Gene clusters for β-lactam antibiotics and control of their expression: why have clusters evolved, and from where did they originate?

While β-lactam compounds were discovered in filamentous fungi, actinomycetes and gram-negative bacteria are also known to produce different types of β-lactams. All β-lactam compounds contain a four-membered β-lactam ring. The structure of their second ring allows these compounds to be classified into penicillins, cephalosporins, clavams, carbapenens or monobactams. Most β-lactams inhibits bacterial cell wall biosynthesis but others behave as β-lactamase inhibitors (e.g., clavulanic acid) and even as antifungal agents (e.g., some clavams). Due to the nature of the second ring in β-lactam molecules, the precursors and biosynthetic pathways of clavams, carbapenems and monobactams differ from those of penicillins and cephalosporins. These last two groups, including cephamycins and cephabacins, are formed from three precursor amino acids that are linked into the α-aminoadipyl-L-cysteinyl-D-valine tripeptide. The first two steps of their biosynthetic pathways are common. The intermediates of these pathways, the characteristics of the enzymes involved, the lack of introns in the genes and bioinformatic analysis suggest that all of them should have evolved from an ancestral gene cluster of bacterial origin, which was surely transferred horizontally in the soil from producer to non-producer microorganisms. The receptor strains acquired fragments of the original bacterial cluster and occasionally inserted new genes into the clusters, which once modified, acquired new functions and gave rise to the final compounds that we know. When the order of genes in the Streptomyces genome is analyzed, the antibiotic gene clusters are highlighted as gene islands in the genome. Nonetheless, the assemblage of the ancestral β-lactam gene cluster remains a matter of speculation. [Int Microbiol 2006; 9(1):9-19

The influence of skull shape modularity on internal skull structures: a 3D-Pilot study using bears

In order to capture the phenotypic variation of the internal skull structures, such as the sinuses or the brain, it is necessary to perform CT scans in a large number of specimens, which is difficult and expensive. Therefore, while the external morphology of the mammalian cranium has been the subject of many morphometric studies, the internal structures of the cranium have been comparatively less studied. Here, we explore how the variation of external shape reflects the morphology of internal structures. We use the family Ursidae (Carnivora, Mammalia) as a case study because bears have a wide variability of cranial morphologies in part associated with different trophic ecologies. To do this, we digitized a set of landmarks in 3D with a Microscribe G2X from the external surface of the cranium in a wide sample of bears. Additionally, the crania of seven bear species were CT-scanned and prepared digitally to visualize the 3D models of the external cranium morphology and of internal structures. Subsequently, we divided the landmarks into two modules, splanchnocranium and neurocranium, and we perform a two-block partial least squares analysis (2B PLS) to explore the intraspecific (static) morphological changes associated with the covariation between them. These morphological changes were visualized using the morphing technique with the 3D models, looking at both the external shape and the internal structures. In addition, we inferred the volume of the sinuses and of the brain in each hypothetical model. Our results show that the first two PLS axes are associated externally with changes in the basicranial angle, face length and cranium height and width. Concerning the internal structures, there are parallel changes in dorso-ventral and medio-lateral expansion of sinuses and brain, accompanied by their corresponding changes in volume. In contrast, the third PLS axis is related to opposite changes in the volume of sinuses and brain. These preliminary results suggest that the opposite relationship between sinuses and brain volumes in the bear cranium is not as evident as expected, at least at intraspecific level.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Conversion of β-carotene into astaxanthin: Two separate enzymes or a bifunctional hydroxylase-ketolase protein?

Astaxanthin is a xanthophyll of great interest in animal nutrition and human health. The market prospect in the nutraceutics industries for this health-protective molecule is very promising. Astaxanthin is synthesized by several bacteria, algae and plants from β-carotene by the sequential action of two enzymes: a β-carotene, 3,3'-hydroxylase that introduces an hydroxyl group at the 3 (and 3') positions of each of the two β-ionone rings of β-carotene, and a β-carotene ketolase that introduces keto groups at carbons 4 and 4' of the β-ionone rings. Astaxanthin is also produced by the yeast-like basidiomycete Xanthophyllomyces dendrorhous. A gene crtS involved in the conversion of β-carotene to astaxanthin has been cloned simultaneously by two research groups. Complementation studies of X. dendrorhous mutants and expression analysis in Mucor circinelloides reveals that the CrtS enzyme is a β-carotene hydroxylase of the P-450 monooxygenase family that converts β-carotene to the hydroxylated derivatives β-cryptoxanthin and zeaxanthin, but it does not form astaxanthin or the ketolated intermediates in this fungus. A bifunctional β-carotene hydroxylase-ketolase activity has been proposed for the CrtS protein. The evidence for and against this hypothesis is analyzed in detail in this review

Proteomics Shows New Faces for the Old Penicillin Producer Penicillium chrysogenum

Fungi comprise a vast group of microorganisms including the Ascomycota (majority of all described fungi), the Basidiomycota (mushrooms or higher fungi), and the Zygomycota and Chytridiomycota (basal or lower fungi) that produce industrially interesting secondary metabolites, such as β-lactam antibiotics. These compounds are one of the most commonly prescribed drugs world-wide. Since Fleming's initial discovery of Penicillium notatum 80 years ago, the role of Penicillium as an antimicrobial source became patent. After the isolation of Penicillium chrysogenum NRRL 1951 six decades ago, classical mutagenesis and screening programs led to the development of industrial strains with increased productivity (at least three orders of magnitude). The new “omics” era has provided the key to understand the underlying mechanisms of the industrial strain improvement process. The review of different proteomics methods applied to P. chrysogenum has revealed that industrial modification of this microorganism was a consequence of a careful rebalancing of several metabolic pathways. In addition, the secretome analysis of P. chrysogenum has opened the door to new industrial applications for this versatile filamentous fungus

Characterization of the iron-regulated desA promoter of Streptomyces pilosus as a system for controlled gene expression in actinomycetes

BACKGROUND: The bioavailability of iron is quite low since it is usually present as insoluble complexes. To solve the bioavailability problem microorganisms have developed highly efficient iron-scavenging systems based on the synthesis of siderophores that have high iron affinity. The systems of iron assimilation in microorganisms are strictly regulated to control the intracellular iron levels since at high concentrations iron is toxic for cells. Streptomyces pilosus synthesizes the siderofore desferrioxamine B. The first step in desferrioxamine biosynthesis is decarboxylation of L-lysine to form cadaverine, a desferrioxamine B precursor. This reaction is catalyzed by the lysine decarboxylase, an enzyme encoded by the desA gene that is repressed by iron. RESULTS: The binding of the DmdR (acronym for divalent metal dependent repressor) to the desA promoter in presence of Fe(2+ )or other divalent ions has been characterized. A 51 bp DNA fragment of the desA promoter containing the 9 bp inverted repeat was sufficient for binding of the DmdR repressor, as observed by the electrophoretic mobility shift assay. The desA mobility shift was prevented by neutralizing DmdR with anti-DmdR antibodies or by chelating the divalent metal in the binding reaction with 2,2'-dipyridyl. Binding to the desA promoter was observed with purified DmdR repressors of Streptomyces coelicolor or Rhodococcus fascians suggesting that there is a common mechanism of iron-regulation in actinomycetes. The complete desA promoter region was coupled using transcriptional fusions to the amy reporter gene (encoding α-amylase) in low copy or multicopy Streptomyces vectors. The iron-regulated desA promoter was induced by addition of the iron chelating agent 2,2'-dipyridyl resulting in a strong expression of the reporter gene. CONCLUSIONS: The iron-regulated desA promoter can be used for inducible expression of genes in Streptomyces species, as shown by de-repression of the promoter when coupled to a reporter gene

Entanglement entropy of an accelerating universe

We have considered the existence of a dependence of the entanglement entropy on the cosmological horizon surface area also in several accelerating models of the current universe both for a quintessence scalar field and for a phantom-energy scenario. It is shown that if a quintessence vacuum cosmic field is considered then, though the case for w>-1 satisfies a second law for entanglement entropy, when w<-1 such a law is violated. It is finally noted that the entanglement entropy and the distinct formulations of cosmic holography share common future surfaces which are optimal screen for the latter descriptions. © 2012 American Physical Society.This work was supported by MICINN under Research Project FIS2008- 06332.Peer Reviewe

Edad y tamaño empresarial y ciclo de vida financiero

The aim of this paper is to analyze whether some of the empirical implications of the Berger and Udell's (1998) financial growth cycle hold. We use a sample of 22.842 observations for year 2003 and test several hypothesis through MANOVA analysis. Results show that companies tend to have different financing structures depending on age and size. The hypothesis about equity is not confirmed, because the older the company is the higher tends to be its value, caused by the increasing reserves. On the other hand, the risk of the company decreases with age. Results related to trade credit, short term credit and financing deficit are mixed, not significant or contrary to the considered hypothesis. El objetivo de este trabajo es examinar si se cumplen algunas de las implicaciones empíricas del ciclo de vida financiero planteado por Berger y Udell (1998). Para ello se utiliza una muestra de 22.842 observaciones para el año 2003 y mediante tests MANOVAs se contrastan diversas hipótesis. Los resultados obtenidos muestran que las empresas tienen distintas estructuras financieras dependiendo de la edad y el tamaño. La tendencia general es que a mayor edad, mayor presencia de fondos propios, ocasionada por el fuerte incremento de los beneficios retenidos, siendo este resultado contrario a la hipótesis planteada. Por otro lado, el riesgo asumido por las empresas disminuye con la edad. La evidencia relativa al empleo de crédito comercial, deuda financiera a corto plazo y déficit financiero es mixta, no significativa, o contraria a las hipótesis planteadas, especialmente para el caso del factor edad.Edad, tamaño, ciclo de vida financiero, asimetría informativa, financiación empresarial. Age, size, financial growth cycle, information asymmetries, corporate finance.

Impact of Vegetable Oil Type on the Rheological and Tribological Behavior of Montmorillonite-Based Oleogels

We formulated and characterized oleogels based on montmorillonite clay and vegetable oils that could serve as eco-friendly semi-solid lubricants. In particular, we studied the influence of the physical-chemical properties of olive, castor, soybean, linseed, and sunflower oils on the rheological, chemical, thermal, and tribological properties of the semi-solid lubricants. We prepared the oleogels via the highly intensive mixing of vegetable oils with clay at a concentration of 30 wt.%. The oleogels exhibited shear-thinning, thixotropy, structural recovery, and gel-like behavior commonly related to that of a three-dimensional network. The results were corroborated via XRD measurements showing the presence of intercalated nanoclay structures well-dispersed in the vegetable oil. Empirical correlations between the content of saturated (SFAs), unsaturated (UFAs), mono-unsaturated (MUFAs) and poly-unsaturated (PUFAs) fatty acids and the plateau modulus of the aerogels were found. From these experimental results, we can conclude that the fatty acid profile of the vegetable oils exerts an important influence on the rheological and tribological properties of resulting clay and vegetable oil oleogelsThis research was supported by FEDER European Programme and Junta de Andalucía, grant number PY20_00751. The authors acknowledge the X-ray Diffraction Service (Universidad de Huelva) for providing full access and assistance in X-ray Diffraction measurement