Regulation of lignocellulose degradation in microorganisms.

Microbial strategies for biomass deconstruction involve an incredible repertoire of enzymatic, structural, and regulatory proteins. From carbohydrate active enzymes to cellulosomes, bacteria, yeast, and filamentous fungi adapt their functional machinery to grow from alternative carbon sources such as lignocellulose and survive starvation. In that context, microbes must be able to sense, bind, degrade, and utilize lignin, cellulose, and hemicelluloses. Nature has developed specialized protein modules, RNA structures, and regulatory systems operating at a genomic, transcription, and translation level. This review briefly summarizes the main regulatory pathways involved in lignocellulose microbial degradation, including carbon catabolite repression; anti-sigma factors; regulatory RNA elements such as small RNAs, antisense RNA, RNA-binding proteins, and selective RNA processing and stabilization; and transcriptional regulators and unfolded protein response. Interplay with global regulators controlling pH response and nitrogen utilization is also revised.

Micromonospora schwarzwaldensis sp. nov., a producer of telomycin, isolated from soil.

A Gram-stain-positive, spore-forming actinomycete strain (HKI0641(T)) was isolated from a soil sample collected in the Black Forest, Germany. During screening for antimicrobial natural products this bacterium was identified as a producer of the antibiotic telomycin. Morphological characteristics and chemotaxonomic data indicated that the strain belonged to the genus Micromonospora. The peptidoglycan of strain HKI0641(T) contained meso-diaminopimelic acid, and the fatty acid profile consisted predominantly of anteiso-C15 : 0, iso-C15 : 0, iso-C16 : 0 and C16 : 0. MK-10(H4), MK-10(H2) and MK-10 were identified as the major menaquinones. To determine the taxonomic positioning of strain HKI0641(T), we computed a binary tanglegram of two rooted phylogenetic trees that were based upon 16S rRNA and gyrB gene sequences. The comparative analysis of the two common classification methods strongly supported the phylogenetic affiliation with the genus Micromonospora, but it also revealed discrepancies in the assignment at the level of the genomic species. 16S rRNA gene sequence analysis identified Micromonospora coxensis DSM 45161(T) (99.1 % sequence similarity) and Micromonospora marina DSM 45555(T) (99.0 %) as the nearest taxonomic neighbours, whereas the gyrB sequence of strain HKI0641(T) indicated a closer relationship to Micromonospora aurantiaca DSM 43813(T) (95.1 %). By means of DNA-DNA hybridization experiments, it was possible to resolve this issue and to clearly differentiate strain HKI0641(T) from other species of the genus Micromonospora. The type strains of the aforementioned species of the genus Micromonospora could be further distinguished from strain HKI0641(T) by several phenotypic properties, such as colony colour, NaCl tolerance and the utilization of carbon sources. The isolate was therefore assigned to a novel species of the genus Micromonospora, for which the name Micromonospora schwarzwaldensis sp. nov. is proposed. The type strain is HKI0641(T) ( = DSM 45708(T) = CIP 110415(T)).

Magnetic Mesoporous Silica Nanoparticles for Drug Delivery Systems: Synthesis, Characterization and Application as Norfloxacin Carrier.

Mesoporous silica nanoparticles, with and without the inclusion of a magnetic core, were hydrothermally synthesized and employed as carrier of the antibiotic norfloxacin (NFX). The antibiotic-loaded materials were prepared by wet impregnation. Differences in drug content (and in further release profile) were directly related to changes in surface area, particle aggregation and hydrophobicity of the solids. The kinetics of NFX release has been studied in batch experiments. In all cases, more than 55% of the antibiotic was quickly desorbed during the first 5 min due to the localization of NFX on the external surface of the nanoparticles. The rest of the drug (situated inside the mesopores) was released through a diffusion-controlled transport and the rate was strongly dependent of the pH, reaching its minimum value at neutral pH. The calculated activation energy confirmed that the release was controlled by a diffusion process. Breaking of H-bonds and electrostatic and hydrophobic interactions appear to be responsible for NFX desorption from the solid surface. Such interactions increase, however, the thermal stability of the drug when the NFX and the carriers are combined. The antimicrobial activities of the drug loaded nanoparticles and the free antibiotic were compared and discussed.

Ganoderma sessile is a fast polysaccharide producer among Ganoderma species.

The selection of fast-growing and high-yield-producing strains is required to satisfy the market demand on fungal food supplements. To that aim, three strains deposited in our collection as G. lucidum and G. oregonense were screened for polysaccharide production and biomass yield. Ganoderma strains deposited as G. lucidum were identified as G. sessile and G. lingzhi by nuc rDNA internal transcribed spacer ITS1-5.8S-ITS2 (ITS) and translation elongation factor 1-α (TEF1-α) phylogenies. The identity of G. oregonense was confirmed by molecular phylogeny and biogeography. Additionally, mycelial antagonism confirmed species differentiation, and strains were further distinguished by morphology and protein profiles. Biomass and polysaccharide yields of G. sessile were clearly different from those of G. lingzhi and G. oregonense in both liquid culture and solid-state fermentation. The maximum polysaccharide yield (4.52 ± 0.83 g L-1) for G. sessile was obtained from submerged cultures at day 9. G. sessile also achieved the highest linear growth in lignocellulosic solid substrates. Consequently, basidiomata were successfully obtained by solid-state fermentation in polypropylene bags, whereas G. lingzhi and G. oregonense mushrooms were not produced in artificial solid substrates. G. sessile, a species frequently collected in America, showed to be a promising polysaccharide producer for the manufacture of dietary supplements.

DNA damaging potential of Ganoderma lucidum extracts.

ETHNOPHARMACOLOGICAL RELEVANCE: Ganoderma lucidum (Lingzhi or Reishi) is a medicinal mushroom historically used in Asian countries to treat a wide variety of diseases and prolong life. In the last years, G. lucidum has been internationally recognized as an effective adjuvant in cancer treatment. Among active components, the most recent research indicates that polysaccharides modulate the immune response favoring the recovery from toxicity of chemo and radiotherapy while triterpenes are cytotoxic to tumoral cells mainly by altering gene expression. Beyond this body of evidence on the efficacy of G. lucidum in cancer treatment, it is not yet understood whether these extracts exert the same mechanisms of action than current antitumoral drugs. AIM OF THE STUDY: In this study, we tested the DNA damaging potential of G. lucidum extracts by the ß-galactosidase biochemical prophage induction assay (BIA) using doxorubicin, a DNA intercalating agent, as a positive control. This assay was traditionally used to screen microbial metabolites towards antitumoral agents. Here, we used this bacterial assay for the first time to assess DNA damage of herbal drugs. RESULTS: After a bioguided assay, only a purified fraction of G. lucidum containing a mixture of C16 and C18:1 fatty acids exerted weak activity which could not be attributed to direct interaction with DNA. At the same concentrations, the induction observed for doxorubicin was clearly contrasting. CONCLUSIONS: The micro BIA assay could be successfully used to demonstrate differences in cellular effects between G. lucidum extracts and doxorubicin. These results showed that G. lucidum extracts display weak DNA damaging potential. Since DNA injury promotes aging and cancer, our results substantiate the traditional use of this mushroom to prolong life.