Production and partial characterization of lipase from Penicillium verrucosum obtained by submerged fermentation of conventional and industrial media

The growing interest in lipase production is related to the potential biotechnological applications that these enzymes present. Current studies on lipase production by submerged fermentation involve the use of agro-industrial residues aiming at increasing economic attractiveness. Based on these aspects, the objective of this work was to investigate lipase production by Penicillium verrucosum in submerged fermentation using a conventional medium based on peptone, yeast extract, NaCl and olive oil, and an industrial medium based on corn steep liquor, Prodex Lac (yeast hydrolysate), NaCl and olive oil, as well as to characterize the crude enzymatic extracts obtained. Kinetics of lipase production was evaluated and the highest enzymatic activities, of 3.15 and 2.22 U.mL-1, were observed when conventional and industrial media were used, respectively. The enzymatic extract showed optimal activity in the range from 30 to 40 °C and at pH 7.0. Although the industrial medium presents economical advantages over the conventional medium, the presence of agro-industrial residues rich in nitrogen and other important nutrients seemed to contribute to a reduction in lipase activity.

Produção de galactooligossacarídeo por Scopulariopis sp.

Os galactooligossacarídeos (GOS) são um grupo de oligossacarídeos não digeríveis (NDOs), resistentes às enzimas digestivas do intestino com efeitos similares ao da fibra. Sua ingestão aumenta seletivamente o crescimento das Bifidobacterium e dos Lactobacilus no intestino. O benéfico da ingestão de galactooligossacarídeos ocorre pelo aumento dessas populações de bifidobactérias no cólon, suprimindo a atividade de bactérias putrefativas e reduzindo a formação de produtos tóxicos por fermentação. A cepa de Scopulariopsis sp. apresentou boa produtividade de β-galactosidase quando crescida em meio de fermentação semissólida. O objetivo deste trabalho foi extrair a enzima β-galactosidase produzida por uma linhagem de Scopulariopsis sp. e avaliar o efeito da temperatura, tempo de reação, concentração de enzima e lactose para produção de GOS. Foram analisadas as temperaturas 35, 45 e 60 °C, os tempos de reação 12, 24 e 48 horas, concentrações de enzima 0,5 a 10 U.mL-1 e 10, 25 e 40% (p/v) de solução de lactose (em 0,1 M de tampão acetato de sódio, pH 5.0). As condições ótimas foram de 40% (p/v), a 45 °C, 10 U.mL-1 de enzima e o melhor tempo foi de 12 horas de reação. Nessas condições, a enzima converteu 20% de lactose em oligossacarídeos (80,8 mg.mL-1 de 4'galactosyl-lactose).

Microparticles obtained by complex coacervation: influence of the type of reticulation and the drying process on the release of the core material

Microparticles obtained by complex coacervation were crosslinked with glutaraldehyde or with transglutaminase and dried using freeze drying or spray drying. Moist samples presented Encapsulation Efficiency (%EE) higher than 96%. The mean diameters ranged from 43.7 ± 3.4 to 96.4 ± 10.3 µm for moist samples, from 38.1 ± 5.36 to 65.2 ± 16.1 µm for dried samples, and from 62.5 ± 7.5 to 106.9 ± 26.1 µm for rehydrated microparticles. The integrity of the particles without crosslinking was maintained when freeze drying was used. After spray drying, only crosslinked samples were able to maintain the wall integrity. Microparticles had a round shape and in the case of dried samples rugged walls apparently without cracks were observed. Core distribution inside the particles was multinuclear and homogeneous and core release was evaluated using anhydrous ethanol. Moist particles crosslinked with glutaraldehyde at the concentration of 1.0 mM.g-1 protein (ptn), were more efficient with respect to the core retention compared to 0.1 mM.g-1 ptn or those crosslinked with transglutaminase (10 U.g-1 ptn). The drying processes had a strong influence on the core release profile reducing the amount released to all dry samples

Production and partial characterization of alkaline polygalacturonase secreted by thermophilic Bacillus sp. SMIA-2 under submerged culture using pectin and corn steep liquor

Polygalacturonase production by the thermophilic Bacillus sp. SMIA-2 cultivated in liquid cultures containing 0.5% (w/v) apple pectin and supplemented with 0.3% (w/v) corn steep liquor, reached its maximum after 36 hours with levels of 39 U.mL-1. The increase in apple pectin and corn steep liquor concentrations in the medium from 0.5 and 0.3%, respectively, to 0.65%, markedly affected the production of polygalacturonase, whose activity increased four times, reaching a maximum of 150.3 U.mL-1. Studies on polygalacturonase characterization revealed that the optimum temperature of this enzyme was between 60-70 °C. Thermostability profile indicated that the enzyme retained about 82 and 63% of its activity at 60 and 70 °C, respectively, after 2 hours of incubation. The optimum pH of the enzyme was found to be 10.0. After incubation of crude enzyme solution at room temperature for 2 hours at pH 8.0, a decrease of about 29% on its original activity was observed. At pH 10.0, the decrease was 25%.

Simultaneous α-amylase and protease production by the soil bacterium Bacillus sp. SMIA-2 under submerged culture using whey protein concentrate and corn steep liquor: compatibility of enzymes with commercial detergents

Protease and α-amylase production by a thermophilic Bacillus sp. SMIA-2 cultivated in liquid cultures containing 0.25% (w/v) starch as a carbon source reached a maximum at 18 hours (47 U.mg-1 Protein) and 36 hours (325 U.mg-1 Protein), respectively. Culture medium supplementation with whey protein concentrate (0.1%, w/v) and corn steep liquor (0.3%, w/v) not only improved the production of both enzymes but also enabled them to be produced simultaneously. Under these conditions, α-amylase and protease production reached a maximum in 18 hours with levels of 401 U.mg-1 protein and 78 U.mg-1 protein, respectively. The compatibility of the enzymes produced with commercial laundry detergent was investigated. In the presence of Campeiro® detergent, α-amylase activity increased while protease activity decreased by about 27%. These enzymes improved the cleaning power of Campeiro® detergent since they were able to remove egg yolk and tomato sauce stains when used in this detergent.

Assessment of fructooligosaccharides production from sucrose in aqueous and aqueous-organic systems using immobilized inulinase from Kluyveromyces marxianus NRRL Y-7571

This work investigated the fructooligosaccharides (FOS) synthesis by immobilized inulinase obtained from Kluyveromyces marxianus NRRL Y-7571 in aqueous and aqueous-organic systems using sucrose as substrate. The sequential strategy of experimental design was used to optimize the FOS conversion in both systems. For the aqueous-organic system, a 2(6-2) fractional design was carried out to evaluate the effects of temperature, sucrose concentration, pH, aqueous/organic ratio, enzyme activity, and polyethylene glycol concentration. For the aqueous system, a central composite design for the enzyme activity and the sucrose concentration was carried out. The highest fructooligosaccharides yield (Y FOS) for the aqueous-organic system was 18.2 ± S0.9 wt%, at 40 ºC, pH 5.0, sucrose concentration of 60% (w/w), enzyme activity of 4 U.mL-1, and aqueous/organic ratio of 25/75 wt%. The highest Y FOS for the aqueous system was 14.6 ± 0.9 wt% at 40 ºC, pH 5.0, sucrose concentration of 60 wt%, and enzyme activity of 4.0 U.mL-1.

Banana waste as substrate for a-amylase production by Bacillus subtilis (CBTK 106) under solid-state fermentation

Bacillus subtilis CBTK 106, isolated from banana wastes, produced high titres of a-amylase when banana fruit stalk was used as substrate in a solid-state fermentation system. The e¤ects of initial moisture content, particle size, cooking time and temperature, pH, incubation temperature, additional nutrients, inoculum size and incubation period on the production of a- amylase were characterised. A maximum yield of 5 345 000 U mg~1 min~1 was recorded when pretreated banana fruit stalk (autoclaved at 121 ¡C for 60 min) was used as substrate with 70% initial moisture content, 400 lm particle size, an initial pH of 7.0, a temperature of 35 ¡C, and additional nutrients (ammonium sulphate/sodium nitrate at 1.0%, beef extract/peptone at 0.5%, glucose/sucrose/starch/maltose at 0.1% and potassium chloride/sodium chloride at 1.0%) in the medium, with an inoculum-to-substrate ratio of 10% (v/w) for 24 h. The enzyme yield was 2.65-fold higher with banana fruit stalk medium compared to wheat bran

Impact of process parameters on chitinase production by an alkalophilic marine Beau6eria bassiana in solid state fermentation

A chitinolytic fungus, Beau6eria bassiana was isolated from marine sediment and significant process parameters influencing chitinase production in solid state fermentation using wheat bran were optimised. The organism was strongly alkalophilic and produced maximum chitinase at pH 9·20. The NaCl and colloidal chitin requirements varied with the type of moistening medium used. Vegetative (mycelial) inoculum was more suitable than conidial inoculum for obtaining maximal enzyme yield. The addition of phosphate and yeast extract resulted in enhancement of chitinase yield. After optimisation, the maximum enzyme yield was 246·6 units g 1 initial dry substrate (U gIDS 1). This is the first report of the production of chitinase from a marine fungus.

Verbal proficiency as fitness indicator: Experimental and comparative research on the evolutionary psychology of language and verbal displays

Summary: Recent research on the evolution of language and verbal displays (e.g., Miller, 1999, 2000a, 2000b, 2002) indicated that language is not only the result of natural selection but serves as a sexually-selected fitness indicator that is an adaptation showing an individual’s suitability as a reproductive mate. Thus, language could be placed within the framework of concepts such as the handicap principle (Zahavi, 1975). There are several reasons for this position: Many linguistic traits are highly heritable (Stromswold, 2001, 2005), while naturally-selected traits are only marginally heritable (Miller, 2000a); men are more prone to verbal displays than women, who in turn judge the displays (Dunbar, 1996; Locke & Bogin, 2006; Lange, in press; Miller, 2000a; Rosenberg & Tunney, 2008); verbal proficiency universally raises especially male status (Brown, 1991); many linguistic features are handicaps (Miller, 2000a) in the Zahavian sense; most literature is produced by men at reproduction-relevant age (Miller, 1999). However, neither an experimental study investigating the causal relation between verbal proficiency and attractiveness, nor a study showing a correlation between markers of literary and mating success existed. In the current studies, it was aimed to fill these gaps. In the first one, I conducted a laboratory experiment. Videos in which an actor and an actress performed verbal self-presentations were the stimuli for counter-sex participants. Content was always alike, but the videos differed on three levels of verbal proficiency. Predictions were, among others, that (1) verbal proficiency increases mate value, but that (2) this applies more to male than to female mate value due to assumed past sex-different selection pressures causing women to be very demanding in mate choice (Trivers, 1972). After running a two-factorial analysis of variance with the variables sex and verbal proficiency as factors, the first hypothesis was supported with high effect size. For the second hypothesis, there was only a trend going in the predicted direction. Furthermore, it became evident that verbal proficiency affects long-term more than short-term mate value. In the second study, verbal proficiency as a menstrual cycle-dependent mate choice criterion was investigated. Basically the same materials as in the former study were used with only marginal changes in the used questionnaire. The hypothesis was that fertile women rate high verbal proficiency in men higher than non-fertile women because of verbal proficiency being a potential indicator of “good genes”. However, no significant result could be obtained in support of the hypothesis in the current study. In the third study, the hypotheses were: (1) most literature is produced by men at reproduction-relevant age. (2) The more works of high literary quality a male writer produces, the more mates and children he has. (3) Lyricists have higher mating success than non-lyric writers because of poetic language being a larger handicap than other forms of language. (4) Writing literature increases a man’s status insofar that his offspring shows a significantly higher male-to-female sex ratio than in the general population, as the Trivers-Willard hypothesis (Trivers & Willard, 1973) applied to literature predicts. In order to test these hypotheses, two famous literary canons were chosen. Extensive biographical research was conducted on the writers’ mating successes. The first hypothesis was confirmed; the second one, controlling for life age, only for number of mates but not entirely regarding number of children. The latter finding was discussed with respect to, among others, the availability of effective contraception especially in the 20th century. The third hypothesis was not satisfactorily supported. The fourth hypothesis was partially supported. For the 20th century part of the German list, the secondary sex ratio differed with high statistical significance from the ratio assumed to be valid for a general population.

Diseño de una vía clínica para la vigilancia de hepatocarcinoma y evaluación económica

El carcinoma Hepatocelular (HCC) representa la sexta causa más frecuente de cáncer, y la tercera causa de muerte relacionada con cáncer en el mundo con aproximadamente 600.000 muertes anuales. En el 70 % de los casos, este se desarrolla en presencia de una enfermedad crónica del hígado como la cirrosis u otras enfermedades inflamatorias, por lo que practicar métodos de tamizaje para su diagnóstico precoz, pudieran establecer un mejor pronóstico. El objetivo de este trabajo es diseñar una vía clínica capaz de homogenizar el proceso de tamizaje de HCC, soportando su realización con la realización de una evaluación económica de esta intervención. Se realiza una búsqueda sistemática de literatura y se propone una vía clínica para la vigilancia de HCC en Colombia. A esta propuesta se aplica una evaluación económica tipo costo-efectividad mediante un modelo de Markov de la intervención propuesta, comparando la aplicación de la vía clínica propuesta frente al manejo actual en 100 pacientes considerados con riesgo (cirrosis, portadores de HBV y/o portadores de HCV) con un horizonte de tiempo de 30 años analizando como desenlace los años de vida salvados (LYS) desde la perspectiva del tercero pagador para Colombia a precios de 2009. El análisis determina una disminución de la mortalidad en un 40%, y un valor ICER de US$ 1,438 por LYS, por lo cual se concluye que resulta costo efectivo la aplicación de esta propuesta de tamizaje. Es necesario realizar una prueba para la aplicación de la vía clínica.

La orientacion profesional integrada en la segunda etapa de E.G.B.

Presentar un modelo de programación con el fin de integrar la orientación profesional en la escuela; presentar la orientación profesional como un proceso de aprendizaje: sistemático y continuado y no una actividad puntual; provocar la conexión entre medio escolar y medio laboral; materializar un programa de seguimiento para orientaciones escolares; demostrar que la implantación de la orientación profesional no supone ningún gasto extra dentro del presupuesto educativo.. La investigación presente, muestra unas pautas sobre la orientación profesional en segundo de educación general básica. Se estructura en varios capítulos: en el primero, se hace una aproximación del tema y se muestran los objetivos, así como integrar la orientación profesional en la escuela. En el segundo capitulo, se hace un análisis de la orientación profesional, señalando el origen y su desarrollo, los factores históricos que favorecieron su aparición, las etapas o el concepto. El tercer capitulo, explica la legislación vigente de la orientación profesional y la presentación de un proyecto. Y por ultimo en el quinto capitulo, aparece la presentación dinámica del proyecto y los diferentes estudios realizados (en formación profesional, a las profesiones, estudio formativo de B.U.P...).. 1) La orientación profesional debe y tiene que estar integrada en los problemas escolares y en la escuela ya que no es un hecho puntual si no un hecho sistemático, un proceso de aprendizaje por lo que se necesita continuidad. 2) Los programas de orientación profesional deben promover la comprensión de si mismo porque el desarrollo profesional se basa en lo bien que los alumnos conocen sus valores actitudes, intereses y necesidades. 3) El desarrollo del programa puede paliar el fracaso escolar ya que supone el descubrir obstáculos, dificultades o problemas por lo que una vez detectados y conocidos se puede buscar soluciones, aminorarlos o desterrarlas. 4) Es un factor motivador del aprendizaje, puesto que conexiona la necesidad de desarrollar, aprender o adquirir conocimientos, hábitos y aptitudes con el desarrollo de un posible trabajo futuro. 5) La orientación profesional es fundamental para el logro de la autorrealización ya que esta tiende a conseguir la plena madurez del sujeto, su personal bienestar y su plena integración en la comunidad.

Carbon addition alters vegetation composition on ex-arable fields

1. Recent changes in European agricultural policy have led to measures to reverse the loss of species-rich grasslands through the creation of new areas on ex-arable land. Ex-arable soils are often characterized by high inorganic nitrogen (N) levels, which lead to the rapid establishment of annual and fast-growing perennial species during the initial phase of habitat creation. The addition of carbon (C) to the soil has been suggested as a countermeasure to reduce plant-available N and alter competitive interactions among plant species. 2. To test the effect of C addition on habitat creation on ex-arable land, an experiment was set up on two recently abandoned fields in Switzerland and on two 6-year-old restoration sites in the UK. Carbon was added as a mixture of either sugar and sawdust or wood chips and sawdust during a period of 2 years. The effects of C addition on soil parameters and vegetation composition were assessed during the period of C additions and 1 year thereafter. 3. Soil nitrate concentrations were reduced at all sites within weeks of the first C addition, and remained low until cessation of the C additions. The overall effect of C addition on vegetation was a reduction in above-ground biomass and cover. At the Swiss sites, the addition of sugar and sawdust led to a relative increase in legume and forb cover and to a decrease in grass cover. The soil N availability, composition of soil micro-organisms and vegetation characteristics continued to be affected after cessation of C additions. 4. Synthesis and applications. The results suggest that C addition in grassland restoration is a useful management method to reduce N availability on ex-arable land. Carbon addition alters the vegetation composition by creating gaps in the vegetation that facilitates the establishment of late-seral plant species, and is most effective when started immediately after the abandonment of arable fields and applied over several years.

Electrochemical and sonoelectrochemical monitoring of indigo reduction by glucose

The reduction of indigo (dispersed in water) to leuco-indigo (dissolved in water) is an important industrial process and investigated here for the case of glucose as an environmentally benign reducing agent. In order to quantitatively follow the formation of leuco-indigo two approaches based on (i) rotating disk voltammetry and (ii) sonovoltammetry are developed. Leuco-indigo, once formed in alkaline solution, is readily monitored at a glassy carbon electrode in the mass transport limit employing hydrodynamic voltammetry. The presence of power ultrasound further improves the leuco-indigo determination due to additional agitation and homogenization effects. While inactive at room temperature, glucose readily reduces indigo in alkaline media at 65 degrees C. In the presence of excess glucose, a surface dissolution kinetics limited process is proposed following the rate law d eta(leuco-indigo)/dt = k x c(OH-) x S-indigo where eta(leuco-indigo) is the amount of leuco-indigo formed, k = 4.1 x 10(-9) m s(-1) (at 65 degrees C, assuming spherical particles of I gm diameter) is the heterogeneous dissolution rate constant,c(OH-) is the concentration of hydroxide, and Sindigo is the reactive surface area. The activation energy for this process in aqueous 0.2 M NaOH is E-A = 64 U mol(-1) consistent with a considerable temperature effects. The redox mediator 1,8-dihydroxyanthraquinone is shown to significantly enhance the reaction rate by catalysing the electron transfer between glucose and solid indigo particles. (c) 2006 Elsevier Ltd. All fights reserved.

A DFT study of propylene polymerization using neutral salicyladiminato nickel(II) and palladium(II) as catalysts

Propylene polymerization using salicyladiminato metal catalalysts has been studied using density functional theory at the B3LYP/LANL2DZ level. In particular, the effects on the reaction mechanisms of changing the metal from Pd(II) to Ni(II) have been investigated. While the reaction mechanisms involving the salicyladiminato Ni(II) catalyst have been found to be similar to those established previously for the salicyladiminato Pd(II) catalyst, the nickel catalyst was found to differentiate the trans-O intermediate from the trans-.N intermediate with an energy difference of 46.63 U mol(-1) significantly more than the palladium catalyst for which the energy difference was calculated as 35.82 kJ mol(-1). The energy difference between the trans-O configuration and the trans-N configuration is decreased significantly when combining a molecule of propylene with the catalyst. For the Ni catalyst, the trans-O isomer is more stable than the trans-N isomer to a greater extent than for Pd, so that the insertion of propylene from 20 is relatively less favoured for Ni than for Pd. It is predicted that the mechanism of isomerization from 20 to 2N through a rotational transition state TS2O2N is more appropriate for the Ni catalyst system. The palladium system shows a larger preference for pi-coordination than its nickel counterpart, although the latter possesses a lower reaction barrier. It was found that the occupation of the trans-O position in the asymmetric salicyladiminato catalyst is also more favored by the alkene as it is by the alkyl so that insertion of the alkene may always start from a particular configuration so that specific products are obtained. (c) 2005 Elsevier B.V. All rights reserved.

Synthesis of isomaltooligosaccharides and oligodextrans by the combined use of dextransucrase and dextranase

Batch syntheses of isomaltooligosaccharides (IMO) from sucrose, using the enzymes dextransucrase and dextranase were performed with the aim of understanding the reaction mechanism and the parameters which affect product characteristics and molecular size. Both activities described for dextransucrase (dextran formation and acceptor reaction) achieved synthesis whilst the hydrolytic activity of dextranase regulated the product molecular size and acceptor availability. Depending on the reaction conditions, the product oligosaccharide mixtures contained mainly sugars (up to 36%) with degrees of polymerization (DP) varying between 10 and 60 together with lower concentrations of both lower and higher molecular weight sugars. Alterations in substrate and dextranase concentrations (50-400 mg ml(-1) and 2.5-46 U ml(-1), respectively) affected the molecular weight of IMO, the reaction rate and the formation of leucrose. This permitted manipulation of the product characteristics. It was found that higher substrate and dextranase concentrations gave rise to products with lower molecular sizes and a dextransucrase:dextranase ratio of 1: 1 or 1:2 appeared to produce a polymer with a molecular weight which is desirable for prebiotic use. (C) 2004 Elsevier Inc. All rights reserved.