Cellulolytic Species Research Articles

Targeted rRNA depletion enables efficient mRNA sequencing in diverse bacterial species and complex co-cultures.

Microbes present one of the most diverse sources of biochemistry in nature, and mRNA sequencing provides a comprehensive view of this biological activity by quantitatively measuring microbial transcriptomes. However, efficient mRNA capture for sequencing presents significant challenges in prokaryotes as mRNAs are not poly-adenylated and typically make up less than 5% of total RNA compared with rRNAs that exceed 80%. Recently developed methods for sequencing bacterial mRNA typically rely on depleting rRNA by tiling large probe sets against rRNAs; however, such approaches are expensive, time-consuming, and challenging to scale to varied bacterial species and complex microbial communities. Therefore, we developed EMBR-seq+, a method that requires fewer than 10 short oligonucleotides per rRNA to achieve up to 99% rRNA depletion in diverse bacterial species. Finally, EMBR-seq+ resulted in a deeper view of the transcriptome, enabling systematic quantification of how microbial interactions result in altering the transcriptional state of bacteria within co-cultures.

Effect of Spirulina Dietary Supplementation in Modifying the Rumen Microbiota of Ewes

Supplementing ruminant diets with microalgae, may prove an effective nutritional strategy to manipulate rumen microbiota. Forty-eight ewes were divided into four homogenous groups (n = 12) according to their fat-corrected milk yield (6%), body weight, age, and days in milk, and were fed individually with concentrate, alfalfa hay, and wheat straw. The concentrate of the control group (CON) had no Spirulina supplementation, while in the treated groups 5 (SP5), 10 (SP10), and 15 g (SP15) of Spirulina were supplemented as an additive in the concentrate. An initial screening using metagenomic next-generation sequencing technology was followed by RT-qPCR analysis for the targeting of specific microbes, which unveiled the main alterations of the rumen microbiota under the Spirulina supplementation levels. The relative abundance of Eubacterium ruminantium and Fibrobacter succinogenes in rumen fluid, as well as Ruminococcus albus in rumen solid fraction, were significantly increased in the SP15 group. Furthermore, the relative abundance of Prevotella brevis was significantly increased in the rumen fluid of the SP5 and SP10 groups. In contrast, the relative abundance of Ruminobacter amylophilus was significantly decreased in the rumen fluid of the SP10 compared to the CON group, while in the solid fraction it was significantly decreased in the SP groups. Moreover, the relative abundance of Selenomonas ruminantium was significantly decreased in the SP5 and SP15 groups, while the relative abundance of Streptococcus bovis was significantly decreased in the SP groups. Consequently, supplementing 15 g Spirulina/ewe/day increased the relative abundance of key cellulolytic species in the rumen, while amylolytic species were reduced only in the solid fraction.

Immobilization techniques improve volumetric hydrogen productivity of Caldicellulosiruptor species in a modified continuous stirred tank reactor

BackgroundCo-cultures and cell immobilization have been used for retaining biomass in a bioreactor, with the aim to improve the volumetric hydrogen productivity (QH2). Caldicellulosiruptor kronotskyensis is a strong cellulolytic species that possesses tāpirin proteins for attaching on lignocellulosic materials. C. owensensis has its reputation as a biofilm former. It was investigated whether continuous co-cultures of these two species with different types of carriers can improve the QH2.ResultsQH2 up to 30 ± 0.2 mmol L−1 h−1 was obtained during pure culture of C. kronotskyensis with combined acrylic fibres and chitosan. In addition, the yield of hydrogen was 2.95 ± 0.1 mol H2 mol−1 sugars at a dilution rate (D) of 0.3 h−1. However, the second-best QH2 26.4 ± 1.9 mmol L−1 h−1 and 25.4 ± 0.6 mmol L−1 h−1 were obtained with a co-culture of C. kronotskyensis and C. owensensis with acrylic fibres only and a pure culture of C. kronotskyensis with acrylic fibres, respectively. Interestingly, the population dynamics revealed that C. kronotskyensis was the dominant species in the biofilm fraction, whereas C. owensensis was the dominant species in the planktonic phase. The highest amount of c-di-GMP (260 ± 27.3 µM at a D of 0.2 h−1) were found with the co-culture of C. kronotskyensis and C. owensensis without a carrier. This could be due to Caldicellulosiruptor producing c-di-GMP as a second messenger for regulation of the biofilms under the high dilution rate (D) to prevent washout.ConclusionsThe cell immobilization strategy using a combination of carriers exhibited a promising approach to enhance the QH2. The QH2 obtained during the continuous culture of C. kronotskyensis with combined acrylic fibres and chitosan gave the highest QH2 among the pure culture and mixed cultures of Caldicellulosiruptor in the current study. Moreover, it was the highest QH2 among all cultures of Caldicellulosiruptor species studied so far.

Internal Transcription Terminators Control Stoichiometry of ABC Transporters in Cellulolytic Clostridia.

ABSTRACTThe extracellular substrate-binding proteins (SBPs) of ATP-binding cassette (ABC) importers tend to be expressed in excess relative to their cognate translocators, but how the stoichiometry of ABC transporters is controlled remains unclear. Here, we elucidated a mechanism contributing to differential gene expression in operons encoding ABC importers by employing cellulolytic Clostridia species, specifically Ruminiclostridium cellulolyticum. We found that there were usually stem-loop structures downstream of SBP genes, which could prematurely terminate the transcription of ABC importers and were putative internal intrinsic terminators, resulting in high transcript levels of upstream SBP genes and low transcript levels of downstream cognate translocator genes. This was determined by their termination efficiencies. Internal terminators had a lower U content in their 3′ U-rich tracts and longer GC-rich stems, which distinguishes them from canonical terminators and potentially endows them with special termination efficiencies. The pairing of U-rich tracts and the formation of unpaired regions in these internal terminators contributed to their folding energies, affecting the stability of their upstream SBP transcripts. Our findings revealed a strategy of internal transcriptional terminators controlling in vivo stoichiometry of their flanking transcripts.IMPORTANCE Operons encoding protein complexes or metabolic pathways usually require fine-tuned gene expression ratios to create and maintain the appropriate stoichiometry for biological functions. In this study, a strategy for controlling differential expression of genes in an operon was proposed by utilizing ABC importers from Ruminiclostridium cellulolyticum. We found that a stem-loop structure is introduced into the intergenic regions of operons encoding ABC importers as the putative internal terminator, which results in the premature termination of transcription. Consequently, the stoichiometric ratio of genes flanking terminators is precisely determined by their termination efficiencies and folding energies at the transcriptional level. Thus, it can be utilized as a promising synthetic biology tool to control the differential expression of genes in an operon.

Screening and identification of novel cellulolytic Trichoderma species from Egyptian habitats

Screening and identification of novel cellulolytic Trichoderma species from Egyptian habitats

Assessing the role of soil microbial communities of natural forest ecosystem

Abstract In forests, edaphic microbial communities are involved in litter decomposition and soil forming processes, with major contribution to humification, especially bacteria and fungi being responsible for the main ecosystem services fulfilled by the soil. Research has been carried out aiming to characterize the structure and diversity of microbial communities in the Rendzic Leptosols (WRB) under natural deciduous forest from Visterna, Babadag Plateau and to assess their contribution to ecosystem services provided by soil. The paper presents the results of quantitative estimations and taxonomic composition of soil and litter communities of heterotrophic bacteria and fungi, identification of cellulolytic species, as well as the microbial biomass and global physiological activities expressed as soil respiration potential. More than a half of bacterial species were common in litter and soil (SI=57.14%) and were represented by dominant species of fluorescent or non-fluorescent pseudomonads and Bacillus subtilis but no similarity was found between the two fungal communities. Fungal populations included cosmopolitan species, such as antagonists and strong cellulolytic representatives of genera Penicillium, Trichoderma, Mortierella, Chaetomium, Epicoccum, Aspergillus. Microbial density and microbial biomass presented the highest values in the litter (684 mg C x kg-1 d.s.) and in surface horizon Am1 of soil profile than in the bottom layers. The highest diversity was found in Am1 horizon (0-5 cm) H’=1.983 bits and ε=0.869 for cellulolytic community. Soil respiration reflected the intense physiological activity of microbiome, with high values associated to numerous effectives of bacteria and fungi especially in surface horizon. Microorganisms identified contribute to formation of soil by recycling of nutrients, cellulose decomposition, the synthesis of stable organic matter (humic acids), aggregation of soil particles, biological control of pathogens by antagonistic activity. They improve plant uptake of water and nutrients by forming symbioses (ectomycorrhizae), thus modelling the structure of vegetation.

Formulation of synthetic bacterial consortia and their evaluation by principal component analysis for lignocellulose rich biomass degradation

As lignocellulosic rich biomasses (LCB) and agriculture wastes are widespread and abundant sources of carbon in nature, their biodegradation for production of biofuel and value added products is gaining impetus worldwide. In the present study, ten synthetic consortia of three different cellulolytic species of Bacillus with Achromobacter xylosoxidans were formulated and evaluated for effective degradation of agro-wastes such as grass straw (GS), grass husk (GH), wheat husk (WH) and corn cob (CC) under two incubation conditions namely shaking and static. The estimations and analyses of patterns of enzyme activities such as endoglucanase, glucoamylase, β-glucosidase, exoglucanase and xylanase over the incubation period of 32 days and principal component analysis (PCA) indicated consortium CM10 comprising of all four bacterial strains was the most promising and active for the degradation of GS. Further investigation of potential of CM10 to degrade GS by employing CMC agar plate based assessment, scanning electron microscopy, thin layer chromatography, native polyacrylamide gel electrophoresis and zymogram analysis confirmed the high cellulolytic potential of consortium CM10. Based on outcome of this study, it is proposed that CM10 could be used for the production of multiple products such as cellulase, reducing sugars and microcrystalline cellulose through biodegradation of LCB.

Four novel strains of cellulolytic symbiotic bacteria isolated and characterized from GI tract of marine fishes of various feeding habits

Abstract Selected marine fishes with different feeding habits were screened for the presence of symbiotic cellulolytic bacteria in their gut. Four cellulolytic species of symbiotic bacteria were isolated from GI tract of marine fishes namely Carangoides praeustus, Filimanus similis, Sardinella longiceps and Sillago sihama. The strains were identified after polyphasic phenotypic and genotypic (16S rRNA gene) characterization as Bacillus subtilis strain TCPC1, Vibrio alginolyticus strain CFSS2C2, Pseudomonas stutzeri strain KSLS4C3 and Klebsiella oxytoca strain MSSC4. (Genbank Accession nos.: JN710380, JN710378, JN710377, JN712301).The results indicated the presence of cellulolytic bacteria in GI tract of marine fishes of carnivorous, phytoplanktivorous and omnivorous feeding habits. Cellulolytic activity was the maximum for B. subtilis strain TCPC1 (0.45 mg glucose ml−1) and V. alginolyticus strain CFSS2C2 (0.24 mg glucose ml−1) at 234 h. While P. stutzeri strain KSLS4C3 showed the maximum utilization (0.22 mg glucose ml−1) from 240 to 258 h. K. oxytoca strain MSSC4 (0.47 mg glucose ml−1) showed three peaks during the study. The maximum rate of cellulose utilization was shown by P. stutzeri strain KSLS4C3 (0.05 mg glucose ml−1 medium h−1) followed by K. oxytoca strain MSSC4 (0.03 mg glucose ml−1 medium h−1).

Age-associated microbiome shows the giant panda lives on hemicelluloses, not on cellulose

The giant panda feeds almost exclusively on bamboo, a diet highly enriched in lignin and cellulose, but is characterized by a digestive tract similar to carnivores. It is still large unknown if and how the giant panda gut microbiota contributes to lignin and cellulose degradation. Here we show the giant pandas’ gut microbiota does not significantly contribute to cellulose and lignin degradation. We found that no operational taxonomic unit had a nearest neighbor identified as a cellulolytic species or strain with a significant higher abundance in juvenile than cubs, a very low abundance of putative lignin and cellulose genes existed in part of analyzing samples but a significant higher abundance of genes involved in starch and hemicellulose degradation in juveniles than cubs. Moreover, a significant lower abundance of putative cellulolytic genes and a significant higher abundance of putative α-amylase and hemicellulase gene families were present in giant pandas than in omnivores or herbivores.

Impacts of environmental exposure on thermal and mycological characteristics of insulation wools

Fieldwork was conducted on 138 energy efficient buildings, to check if building site practices induced exposure to weather of thermal insulation. In nearly 80% of cases, insulation was exposed to weather due to insufficient protection. Therefore insulating wools were exposed, divided into three parts (control, exposure under shelter, exposure to weather). After the exposure period these samples were submitted to thermal conductivity measurements and infrared spectroscopy. A numerical study was carried out to evaluate the influence of weather conditions on thermal performance. Identifications of molds were realized by direct cultures of each sample. Mycological analyses provided the most significant results, showing a significant effect of exposure condition, with possible consequences in human health (toxinogenic, allergenic or pathogenic species) and material properties (cellulolytic species). Infrared spectrometry also showed some changes in sensitivity to water. These results show the interest to study in more detail the variations in the sensitivity, to moisture and mold, of insulation material throughout its ageing on site, and the interactions with the practices of the building process.

Evolutionary Relationships and Taxa-Specific Conserved Signature Indels Among Cellulases of Archaea, Bacteria, and Eukarya

The cellulases from different cellulolytic organisms have evolutionary relationships, which range from single-celled prokaryotes to the complex eukaryotes of the living world. This in silico analysis revealed the presence of a conserved cellulase domain along with evolutionary relationships among cellulases from several species of Archaea, Bacteria, and Eukarya. The amino acid sequences of cellulases from Archaea and Bacteria showed closer identity with their domain or phylum members that provided insights into convergent and divergent evolution of cellulases from other enzymes with different substrate specificities. Evolutionary relatedness was also observed in phylogenetic trees among a number of cellulase sequences of diverse taxa. In cellulases, propensity for alanine, glycine, leucine, serine, and threonine was high, but low for cysteine, histidine, and methionine. Catalytic aspartic acid had a higher propensity than glutamic acid, and both were involved in regular expression patterns. Characteristic group and multigroup-specific conserved signature indels located in the catalytic domains of cellulases were observed that further clarified evolutionary relationships. These indels can be distinctive molecular tools for understanding phylogeny and identification of unknown cellulolytic species of common evolutionary descent in different environments.

Overlap in substrate utilisation and spatial exclusion in some microfungi which act as early cellulose colonisers in a Mediterranean environment

A trapping system was set up to isolate culturable cellulolytic fungi at the soil-leaf litter interface in an area of Mediterranean maquis located in southern Italy. Seven cellulolytic taxa were isolated and cultured to represent the pioneers in the primordial phase of cellulose colonisation. The functional diversity of fungal isolates was analysed using a phenotype microarray technique to generate a profile of their functional traits. The extent of the overlap in substrate utilisation by the various species was subsequently determined.Some of the species that acted as key cellulose decomposers are considered globally distributed air-borne contaminants. Most of the cellulolytic species that were isolated during the experiment are also well known for their competitive abilities. Our study showed that at an early stage of cellulose colonisation a few fungal species with potential overlap in the overall metabolic function were able to establish themselves. The species that prevailed at the onset of cellulose colonisation did not co-occur in the field sites. A sort of “founder effect” could be hypothesised for microfungi when colonising sterile cellulose. This would suggest that scale (i.e. the spatial component) exerts a strong influence on the causative mechanisms that link genetic fungal diversity to specific ecosystem functions.

Контамінація та ушкодження мікроміцетами зерна та кормів

Контамінація та ушкодження мікроміцетами зерна та кормів

Structure features of GH10 xylanase from <italic>Caldicellulosiruptor bescii</italic>: implication for its thermophilic adaption and substrate binding preference

Caldicellulosiruptor bescii is the most thermophilic cellulolytic species of organisms known to date. In our previous study, GH10 xylanase CbXyn10B from C. bescii displayed outstanding hydrolytic activity toward various xylans at high temperatures. To understand the structural basis for this protein's catalysis and thermostability, we solved the crystal structures of CbXyn10B and its complexes with xylooligosaccharides. These structural models were used to guide comparison with its mesophilic counterpart PbXyn10B. A distinctive structural feature is that thermophilic CbXyn10B presents a relatively stable interaction between the extended loops L7 and L8 in the catalytic cleft by an extensive hydrogen bonding network, which is mediated by Lys306, Arg314 and three well-ordered water molecules. Moreover, a unique aromatic cluster consisting of Try17, Phe20, Phe21, and Phe337 may enhance the interaction between the N- and C- terminus. Targeted mutagenesis demonstrated that these interactions substantially contribute to enzyme stabilization, as indicated by a considerable decrease in the melting temperature (Tm) of CbXyn10B by substituting critical residues with Ala. Therefore, it was shown that not only the aromatic interaction connecting protein termini but also the extensive hydrogen bonding network formed between surface loops could restrict the local structural flexibility and contribute significantly to the overall stability of enzymes. Furthermore, the xylooligosaccharides were found to tightly bind to the glycone subsites of xylanase, indicating higher affinities at these subsites and reflecting its substrate binding preference. Our results suggest that CbXyn10B is stabilized with distinct rigidity at the catalytic cleft as well as the terminal regions, which provides insights into the evolutionary strategy for accommodating the functional needs of GH10 enzymes to high temperature.

Characterization of dominant and cellulolytic bacterial communities along the gut of silver carp Hypophthalmichthys molitrix during cyanobacterial blooms

Silver carp is one of the most important planktivorous fish in Chinese aquaculture and plays a significant role controlling cyanobacterial blooms. A balanced gut microbiota is crucial for growth and health of the host because of its important roles in immune defense, digestion of complex carbohydrates, and production of enterocytes. In our study, the dominant bacterial and cellulolytic bacterial (Clostridium I, Clostridium III, Clostridium XIVab, and Fibrobacter) communities in the contents and mucus of the silver carp gut (foregut, midgut, and hindgut) were analyzed by denaturing gradient gel electrophoresis and quantitative polymerase chain reaction (qPCR) analyses. The results revealed that the dominant and cellulolytic bacterial communities were significantly different among gut regions as well as in contents and mucus. Bacterial diversity and richness in contents and mucus increased along the gut and were higher in contents than those in local mucus. A sequence analysis of gut samples exhibited the conservative phylotypes of Proteobacteria, Actinobacteria, and Firmicutes. The gut of silver carp harbored an abundance of cellulolytic bacteria, particularly Clostridium XIV ab. The foregut segment had the highest proportions of the four cellulolytic bacteria, followed by the midgut and hindgut. However, the proportions of cellulolytic species in the silver carp gut was much lower than those in the terrestrial vertebrate gastrointestinal tract. We conclude that gut bacteria could help silver carp obtain energy from cyanobacteria, which may be why silver carp can maintain high growth rates during cyanobacterial blooms.

APPLICATION OF DNA TECHNIQUES FOR IDENTIFICATION OF FUNGAL COMMUNITIES COLONIZING BOOK OF EGYPT VOLUMES V PLANCHES ANTIQUITIES

This study represents both a study case about the fungi associated with a description ofEgypt volumes V Planches Antiquities in the Central library of Islamic manuscripts in Cairo,were used for identification of isolates from these brownish area from a description of EgyptThe plate method with a DNA was used for identification of fungi. The two internal transcribedspacers and the 5.8S gene (ITS1-5.8S- ITS2) from the nuclear ribosomal DNA wereamplified. The results show that the most dominant fungi isolated from the examined oldpaper samples belong to Aspergillus spp. (niger – flavus – fumigates and – terreus). In thepresent study, culture-independent molecular methods were applied to identify fungicommunities colonizing paper samples. Some of these fungi are cellulolytic species. Purefungal strains were used to constitute a marker for further comparative investigations of adescription of Egypt volumes V Planches Antiquities. This study represents both a study case about the fungi associated with a description ofEgypt volumes V Planches Antiquities in the Central library of Islamic manuscripts in Cairo,were used for identification of isolates from these brownish area from a description of EgyptThe plate method with a DNA was used for identification of fungi. The two internal transcribedspacers and the 5.8S gene (ITS1-5.8S- ITS2) from the nuclear ribosomal DNA wereamplified. The results show that the most dominant fungi isolated from the examined oldpaper samples belong to Aspergillus spp. (niger – flavus – fumigates and – terreus). In thepresent study, culture-independent molecular methods were applied to identify fungicommunities colonizing paper samples. Some of these fungi are cellulolytic species. Purefungal strains were used to constitute a marker for further comparative investigations of adescription of Egypt volumes V Planches Antiquities.

Biomass utilization by gut microbiomes.

Mammals rely entirely on symbiotic microorganisms within their digestive tract to gain energy from plant biomass that is resistant to mammalian digestive enzymes. Especially in herbivorous animals, specialized organs (the rumen, cecum, and colon) have evolved that allow highly efficient fermentation of ingested plant biomass by complex anaerobic microbial communities. We consider here the two most intensively studied, representative gut microbial communities involved in degradation of plant fiber: those of the rumen and the human large intestine. These communities are dominated by bacteria belonging to the Firmicutes and Bacteroidetes phyla. In Firmicutes, degradative capacity is largely restricted to the cell surface and involves elaborate cellulosome complexes in specialized cellulolytic species. By contrast, in the Bacteroidetes, utilization of soluble polysaccharides, encoded by gene clusters (PULs), entails outer membrane binding proteins, and degradation is largely periplasmic or intracellular. Biomass degradation involves complex interplay between these distinct groups of bacteria as well as (in the rumen) eukaryotic microorganisms.

Effects of nitrogen addition and litter properties on litter decomposition and enzyme activities of individual fungi

Litter decomposition is an important process of C and N cycling in the soil. Variation in the response of litter decomposition to nitrogen (N) addition (positive, negative or neutral) has been observed in many field studies. However, mechanism about variability in individual fungal species response to N addition has not yet been well demonstrated in the literature. Therefore, the objective of this study was to investigate the effects of N addition and litter chemistry properties on litter decomposition and enzyme activities of individual fungi. Three fungal species (Penicillium, Aspergillus, and Trichoderma) were isolated from a subtropical mixed forest soil. An incubation experiment was conducted using the individual fungi with two types of litter (leaf of Pinus massoniana and needle of Cryptocarya chinensis) and different N addition levels (0, 50 and 100 for N-deficient treatments, and 500 and 1000μgN for N-excessive treatments). Cumulative CO2-C, enzyme activities, and lignin and cellulose loss were measured during the incubation period of 60 days. Litter decomposition and enzyme activities significantly varied with the fungal species, while the N addition and litter types greatly affected fungal enzyme activities. The N treatments significantly increased lignin-rich needle decomposition by lignocellulose decomposers (Penicillium and Aspergillus) but did not affect their leaf decomposition. On the contrary, The N treatments stimulated leaf decomposition by cellulolytic species (Trichoderma) but did not affect its needle decomposition. Correlation analysis showed that lignin in the litter was the key component to affect litter decomposition. Activities of N-acetyl-β-glucosaminidase and phenol oxidase were both positively correlated to litter decomposition. The fungi (Penicillium and Aspergillus) with higher production of N-acetyl-β-glucosaminidase showed higher litter decomposition ability. The low N addition levels stimulated Penicillium and Aspergillus litter decomposition, but they still required more N source (e.g., litter N source) to support decomposition. Depressed fungal litter N uptake (lower N-acetyl-β-glucosaminidase activities) only occurred at the highest N addition level. Litter decomposition of Trichoderma depended more on external N and its litter decomposition capability was the lowest among the three species.

Fungal communities colonising empty Cepaea hortensis shells differ according to litter type

Soil and litter fungi can colonise and decompose many natural materials, including highly resilient proteinaceous compounds of animal origin. The shells of terrestrial gastropods are formed from such a compound (conchiolin) combined with inorganic calcium carbonate. In this study, we investigated fungal communities colonising empty shells of the common terrestrial gastropod Cepaea hortensis . Shells were exposed on the surface of litter from four different forest types (alder alluvial, oak-hornbeam, peat-bog pine and scree forest) and the fungi were surveyed and identified in four 3-month periods. We found 27 fungal species, one fungus forming mycelial cords, seven types of sterile mycelium and streptomycetes colonising the shells. The most frequent fungal species identified were common soil fungi. Multivariate analysis revealed a significant effect of the litter type on the fungal community. Humidity and pH at the locality are likely to be more important for fungal communities than the proteinaceous material of the shell. • First systematic study of microfungi on terrestrial gastropod shells. • Significant effect of time and litter type. • Fungi found on gastropod shells mostly known as chitinolytic or cellulolytic species. • Little effect of the polymer conchiolin on composition of fungal communities.

Enzyme activities of aerobic lignocellulolytic bacteria isolated from wet tropical forest soils

Lignocellulolytic bacteria have promised to be a fruitful source of new enzymes for next-generation lignocellulosic biofuel production. Puerto Rican tropical forest soils were targeted because the resident microbes decompose biomass quickly and to near-completion. Isolates were initially screened based on growth on cellulose or lignin in minimal media. 75 Isolates were further tested for the following lignocellulolytic enzyme activities: phenol oxidase, peroxidase, β-d-glucosidase, cellobiohydrolase, β-xylopyranosidase, chitinase, CMCase, and xylanase. Cellulose-derived isolates possessed elevated β-d-glucosidase, CMCase, and cellobiohydrolase activity but depressed phenol oxidase and peroxidase activity, while the contrary was true of lignin isolates, suggesting that these bacteria are specialized to subsist on cellulose or lignin. Cellobiohydrolase and phenol oxidase activity rates could classify lignin and cellulose isolates with 61% accuracy, which demonstrates the utility of model degradation assays. Based on 16S rRNA gene sequencing, all isolates belonged to phyla dominant in the Puerto Rican soils, Proteobacteria, Firmicutes, and Actinobacteria, suggesting that many dominant taxa are capable of the rapid lignocellulose degradation characteristic of these soils. The isolated genera Aquitalea, Bacillus, Burkholderia, Cupriavidus, Gordonia, and Paenibacillus represent rarely or never before studied lignolytic or cellulolytic species and were undetected by metagenomic analysis of the soils. The study revealed a relationship between phylogeny and lignocellulose-degrading potential, supported by Kruskal–Wallis statistics which showed that enzyme activities of cultivated phyla and genera were different enough to be considered representatives of distinct populations. This can better inform future experiments and enzyme discovery efforts.