of these enzymes, many prokaryotes and eukaryotes have been exploited to produce cellulase and xylanases in good yields, yet yeasts have rarely been explored for their plant-cell-wall-degrading activities. This review is focused on summarizing reports about cellulolytic and xylanolytic yeasts, their properties, and their biotechnological applications.

Deletion of AA9 Lytic Polysaccharide Monooxygenases Impacts A. nidulans Secretome and Growth on Lignocellulose.

ABSTRACTLytic polysaccharide monooxygenases (LPMOs) are oxidative enzymes found in viruses, archaea, and bacteria as well as eukaryotes, such as fungi, algae and insects, actively contributing to the degradation of different polysaccharides. In Aspergillus nidulans, LPMOs from family AA9 (AnLPMO9s), along with an AA3 cellobiose dehydrogenase (AnCDH1), are cosecreted upon growth on crystalline cellulose and lignocellulosic substrates, indicating their role in the degradation of plant cell wall components. Functional analysis revealed that three target LPMO9s (AnLPMO9C, AnLPMO9F and AnLPMO9G) correspond to cellulose-active enzymes with distinct regioselectivity and activity on cellulose with different proportions of crystalline and amorphous regions. AnLPMO9s deletion and overexpression studies corroborate functional data. The abundantly secreted AnLPMO9F is a major component of the extracellular cellulolytic system, while AnLPMO9G was less abundant and constantly secreted, and acts preferentially on crystalline regions of cellulose, uniquely displaying activity on highly crystalline algae cellulose. Single or double deletion of AnLPMO9s resulted in about 25% reduction in fungal growth on sugarcane straw but not on Avicel, demonstrating the contribution of LPMO9s for the saprophytic fungal lifestyle relies on the degradation of complex lignocellulosic substrates. Although the deletion of AnCDH1 slightly reduced the cellulolytic activity, it did not affect fungal growth indicating the existence of alternative electron donors to LPMOs. Additionally, double or triple knockouts of these enzymes had no accumulative deleterious effect on the cellulolytic activity nor on fungal growth, regardless of the deleted gene. Overexpression of AnLPMO9s in a cellulose-induced secretome background confirmed the importance and applicability of AnLPMO9G to improve lignocellulose saccharification.IMPORTANCE Fungal lytic polysaccharide monooxygenases (LPMOs) are copper-dependent enzymes that boost plant biomass degradation in combination with glycoside hydrolases. Secretion of LPMO9s arsenal by Aspergillus nidulans is influenced by the substrate and time of induction. These findings along with the biochemical characterization of novel fungal LPMO9s have implications on our understanding of their concerted action, allowing rational engineering of fungal strains for biotechnological applications such as plant biomass degradation. Additionally, the role of oxidative players in fungal growth on plant biomass was evaluated by deletion and overexpression experiments using a model fungal system.

Discovery, characterization, and metabolic engineering of Rieske non-heme iron monooxygenases for guaiacol O-demethylation

Aryl- O -demethylation is a common rate-limiting step in the catabolism of lignin-related compounds, including guaiacol. Here we used randomly barcoded transposon insertion sequencing (RB-TnSeq) in the bacterium Novosphingobium aromaticivorans to identify a Rieske-type guaiacol O -demethylase, GdmA. Similarity searches identified GdmA homologs in other bacteria, along with candidate reductase partners, denoted GdmB. GdmAB combinations were biochemically characterized for activity with several lignin-related substrates. Structural and sequence comparisons of vanillate- and guaiacol-specific O- demethylase active sites revealed conserved hallmarks of substrate specificity. GdmAB combinations were also evaluated in Pseudomonas putida KT2440, which does not natively utilize guaiacol. GdmAB from Cupriavidus necator N-1 demonstrated the highest rate of guaiacol turnover in vitro and in engineered P. putida strains and notably higher catalytic efficiency than a cytochrome P450 system (GcoAB) and the vanillate Rieske-type O -demethylase from P. putida (VanAB). The GdmAB O -demethylases described here expand the suite of options for microbial conversion of a model lignin-derived substrate. • A Rieske monooxygenase, GdmA, demethylates guaiacol in N. aromaticivorans DSM12444 • GdmA homologs and associated GdmB reductases display differences in specific activity • Structural comparison of O -demethylase active sites illuminates substrate specificity • Expression of GdmAB in engineered P. putida KT2440 enables growth on guaiacol Lignin is an abundant heterogenous component of plant cell walls, and its valorization is key to enable a profitable and sustainable global biofuels industry. A major challenge to convert lignin to chemicals is overcoming the inherent heterogeneity of deconstructed lignin polymers, and in recent years, microbial conversion has emerged as a means to produce single target compounds from mixtures of aromatic compounds. This work reports the discovery, characterization, and engineering of an enzyme system that can efficiently demethylate a lignin-related compound, 2-methoxy-phenol (commonly known as guaiacol). Aromatic O -demethylation is a common rate-limiting step in aromatic catabolic pathways and, thus, a prime target for discovery, engineering, and evolution of new enzymes. Going forward, biocatalysis development like that reported here may ultimately enable an integrated chemocatalytic and biocatalytic valorization approach for a major component of plant cell walls. Bacterial catabolism of lignin-related aromatic compounds is often limited by aryl- O -demethylation. This study identified a Rieske-type monooxygenase, GdmA, that catalyzes O- demethylation of guaiacol in Novosphingobium aromaticivorans. Detailed characterization of this enzyme, along with GdmA homologs and associated reductase partners (GdmB), identified differences in specific activity and substrate preference. Heterologous expression of GdmAB in Pseudomonas putida enabled growth on guaiacol. GdmAB O -demethylases expand the suite of options for microbial conversion of a model lignin-derived substrate.

Spartina argentinensis valorization and process optimization for enhanced production of hydrolytic enzymes by filamentous fungus

Hemicellulose is a major component of plant cell walls and xylan is the most predominant polysaccharide. Xylan degrading enzymes integrate the xylanolytic system. Xylanolytic enzymes were produced by fermentation by Aspergillus niger and Thermomyces lanuginosus grown on grass considered unsuitable for farming called Spartina argentinensis . Significant parameters: type of fermentation used (SmF or SSF), type of leaves of S. argentinensis (green or senescent) and conidia final concentration for xylanase production were screened and optimized. The main results showed that the highest levels of xylanolytic enzyme production were obtained by 1 × 10 5 conidia/mL of A. niger in SmF at 96 h, 30ºC, with a mixture of 20.20% of senescent and 79.80% of green leaves. The xylanase specific activity obtained was 62 U/mg, higher than the activity obtained (23 U/mg) in previous work ( Taddia et al., 2019 ), and the concentration of xylanolytic production over that of glucanase activity was maximized five times. The optimized enzymatic extract obtained was characterized by LC-MS and HPLC of carbohydrates. Six enzymes were identified as constituents of the xylanolytic complex and seven carbohydrates. Moreover, the xylanolytic enzyme extract was stable for 30 days at 20 °C. Thus, S. argentinensis can be used within the framework of a circular economy, rendering a synergistic combination of the xylanolytic enzymes with industrial applications. • Lignocellulosic biomass; Spartina argentinensis was utilized in the fermentation. • Enzyme statistical design production was optimized. • Maximum xylanase specific activity of 61 U/mL was obtained. • The enzymatic extract maintained 100% its activity after 30 days at 20°C.

Oryza sativa PECTIN DEFECTIVE TAPETUM1 affects anther development through a pectin-mediated signaling pathway in rice.

Galacturonosyltransferase (GalAT) is required for the synthesis of pectin, an important component of plant cell walls that is also involved in signal transduction. Here, we describe the rice (Oryza sativa) male-sterile mutant O. sativa pectin-defective tapetum1 (ospdt1), in which GalAT is mutated. The ospdt1 mutant exhibited premature programmed cell death (PCD) of the tapetum and disordered pollen walls, resulting in aborted pollen grains. Pectin distribution in the anther sac was comparable between the mutant and the wild-type, suggesting that the structural pectin was not dramatically affected in ospdt1. Wall-associated kinases are necessary for the signal transduction of pectin, and the intracellular distribution of O. sativa indica WALL-ASSOCIATED KINASE1 (OsiWAK1), which binds pectic polysaccharides to its extracellular domain, was affected in ospdt1. OsiWAK1 RNA interference lines exhibited earlier tapetal PCD, similar to ospdt1. Furthermore, overexpression of OsiWAK1 in ospdt1 lines partially rescued the defects observed in ospdt1, suggesting that OsiWAK1 plays pivotal roles in the function of OsPDT1. These results suggest that the mutation of OsPDT1 does not dramatically affect structural pectin but affects components of the pectin-mediated signaling pathway, such as OsiWAK1, and causes male sterility.

Exploring the impact of Verticillium wilt disease on the mechanical properties of elementary flax (Linum usitatissimum L.) fibres

Verticillium wilt is a disease caused by the fungus Verticillium dahliae (V. dahliae) which is widespread in flax (Linum usitatissimum L.) cultures. V. dahliae negatively impacts the fibre yield with up to 60% loss at the scutching step of fibre transformation. Yet, little is known about the consequences of V. dahliae on the mechanical properties of flax fibres. In this study, we investigated the tensile characterisation of elementary flax fibres impacted by V. dahlia. Using elementary tensile tests, we observed an important decrease in the mechanical properties of the studied flax fibres, with a 32% decrease of the tensile strength at break and a 15% decrease of the strain at break. Further investigation of the flax fibres cell wall using atomic force microscopy (AFM) in peak-force non-quantitative mechanical mapping mode, showed a 23% decrease in the longitudinal indentation modulus of flax fibre cell walls, measured directly in their cross section. This is arguably due to the important enzymatic arsenal of V. dahliae revealed by a bioinformatic analysis of secreted carbohydrate-active enzymes targeting plant cell wall components. The coupling of this analysis with electron microscopy observations and sugar analysis highlighted the degradation mechanisms of this fungus which significantly affects the mechanical performance of flax fibres

Electron ionization mass spectrometric fragmentation and multiple reaction monitoring quantification of ferulic and p-coumaric acid trimethylsilyl derivatives in deposited atmospheric particles.

Ferulic and p-coumaric acids are important biological and structural components of plant cell walls and possess antioxidant and antimicrobial properties. These phenolic acids are widespread in environmental samples. However, when they are present at very low concentrations or in very complex lipid extracts, their identification and quantification can be challenging. The electron ionization mass spectrometry (EI-MS) fragmentation pathways of ferulic and p-coumaric acid trimethylsilyl (TMS) derivatives were investigated. These pathways were deduced by (i) low-energy collision-induced dissociation (CID) gas chromatography (GC)/EI-MS/MS, (ii) accurate mass measurement, and (iii) 13 C labelling. These compounds were then characterized and quantified in multiple reaction monitoring (MRM) mode in total lipid extracts of deposited atmospheric particles using highly specific transitions based on the main fragmentation pathways elucidated. Low-energy CID-MS/MS analyses, accurate mass measurement and 13 C labelling enabled us to elucidate EI-MS fragmentations of ferulic and p-coumaric acid TMS derivatives. Some specific fragmentations proved useful for subsequent characterization and quantification of these compounds. As an application of some of the described fragmentations, trace amounts of these phenolic acids were characterized and quantified in MRM mode in wet- and dry-deposited atmospheric particles containing low proportions of organic matter. EI-MS fragmentations of ferulic and p-coumaric acid TMS derivatives exhibit specific fragment ions that can be very useful for the quantification of trace amounts of both phenolic acids in environmental samples.

Screening cellulose synthesis related genes of EgrEXP and EgrHEX in Eucalyptus grandis

Eucalyptus (including Eucalyptus grandis) is an excellent wood forest tree species that provides a large number of plant fiber raw materials for the paper and timber industries. Cellulose, an essential structural component in plant cell walls, is a renewable biomass resource that plays a very important role in nature. There is still a lack of research on the role of gene regulation in cellulose synthesis. To study the genes of cellulose synthesis, the wood chemical indexes of Eucalyptus grandis were analyzed by taking three different parts from the main stem of Eucalyptus grandis as raw materials. The results showed that the cellulose content in the middle of the trunk was significantly higher than that at the chest diameter and at the upper part of the trunk. A total of 296 differentially expressed genes (DEGs) were obtained from the three site by transcriptome, and 19 key candidate genes were related to the synthesis of cellulose in Eucalyptus grandis. EgrEXP1 and EgrHEX4 were overexpressed in 84 K poplar, the content of cellulose and lignin in genetically modified plants was significantly higher than that of wild type 84 K poplar. Also, the average plant height and average root count were significantly higher than those of control plants, and the average diameter of the middle and stem bases were significantly larger than those of control plants. In this study, the genes related to cellulose synthesis in Eucalyptus grandis are studied, which serve as a strong foundation for understanding the molecular regulation of cellulose synthesis in plants.

Isolation and characterization of microorganisms capable of cleaving the ether bond of 2-phenoxyacetophenone

Lignin is a heterogeneous aromatic polymer and major component of plant cell walls. The β-O-4 alkyl aryl ether is the most abundant linkage within lignin. Given that lignin is effectively degraded on earth, as yet unknown ether bond–cleaving microorganisms could still exist in nature. In this study, we searched for microorganisms that transform 2-phenoxyacetophenone (2-PAP), a model compound for the β-O-4 linkage in lignin, by monitoring ether bond cleavage. We first isolated microorganisms that grew on medium including humic acid (soil-derived organic compound) as a carbon source. The isolated microorganisms were subsequently subjected to colorimetric assay for 2-PAP ether bond–cleaving activity; cells of the isolated strains were incubated with 2-PAP, and strains producing phenol via ether bond cleavage were selected using phenol-sensitive Gibbs reagent. This screening procedure enabled the isolation of various 2-PAP–transforming microorganisms, including 7 bacteria (genera: Acinetobacter, Cupriavidus, Nocardioides, or Streptomyces) and 1 fungus (genus: Penicillium). To our knowledge, these are the first microorganisms demonstrated to cleave the ether bond of 2-PAP. One Gram-negative bacterium, Acinetobacter sp. TUS-SO1, was characterized in detail. HPLC and GC–MS analyses revealed that strain TUS-SO1 oxidatively and selectively cleaves the ether bond of 2-PAP to produce phenol and benzoate. These results indicate that the transformation mechanism differs from that involved in reductive β-etherase, which has been well studied. Furthermore, strain TUS-SO1 efficiently transformed 2-PAP; glucose-grown TUS-SO1 cells converted 1 mM 2-PAP within only 12 h. These microorganisms might play important roles in the degradation of lignin-related compounds in nature.

Broad Specific Xyloglucan:Xyloglucosyl Transferases Are Formidable Players in the Re-Modelling of Plant Cell Wall Structures.

Plant xyloglucan:xyloglucosyl transferases, known as xyloglucan endo-transglycosylases (XETs) are the key players that underlie plant cell wall dynamics and mechanics. These fundamental roles are central for the assembly and modifications of cell walls during embryogenesis, vegetative and reproductive growth, and adaptations to living environments under biotic and abiotic (environmental) stresses. XET enzymes (EC 2.4.1.207) have the β-sandwich architecture and the β-jelly-roll topology, and are classified in the glycoside hydrolase family 16 based on their evolutionary history. XET enzymes catalyse transglycosylation reactions with xyloglucan (XG)-derived and other than XG-derived donors and acceptors, and this poly-specificity originates from the structural plasticity and evolutionary diversification that has evolved through expansion and duplication. In phyletic groups, XETs form the gene families that are differentially expressed in organs and tissues in time- and space-dependent manners, and in response to environmental conditions. Here, we examine higher plant XET enzymes and dissect how their exclusively carbohydrate-linked transglycosylation catalytic function inter-connects complex plant cell wall components. Further, we discuss progress in technologies that advance the knowledge of plant cell walls and how this knowledge defines the roles of XETs. We construe that the broad specificity of the plant XETs underscores their roles in continuous cell wall restructuring and re-modelling.

Evolutionary Dynamics of Host Organs for Microbial Symbiosis in Tortoise Leaf Beetles (Coleoptera: Chrysomelidae: Cassidinae).

ABSTRACTDiverse insects host specific microbial symbionts that play important roles for their growth, survival, and reproduction. They often develop specialized symbiotic organs for harboring the microbial partners. While such intimate associations tend to be stably maintained over evolutionary time, the microbial symbionts may have been lost or replaced occasionally. How symbiont acquisitions, replacements, and losses are linked to the development of the host’s symbiotic organs is an important but poorly understood aspect of microbial symbioses. Cassidine leaf beetles are associated with a specific gammaproteobacterial lineage, Stammera, whose reduced genome is streamlined for producing pectin-degrading enzymes to assist the host’s digestion of food plants. We investigated the symbiotic system of 24 Japanese cassidine species and found that (i) most species harbored Stammera within paired symbiotic organs located at the foregut-midgut junction, (ii) the host phylogeny was largely congruent with the symbiont phylogeny, indicating stable host-symbiont association over evolutionary time, (iii) meanwhile, the symbiont was not detected in three distinct host lineages, uncovering recurrent losses of the ancient microbial mutualist, (iv) the symbiotic organs were vestigial but present in the symbiont-free lineages, indicating evolutionary persistence of the symbiotic organs even in the absence of the symbiont, and (v) the number of the symbiotic organs was polymorphic among the cassidine species, either two or four, unveiling a dynamic evolution of the host organs for symbiosis. These findings are discussed as to what molecular mechanisms and evolutionary trajectories underpin the recurrent symbiont losses and the morphogenesis of the symbiotic organs in the herbivorous insect group.

The preparation of an insulator material using renewable sources and application of palm oil tree – A Review

Excessive heat will cause discomfort while doing daily life even the use of electrical items such as fans and air conditioner are not entirely helpful. This is because the building does not have a layer to absorb the heat. Then, the best initiative to lessen the heat in building is by adding thermal insulation material into the building material. Past researcher shown most thermal insulators used are fiberglass, mineral wool, polystyrene and polyurethane foam. However, there are some flaws with the thermal insulator that cause new ideas to emerge namely cellulose. Cellulose is an insoluble substance that is a major component of plant cell walls and vegetable fibres such as cotton. Normally cellulose can be found in the plants. By cause to the increasing surplus of oil palm trees then the use of cellulose in oil palm is the best. This paper review discusses about preparation of an insulator material using renewable sources and application of palm oil tree.

Down-regulation of OsMYB103L distinctively alters beta-1,4-glucan polymerization and cellulose microfibers assembly for enhanced biomass enzymatic saccharification in rice

BackgroundAs a major component of plant cell walls, cellulose provides the most abundant biomass resource convertible for biofuels. Since cellulose crystallinity and polymerization have been characterized as two major features accounting for lignocellulose recalcitrance against biomass enzymatic saccharification, genetic engineering of cellulose biosynthesis is increasingly considered as a promising solution in bioenergy crops. Although several transcription factors have been identified to regulate cellulose biosynthesis and plant cell wall formation, much remains unknown about its potential roles for genetic improvement of lignocellulose recalcitrance.ResultsIn this study, we identified a novel rice mutant (Osfc9/myb103) encoded a R2R3-MYB transcription factor, and meanwhile generated OsMYB103L-RNAi-silenced transgenic lines. We determined significantly reduced cellulose levels with other major wall polymers (hemicellulose, lignin) slightly altered in mature rice straws of the myb103 mutant and RNAi line, compared to their wild type (NPB). Notably, the rice mutant and RNAi line were of significantly reduced cellulose features (crystalline index/CrI, degree of polymerization/DP) and distinct cellulose nanofibers assembly. These alterations consequently improved lignocellulose recalcitrance for significantly enhanced biomass enzymatic saccharification by 10–28% at p < 0.01 levels (n = 3) after liquid hot water and chemical (1% H2SO4, 1% NaOH) pretreatments with mature rice straws. In addition, integrated RNA sequencing with DNA affinity purification sequencing (DAP-seq) analyses revealed that the OsMYB103L might specifically mediate cellulose biosynthesis and deposition by regulating OsCesAs and other genes associated with microfibril assembly.ConclusionsThis study has demonstrated that down-regulation of OsMYB103L could specifically improve cellulose features and cellulose nanofibers assembly to significantly enhance biomass enzymatic saccharification under green-like and mild chemical pretreatments in rice. It has not only indicated a powerful strategy for genetic modification of plant cell walls in bioenergy crops, but also provided insights into transcriptional regulation of cellulose biosynthesis in plants.

Genomic insight of two indigenous probiotics Lactobacillus plantarum Dad-13 and Lactobacillus plantarum Mut-7 from different origins of Indonesian fermented foods

Abstract. Suroto DA, Hasan PN, Rahayu ES. 2021. Genomic insight of two indigenous probiotics Lactobacillus plantarum Dad-13 and Lactobacillus plantarum Mut-7 from different origins of Indonesian fermented foods. Biodiversitas 22: 5491-5500. Lactobacillus plantarum Dad-13 and Lactobacillus plantarum Mut-7 are probiotics isolated from traditional fermented foods. In the present study two strains of L. plantarum were sequenced using Illumina NovaSeq PE150 platform and in silico analysis was performed using RAST webserver to unravel the functional metabolic between two strains. The L. plantarum Dad-13 and Mut-7 genome sizes were 3.25 and 3.98 Mb with GC content of them being almost similar i.e. 44.4 and 44.3%, respectively. Two lactobacilli also had some differences in their metabolic genes, particularly in carbohydrate subsystem. L. plantarum Mut-7 had genes related to the metabolisms of plant cell wall components such as dextrin, arabinose, xylose indicated the adaptation to their environmental origins (fermented dried cassava), while those genes were not observed in L. plantarum Dad-13 which was isolated from fermented buffalo’s milk (dadih). Further analysis using BAGEL4 webserver exhibited their different potential to synthesize bacteriocins, L. plantarum Dad-13 was predicted to produce more types of bacteriocins than L. plantarum Mut-7. This investigation revealed that L. plantarum Dad-13 and L. plantarum Mut-7 have a great potential related with their properties as probiotics.

Bioactive Carbohydrate Polymers-Between Myth and Reality.

Polysaccharides are complex macromolecules long regarded as energetic storage resources or as components of plant and fungal cell walls. They have also been described as plant mucilages or microbial exopolysaccharides. The development of glycosciences has led to a partial and difficult deciphering of their other biological functions in living organisms. The objectives of glycobiochemistry and glycobiology are currently to correlate some structural features of polysaccharides with some biological responses in the producing organisms or in another one. In this context, the literature focusing on bioactive polysaccharides has increased exponentially during the last two decades, being sometimes very optimistic for some new applications of bioactive polysaccharides, notably in the medical field. Therefore, this review aims to examine bioactive polysaccharide, taking a critical look of the different biological activities reported by authors and the reality of the market. It focuses also on the chemical, biochemical, enzymatic, and physical modifications of these biopolymers to optimize their potential as bioactive agents.

Genome-wide association study for lignocellulosic compounds and fermentable sugar in rice straw.

Cellulose and lignin are the two main components of secondary plant cell walls with substantial impact on stalk in the field and on straw during industrial processing. The amount of fermentable sugar that can be accessed is another important parameter affecting various industrial applications. In the present study, genetic variability of rice (Oryza sativa L.) genotypes for cellulose, lignin, and fermentable sugars contents was analyzed in rice straw. A genome-wide association study of 33,484 single nucleotide polymorphisms (SNPs) with a minor allele frequency (MAF)>0.05 was performed. The genome-wide association study identified seven, three, and three genomic regions to be significantly associated with cellulose, lignin, and fermentable sugar contents, respectively. Candidate genes in the associated genomic regions were enzymes mainly involved in cell wall metabolism. Novel SNP markers associated with cellulose were tagged to GH16, peroxidase, GT6, GT8, and CSLD2. For lignin content, Villin protein, OsWAK1/50/52/53, and GH16 were identified. For fermentable sugar content, UTP-glucose-1-phosphate uridylyltransferase, BRASSINOSTEROID INSENSITIVE 1, and receptor-like protein kinase 5 were found. The results of this study should improve our understanding of the genetic basis of the factors that might be involved in biosynthesis, turnover, and modification of major cell wall components and saccharides in rice straw.

Short-term cellulose addition decreases microbial diversity and network complexity in an Ultisol following 32-year fertilization

Cellulose is an essential component of plant cell walls, and one of the major constituents of soil organic matter. Decomposition of cellulose, mediated by microorganisms, is critical to the sustainable development of arable soils. However, how exogenous cellulose addition, as a surrogate for organic material amendment, influences the diversity and community compositions of soil microorganisms and their network complexity remains largely unknown. To bridge this knowledge gap, we conducted a microcosm experiment incubated with soils that had been subjected to long-term mineral or organic fertilization. Cellulose addition increased the cumulative CO2 emission in all treatments, with the highest value found in the pig manure amended soils. Cellulose addition significantly reduced the alpha diversity of bacteria and fungi across all fertilization treatments, with a more pronounced effect observed for fungi. Moreover, cellulose addition strongly altered bacterial community structure by increasing the relative abundance of copiotrophic bacteria such as Actinobacteria and Proteobacteria while reducing that of oligotrophic bacteria (e.g., Chloroflexi). Compared with no-fertilizer treatment, the influence of cellulose addition on bacterial and fungal community structure was stronger in long-term mineral or organic fertilized soils. Cellulose addition altered the relative abundance of bacterial and fungal functional groups by generally enriching microbes involved in chemoheterotrophy and suppressing those involved in methylotrophy, pathotroph, saprotroph-symbiotroph, and saprotroph-pathotroph. In addition, cellulose addition decreased soil microbial complexity and the number of potential keystone species, but did not change the key role of Chloroflexi in soil microbial network. Overall, our results demonstrated that cellulose addition decreased soil microbial diversity and network complexity, with possible short-term negative consequences for ecosystem functioning in Ultisols.

Isolation and screening of local cellulolytic fungi from the digestive tract larvae of Oryctes rhinoceros L., North Sumatra

Cellulose, which is the main component of plant cell walls from higher plants, has been studied from different aspects. It is insoluble in a wide variety of solvents and is resistant to various chemicals treatments. Fungi are a group of cellulose-degrading microbes and plays major role in recycling of lignocellulosic material in nature. This study aimed to obtain cellulolytic fungi from the digestive tract of Oryctes rhinoceros L. larvae and to determine cellulolytic activity. Isolation and screening of cellulolytic fungi in the digestive tract of insects were carried out with specific medium Carboxymethyl Cellulose (CMC) and the Congo Red method to obtain potential cellulolytic isolates. Eleven fungal isolates showed positive results as cellulolytic fungi. The highest cellulolytic activity was obtained from isolate F05L with a cellulolytic index of 0.90 and isolate F10L of 0.66. The smallest cellulolytic activity was obtained from isolate F02L with a cellulolytic index of 0.14. All isolates would be identified to the species level and analyzed its potential applications. Our result can provide in addition to the environmental and industrial fields, cellulolytic fungi can a solution to the problem of pollution, namely reducing the amount of cellulose waste, and can be added value to the use of waste into processed organic fertilizers to be able to provide solutions to the problem of organic waste degradation.

Water-accessibility of interfibrillar spaces in spruce wood cell walls

Water interactions and accessibility of the nanoscale components of plant cell walls influence their properties and processability in relation to many applications. We investigated the water-accessibility of nanoscale pores within the fibrillar structures of unmodified Norway spruce cell walls by small-angle neutron scattering (SANS) and Fourier-transform infra-red (FTIR) spectroscopy. The different sensitivity of SANS to hydrogenated (hbox {H}_2hbox {O}) and deuterated water (hbox {D}_2hbox {O}) was utilized to follow the exchange kinetics of water among cellulose microfibrils. FTIR spectroscopy was used to study the time-dependent re-exchange of OD groups to OH in wood samples transferred from liquid hbox {D}_2hbox {O} to hbox {H}_2hbox {O}. In addition, the effects of drying on the nanoscale structure and its water-accessibility were addressed by comparing SANS results and the kinetics of water exchange between never-dried and dried/rewetted wood samples. The results of the kinetic analyses allowed to identify two processes with different timescales. The diffusion-driven exchange of water in the spaces between microfibrils, which was observed with both SANS and FTIR, takes place within minutes and rather homogeneously. The second, slower process appeared only in the OD/OH re-exchange followed by FTIR, and it still continued after several weeks of immersion in hbox {H}_2hbox {O}. SANS could not detect any significant difference between the never-dried and dried/rewetted samples, whereas FTIR revealed a small portion of OD groups that resisted the re-exchange and this portion became larger with drying.Graphic abstract

Environmental role of aromatic carboxylesterases

The carboxylesterases (EC 3.1.1.x) are widely distributed and form an important yet diverse group of hydrolases catalysing the ester bond cleavage in a variety of substrates. Besides acting on plant cell wall components like cutin, tannin and feruloyl esters, they are often the first line of defence to metabolize drugs, xenobiotics, pesticides, insecticides and plastic. But for the promiscuity of some carboxylesterases and cutinases, very few enzymes act exclusively on aromatic carboxylic acid esters. Infrequent occurrence of aromatic carboxylesterases suggests that aromatic carboxylesters are inherently more difficult to hydrolyse than the regular carboxylesters because of both steric and polar effects. Naturally occurring aromatic carboxylesters were rare before the anthropogenic activity augmented their environmental presence and diversity. An appraisal of the literature shows that the hydrolysis of aromatic carboxylic esters is a uniquely difficult endeavour and hence deserves special attention. Enzymes to hydrolyse such esters are evolving rapidly in nature. Very few such enzymes are known and they often display much lower catalytic efficiencies. Obviously, the esters of aromatic carboxylic acids, including polyethylene terephthalate waste, pose an environmental challenge. In this review, we highlight the uniqueness of aromatic carboxylesters and then underscore the importance of relevant carboxylesterases.