Differential adaptation of membranes of two osmotolerant fungi,Aspergillus chevalieri and Penicillium expansum to high sucrose concentrations

Aspergillus chevalieri and Penicillium expansum were able to tolerate sucrose concentrations in the growth media up to 80% (w/v). At 50% sucrose the growth rate is approximately 1.4 and 1.2 times, respectively, higher than in the control. While at 80% sucrose it drops to 35% and 45% of the control level for both fungi. Lipids and proteins in plasma membranes increased with increasing sucrose concentrations in the growth medium. Phospholipid content in membranes of both organisms being also increased, phosphatidyl glycerol was the major detected phospholipid and represented the highest increase. The fatty acid composition of fraction enriched plasma membrane of both fungi changed when they were grown in high sucrose concentrations. Some fatty acids which had not been detected in control cultures were present and the proportions of other fatty acids changed. At 50% sucrose the unsaturation index of membranes decreased by 20-25% in both fungi, indicating that the plasma membrane is less fluid at this concentration. At 80% sucrose a similar trend was observed for P. expansum but for A. chevalieri the unsaturation index was little changed compared with the control. The fluorescence polarization values of 1,6-diphenyl 1,3,5-hexatriene (DPH) in membranes of both fungi grown at 80% sucrose increased, indicating a decrease in membrane fluidity. At 50% sucrose the increase in saturation of membrane fatty acids would tend to reduce membrane fluidity but in A. chevalieri at 80% sucrose fatty acids did not become more saturated. In this case the marked increase in sterols at this sucrose concentration may be responsible for low membrane fluidity.     

Novel Nucleoside Analogues: First Synthesis of Pyridine-4-Thioglycosides and Their Cytotoxic Evaluation

The reaction of sodium 2,2-dicyanoethene-1,1-bis(thiolate) with 2-cyano-N-arylacetamides afforded sodium pyridine-4-thiolates, coupling of the latters with 2,3,4,6-tetra-O-acetyl-D-gluco- and D-galactopyranosyl bromides, respectively, afforded new pyridine-4-thioglycosides. Ammonolysis of the latter compounds afforded the free thioglycosides. The antitumor activities of the synthesized compounds were tested against human tumor cell lines; lung (A549), colon (HCT116), liver (HEPG2), and prostate (PC3).     

Biodegradation of aliphatic homopolyesters and aliphatic-aromatic copolyesters by anaerobic microorganisms

The anaerobic degradability of natural and synthetic polyesters is investigated applying microbial consortia (3 sludges, 1 sediment) as well as individual strains isolated for this purpose. In contrast to aerobic conditions, the natural homopolyester polyhydroxybutyrate (PHB) degrades faster than the copolyester poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV). For the synthetic polyester poly(epsilon-caroplacton) (PCL), microbial degradation in the absence of oxygen could be clearly demonstrated; however, the degradation rate is significantly lower than for PHB and PHBV. Other synthetic polyesters such as poly(trimethylene adipate) (SP3/6), poly(tetramethylene adipate) (SP4/6), and aliphatic-aromatic copolyesters from 1,4-butanediol, terephthalic acid, and adipic acid (BTA-copolymers) exhibit only very low anaerobic microbial susceptibility. A copolyester with high amount of terephthalic acid (BTA 40:60) resisted the anaerobic breakdown even under thermophilic conditions and/or when blended with starch. For the synthetic polymers, a number of individual anaerobic strain could be isolated which are able to depolymerize the polymers and selected strains where identified as new species of the genus Clostridium or Propionispora. Their distinguished degradation patterns point to the involvement of different degrading enzymes which are specialized to depolymerize either the natural polyhydroxyalkanoates (e.g., PHB), the synthetic polyester PCL, or other synthetic aliphatic polyesters such as SP3/6. It can be supposed that these enzymes exhibit comparable characteristics as those described to be responsible for aerobic polyester degradation (lipases, cutinases, and PHB-depolymerases).     

MICAL-like1 mediates epidermal growth factor receptor endocytosis

Small GTPase Rabs are required for membrane protein sorting/delivery to precise membrane domains. Rab13 regulates epithelial tight junction assembly and polarized membrane transport. Here we report that Molecule Interacting with CasL (MICAL)-like1 (MICAL-L1) interacts with GTP-Rab13 and shares a similar domain organization with MICAL. MICAL-L1 has a calponin homology (CH), LIM, proline rich and coiled-coil domains. It is associated with late endosomes. Time-lapse video microscopy shows that green fluorescent protein-Rab7 and mcherry-MICAL-L1 are present within vesicles that move rapidly in the cytoplasm. Depletion of MICAL-L1 by short hairpin RNA does not alter the distribution of a late endosome/lysosome-associated protein but affects the trafficking of epidermal growth factor receptor (EGFR). Overexpression of MICAL-L1 leads to the accumulation of EGFR in the late endosomal compartment. In contrast, knocking down MICAL-L1 results in the distribution of internalized EGFR in vesicles spread throughout the cytoplasm and promotes its degradation. Our data suggest that the N-terminal CH domain associates with the C-terminal Rab13 binding domain (RBD) of MICAL-L1. The binding of Rab13 to RBD disrupts the CH/RBD interaction, and may induce a conformational change in MICAL-L1, promoting its activation. Our results provide novel insights into the MICAL-L1/Rab protein complex that can regulate EGFR trafficking at late endocytic pathways.     

Degradation of natural and synthetic polyesters under anaerobic conditions

Often, degradability under anaerobic conditions is desirable for plastics claimed to be biodegradable, e.g. in anaerobic biowaste treatment plants, landfills and in natural anaerobic sediments. The biodegradation of the natural polyesters poly(beta-hydroxybutyrate) (PHB), poly(beta-hydroxybutyrate-co-11.6%-beta-hydroxyvalerate) (PHBV) and the synthetic polyester poly(epsilon-caprolactone) (PCL) was studied in two anaerobic sludges and individual polyester degrading anaerobic strains were isolated, characterized and used for degradation experiments under controlled laboratory conditions. Incubation of PHB and PHBV films in two anaerobic sludges exhibited significant degradation in a time scale of 6-10 weeks monitored by weight loss and biogas formation. In contrast to aerobic conditions, PHB was degraded anaerobically more rapidly than the copolyester PHBV, when tested with either mixed cultures or a single strained isolate. PCL tends to degrade slower than the natural polyesters PHB and PHBV. Four PHB and PCL degrading isolates were taxonomically identified and are obviously new species belonging to the genus Clostridium group I. The depolymerizing enzyme systems of PHB and PCL degrading isolates are supposed to be different. Using one isolated strain in an optimized laboratory degradation test with PHB powder, the degradation time was drastically reduced compared to the degradation in sludges (2 days vs. 6-10 weeks).     

Growth, zearalenone production and some metabolic activities of Fusarium moniliforme under salt stress

With a increasing salinity, a decrease in the growth rate of Fusarium moniliforme was observed. The percentage of germinated conidia decreased with increasing salinity (% germination ranged from 80.3% at 0.0% NaCl to 0.0% at 15% NaCl). Concentration of 12.5% NaCl produced the highest chlamydospore-like structures. Concentration of 5% NaCl increased zearalenone production which decreased with increasing salt stress. Escherichia coli was more tolerant to the toxin than Bacillus subtilis. The total amount of lipids produced by F. moniliforme changed with increasing the concentration of NaCl in the growth medium. The genomic DNA of the control and treated samples showed a common band of more than 20 kilobases. Similar RAPD-PCR patterns were also produced.     

Antibacterial and Anticancer activity of ε -poly-L-lysine (ε -PL) produced by a marine Bacillus subtilis sp

A marine Bacillus subtilis SDNS was isolated from sea water in Alexandria and identified using 16S rDNA sequence analysis. The bacterium produced a compound active against a number of gram negativeve bacteria. Moreover, the anticancer activity of this bacterium was tested against three different human cell lines (Hela S3, HepG2 and CaCo). The highest inhibition activity was recorded against Hela S3 cell line (77.2%), while almost no activity was recorded towards CaCo cell line. HPLC and TLC analyses supported evidence that Bacillus subtilis SDNS product is ?;-poly-L-lysine. To achieve maximum production, Plackett-Burman experimental design was applied. A 1.5 fold increase was observed when Bacillus subtilis SDNS was grown in optimized medium composed of g/l: (NH(4) )(2) SO(4) , 15; K(2) HPO(4) , 0.3; KH(2) PO(4) , 2; MgSO(4) · 7 H(2) O, 1; ZnSO(4) · 7 H(2) O, 0; FeSO(4) · 7 H(2) O, 0.03; glucose, 25; yeast extract, 1, pH 6.8. Under optimized culture condition, a product value of 76.3 mg/l could be obtained. According to available literature, this is the first announcement for the production of ?;-poly-L-lysine (?;-PL) by a member of genus Bacillus. (? 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim).