CAZyChip: Dynamic assessment of exploration of glycoside hydrolases in microbial diversity ecosystems.
Ontology highlight
ABSTRACT:
ORGANISM(S): Bacteria
PROVIDER: GSE80173 | GEO | 2016/12/01
SECONDARY ACCESSION(S): PRJNA318173
REPOSITORIES: GEO
Similar Datasets
Project description:Few aerobic hyperthermophiles degrade polysaccharides. We describe the genome-enabled enrichment and isolation of an aerobic hyperthermophile, Fervidibacter sacchari, which was originally ascribed to candidate phylum Fervidibacteria. F. sacchari uses polysaccharides and monosaccharides as sole carbon sources from 65-87.5 °C, and its genome encodes 117 glycoside hydrolases (GHs) spanning 49 GH families, including 31 homologs of understudied GH109, GH177, and GH179 domains. Here, we analyzed the transcriptomes of F. sacchari cells grown on eight different sole carbon and energy sources (beta-glucan, chondroitin sulfate, corn stover, gellan gum, locust bean gum, starch, xanthan gum, and xyloglucan) to link glycoside hydrolase substrate to function, as well as identify potential regulatory mechanisms. These data will provide preliminary characterization of novel carbohydrate-active enzymes at high temperatures.
2023-12-31 | GSE249938 | GEO
Project description:BackgroundMicroorganisms constitute a reservoir of enzymes involved in environmental carbon cycling and degradation of plant polysaccharides through their production of a vast variety of Glycoside Hydrolases (GH). The CAZyChip was developed to allow a rapid characterization at transcriptomic level of these GHs and to identify enzymes acting on hydrolysis of polysaccharides or glycans.ResultsThis DNA biochip contains the signature of 55,220 bacterial GHs available in the CAZy database. Probes were designed using two softwares, and microarrays were directly synthesized using the in situ ink-jet technology. CAZyChip specificity and reproducibility was validated by hybridization of known GHs RNA extracted from recombinant E. coli strains, which were previously identified by a functional metagenomic approach. The GHs arsenal was also studied in bioprocess conditions using rumen derived microbiota.ConclusionsThe CAZyChip appears to be a user friendly tool for profiling the expression of a large variety of GHs. It can be used to study temporal variations of functional diversity, thereby facilitating the identification of new efficient candidates for enzymatic conversions from various ecosystems.
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Project description:Across many environments microbial glycoside hydrolases support the enzymatic processing of carbohydrates, a critical function in many ecosystems. Little is known about how the microbial composition of a community and the potential for carbohydrate processing relate to each other. Here, using 1,934 metagenomic datasets, we linked changes in community composition to variation of potential for carbohydrate processing across environments. We were able to show that each ecosystem-type displays a specific potential for carbohydrate utilization. Most of this potential was associated with just 77 bacterial genera. The GH content in bacterial genera is best described by their taxonomic affiliation. Across metagenomes, fluctuations of the microbial community structure and GH potential for carbohydrate utilization were correlated. Our analysis reveals that both deterministic and stochastic processes contribute to the assembly of complex microbial communities.
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Project description:During the last decades, the impact of hyperthermophiles and their enzymes has been intensively investigated for implementation in various high-temperature biotechnological processes. Biocatalysts of hyperthermophiles have proven to show extremely high thermo-activities and thermo-stabilities and are identified as suitable candidates for numerous industrial processes with harsh conditions, including the process of an efficient plant biomass pretreatment and conversion. Already-characterized archaea-originated glycoside hydrolases (GHs) have shown highly impressive features and numerous enzyme characterizations indicated that these biocatalysts show maximum activities at a higher temperature range compared to bacterial ones. However, compared to bacterial biomass-degrading enzymes, the number of characterized archaeal ones remains low. To discover new promising archaeal GH candidates, it is necessary to study in detail the microbiology and enzymology of extremely high-temperature habitats, ranging from terrestrial to marine hydrothermal systems. State-of-the art technologies such as sequencing of genomes and metagenomes and automated binning of genomes out of metagenomes, combined with classical microbiological culture-dependent approaches, have been successfully performed to detect novel promising biomass-degrading hyperthermozymes. In this review, we will focus on the detection, characterization and similarities of archaeal GHs and their unique characteristics. The potential of hyperthermozymes and their impact on high-temperature industrial applications have not yet been exhausted.
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Project description:Allylic cyclitols were investigated as covalent inhibitors of glycoside hydrolases by chemical, enzymatic, proteomic, and computational methods. This approach was inspired by the C7 cyclitol natural product streptol glucoside, which features a potential carbohydrate leaving group in the 4-position (carbohydrate numbering). To test this hypothesis, carbocyclic inhibitors with leaving groups in the 4- and 6- positions were prepared. The results of enzyme kinetics analyses demonstrated that dinitrophenyl ethers covalently inhibit α-glucosidases of the GH13 family without reactivation. The labeled enzyme was studied by proteomics, and the active site residue Asp214 was identified as modified. Additionally, computational studies, including enzyme homology modeling and density functional theory (DFT) calculations, further delineate the electronic and structural requirements for activity. This study demonstrates that previously unexplored 4- and 6-positions can be exploited for successful inhibitor design.
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Project description:Chronic wounds will impact 2% of the United States population at some point in their life. These wounds are often associated with a reoccurring, chronic infection caused by a community of microorganisms encased in an extracellular polymeric substance (EPS), or a biofilm. Biofilm-associated microbes can exhibit tolerance to antibiotics, which has prompted researchers to investigate therapeutics that improve antibiotic efficacy. Glycoside hydrolases (GHs), enzymes that target the polysaccharide linkages within the EPS, are one potential adjunctive therapy. In order to develop GH-based therapeutics, it is imperative that we understand whether the composition of biofilm EPS changes based on the environment and/or presence of other microbes. Here, we utilized ?-amylase and cellulase to target the polysaccharides within the EPS of mono- and dual-species Pseudomonas aeruginosa and Staphylococcus aureus biofilms in three different models that vary in clinical relevancy. We show that biofilms established in an in vitro well-plate model are not strongly adhered to the polystyrene surface and do not accurately reflect the GH efficacy seen with biofilms grown in vivo. However, dispersal efficacy in an in vitro wound microcosm model was more reflective of that seen in a murine wound model. We also saw a striking loss of efficacy for cellulase to disperse S. aureus in both mono- and dual species biofilms grown in the wound models, suggesting that EPS constituents may be altered depending on the environment.
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Project description:The persistent nature of chronic wounds leaves them highly susceptible to invasion by a variety of pathogens that have the ability to construct an extracellular polymeric substance (EPS). This EPS makes the bacterial population, or biofilm, up to 1,000-fold more antibiotic tolerant than planktonic cells and makes wound healing extremely difficult. Thus, compounds which have the ability to degrade biofilms, but not host tissue components, are highly sought after for clinical applications. In this study, we examined the efficacy of two glycoside hydrolases, ?-amylase and cellulase, which break down complex polysaccharides, to effectively disrupt Staphylococcus aureus and Pseudomonas aeruginosa monoculture and coculture biofilms. We hypothesized that glycoside hydrolase therapy would significantly reduce EPS biomass and convert bacteria to their planktonic state, leaving them more susceptible to conventional antimicrobials. Treatment of S. aureus and P. aeruginosa biofilms, grown in vitro and in vivo, with solutions of ?-amylase and cellulase resulted in significant reductions in biomass, dissolution of the biofilm, and an increase in the effectiveness of subsequent antibiotic treatments. These data suggest that glycoside hydrolase therapy represents a potential safe, effective, and new avenue of treatment for biofilm-related infections.
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Project description:The cell wall is among the first plant structures encountered by necrotrophic fungal pathogens, such as Botrytis cinerea. The composition of plant cell walls varies depending on the species, type of cell or tissue, and stage of development. Cell walls are important reservoirs of energy-rich sugars for pathogens, but also are barriers that impair colonization of host tissues. Growing fungal hyphae secrete enzymes that hydrolyze cell wall polysaccharides. Degradation of wall polysaccharides provides nutrients for the pathogen and improves the access of secreted Botrytis enzymes to all host cell wall targets and cytoplasmic constituents. Destruction of host cell walls results in tissue maceration, a hallmark of diseases caused by Botrytis. The Botrytis genome encodes 1,155 predicted carbohydrate-active enzyme (CAZy) genes; products of 275 are potentially secreted. Transcriptome sequencing identified Botrytis CAZy genes expressed during infections of lettuce leaves, ripe tomato fruit and grape berries. On all three hosts, Botrytis expresses a common group of 229 predicted CAZy genes including 28 pectin-modifying enzymes, 21 hemicellulose-modifying proteins, 18 enzymes targeting pectin and hemicellulose side-branches, and 16 enzymes that may degrade cellulose. Pectin polysaccharides are abundant in grape and tomato cell walls, but lettuce leaf walls are predominantly hemicelluloses and cellulose. These results suggest that Botrytis targets similar wall polysaccharide networks; e.g., pectins, on leaves and fruit, but also attacks unique host wall polysaccharide substrates The diversity of the Botrytis CAZy proteins may be partly responsible for its wide host range.
2014-09-24 | GSE57586 | GEO
Project description:The cell wall is among the first plant structures encountered by necrotrophic fungal pathogens, such as Botrytis cinerea. The composition of plant cell walls varies depending on the species, type of cell or tissue, and stage of development. Cell walls are important reservoirs of energy-rich sugars for pathogens, but also are barriers that impair colonization of host tissues. Growing fungal hyphae secrete enzymes that hydrolyze cell wall polysaccharides. Degradation of wall polysaccharides provides nutrients for the pathogen and improves the access of secreted Botrytis enzymes to all host cell wall targets and cytoplasmic constituents. Destruction of host cell walls results in tissue maceration, a hallmark of diseases caused by Botrytis. The Botrytis genome encodes 1,155 predicted carbohydrate-active enzyme (CAZy) genes; products of 275 are potentially secreted. Transcriptome sequencing identified Botrytis CAZy genes expressed during infections of lettuce leaves, ripe tomato fruit and grape berries. On all three hosts, Botrytis expresses a common group of 229 predicted CAZy genes including 28 pectin-modifying enzymes, 21 hemicellulose-modifying proteins, 18 enzymes targeting pectin and hemicellulose side-branches, and 16 enzymes that may degrade cellulose. Pectin polysaccharides are abundant in grape and tomato cell walls, but lettuce leaf walls are predominantly hemicelluloses and cellulose. These results suggest that Botrytis targets similar wall polysaccharide networks; e.g., pectins, on leaves and fruit, but also attacks unique host wall polysaccharide substrates The diversity of the Botrytis CAZy proteins may be partly responsible for its wide host range.
2014-09-24 | GSE57587 | GEO
Project description:The identification of glycoside hydrolases (GHs) for efficient polysaccharide deconstruction is essential for the development of biofuels. Here, we investigate the potential of sequential HMM-profile identification for the rapid and precise identification of the multi-domain architecture of GHs from various datasets. First, as a validation, we successfully reannotated >98% of the biochemically characterized enzymes listed on the CAZy database. Next, we analyzed the 43 million non-redundant sequences from the M5nr data and identified 322,068 unique GHs. Finally, we searched 129 assembled metagenomes retrieved from MG-RAST for environmental GHs and identified 160,790 additional enzymes. Although most identified sequences corresponded to single domain enzymes, many contained several domains, including known accessory domains and some domains never identified in association with GH. Several sequences displayed multiple catalytic domains and few of these potential multi-activity proteins combined potentially synergistic domains. Finally, we produced and confirmed the biochemical activities of a GH5-GH10 cellulase-xylanase and a GH11-CE4 xylanase-esterase. Globally, this "gene to enzyme pipeline" provides a rationale for mining large datasets in order to identify new catalysts combining unique properties for the efficient deconstruction of polysaccharides.
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