Lignocellulose-Adapted Endo-Cellulase Producing Streptomyces Strains for Bioconversion of Cellulose-Based Materials

Twenty-four Actinobacteria strains, isolated from Arundo donax, Eucalyptus camaldulensis and Populus nigra biomass during natural biodegradation and with potential enzymatic activities specific for the degradation of lignocellulosic materials, were identified by a polyphasic approach. All strains belonged to the genus Streptomyces (S.) and in particular, the most highly represented species was Streptomyces argenteolus representing 50% of strains, while 8 strains were identified as Streptomyces flavogriseus (synonym S. flavovirens) and Streptomyces fimicarius (synonyms Streptomyces acrimycini, Streptomyces baarnensis, Streptomyces caviscabies, and Streptomyces flavofuscus), and the other four strains belonged to the species Streptomyces drozdowiczii, Streptomyces rubrogriseus, Streptomyces albolongus, and Streptomyces ambofaciens. Moreover, all Streptomyces strains, tested for endo and exo-cellulase, cellobiase, xylanase, pectinase, ligninase, peroxidase, and laccase activities using qualitative and semi-quantitative methods on solid growth medium, exhibited multiple enzymatic activities (from three to six). The 24 strains were further screened for endo-cellulase activity in liquid growth medium and the four best endo-cellulase producers (S. argenteolus AE58P, S. argenteolus AE710A, S. argenteolus AE82P, and S. argenteolus AP51A) were subjected to partial characterization and their enzymatic crude extracts adopted to perform saccharification experiments on A. donax pretreated biomass. The degree of cellulose and xylan hydrolysis was evaluated by determining the kinetics of glucose and xylose release during 72 h incubation at 50°C from the pretreated biomass in the presence of cellulose degrading enzymes (cellulase and β-glucosidase) and xylan related activities (xylanase and β-xylosidase). The experiments were carried out utilizing the endo-cellulase activities from the selected S. argenteolus strains supplemented with commercial β-gucosidase and xylanase preparations from Genencore (Accellerase BG and Accellerase XY). Cellulose and xylan conversion, when conducted using commercial (hemi)cellulases, gave glucose and xylose yields of 30.17 and 68.9%, respectively. The replacement of the cellulolytic preparation from Genencor (Accellerase 1500), with the endo-cellulase from S. argenteolus AE58P resulted in almost 76% of the glucose yield obtained in the presence of the commercial counterpart. Due to the promising results obtained by using the enzymatic crude extracts from S. argenteolus AE58P in the pretreated A. donax saccharification experiments, the proteins putatively responsible for endo-cellulase activity in this strain were identified by proteomics. Several proteins were confidently identified in different Streptomyces spp., eight of which belong to the class of Carbohydrate active enzymes. Overall results highlighted the biotechnological potential of S. argenteolus AE58P being an interesting candidate biocatalyst-producing bacterium for lignocellulose conversion and production of biochemicals and bioenergy

Microbial characterization of sourdough for sweet baked products in the Campania region (southern Italy) by a polyphasic approach

The microflora of nine sourdoughs used for sweet bakery products underwent preliminary microbiological characterization using lactic acid bacteria (LAB) and yeast enumeration. Five sourdough samples were submitted for microbial identification by culture-dependent techniques employing 16S and 26S rRNA genes sequencing, as well as a culture-independent technique using PCR-DGGE analysis. The LAB species isolated belonged principally to facultative heterofermentative Lactobacillus spp., Leuconostoc spp., and Lactococcus spp. Yeast strains were identified as Saccharomyces cerevisiae, with one exception represented by a strain belonging to Metschnikowia pulcherrima. PCR-DGGE analysis allowed the identification of Streptococcus thermophilus, Lactobacillus sakei, Weissella groceries and Lactobacillus sanfranciscensis among lactic acid bacteria and Saccharomyces cerevisiae and Metschnikowia pulcherrima among yeasts. This polyphasic approach highlighted different levels of biodiversity, from two to eight different typical LAB species, always associated to Saccharomyces cerevisiae, that could be selected to be specifically used in naturally fermented brioche and cornetto preparation

Valorisation of hydrothermal liquefaction wastewater in agriculture: effects on tobacco plants and rhizosphere microbiota

Industrial wastewater obtained from hydrothermal liquefaction (HTL-WW) of food wastes for biofuels production could represent a source of crop nutrients since it is characterized by a high amount of organic and inorganic compounds. In the present work, the potential use of HTL-WW as irrigation water for industrial crops was investigated. The composition of the HTL-WW was rich in nitrogen, phosphorus, and potassium with high level of organic carbon. A pot experiment with Nicotiana tabacum L. plants was conducted using diluted wastewater to reduce the concentration of some chemical elements below the official accepted threshold values. Plants were grown in the greenhouse under controlled conditions for 21 days and irrigated with diluted HTL-WW every 24 hours. Soils and plants were sampled every seven days to evaluate, over time, the effect of wastewater irrigation both on soil microbial populations, through high-throughput sequencing, and plant growth parameters, through the measurement of different biometric indices. Metagenomic results highlighted that, in the HTL-WW treated rhizosphere, the microbial populations shifted via their mechanisms of adaptation to the new environmental conditions, establishing a new balance among bacterial and fungal communities. Identification of microbial taxa occurring in the rhizosphere of tobacco plants during the experiment highlighted that the HTL-WW application improved the growth of Micrococcaceae, Nocardiaceae and Nectriaceae, which included key species for denitrification, organic compounds degradation and plant growth promotion. As a result, irrigation with HTL-WW improved the overall performance of tobacco plants which showed higher leaf greenness and increased number of flowers compared to irrigated control plants. Overall, these results demonstrate the potential feasibility of using of HTL-WW in irrigated agriculture

Anaerobic Process for Bioenergy Recovery From Dairy Waste: Meta-Analysis and Enumeration of Microbial Community Related to Intermediates Production

Dairy wastes are widely studied for the hydrogen and methane production, otherwise the changes in microbial communities related to intermediate valuable products was not deeply investigated. Culture independent techniques are useful tools for exploring microbial communities in engineered system having new insights into their structure and function as well as potential industrial application. The deep knowledge of the microbiota involved in the anaerobic process of specific waste and by-products represents an essential step to better understand the entire process and the relation of each microbial population with biochemical intermediates and final products. Therefore, this study investigated the microbial communities involved in the laboratory-scale anaerobic digestion of a mixture of mozzarella cheese whey and buttermilk amended with 5% w/v of industrial animal manure pellets. Culture-independent methods by employing high-throughput sequencing and microbial enumerations highlighted that lactic acid bacteria, such as Lactobacillaceae and Streptococcaceae dominated the beginning of the process until about day 14 when a relevant increase in hydrogen production (more than 10 ml H2 gVS-1 from days 13 to 14) was observed. Furthermore, during incubation a gradual decrease of lactic acid bacteria was detected with a simultaneous increase of Clostridia, such as Clostridiaceae and Tissierellaceae families. Moreover, archaeal populations in the biosystem were strongly related to inoculum since the non-inoculated samples of the dairy waste mixture had a relative abundance of archaea less than 0.1%; whereas, in the inoculated samples of the same mixture several archaeal genera were identified. Among methanogenic archaea, Methanoculleus was the dominant genus during all the process especially when the methane production occurred, and its relative abundance increased up to 99% at the end of the incubation time highlighting that methane was formed from dairy wastes primarily by the hydrogenotrophic pathway in the reactors

Bio-Based Succinate Production from Arundo donax Hydrolysate with the New Natural Succinic Acid-Producing Strain Basfia succiniciproducens BPP7

n/

Characterization in the archaeological excavation site of heterotrophic bacteria and fungi of deteriorated wall painting of Herculaneum in Italy.

Microbiological characterization of frescos in four different famous locations of excavation sites of Herculaneum (the Collegio degli Augustali, Casa del Colonnato Tuscanico, Casa dello Scheletro and Casa del Gran Portale), were carried out. The use of infrared thermography allowed us to detect sample points on frescos with greatest moisture not visible to the naked eye, resulting in structural damage. The microclimatic conditions provided perfect habitat for bacteria, especially of spore forming and mould. In fact, heterotrophic bacteria were prevalent in all wall paintings monitored, whereas fungi were also detected but at lower levels. Cultural–based methods allowed to obtain a total of 48 bacteria and 23 mould isolates molecularly identified by 16S and 26S rRNA partial sequence analysis. With this approach, Bacillus-related species (B. cereus/B. thuringiensis group, B. simplex/B.muralis group, B. megaterium and B. subtilis) were isolated in all sample points analysed with the exception of the Casa dello Scheletro in which Micrococcus luteus/Arthrobacter sp./Variovorax sp. group and Streptomyces fragilis were found. Mould isolates were closest related to different genera in which predominated Aspergillus, Penicillium and Fusarium together with the unusual genera as Microascus and Coprinus. Sequencing of the 16S ribosomal DNAs, selected on the basis of DGGE profiling, enabled detection of bacterial species closest related to Microbacterium groups in all sample points analysed, also associated with Brevibacterium, Streptomyces and Stenotrophomonas

Comparative assessment of autochthonous bacterial and fungal communities and microbial biomarkers of polluted agricultural soils of the Terra dei Fuochi

Organic and inorganic xenobiotic compounds can affect the potential ecological function of the soil, altering its biodiversity. Therefore, the response of microbial communities to environmental pollution is a critical issue in soil ecology. Here, a high-throughput sequencing approach was used to investigate the indigenous bacterial and fungal community structure as well as the impact of pollutants on their diversity and richness in contaminated and noncontaminated soils of a National Interest Priority Site of Campania Region (Italy) called “Terra dei Fuochi”. The microbial populations shifted in the polluted soils via their mechanism of adaptation to contamination, establishing a new balance among prokaryotic and eukaryotic populations. Statistical analyses showed that the indigenous microbial communities were most strongly affected by contamination rather than by site of origin. Overabundant taxa and Actinobacteria were identified as sensitive biomarkers for assessing soil pollution and could provide general information on the health of the environment. This study has important implications for microbial ecology in contaminated environments, increasing our knowledge of the capacity of natural ecosystems to develop microbiota adapted to polluted soil in sites with high agricultural potential and providing a possible approach for modeling pollution indicators for bioremediation purposes

Compost and microbial biostimulant applications improve plant growth and soil biological fertility of a grass-based phytostabilization system

In this work, a grass-based phytoremediation system integrated with an organic amendment and biostimulants was evaluated for remediating contaminated sites. Plant growth and biological fertility were monitored to assess the efficacy of a vegetative cap used as a safety measure to reduce sanitary and environmental risks of industrially contaminated soils and soil-washing sludges. Both matrices were potentially contaminated with Pb and Zn with an ecological risk index from low to moderate. According to potentially toxic elements (PTEs) bioaccessibility tests, the exposure to the released fine particulate matter may cause serious risks to human beings, in particular to children. The grass mixture was well adapted to both the substrates and a low PTEs mobility was detected, thus, reducing the leaching risk to ground water sources. Compost addition augmented significantly nitrogenase reductase (nifH) and ammonia monooxygenase (amoA) gene expression abundance in both substrates. Furthermore, a positive interaction between compost fertilization and a Trichoderma-based biostimulant inoculation was recorded in sludges resulting in a significant stimulation of nitrogen-fixing and ammonia-oxidizing bacteria. The application of compost and biostimulant increased soil fertility and plant growth. Furthermore, there was a slight reduction in PTE bioaccessibility, thus, improving the efficiency of the phytostabilization, limiting the resuspension and dispersion of the health-risk soil particulate

Biostimulant activity of azotobacter chroococcum and trichoderma harzianum in durum wheat under water and nitrogen deficiency

Biostimulants hold great potential for developing integrated sustainable agriculture systems. The rhizobacteria Azotobacter chroococcum strain 76A and the fungus Trichoderma harzianum strain T22, with demonstrated biostimulant activity in previous systems, were evaluated in Triticum durum cv Creso for their ability to enhance growth and tolerance to drought stress. Growth and drought tolerance were evaluated in conditions of low and high soil nitrogen, with two levels of water stress. T. harzianum increased plant growth (+16%) under control conditions and tolerance to moderate drought stress (+52%) under optimal fertilization, while A. chroococcum conferred a growth penalty (−28%) in well-watered conditions under suboptimal fertilization and increased tolerance only under extreme drought stress (+15%). This growth penalty was ameliorated by nitrogen fertilization. T. harzianum abundance was found to be positively correlated to extreme soil drying, whereas A. chroococcum-induced tolerance was dependent on soil nitrogen availability. These results indicate that while biostimulants may enhance growth and stress tolerance, nutrient availability soil and environmental conditions heavily influence these responses. These interactions should be considered when designing biostimulant products targeted to specific cultural conditions