Scale-up of PUF-immobilized fungal chitosanase–lipase preparation production

Mucor circinelloides IBT-83 mycelium that exhibits both lipolytic (A_L) and chitosanolytic (A_CH) activities was immobilized into polyurethane foam in a 30?L laboratory fermenter. The process of immobilization was investigated in terms of the carrier porosity, its type, amount, and shape, location inside the fermenter, mixing, and aeration parameters during the culture, as well as downstream processing operations. The selected conditions allowed for immobilization of approximately 7?g of defatted and dried mycelium in 1?g of carrier, i.e., seven times more than achievable in 1?L shake-flasks. Enzymatic preparation obtained by this method exhibited both the chitosanolytic (A_CH 432.5?±?6.8?unit/g) and lipolytic (A_L 150.0?±?9.3?U/g) activities. The immobilized preparation was successfully used in chitosan hydrolysis to produce chitooligosaccharides and low molecular weight chitosan, as well as in waste fats degradation and in esters synthesis in nonaqueous media. It was found that the half-life of immobilized preparations stored at room temperature is on average of 200 days.     

Do flocks of great cormorants and goosanders avoid spatial overlap in foraging habitat during the non-breeding season?

Species distribution, ecology, and behaviour are shaped, amongst other things, by interspecific, antagonistic interactions, and this phenomenon is particularly noticeable among predators. We studied the spatial co-distribution of two top piscivorous bird species foraging on inland waters outside breeding season. We considered the hypothesis that goosanders, Mergus merganser, and great cormorants, Phalacrocorax carbo, avoid foraging in close proximity to each other. Data collected on five river-reservoir systems in the Western Carpathians (Poland and Slovakia) during two periods (2014–2015 and 2015–2016) showed that goosander numbers reduced significantly and their foraging areas changed when large flocks of cormorants arrived and began foraging. We also found that inter-flock distances were greatest between flocks of goosanders and cormorants, suggesting that the former species avoided the waters occupied by the latter. Distribution of flocks of both species was additionally determined by the location of foraging place in river-reservoir system, weather, and presence of other piscivorous birds (e.g. grebes) and raptors (e.g. eagles). Together with the results of research in adjacent Bohemia, this study suggests that competition between cormorants and goosanders may arise when bodies of water suitable for piscivorous foraging are scattered and limited in number, as in the mountainous areas of Central Europe.     

Acorns of invasive Northern Red Oak (<Emphasis Type="Italic">Quercus rubra</Emphasis>) in Europe are larval hosts for moths and beetles

In their first phase of expanding into new areas, invasive plants often take advantage of the inability of existing herbivores and pathogenic species to exploit them. However, in the longer term local enemies may adapt to using these invasive species as a food source. This study assesses the use of mature acorns of two oak species in Europe (the native Pedunculate Oak Quercus robur and the invasive Northern Red Oak Quercus rubra) by moths Cydia fagiglandana and Cydia splendana and beetles Curculio spp. We show that acorns of invasive oak species can be equally attractive to C. splendana but only partially so to C. fagiglandana where infestation rates where significantly lower (approximately half) compared to the native oak. The infestation by Curculio beetles of Northern Red Oak was marginal, less than 1% of the rate in the native oak species. The larval final weights did not differ significantly between host species, but emergence of C. splendana and Curculio spp. took significantly longer in acorns of Northern Red Oak. It is likely that C. fagiglandana and C. splendana have increased their niche breadths by exploiting invasive oak species and avoiding competition with the Curculio weevils. Furthermore, the occurrence of Northern Red Oak could stabilize food resources during years when native oak species have poor acorn crops.     

Modulation of MRP1 protein transport by plant, and synthetically modified flavonoids

The influence of novel synthetic and plant origin flavonoids on activity of multidrug resistance-associated protein (MRP1) was investigated in human erythrocytes used as a cell model expressing MRP1 in plasma membrane. The fluorescent probe, BCPCF (2', 7'-bis-(3-carboxy-propyl)-5-(and-6)-carboxyfluorescein), was applied as a substrate for MRP1 multidrug resistance transporter. The effect of compounds belonging to different classes of natural flavonoids: flavone, flavonol, isoflavones and flavanolignan was compared with action of new synthetic derivatives of genistein. Most of the flavonoids showed strong or moderate ability to inhibit transport carried out by MRP1. Inhibitory properties of flavonoids were compared to the effects of indomethacin, probenecid and MK-571 known as MRP1 inhibitors. Studying the influence of new synthetic genistein derivatives on BCPCF transport we have found that the presence of hydrophobic groups substituting hydrogen of hydroxyl group at the position 4' in ring B of isoflavone is more important for inhibitory properties than hydrophobic substitution at the position 7 in ring A. In case of naturally occurring isoflavones the replacement of hydrogen at position 4' by hydrophobic ring structure seems also to be favourable for inhibition potency.     

Influence of restoration on arbuscular mycorrhiza of Biscutella laevigata L. (Brassicaceae) and Plantago lanceolata L. (Plantaginaceae) from calamine spoil mounds

The arbuscular mycorrhizal status of two plant species, Biscutella laevigata L. and Plantago lanceolata L., was investigated on calamine spoil mounds in Boles?aw (southern Poland). Although B. laevigata is a member of the Brassicaceae, a family generally accepted as non-mycorrhizal, this species formed AM symbioses on both heavy metal-contaminated and non-contaminated sites. Besides vesicles and coils, arbuscules were also observed, especially in roots collected prior to seed maturity. Relative mycorrhizal root length and relative arbuscular richness were usually much higher in P. lanceolata than in B. laevigata but not absolute arbuscule richness. Roots of P. lanceolata showed higher colonisation than B. laevigata. Although roots were collected from plants in close proximity, no correlation in mycorrhizal parameters was found between the two species.     

Pyruvate carboxylase is an essential protein in the assembly of yeast peroxisomal oligomeric alcohol oxidase

Hansenula polymorpha ass3 mutants are characterized by the accumulation of inactive alcohol oxidase (AO) monomers in the cytosol, whereas other peroxisomal matrix proteins are normally activated and sorted to peroxisomes. These mutants also have a glutamate or aspartate requirement on minimal media. Cloning of the corresponding gene resulted in the isolation of the H. polymorpha PYC gene that encodes pyruvate carboxylase (HpPyc1p). HpPyc1p is a cytosolic, anapleurotic enzyme that replenishes the tricarboxylic acid cycle with oxaloacetate. The absence of this enzyme can be compensated by addition of aspartate or glutamate to the growth media. We show that HpPyc1p protein but not the enzyme activity is essential for import and assembly of AO. Similar results were obtained in the related yeast Pichia pastoris. In vitro studies revealed that HpPyc1p has affinity for FAD and is capable to physically interact with AO protein. These data suggest that in methylotrophic yeast pyruvate carboxylase plays a dual role in that, besides its well-characterized metabolic function as anapleurotic enzyme, the protein fulfils a specific role in the AO sorting and assembly process, possibly by mediating FAD-binding to AO monomers.     

In vitro effect of leptin on steroids' secretion by FSH- and LH-treated porcine small, medium and large preovulatory follicles

The role of exogenous leptin in the follicular steroidogenesis in pigs has not been fully elucidated and available data are controversial. In the current study porcine follicles were recovered from ovaries during early, middle, and preovulatory stages of the follicular phase of the estrous cycle. Follicles were cultured in the presence of the recombinant ovine leptin (oLEP) with or without LH (100 ng/ml) or FSH (100 ng/ml). Medium estradiol (E(2)), testosterone (T) and progesterone (P(4)) concentrations were determined after 48h of culture. Leptin at a dose of 2 ng/ml had no effect on basal E(2) and T secretion by small and medium follicles but decreased E(2) secretion by large follicles. Significant synergistic action of FSH and leptin resulting in a 2 - 5 fold stimulation of E(2) secretion by small and medium follicles was observed. The aromatase inhibitor, CGS 16949A augmented T secretion and inhibited E(2) secretion by control and FSH-treated medium follicles. In FSH and leptin-treated follicles, the inhibitory action of CGS 16949A on E(2) secretion was observed. However, there was no augmentation of T secretion. In leptin-treated follicles the stimulatory action on P(4) secretion was observed only during the preovulatory stage. In these follicles, significant synergistic action of leptin with LH on P(4) secretion was also noted. These results indicate that there is a maturation-dependent action of leptin on both E(2) and P(4) secretion. They also suggest a synergistic action of leptin and FSH on E(2) secretion by small and medium follicles as well as leptin and LH on P(4) secretion by large follicles in pigs.