Screening, isolation, and characterization of glycosyl-hydrolase-producing fungi from desert halophyte plants

Fungal strains naturally occurring on the wood and leaves of the salt-excreting desert tree Tamarix were isolated and characterized for their ability to produce cellulose- and starch- degrading enzymes. Of the 100 isolates, six fungal species were identified by ITS1 sequence analysis. No significant differences were observed among taxa isolated from wood samples of different Tamarix species, while highly salt-tolerant forms related to the genus Scopulariopsis (an anamorphic ascomycete) occurred only on the phylloplane of T. aphylla. All strains had cellulase and amylase activities, but the production of these enzymes was highest in strain D, a Schizophyllum-commune- related form. This strain, when grown on pretreated Tamarix biomass, produced an enzymatic complex containing levels of filter paperase (414 ± 16 IU/ml) that were higher than those of other S. commune strains. The enzyme complex was used to hydrolyze different lignocellulosic substrates, resulting in a saccharification rate of pretreated milk thistle (73.5 ± 1.2 %) that was only 10 % lower than that obtained with commercial cellulases. Our results support the use of Tamarix biomass as a useful source of cellulolytic and amylolytic fungi and as a good feedstock for the economical production of commercially relevant cellulases and amylases. [Int Microbiol 2014; 17(1):41-48]Keywords: Schizophyllum commune · Tamarix ssp. · cellulase activity · amylase activit

Degradation of tetracyclines and sulfonamides by stevensite- and biochar-immobilized laccase systems and impact on residual antibiotic activity

BACKGROUND: Stevensite and biochar were investigated to covalently immobilize laccase from Myceliophthora thermophila (MtL) and Pleurotus eryngii (PeL) through the sequential application of aminopropyltriethoxysilane and glutaraldehyde. The immobilized preparations were tested to remove three tetracyclines and six sulfonamides at 0.1 mmol L−1 of each antibiotic. Degradation experiments were conducted both in the absence and in the presence (0.2 mmol L−1) of ABTS, 1-hydroxybenzotriazol (HBT), syringaldehyde or violuric acid. The residual antibiotic activity was tested towards five bacterial species and a bacterial consortium from wastewater. RESULTS: Higher values of activity yields (74% and 70.3%) and catalytic capabilities (1426 and 1405 IU g−1) were obtained with PeL on stevensite and biochar than with MtL. Stevensite enabled higher reusability and storage stability than biochar. Best removals of tetracyclines and sulfonamides were obtained with immobilized-laccase systems coupled to ABTS or syringaldehyde. Immobilized-laccase/ABTS systems removed 100% of tetracyclines while only chlortetracycline was completely removed in the presence of syringaldehyde. With ABTS, the most effectively removed sulfonamides were sulfathiazole and sulfadiazine (up to 100% and 54%), while syringaldehyde best supported the removal of sulfanilamide, sulfamethazine and sulfamethoxazole (up to 42%, 45% and 46%, respectively). In some cases, an effective antibiotics removal led to either low or no residual antibiotic activity. CONCLUSION: MtL and PeL were immobilized successfully on biochar and stevensite. The addition of either ABTS or syringaldehyde enhanced significant removals, up to 100%, of tetracyclines and sulfonamides by the immobilized laccase systems. Noteworthy, biochar-immobilized laccases/ABTS led to complete suppression of the antibiotic activity of tetracyclines. © 2018 Society of Chemical Industry

Implications of polluted soil biostimulation and bioaugmentation with spent mushroom substrate (Agaricus bisporus) on the microbial community and polycyclic aromatic hydrocarbons biodegradation

Different applications of spent Agaricus bisporus substrate (SAS), a widespread agro-industrial waste, were investigated with respect to the remediation of a historically polluted soil with Polycyclic Aromatic Hydrocarbons (PAH). In one treatment, the waste was sterilized (SSAS) prior to its application in order to assess its ability to biostimulate, as an organic amendment, the resident soil microbiota and ensuing contaminant degradation. For the other treatments, two bioaugmentation approaches were investigated; the first involved the use of the waste itself and thus implied the application of A. bisporus and the inherent microbiota of the waste. In the second treatment, SAS was sterilized and inoculated again with the fungus to assess its ability to act as a fungal carrier. All these treatments were compared with natural attenuation in terms of their impact on soil heterotrophic and PAH-degrading bacteria, fungal growth, biodiversity of soil microbiota and ability to affect PAH bioavailability and ensuing degradation and detoxification. Results clearly showed that historically PAH contaminated soil was not amenable to natural attenuation. Conversely, the addition of sterilized spent A. bisporus substrate to the soil stimulated resident soil bacteria with ensuing high removals of 3-ring PAH. Both augmentation treatments were more effective in removing highly condensed PAH, some of which known to possess a significant carcinogenic activity. Regardless of the mode of application, the present results strongly support the adequacy of SAS for environmental remediation purposes and open the way to an attractive recycling option of this wasteThis work was financially supported by the Ministry of Science and Innovation of Spain (Project CTM2009-13140-C02-02

Rapid bioassay for evaluating enzyme production in fungal isolates from environmental sources

Fungal hydrolytic enzymes have a great potential due to the rapid development of enzyme technology and their industrial applications. A fluorimetric microplate assay has been developed for measuring activity of β-D-exoglucanase, β-D-glucosidase, α-D-glucosidase, β-N-acetyl-hexosaminidase, β-D-xylosidase in diluted culture broth samples. The substrates used are conjugates of the highly fluorescent compounds 4-methylumbelliferone (MUB) and thus product formation can be measured directly in the microplate without previous extraction and purification of the product. The developed system was evaluated on eight fungal strains isolated from shrub species (Tamarix) that can be used in phytoremediation. Depending upon the enzyme, each species exhibited different levels of enzymatic activities as well as different production profiles. The fluorimetric method could constitute an effective alternative to the pectrophotometric method to screen hydrolase-producing microbial strains

Cultivating conditions optimization of the anaerobic digestion of corn ethanol distillery residuals using response surface methodology.

AbstractThis study investigated the individual and interactive effects of three factors — temperature, inoculum/substrate ratio (ISR) and inoculum typology — on the anaerobic digestion of corn ethanol distillery wastewater. Biochemical methane potential assays planned with factorial design with two independent quantitative variables on three levels (ISR: 1:1, 2:1 and 3:1; temperature: 30°C, 33.5°C, 37°C) and one independent qualitative variable (inoculum type: suspended, granular, mixed) have been performed. Response Surface Methodology has been used to study the effect of the factors with the aim of maximizing the specific methane yields (YCH4) obtainable with this substrate. The results show that all three investigated factors influence in a significant matter the YCH4, the ISR having the strongest effect on it. The temperature has significant influence on the YCH4 only in combination with high ISR values. The optimal conditions for the maximum YCH4 (551 mL CH4 g−1 VSadded) have been found at 37°C operating temperature, ISR=3:1 and using granular inoculum. These conditions gave rise to a 4-fold increase of YCH4 with respect to the worst combination of factors (YCH4=129 mL g−1 VSadded for the suspended inoculum type, at 30°C and ISR=1:1). The results improve the knowledge on the digestion of this substrate, providing information for successful process up-scaling

Trematocine, a Novel Antimicrobial Peptide from the Antarctic Fish Trematomus bernacchii: Identification and Biological Activity

Antimicrobial peptides (AMPs) are short peptides active against a wide range of pathogens and, therefore, they are considered a useful alternative to conventional antibiotics. We have identified a new AMP in a transcriptome derived from the Antarctic fish Trematomus bernacchii. This peptide, named Trematocine, has been investigated for its expression both at the basal level and after in vivo immunization with an endemic Antarctic bacterium (Psychrobacter sp. TAD1). Results agree with the expected behavior of a fish innate immune component, therefore we decided to synthesize the putative mature sequence of Trematocine to determine the structure, the interaction with biological membranes, and the biological activity. We showed that Trematocine folds into a \u3b1-helical structure in the presence of both zwitterionic and anionic charged vesicles. We demonstrated that Trematocine has a highly specific interaction with anionic charged vesicles and that it can kill Gram-negative bacteria, possibly via a carpet like mechanism. Moreover, Trematocine showed minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values against selected Gram-positive and Gram-negative bacteria similar to other AMPs isolated from Antarctic fishes. The peptide is a possible candidate for a new drug as it does not show any haemolytic or cytotoxic activity against mammalian cells at the concentration needed to kill the tested bacteria