Hydrolytic Potential Research Articles

Microwave facilitated bacterial liquefaction of sago biomass for efficient biomethane production: Energy and cost assessment

Sago has been considered to be a major staple food throughout the world for centuries. Thus, the sago processing industry also contributes to economic growth. However, waste disposal from this industry is an environmental concern and is addressed in this study. The generated solid waste sago is a starchy-rich substrate that is underutilized and can be exploited for bioenergy production. The present study focused on improving the hydrolytic potential of sago biomass during anaerobic digestion by implementing the combined microwave and bacterial pretreatment techniques. The microwave liquefaction yields the maximum solubilization of 11 % at the microwave power of 720 watts and the time of 11 min. Incorporation of bacterial pretreatment significantly enhances COD solubilization from 11 % to 31.6 %. This combined pretreatment of sago biomass shows the biomethane generation of 0.223 L/gCOD compared to the microwave (0.106 L/gCOD) and control sample (0.044 L/gCOD). Energy and cost analysis of combinative pretreatment shows a positive net energy of 0.97 kWh/kg and a net cost of -1.63 USD/ton. Thus, this combined pretreatment can be effectively adapted on an industrial scale with further improvements.

Bioprospecting for moderately halophilic eubacteria for potential biotechnological applications from Sambhar Lake, Rajasthan, India

Sambhar Lake an athalassohaline habitat located at 27°58′N 75°55′E, Rajasthan, India is a major source of salt production in the country. From surface lake water, soil, and shore sediments, fifty-nine moderate halophiles were isolated which were subsequently grouped according to shape, colony characteristics, and staining into twenty-two isolates. Fourier transform infrared spectroscopy profiling identifiedthese isolates as eubacterial with characteristic C=O stretching of ester functional groups. Observations further indicatedsimilarity within some Halomonasisolates indicating potential phylogenetic lineages. The FASTA sequences obtained after sequencing with universal bacterial primers were processed for phylogenetic analysis. Predominantly Gram-positive genera like Alkalibacillus, Amphibacillus, Marinococcus, Piscibacillus, Planococcus, Salinicoccus, Staphylococcusand Virgibacilluswith only two Gram-negative strains of Halomonaswere identified. The genus Amphibacilluswas recognizedfor the first time in the study of Sambhar Lake. Despite being moderately halophilic, several isolates exhibited high salt tolerance with growth in 25% salt. All isolates were mesophilic with growth observed between 18-42°C which matches the temperature profile of the region. Analysis of hydrolytic potential identified eighteen isolates as protease producers, thirteen as lipase producers, and ten as cellulase-producing strains. Further evaluation showed the dominance of C10:0, C12:0, C13:0, C14:0, C15:0, C16:0, C17:0, C18:0, C18:1 FAMEs, among which presence of C16:0 and C18:1 fatty acid indicated probable antimicrobial potentials of these strains.

Comparative genomic analysis of Planctomycetota potential for polysaccharide degradation identifies biotechnologically relevant microbes

BackgroundMembers of the Planctomycetota phylum harbour an outstanding potential for carbohydrate degradation given the abundance and diversity of carbohydrate-active enzymes (CAZymes) encoded in their genomes. However, mainly members of the Planctomycetia class have been characterised up to now, and little is known about the degrading capacities of the other Planctomycetota. Here, we present a comprehensive comparative analysis of all available planctomycetotal genome representatives and detail encoded carbohydrolytic potential across phylogenetic groups and different habitats.ResultsOur in-depth characterisation of the available planctomycetotal genomic resources increases our knowledge of the carbohydrolytic capacities of Planctomycetota. We show that this single phylum encompasses a wide variety of the currently known CAZyme diversity assigned to glycoside hydrolase families and that many members encode a versatile enzymatic machinery towards complex carbohydrate degradation, including lignocellulose. We highlight members of the Isosphaerales, Pirellulales, Sedimentisphaerales and Tepidisphaerales orders as having the highest encoded hydrolytic potential of the Planctomycetota. Furthermore, members of a yet uncultivated group affiliated to the Phycisphaerales order could represent an interesting source of novel lytic polysaccharide monooxygenases to boost lignocellulose degradation. Surprisingly, many Planctomycetota from anaerobic digestion reactors encode CAZymes targeting algal polysaccharides – this opens new perspectives for algal biomass valorisation in biogas processes.ConclusionsOur study provides a new perspective on planctomycetotal carbohydrolytic potential, highlighting distinct phylogenetic groups which could provide a wealth of diverse, potentially novel CAZymes of industrial interest.

PURIFICATION AND CHARACTERIZATION OF AMYLASE PRODUCED FROM PROBIOTIC LACTOBACILLUS PLANTARUM CS FOR INDUSTRIAL APPLICATIONS

Previous studies have demonstrated that probiotic Lactobacillus plantarum CS was able to generate an appreciable amount of extracellular amylase, hence the need to purify and characterize it. The aim of the study was to purify and characterize crude amylase from probiotic Lactobacillus plantarum CS for its industrial applications Three purification steps including ammonium sulphate precipitation, ion exchange chromatography on carboxymethyl sephadex and gel filtration on Sephadex G-75 were utilized. The homogeneity of the purified enzyme was confirmed using sodium deodocyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The purified amylase was characterized on different parameters including substrates hydrolyses, pH and temperature activity and stability profiles. The general purification elution profile revealed two different peaks of amylase activities with outstanding one having a molecular weight of 59.7kDa. Its purification fold was 4.0 with specific activity of 16.44U/mg protein and enzyme yield of 3%. Temperature optimal activity and stability was at 400C and 7.5 for pH activity and stability. Mangenese (Mn2+) (135.17%), tween 80 (128.30%) and some food condiments garlic, thyme, ginger, and tumeric) significantly (p> 0.05) enhanced amylase activity (≥262.40%). However, selenium (Se4+) and hydrogen peroxide (H2O2) were observed to have greatest inhibiting effect (≥30.9%) on the enzyme. Substrate hydrolysis profiles showed that the amylase hydrolyzed all the test starchy substrates with the highest hydrolytic potential on indigenous sweet potato starch (Km value/ Vmax of 1.33mg/ml/ 7.89ml). The rate of hydrolysis of other test substrates had yam> rice>cassava>corn with km values ≤ 4.0mg/ml and Vmax ≤ 25ml. The obtained results gave an insight that amylase produced from Lactobacillus plantarum CS met with the possessed properties suitable for any industrial application especially in food

Rich microbial and depolymerising diversity in Antarctic krill gut.

With almost a quadrillion individuals, the Antarctic krill processes five million tons of organic carbon every day during austral summer. This high carbon flux requires a broad range of hydrolytic enzymes to decompose the diverse food-derived biopolymers. While krill itself possesses numerous such enzymes, it is unclear, to what extent the endogenous microbiota contribute to the hydrolytic potential of the gut environment. Here we applied amplicon sequencing, shotgun metagenomics, cultivation, and physiological assays to characterize the krill gut microbiota. The broad bacterial diversity (273 families, 919 genera, and 2,309 species) also included a complex potentially anaerobic sub-community. Plate-based assays with 198 isolated pure cultures revealed widespread capacities to utilize lipids (e.g., tributyrin), followed by proteins (casein) and to a lesser extent by polysaccharides (e.g., alginate and chitin). While most isolates affiliated with the genera Pseudoalteromonas and Psychrobacter, also Rubritalea spp. (Verrucomicrobia) were observed. The krill gut microbiota growing on marine broth agar plates possess 13,012 predicted hydrolyses; 15-fold more than previously predicted from a transcriptome-proteome compendium of krill. Cultivation-independent and -dependent approaches indicated members of the families Flavobacteriaceae and Pseudoalteromonadaceae to dominate the capacities for lipid/protein hydrolysis and to provide a plethora of carbohydrate-active enzymes, sulfatases, and laminarin- or porphyrin-depolymerizing hydrolases. Notably, also the potential to hydrolyze plastics such as polyethylene terephthalate and polylactatide was observed, affiliating mostly with Moraxellaceae. Overall, this study shows extensive microbial diversity in the krill gut, and suggests that the microbiota likely play a significant role in the nutrient acquisition of the krill by enriching its hydrolytic enzyme repertoire.IMPORTANCEThe Antarctic krill (Euphausia superba) is a keystone species of the Antarctic marine food web, connecting the productivity of phyto- and zooplankton with the nutrition of the higher trophic levels. Accordingly, krill significantly contributes to biomass turnover, requiring the decomposition of seasonally varying plankton-derived biopolymers. This study highlights the likely role of the krill gut microbiota in this ecosystem function by revealing the great number of diverse hydrolases that microbes contribute to the krill gut environment. The here resolved repertoire of hydrolytic enzymes could contribute to the overall nutritional resilience of krill and to the general organic matter cycling under changing environmental conditions in the Antarctic sea water. Furthermore, the krill gut microbiome could serve as a valuable resource of cold-adapted hydrolytic enzymes for diverse biotechnological applications.

Harnessing the anti-cancer potential of linamarin: A computational study on design and hydrolysis mechanisms of its derivatives

Cassava extracts containing cyanogenic compounds demonstrate anticancer properties. The cyanogenic glucoside linamarin found abundantly in cassava can release hydrogen cyanide (HCN) upon hydrolysis, a potent cytotoxin. However, linamarin's hydrolysis mechanism by human enzymes is poorly delineated and constitutes a bottleneck for therapeutic development. This study aimed to investigate linamarin's hydrolysis mechanism by human β-glucosidase and identify structural derivatives with enhanced hydrolytic potential using density functional theory calculations. Results revealed α-anomeric derivatives as promising, with leaving group ability and steric bulk strongly governing hydrolysability. We identified several linamarin analogs with predicted rapid hydrolysis kinetics that may enable swift cytotoxic HCN release against cancer cells. This investigation enriches understanding of cyanogenic glycoside reactivity to facilitate their development as targeted antineoplastic agents. The identified derivatives set the groundwork for experimental evaluation of enhanced linamarin-inspired compounds as innovative cancer therapeutics.

The potential of in-house pectinolytic enzymes for industrial application

Pectinases are one of the essential groups of enzymes in the field of biotechnology and have applications in various industries including food, textile, paper industries, and waste management. Pectinases, like other industrial enzymes, are usually expensive. This research aimed at producing in-house pectinase from waste fruits, by an inductive mechanism. The study specifically optimized the production of in-house pectinases from mango and pawpaw peels used as substrates and also demonstrated the hydrolytic potential of the substrate-based in-house pectinase enzymes through viscosity analysis. The mango and pawpaw peels obtained from Blue Skies Co. Ltd. were oven-dried and milled to sieve particle sizes depending on the mode of the fermentation process. Measured amounts of the substrates were put into 50 ml Falcon tubes and subjected to both solid-state and submerged fermentation processes. The substrates were inoculated with one milliliter of Aspergillus niger isolate suspension and incubated at 30 °C for 2, 4, 6, 8, and 10 days under both fermentation processes. The crude enzyme extracts (in-house pectinolytic enzymes) obtained from the mango and pawpaw peels of both fermentation processes were used as enzyme sources for the hydrolysis of 1% w/v pectin solution prepared in phosphate buffer, pH 6.8 at 40 °C in viscosity analysis. The viscosities of the various treatments were recorded at different time intervals of 10, 20, and 30 min. Based on the viscosity analysis, it was observed that pectinolytic enzymes from pawpaw peels showed higher pectinolytic potential relative to the mango counterpart for both fermentation processes. However, there were consistent optima at day 4 for the mango peels and day 6 for the pawpaw peels using the submerged fermentation process. Also, there was a linear correlation between the pectinolytic activity and incubation period. The results obtained from this study show a higher potential to produce pectinases from agro-wastes such as mango and pawpaw peels for industrial purposes using an appropriate fermentation process.

Nocardioides okcheonensis sp. nov., isolated from riverside soil.

An actinobacterial strain designated MMS20-HV4-12T, displaying a high hydrolytic potential for various substrates, was isolated from a riverside soil sample and characterized by polyphasic taxonomic analysis. Growth occurred at 10-37 °C (optimum, 30°C), with NaCl concentrations of 0-4 % (optimum, 0 %) and at pH 7-9 (optimum, pH 8). MMS20-HV4-12T was catalase-positive, oxidase-negative, rod-shaped and formed creamy white-coloured colonies. Based on the results of 16S rRNA gene sequence analysis, MMS20-HV4-12T was found to be mostly related to the type strains of Nocardioides alpinus (98.3 % sequence similarity), Nocardioides furvisabuli (98.1 %) and Nocardioides zeicaulis (98.0 %). MMS20-HV4-12T showed optimal growth on Reaoner's 2A agar, forming white-coloured colonies. The diagnostic polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol, the major fatty acids were iso-C16 : 0, C17 : 1 ω8c and 10-methyl-C17 : 0, the major isoprenoid quinone was MK-8(H4), the diagnostic cell-wall sugar was galactose, and the cell-wall diamino acid was ll-diaminopimelic acid. The genome of MMS20-HV4-12T was 4.47 Mbp in size with a G+C content of 72.9 mol%. The genome based analysis indicated low relatedness between MMS20-HV4-12T and all compared species of Nocardioides, as the highest digital DNA-DNA hybridization and the orthologous average nucleotide identity values were 26.8 and 83.8% respectively. Based on genotypic, phenotypic and phylogenomic characterization, MMS20-HV4-12T evidently represents a novel species of genus Nocardioides, for which the name Nocardioides okcheonensis sp. nov. (type strain=MMS20-HV4-12T=KCTC 49651T=LMG 32360T) is proposed.

A holobiont approach towards polysaccharide degradation by the highly compartmentalised gut system of the soil-feeding higher termite Labiotermes labralis

BackgroundTermites are among the most successful insects on Earth and can feed on a broad range of organic matter at various stages of decomposition. The termite gut system is often referred to as a micro-reactor and is a complex structure consisting of several components. It includes the host, its gut microbiome and fungal gardens, in the case of fungi-growing higher termites. The digestive tract of soil-feeding higher termites is characterised by radial and axial gradients of physicochemical parameters (e.g. pH, O2 and H2 partial pressure), and also differs in the density and structure of residing microbial communities. Although soil-feeding termites account for 60% of the known termite species, their biomass degradation strategies are far less known compared to their wood-feeding counterparts.ResultsIn this work, we applied an integrative multi-omics approach for the first time at the holobiont level to study the highly compartmentalised gut system of the soil-feeding higher termite Labiotermes labralis. We relied on 16S rRNA gene community profiling, metagenomics and (meta)transcriptomics to uncover the distribution of functional roles, in particular those related to carbohydrate hydrolysis, across different gut compartments and among the members of the bacterial community and the host itself. We showed that the Labiotermes gut was dominated by members of the Firmicutes phylum, whose abundance gradually decreased towards the posterior segments of the hindgut, in favour of Bacteroidetes, Proteobacteria and Verrucomicrobia. Contrary to expectations, we observed that L. labralis gut microbes expressed a high diversity of carbohydrate active enzymes involved in cellulose and hemicelluloses degradation, making the soil-feeding termite gut a unique reservoir of lignocellulolytic enzymes with considerable biotechnological potential. We also evidenced that the host cellulases have different phylogenetic origins and structures, which is possibly translated into their different specificities towards cellulose. From an ecological perspective, we could speculate that the capacity to feed on distinct polymorphs of cellulose retained in soil might have enabled this termite species to widely colonise the different habitats of the Amazon basin.ConclusionsOur study provides interesting insights into the distribution of the hydrolytic potential of the highly compartmentalised higher termite gut. The large number of expressed enzymes targeting the different lignocellulose components make the Labiotermes worker gut a relevant lignocellulose-valorising model to mimic by biomass conversion industries.

Efficiency of an alkaline, thermostable, detergent compatible, and organic solvent tolerant lipase with hydrolytic potential in biotreatment of wastewater

Discharging the tannery wastewater into the environment is a serious challenge worldwide due to the release of severe recalcitrant pollutants such as oil compounds and organic materials. The biological treatment through enzymatic hydrolysis is a cheap and eco-friendly method for eliminating fatty substances from wastewater. In this context, lipases can be utilized for bio-treatment of wastewater in multifaceted industrial applications. To overcome the limitations in removing pollutants in the effluent, we aimed to identify a novel robust stable lipase (PersiLipase1) from metagenomic data of tannery wastewater for effective bio-degradation of the oily wastewater pollution. The lipase displayed remarkable thermostability and maintained over 81 % of its activity at 60 °C.After prolonged incubation for 35 days at 60°C, the PersiLipase1 still maintained 53.9 % of its activity. The enzyme also retained over 67 % of its activity in a wide range of pH (4.0 to 9.0). In addition, PersiLipase1 demonstrated considerable tolerance toward metal ions and organic solvents (e.g., retaining >70% activity after the addition of 100 mM of chemicals). Hydrolysis of olive oil and sheep fat by this enzyme showed 100 % efficiency. Furthermore, the PersiLipase1 proved to be efficient for biotreatment of oil and grease from tannery wastewater with the hydrolysis efficiency of 90.76 % ± 0.88. These results demonstrated that the metagenome-derived PersiLipase1 from tannery wastewater has a promising potential for the biodegradation and management of oily wastewater pollution.

Optimization Studies on Extracellular Protease Production by Aspergillus niger and Aspergillus terreus Using Skim Milk Casein as Substrate

Aim: Proteases are proteolytic enzymes that have a wide range of applications in the industrial sectors. This study investigated the production and optimization of protease by protease-producing fungi, (Aspergillus niger and A. terreus) using skim milk casein as substrate.
 Materials and Methods: Aspergillus terreus and A. niger obtained from the Department of Plant Science and Biotechnology, University of Jos, Nigeria were screened for extracellular protease on skim milk agar medium. Optimization studies across different parameters: temperatures (30-80oC), pH (3-10), substrate Concentration (0.25-2 %), and incubation period (6 days) using submerged fermentation were done for maximum protease activity.
 Results: The maximum temperature of protease activity was 50oC (116.5 IU/ml) for Aspergillus niger and 40oC (121.86 IU/ml) for Aspergillus terreus. The Optimum pH for protease activity was pH 9 (106.85 IU/ml) for Aspergillus niger and pH 8 (107.90 IU/ml) for Aspergillus terreus. Optimum protease activity (29.36 IU/ml) at an incubation period of 48 hours was recorded for Aspergillus niger and 72 hours (182.2 IU/ml) for Aspergillus terreus. At 2% substrate concentration a maximum activity of 87.21 IU/ml and 76.5 IU/ml was recorded for Aspergillus niger and A. terreus, respectively.
 Conclusion: The test fungi Aspergillus niger and Aspergillus terreus exhibited maximum hydrolytic potential for protease production which has a vast application in different industries.

Activated sludge microbial communities and hydrolytic potential in a full-scale SBR system treating landfill leachate

Landfill leachate, due to its recalcitrant nature and toxicity, poses a serious environmental threat, which requires the implementation of effective treatment processes. In this work, a full-scale treatment system consisting of two Sequencing Batch Reactors (SBRs) was used for the processing of landfill leachate of intermediate to mature age (BOD/COD ratio of 0.16). Biosystem operation resulted in BOD5, COD and TKN removal efficiencies of 81%, 39% and 76%, respectively, whereas the low residual NO3 --N concentration in the effluent (4.01 ± 0.10 mg/L) was indicative of the efficient denitrification process. Assessment of hydrolytic potential of activated sludge revealed high endocellular and extracellular lipase activities, which reached values up to 206 and 141 U/g protein respectively, possibly as the consequence of plastics degradation during maturation process. Implementation of Illumina sequencing indicated the predominance of Alphaproteobacteria, accompanied by members of Bacteroidetes, Betaproteobacteria and Chloroflexi. Paracoccus was the predominant genus identified, followed by representatives of the genera Bellilinea, Flavobacterium, Thauera and Truepera. Nitrosomonas was the major ammonia-oxidizing bacterium (AOB), while nitrite oxidation was mainly achieved by the uncultured nitrite-oxidizing bacterium (NOB) Candidatus Nitrotoga.

What lies on macroalgal surface: diversity of polysaccharide degraders in culturable epiphytic bacteria

Macroalgal surface constitutes a peculiar ecological niche and an advantageous substratum for microorganisms able to degrade the wide diversity of algal glycans. The degrading enzymatic activities of macroalgal epiphytes are of paramount interest for the industrial by-product sector and biomass resource applications. We characterized the polysaccharide hydrolytic profile of bacterial isolates obtained from three macroalgal species: the red macroalgae Asparagopsis taxiformis and Sphaerococcus coronopifolius (Rhodophyceae) and the brown Halopteris scoparia (Phaeophyceae), sampled in South Portugal. Bacterial enrichment cultures supplemented with chlorinated aliphatic compounds, typically released by marine algae, were established using as inoculum the decaying biomass of the three macroalgae, obtaining a collection of 634 bacterial strains. Although collected from the same site and exposed to the same seawater seeding microbiota, macroalgal cultivable bacterial communities in terms of functional and phylogenetic diversity showed host specificity. Isolates were tested for the hydrolysis of starch, pectin, alginate and agar, exhibiting a different hydrolytic potential according to their host: A. taxiformis showed the highest percentage of active isolates (91%), followed by S. coronopifolius (54%) and H. scoparia (46%). Only 30% of the isolates were able to degrade starch, while the other polymers were degraded by 55–58% of the isolates. Interestingly, several isolates showed promiscuous capacities to hydrolyze more than one polysaccharide. The isolate functional fingerprint was statistically correlated to bacterial phylogeny, host species and enrichment medium. In conclusion, this work depicts macroalgae as holobionts with an associated microbiota of interest for blue biotechnologies, suggesting isolation strategies and bacterial targets for polysaccharidases’ discovery.

Isolation and molecular identification of cellulose/hemicellulose degrading bacteria from agricultural compost and determination of their hydrolytic potential

Nine cellulolytic bacteria were isolated from agricultural residues compost on selective culture medium amended with cellulose (filter paper) as the sole carbon source. Five of the nine isolates (CX2, CX4, CX5, CX7 and CX9) were able to degrade both cellulose (CMC) and xylan in congo red agar plates. The five isolates were able to depolymerize the agricultural wastes, rice straw and sugarcane bagasse. CMC-ase and xylanase production by the five isolates were determined on rice straw and sugarcane bagasse. CX2 and CX9 exhibited the highest CMC-ase and xylanase production on both rice straw and sugarcane bagasse. Among the isolated strains, CX9 produced the highest CMC-ase activity (71 pKat ml−1) on rice straw, whereas CX9 produced the highest xylanase activity (7.83 nKat ml−1) on sugarcane bagasse. In general, higher CMC-ase and xylanase production was obtained on rice straw compared with sugarcane bagasse. According to the sequences of the 16 s rRNA genes, the five isolates were identified as: Paracoccus kondratievae strain GB (CX2 and CX4), Paracoccus communis (CX5), Bacillus australimaris (CX7) and Bacillus pumilus (CX9). The sequences of the 16S rRNA genes of five isolates were submitted to the gene bank.

Matrix metalloproteinase-9 activity in ewes' milk and its relationship to somatic cell counts

The presence and the activity of gelatinase-B/matrix metalloproteinase 9 (MMP-9) in ewes' milk was evaluated with a particular focus on the possible correlation with the somatic cell count (SCC). Forty five individual milk samples obtained from lacaune sheep, previously analysed for SCC with values ranging from 5.0 × 104 to 1.6 × 107 cells mL−1, gave overall gelatinolytic activity between 0.53 ± 0.05 and 4.53 ± 0.55 fluorescence units min−1 mg−1 protein. Zymography showed that the variation in the gelatinolytic activity depended on variations in the specific activity of MMP-9; no significant differences in MMP-2 (gelatinase-A) activity were observed. Pearson's correlation calculations gave a fair correlation between the presence of somatic cells and the enzymatic activity. MMP-9 in the context of proteolytic activities related to the presence of somatic cells acquires greater importance taking into account the hydrolytic potential of MMP-9 versus casein, evaluated in this study by exploiting a molecular docking approach.

Characterization of Cellulose-Degrading Bacteria Isolated from Soil and the Optimization of Their Culture Conditions for Cellulase Production

The characterization of bacteria with hydrolytic potential significantly contributes to the industries. Six cellulose-degrading bacteria were isolated from mixture soil samples collected at Kingfisher Lake and the University of Manitoba campus by Congo red method using carboxymethyl cellulose agar medium and identified as Paenarthrobacter sp. MKAL1, Hymenobacter sp. MKAL2, Mycobacterium sp. MKAL3, Stenotrophomonas sp. MKAL4, Chryseobacterium sp. MKAL5, and Bacillus sp. MKAL6. Their cellulase production was optimized by controlling different environmental and nutritional factors such as pH, temperature, incubation period, substrate concentration, nitrogen, and carbon sources using the dinitrosalicylic acid and response surface methods. Except for Paenarthrobacter sp. MKAL1, all strains are motile. Only Bacillus sp. MKAL6 was non-salt-tolerant and showed gelatinase activity. Sucrose enhanced higher cellulase activity of 78.87 ± 4.71 to 190.30 ± 6.42 U/mL in these strains at their optimum pH (5-6) and temperature (35-40°C). The molecular weights of these cellulases were about 25kDa. These bacterial strains could be promising biocatalysts for converting cellulose into glucose for industrial purposes.

Evaluating the xerophilic potential of moulds on selected egg tempera paints on glass and wooden supports using fluorescent microscopy

• 11 fungal isolates from cultural heritage institutions’ interiors were screened for xerophilic trait. • Specially designed incubators were constructed to hold specific RH levels of 55%, 63% and 74.% • Fungal growth was monitored at low relative humidity on wood and glass supports painted by traditional artists’ paints. • Effects of pigments, support materials and strain variability are discussed. Even though contamination of painted artwork by xerophilic moulds frequently causes aesthetical, physical and/or biochemical biodeterioration, mould growth on paints, prepared from assorted traditional artists’ pigments, has yet to be systematically evaluated especially with regard to low relative humidity (RH) levels and painted support materials. Therefore, we investigated 11 fungal strains isolated mostly from cultural heritage institutions’ interiors for their potential to grow on egg tempera paint films prepared with different colouring agents and applied on wooden and glass supports which were maintained in monoculture in specially designed incubators at three different RH levels of 55%, 63% and 74%. The growth rate of mould over the surface was assessed using fluorescent microscopy after Calcofluor White staining. Additionally, these stains were screened for their xerophilic and hydrolytic potential using standard microbiological assays. Results show that when comparing growth rates on egg tempera paint films, 6 isolates grew exclusively on wood, exemplifying the greater susceptibility of this supporting material to mould attack. Prussian blue paint also stimulated the growth of 6 isolates, and the maximum overall expansion (38%) was observed on Prussian blue painted wood. RH was the key factor limiting growth, and at RH of 55% only a slight growth of 2 isolates was observed on Prussian blue painted wood. On the same samples incubated at RH of 63%, 10 isolates exhibited a moderate to strong growth and 4 of these showed an additional increase in growth at 74% RH. Paints consisting of artists’ pigments carmine lake or lead white in general completely prevented the development of moulds. Nevertheless, tolerance was species/strain dependant and the growths of isolates Cladosporium halotolerans EXF-15,333, Aspergillus niger EXF-14,897 and Aspergillus creber EXF-15,148 on lead white paint (containing ions and salts of heavy metal lead (Pb +2 )) even exceeds 11%. Standard microbiological tests showed that all stains had hydrolytic potential and proved positive for xerophilic trait, nevertheless their ability to develop on egg tempera paint films was mostly dependant of very specific conditions.

Extracellular hydrolytic potential drives microbiome shifts during anaerobic co-digestion of sewage sludge and food waste

Bacterial community structure and dynamics in anaerobic digesters are primarily influenced by feedstock composition. It is therefore important to unveil microbial traits that explain microbiome variations in response to substrate changes. Here, gene and genome-centric metagenomics were used to examine microbiome dynamics in four laboratory-scale reactors, in which sewage sludge was co-digested with increasing amounts of food waste. A co-occurrence network revealed microbiome shifts in response to changes in substrate composition and concentration. Food waste concentration correlated with extracellular enzymes and metagenome-assembled genomes (MAGs) involved in the degradation of complex carbohydrates commonly found in fruits and plant cell walls as well as with the abundance of hydrolytic MAGs. A key role was attributed to Proteiniphillum for being the only bacteria that encoded the complete pectin degradation pathway. These results suggest that changes of feedstock composition establish new microbial niches for bacteria with the capacity to degrade newly added substrates.

Phenotype Switching in Metal-Tolerant Bacteria Isolated from a Hyperaccumulator Plant.

Simple SummaryAzolla filiculoides L. is an aquatic fern with the potential for degradation and accumulation of pollutants. It is accompanied by microorganisms (a microbiome) that may participate in these processes. Microorganisms showing specific phenotypes may promote plant growth in the presence of pollutants. We intended to identify such beneficial strains by studying their potential for the degradation of given organic compounds and the production of hydrolytic enzymes and phytohormones under heavy metal stress (Pb, Cd, Cr (VI), Ni, Ag, and Au). We found 10 isolates displaying varying phenotypes depending on the stress factor. The most efficient was Delftia sp., which showed potential for both degradation of organics and plant growth promotion. Other strains were more efficient at metabolizing organics or exhibited enzymatic responses in the presence of the studied metals. These identified phenotypes made all strains beneficial in both supporting plants in unfavorable conditions and degradation of organic compounds. A biopreparation containing these strains may be valuable as both a biofertilizer and a bioremediation agent.As an adaptation to unfavorable conditions, microorganisms may represent different phenotypes. Azolla filiculoides L. is a hyperaccumulator of pollutants, but the functions of its microbiome have not been well recognized to date. We aimed to reveal the potential of the microbiome for degradation of organic compounds, as well as its potential to promote plant growth in the presence of heavy metals. We applied the BiologTM Phenotypic Microarrays platform to study the potential of the microbiome for the degradation of 96 carbon compounds and stress factors and assayed the hydrolytic potential and auxin production by the microorganisms in the presence of Pb, Cd, Cr (VI), Ni, Ag, and Au. We found various phenotype changes depending on the stress factor, suggesting a possible dual function of the studied microorganisms, i.e., in bioremediation and as a biofertilizer for plant growth promotion. Delftia sp., Staphylococcus sp. and Microbacterium sp. exhibited high efficacy in metabolizing organic compounds. Delftia sp., Achromobacter sp. and Agrobacterium sp. were efficient in enzymatic responses and were characterized by metal tolerant. Since each strain exhibited individual phenotype changes due to the studied stresses, they may all be beneficial as both biofertilizers and bioremediation agents, especially when combined in one biopreparation.

Enzymatic Pretreatment with Laccases from Lentinus sajor-caju Induces Structural Modification in Lignin and Enhances the Digestibility of Tropical Forage Grass (Panicum maximum) Grown under Future Climate Conditions.

Since laccase acts specifically in lignin, the major contributor to biomass recalcitrance, this biocatalyst represents an important alternative to the pretreatment of lignocellulosic biomass. Therefore, this study investigates the laccase pretreatment and climate change effects on the hydrolytic performance of Panicum maximum. Through a Trop-T-FACE system, P. maximum grew under current (Control (C)) and future climate conditions: elevated temperature (2 °C more than the ambient canopy temperature) combined with elevated atmospheric CO2 concentration(600 μmol mol−1), name as eT+eC. Pretreatment using a laccase-rich crude extract from Lentinus sajor caju was optimized through statistical strategies, resulting in an increase in the sugar yield of P. maximum biomass (up to 57%) comparing to non-treated biomass and enabling hydrolysis at higher solid loading, achieving up to 26 g L−1. These increments are related to lignin removal (up to 46%) and lignin hydrophilization catalyzed by laccase. Results from SEM, CLSM, FTIR, and GC-MS supported the laccase-catalyzed lignin removal. Moreover, laccase mitigates climate effects, and no significant differences in hydrolytic potential were found between C and eT+eC groups. This study shows that crude laccase pretreatment is a potential and sustainable method for biorefinery solutions and helped establish P. maximum as a promising energy crop.