Effect of culture medium and cryoprotectants on the growth and survival of probiotic lactobacilli during freeze drying

To investigate the effects of the medium and cryoprotective agents used on the growth and survival of Lactobacillus plantarum and Lactobacillus rhamnosus GG during freeze drying. A complex medium was developed consisting primarily of glucose, yeast extract and vegetable-derived peptone. Trehalose, sucrose and sorbitol were examined for their ability to protect the cells during freeze drying. Using standardized amount of cells and the optimized freeze drying media, the effect of the growth medium on cell survival during freeze drying was investigated. The results showed that glucose and yeast extract were the most important growth factors, while sucrose offered better protection than trehalose and sorbitol during freeze drying. When the cells were grown under carbon limiting conditions, their survival during freeze drying was significantly decreased. A clear relationship was observed between cell growth and the ability of the cells to survive during the freeze drying process. The survival of probiotic strains during freeze drying was shown to be dependent on the cryoprotectant used and the growth medium.

New Culture Medium to Xanthan Production by Xanthomonas campestris pv. campestris

Xanthan is a important biopolymer for commercial purpose and it is produced in two stages by Xanthomonas campestris. In the first one, the bacterium is cultivated in the complex medium enriched in nitrogen and the biomass produced is used as inoculum for the next stage in which the gum is produced in another medium. In this work a new medium for the first stage is proposed in place of currently used YM medium. Different formulated growth media were studied and the correspondent biomass produced was analysed as inoculum for the second stage. The inoculum and gum were produced by batch process in shaker at 27A degrees C in pH 6.0 and at 30A degrees C in pH 7.0, respectively. The gum was precipitated with ethanol (3:1 v/v). The dryed biomass and xathan gum produced were determined by drying in oven at 105 and 40A degrees C, respectively. The viscosity of the fermentation broth and 1% gum solution in water were determined in Brookfield viscometer. The formulated medium presented the increase in gum production (30%), broth (136%) and 1% gum solution viscosity (60%) compared to YM, besides the inferior cost. The results showed the importance of the quality of the inoculum from the first stage of the culture which influenced on the gum viscosity in the second stage.

Presence of Nicotinamide Adenine Dinucleotide–Independent Haemophilus paragallinarum in Mexico

Two isolates of Haemophilus paragallinarum were obtained from a layer chicken in Mexico. The isolates were confirmed as H. paragallinarum by polymerase chain reaction and conventional biochemical identification. The isolates were nicotinamide adenine dinucleotide (NAD) independent—growing on blood agar without the need of a nurse colony as well as on a complex medium that lacked both NAD and chicken serum. Both isolates were pathogenic, causing the typical clinical signs of infectious coryza in susceptible chickens. One isolate was Page serovar B/Kume serovar B-1 and the other isolate was Page serovar C/Kume serovar C-2. The isolates were associated with a field outbreak that involved an egg drop of 20% over a 3 wk period and a doubling of weekly mortality (from 0.1% to 0.2%). This is the first report of NAD-independent H. paragallinarum outside South Africa and is the first time that NADindependent H. paragallinarum of serovar B has been reported. Abbreviations: NAD ¼ nicotinamide adenine dinucleotide; NAM ¼ nicotinamide; PCR ¼ polymerase chain reaction; TM ¼ complete growth medium without chicken serum or nicotinamide adenine dinucleotide; TM/SN ¼ complete growth medium that contains both chicken serum and nicotinamide adenine dinucleotide

The white-rot fungi Phlebia radiata and Dichomitus squalens in wood-based cultures: expression of laccases, lignin peroxidases, and oxalate decarboxylase

Basidiomycetous white-rot fungi are the only organisms that can efficiently decompose all the components of wood. Moreover, white-rot fungi possess the ability to mineralize recalcitrant lignin polymer with their extracellular, oxidative lignin-modifying enzymes (LMEs), i.e. laccase, lignin peroxidase (LiP), manganese peroxidase (MnP), and versatile peroxidase (VP). Within one white-rot fungal species LMEs are typically present as several isozymes encoded by multiple genes. This study focused on two effi cient lignin-degrading white-rot fungal species, Phlebia radiata and Dichomitus squalens. Molecular level knowledge of the LMEs of the Finnish isolate P. radiata FBCC43 (79, ATCC 64658) was complemented with cloning and characterization of a new laccase (Pr-lac2), two new LiP-encoding genes (Pr-lip1, Pr-lip4), and Pr-lip3 gene that has been previously described only at cDNAlevel. Also, two laccase-encoding genes (Ds-lac3, Ds-lac4) of D. squalens were cloned and characterized for the first time. Phylogenetic analysis revealed close evolutionary relationships between the P. radiata LiP isozymes. Distinct protein phylogeny for both P. radiata and D. squalens laccases suggested different physiological functions for the corresponding enzymes. Supplementation of P. radiata liquid culture medium with excess Cu2+ notably increased laccase activity and good fungal growth was achieved in complex medium rich with organic nitrogen. Wood is the natural substrate of lignin-degrading white-rot fungi, supporting production of enzymes and metabolites needed for fungal growth and the breakdown of lignocellulose. In this work, emphasis was on solid-state wood or wood-containing cultures that mimic the natural growth conditions of white-rot fungi. Transcript analyses showed that wood promoted expression of all the presently known LME-encoding genes of P. radiata and laccase-encoding genes of D. squalens. Expression of the studied individual LME-encoding genes of P. radiata and D. squalens was unequal in transcript quantities and apparently time-dependent, thus suggesting the importance of several distinct LMEs within one fungal species. In addition to LMEs, white-rot fungi secrete other compounds that are important in decomposition of wood and lignin. One of these compounds is oxalic acid, which is a common metabolite of wood-rotting fungi. Fungi produce also oxalic-acid degrading enzymes of which the most widespread is oxalate decarboxylase (ODC). However, the role of ODC in fungi is still ambiguous with propositions from regulation of intra and extracellular oxalic acid levels to a function in primary growth and concomitant production of ATP. In this study, intracellular ODC activity was detected in four white-rot fungal species, and D. squalens showed the highest ODC activity upon exposure to oxalic acid. Oxalic acid was the most common organic acid secreted by the ODC-positive white-rot fungi and the only organic acid detected in wood cultures. The ODC-encoding gene Ds-odc was cloned from two strains of D. squalens showing the first characterization of an odc-gene from a white-rot polypore species. Biochemical properties of the D. squalens ODC resembled those described for other basidiomycete ODCs. However, the translated amino acid sequence of Ds-odc has a novel N-terminal primary structure with a repetitive Ala-Ser-rich region of ca 60 amino acid residues in length. Expression of the Ds-odc transcripts suggested a constitutive metabolic role for the corresponding ODC enzyme. According to the results, it is proposed that ODC may have an essential implication for the growth and basic metabolism of wood-decaying fungi.

One-Carbon Metabolic Pathway Rewiring in Escherichia coli Reveals an Evolutionary Advantage of 10-Formyltetrahydrofolate Synthetase (Fhs) in Survival under Hypoxia

In cells, N-10-formyltetrahydrofolate (N-10-fTHF) is required for formylation of eubacterial/organellar initiator tRNA and purine nucleotide biosynthesis. Biosynthesis of N-10-fTHF is catalyzed by 5,10-methylene-tetrahydrofolate dehydrogenase/cyclohydrolase (FolD) and/or 10-formyltetrahydrofolate synthetase (Fhs). All eubacteria possess FolD, but some possess both FolD and Fhs. However, the reasons for possessing Fhs in addition to FolD have remained unclear. We used Escherichia coli, which naturally lacks fhs, as our model. We show that in E. coli, the essential function of folD could be replaced by Clostridium perfringens fhs when it was provided on a medium-copy-number plasmid or integrated as a single-copy gene in the chromosome. The fhs-supported folD deletion (Delta folD) strains grow well in a complex medium. However, these strains require purines and glycine as supplements for growth in M9 minimal medium. The in vivo levels of N-10-fTHF in the Delta folD strain (supported by plasmid-borne fhs) were limiting despite the high capacity of the available Fhs to synthesize N-10-fTHF in vitro. Auxotrophy for purines could be alleviated by supplementing formate to the medium, and that for glycine was alleviated by engineering THF import into the cells. The Delta folD strain (harboring fhs on the chromosome) showed a high NADP(+)-to-NADPH ratio and hypersensitivity to trimethoprim. The presence of fhs in E. coli was disadvantageous for its aerobic growth. However, under hypoxia, E. coli strains harboring fhs outcompeted those lacking it. The computational analysis revealed a predominant natural occurrence of fhs in anaerobic and facultative anaerobic bacteria.

复杂介质边界元法数值模拟研究

A major impetus to study the rough surface and complex structure in near surface model is because accuracy of seismic observation and geophysical prospecting can be improved. Wave theory study about fluid-satuated porous media has important significance for some scientific problems, such as explore underground resources, study of earth's internal structure, and structure response of multi-phase porous soil under dynamic and seismic effect. Seismic wave numerical modeling is one of the effective methods which understand seismic propagation rules in complex media. As a numerical simulation method, boundary element methods had been widely used in seismic wave field study. This paper mainly studies randomly rough surface scattering which used some approximation solutions based on boundary element method. In addition, I developed a boundary element solution for fluid saturated porous media. In this paper, we used boundary element methods which based on integral expression of wave equation to study the free rough surface scattering effects of Kirchhoff approximation method, Perturbation approximation method, Rytov approximation method and Born series approximation method. Gaussian spectrum model of randomly rough surfaces was chosen as the benchmark model. The approximation methods result were compared with exact results which obtained by boundary element methods, we study that the above approximation methods were applicable how rough surfaces and it is founded that this depends on and ( here is the wavenumber of the incident field, is the RMS height and is the surface correlation length ). In general, Kirchhoff approximation which ignores multiple scatterings between any two surface points has been considered valid for the large-scale roughness components. Perturbation theory based on Taylor series expansion is valid for the small-scale roughness components, as and are .Tests with the Gaussian topographies show that the Rytov approximation methods improves the Kirchhoff approximation in both amplitude and phase but at the cost of an extra treatment of transformation for the wave fields. The realistic methods for the multiscale surfaces come with the Born series approximation and the second-order Born series approximation might be sufficient to guarantee the accuracy of randomly rough surfaces. It could be an appropriate choice that a complex rough surface can be divided into large-, medium-, and small-scale roughness components with their scattering features be studied by the Kirchhoff or Rytov phase approximations, the Born series approximation, and the perturbation theory, respectively. For this purpose, it is important to select appropriate parameters that separate these different scale roughness components to guarantee the divided surfaces satisfy the physical assumptions of the used approximations, respectively. In addition, in this paper, the boundary element methods are used for solving the porous elastic wave propagation and carry out the numerical simulation. Based on the fluid-saturated porous model, this paper analyses and presents the dynamic equation of elastic wave propagation and boundary integral equation formulation of fluid saturated porous media in frequency domain. The fundamental solutions of the elastic wave equations are obtained according to the similarity between thermoelasticity and poroelasticity. At last, the numerical simulation of the elastic wave propagation in the two-phase isotropic media is carried out by using the boundary element method. The results show that a slow quasi P-wave can be seen in both solid and fluid wave-field synthetic seismograms. The boundary element method is effective and feasible.

波动方程辛几何算法三维叠前深度偏移及其并行实现

Oil and scientific groups have been focusing on the 3D wave equation prestack depth migration since it can solve the complex problems of the geologic structure accurately and maintain the wave information, which is propitious to lithology imaging. The symplectic method was brought up by Feng Kang firstly in 1984 and became the hotspot of numerical computation study. It will be widely applied in many scientific field of necessity because of its great virtue in scientific sense. This paper combines the Symplectic method and the 3-D wave equation prestack depth migration to bring up an effectual numerical computation method of wave field extrapolatation technique under the scientific background mentioned above. At the base of deep analysis of computation method and the performance of PC cluster, a seismic prestack depth migration flow considering the virtue of both seismic migration method and Pc cluster has formatted. The software, named 3D Wave Equation Prestack Depth Migration of Symplectic Method, which is based on the flow, has been enrolled in the National Bureau of Copyright (No. 0013767). Dagang and Daqing Oil Field have now put it into use in the field data processing. In this paper, the one way wave equation operator is decompounded into a phase shift operator and a time shift operator and the correct item with high rank Symplectic method when approaching E exponent. After reviewing eliminating alias frequency of operator, computing the maximum angle of migration and the imaging condition, we present the test result of impulse response of the Symplectic method. Taking the imaging results of the SEG/EAGE salt and overthrust models for example and seeing about the imaging ability with complex geologic structure of our software system, the paper has discussed the effect of the selection of imaging parameters and the effectuation on the migration result of the seismic wavelet and compared the 2-D and 3-D prestack depth migration result of the salt mode. We also present the test result of impulse response with the overthrust model. The imaging result of the two international models indicates that the Symplectic method of 3-D prestack depth migration accommodates great transversal velocity variation and complex geologic structure. The huge computing cost is the key obstruction that 3-D prestack depth migration wave equation cannot be adopted by oil industry. After deep analysis of prestack depth migration flow and the character of PC cluster ,the paper put forward :i)parallel algorithms in shot and frequency domain of the common shot gather 3-D wave equation prestack migration; ii)the optimized setting scheme of breakpoint in field data processing; iii)dynamic and static load balance among the nodes of the PC cluster in the 3-D prestack depth migration. It has been proven that computation periods of the 3-D prestack depth migration imaging are greatly shortened given that adopting the computing method mentioned in the paper. In addition,considering the 3-D wave equation prestack depth migration flow in complex medium and examples of the field data processing, the paper put the emphasis on: i)seismic data relative preprocessing, ii) 2.5D prestack depth migration velocity analysis, iii)3D prestack depth migration. The result of field data processing shows satisfied application ability of the flow put forward in the paper.

双聚焦偏移速度估计与可分表示叠前弹性反演

In exploration seismology, the geologic target of oil and gas reservoir in complex medium request the high accuracy image of the structure and lithology of the medium. So the study of the prestack image and the elastic inversion of seismic wave in the complex medium come to the leading edge. The seismic response measured at the surface carries two fundamental pieces of information: the propagation effects of the medium and the reflections from the different layer boundaries in the medium. The propagation represent the low-wavenumber component of the medium, it is so-called the trend or macro layering, whereas the reflections represent the high-wavenumber component of the medium, it is called the detailed or fine layering. The result of migration velocity analysis is the resolution of the low-wavenumber component of the medium, but the prestack elastic inversion provided the resolution of the high-wavvenumber component the medium. In the dissertation, the two aspects about the migration velocity estimation and the elastic inversion have been studied.Firstly, any migration velocity analysis methods must include two basic elements: the criterion that tell us how to know whether the model parameters are correct and the updating that tell us how to update the model parameters when they are incorrect, which are effected on the properties and efficiency of the velocity estimation method. In the dissertation, a migration velocity analysis method based on the CFP technology has been presented in which the strategy of the top-down layer stripping approach are adapted to avoid the difficult of the selecting reduce .The proposed method has a advantage that the travel time errors obtained from the DTS panel are defined directly in time which is the difference with the method based on common image gather in which the residual curvature measured in depth should be converted to travel time errors.In the proposed migration velocity analysis method, the four aspects have been improved as follow:? The new parameterization of velocity model is provided in which the boundaries of layers are interpolated with the cubic spline of the control location and the velocity with a layer may change along with lateral position but the value is calculated as a segmented linear function of the velocity of the lateral control points. The proposed parameterization is suitable to updating procedure.? The analytical formulas to represent the travel time errors and the model parameters updates in the t-p domain are derived under local lateral homogeneous. The velocity estimations are iteratively computed as parametric inversion. The zero differential time shift in the DTS panel for each layer show the convergence of the velocity estimation.? The method of building initial model using the priori information is provided to improve the efficiency of velocity analysis. In the proposed method, Picking interesting events in the stacked section to define the boundaries of the layers and the results of conventional velocity analysis are used to define the velocity value of the layers? An interactive integrate software environment with the migration velocity analysis and prestack migration is built.The proposed method is firstly used to the synthetic data. The results of velocity estimation show both properties and efficiency of the velocity estimation are very good.The proposed method is also used to the field data which is the marine data set. In this example, the prestack and poststack depth migration of the data are completed using the different velocity models built with different method. The comparison between them shows that the model from the proposed method is better and improves obviously the quality of migration.In terms of the theoretical method of expressing a multi-variable function by products of single-variable functions which is suggested by Song Jian (2001), the separable expression of one-way wave operator has been studied. A optimization approximation with separable expression of the one-way wave operator is presented which easily deal with the lateral change of velocity in space and wave number domain respectively and has good approach accuracy. A new prestack depth migration algorithm based on the optimization approximation separable expression is developed and used to testing the results of velocity estimation.Secondly, according to the theory of the seismic wave reflection and transmission, the change of the amplitude via the incident angle is related to the elasticity of medium in the subsurface two-side. In the conventional inversion with poststack datum, only the information of the reflection operator at the zero incident angles can be used. If the more robust resolutions are requested, the amplitudes of all incident angles should be used.A natural separable expression of the reflection/transmission operator is represented, which is the sum of the products of two group functions. One group function vary with phase space whereas other group function is related to elastic parameters of the medium and geological structure.By employing the natural separable expression of the reflection/transmission operator, the method of seismic wave modeling with the one-way wave equation is developed to model the primary reflected waves, it is adapt to a certain extent heterogeneous media and confirms the accuracy of AVA of the reflections when the incident angle is less than 45'. The computational efficiency of the scheme is greatly high.The natural separable expression of the reflection/transmission operator is also used to construct prestack elastic inversion algorithm. Being different from the AVO analysis and inversion in which the angle gathers formed during the prstack migration are used, the proposed algorithm construct a linear equations during the prestack migration by the separable expression of the reflection/transmission operator. The unknowns of the linear equations are related to the elasticity of the medium, so the resolutions of them provided the elastic information of the medium.The proposed method of inversion is the same as AVO inversion in , the difference between them is only the method processing the amplitude via the incident angle and computational domain.

Stress tolerance and virulence of insect-pathogenic fungi are determined by environmental conditions during conidial formation

The virulence to insects and tolerance to heat and UV-B radiation of conidia of entomopathogenic fungi are greatly influenced by physical, chemical, and nutritional conditions during mycelial growth. This is evidenced, for example, by the stress phenotypes of Metarhizium robertsii produced on various substrates. Conidia from minimal medium (Czapek's medium without sucrose), complex medium, and insect (Lepidoptera and Coleoptera) cadavers had high, moderate, and poor tolerance to UV-B radiation, respectively. Furthermore, conidia from minimal medium germinated faster and had increased heat tolerance and were more virulent to insects than those from complex medium. Low water-activity or alkaline culture conditions also resulted in production of conidia with high tolerance to heat or UV-B radiation. Conidia produced on complex media exhibited lower stress tolerance, whereas those from complex media supplemented with NaCl or KCl (to reduce water activity) were more tolerant to heat and UV-B than those from the unmodified complex medium. Osmotic and nutritive stresses resulted in production of conidia with a robust stress phenotype, but also were associated with low conidial yield. Physical conditions such as growth under illumination, hypoxic conditions, and heat shock before conidial production also induced both higher UV-B and heat tolerance; but conidial production was not decreased. In conclusion, physical and chemical parameters, as well as nutrition source, can induce great variability in conidial tolerance to stress for entomopathogenic fungi. Implications are discussed in relation to the ecology of entomopathogenic fungi in the field, and to their use for biological control. This review will cover recent technologies on improving stress tolerance of entomopathogenic fungi for biological control of insects.

Effect of growth time on the surface and adhesion properties of Lactobacillus rhamnosus GG

Aims: To investigate the changes in the surface properties of Lactobacillus rhamnosus GG during growth, and relate them with the ability of the Lactobacillus cells to adhere to Caco-2 cells. Methods and Results: Lactobacillus rhamnosus GG was grown in complex medium, and cell samples taken at four time points and freeze dried. Untreated and trypsin treated freeze dried samples were analysed for their composition using SDS-PAGE analysis and Fourier transform infrared spectroscopy (FTIR), hydrophobicity and zeta potential, and for their ability to adhere to Caco-2 cells. The results suggested that in the case of early exponential phase samples (4 and 8 h), the net surface properties, i.e. hydrophobicity and charge, were determined to a large extent by anionic hydrophilic components, whereas in the case of stationary phase samples (13 and 26 h), hydrophobic proteins seemed to play the biggest role. Considerable differences were also observed between the ability of the different samples to adhere to Caco-2 cells; maximum adhesion was observed for the early stationary phase sample (13 h). The results suggested that the adhesion to Caco-2 cells was influenced by both proteins and non-proteinaceous compounds present on the surface of the Lactobacillus cells. Conclusion: The surface properties of Lact. rhamnosus GG changed during growth, which in return affected the ability of the Lactobacillus cells to adhere to Caco-2 cells. Significance and Impact of the Study: The levels of adhesion of Lactobacillus cells to Caco-2 cells were influenced by the growth time and reflected changes on the bacterial surface. This study provides critical information on the physicochemical factors that influence bacterial adhesion to intestinal cells.

Evaluation of different media for the production of cephalosporins by Streptomyces clavuligerus ATCC 27064

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Adição de precursores em meio complexo solúvel para a produção de Cefamicina C por Streptomyces clavuligerus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Modelagem sísmica por diferenças finitas em meios bidimensionais com difratores

Este trabalho tem por objetivo a modelagem sísmica em meios com fortes descontinuidades de propriedades físicas, com ênfase na existência de difrações. Como parte deste estudo foi feita a análise numérica visando as condições de estabilidade e de fronteiras utilizadas no cálculo do campo de ondas sísmicas. Para a validação do programa de diferenças finitas foi feita a comparação cinemática com a Teoria do Raio para um modelo simples. O motivo deste estudo é ter uma melhor compreensão e controle sobre os problemas de modelagem, visando contribuir para a solução de problemas na interpretação de dados sísmicos. Segundo vários autores na literatura geológica, Derby (1877), Evans (1906), Paiva (1929) e Moura (1938). A Bacia do Amazonas é constituída por rochas sedimentares depositadas desde o Ordoviciano até o recente, atingindo espessuras da ordem de 5 km. Os corpos de diabásio, presentes entre os sedimentos paleozóicos, estão dispostos na forma de soleiras, alcançando espessuras de centenas de metros, perfazendo um volume total de 90.000 quilômetros cúbicos. A ocorrência de tais estruturas é responsável pela existência de reflexões múltiplas durante a propagação da onda sísmica, impossibilitando uma melhor interpretação dos horizontes refletores que se encontram abaixo das soleiras. Para representar situações geológicas desse tipo foi usado um modelo acústico de velocidades. Para o cálculo dos sismogramas foi utilizado um programa de diferenças finitas com aproximação de quarta ordem da equação da onda acústica no espaço e no tempo. As aplicações dos métodos de diferenças finitas para o estudo de propagação de ondas sísmicas têm melhorado a compreensão sobre a propagação em meios onde existem heterogeneidades significativas, tendo como resultado boa resolução na interpretação dos eventos de reflexão sísmica em áreas de interesse. Como resultado dos experimentos numéricos realizados em meio de geologia complexa, foi observado a influência significativa das múltiplas devido a camada de alta velocidade, o que faz com que haja maior perda de energia dificultando a interpretação dos alvos. Por esta razão recomendo a integração de dados de superfície com os dados de poço, com o objetivo de se obter uma melhor imagem dos alvos abaixo das soleiras de diabásio.

Production and characterization of poly-(3-hydroxybutyrate) from recombinant Escherichia coli grown on cheap renewable carbon substrates

Although the biopolymer poly-(3-hydroxybutyrate), P[3HB], presents physicochemical properties that make it an alternative material to conventional plastics, its biotechnological production is quite expensive. As carbon substrates contribute greatly to P[3HB] production cost, the utilization of a cheaper carbon substrate and less demanding micro-organisms should decrease its cost. In the present study a 23 factorial experimental design was applied, aiming to evaluate the effects of using hydrolysed corn starch (HCS) and soybean oil (SBO) as carbon substrates, and cheese whey (CW) supplementation in the mineral medium (MM) on the responses, cell dried weigh (DCW), percentage P[3HB] and mass P[3HB] by recombinant Escherichia coli strains JM101 and DH10B, containing the P[3HB] synthase genes from Cupriavidus necator (ex-Ralstonia eutropha). The analysis of effects indicated that the substrates and the supplement and their interactions had positive effect on CDW. Statistically generated equations showed that, at the highest concentrations of HCS, SO and CW, theoretically it should be possible to produce about 2 g L(1) DCW, accumulating 50% P[3HB], in both strains. To complement this study, the strain that presented the best results was cultivated in MM added to HCS, SBO and CW ( in best composition observed) and complex medium (CM) to compare the obtained P[3HB] in terms of physicochemical parameters. The obtained results showed that the P[3HB] production in MM (1.29 g L(-1)) was approximately 20% lower than in CM (1.63 g L(-1)); however, this difference can be compensated by the lower cost of the MM achieved by the use of cheap renewable carbon sources. Moreover, using differential scanning calorimetry and thermogravimetry analyses, it was observed that the polymer produced in MM was the one which presented physicochemical properties (Tg and Tf) that were more similar to those found in the literature for P[3HB].

Adattamento e acclimatazione a diverse temperature di lieviti psicrofili obbligati e facoltativi e di lieviti mesofili. Studio della produzione di acidi grassi polinsaturi omega-3 e omega-6 per via fermentativa.

Adaptation and acclimation to different temperatures of obligate psychrophilic, facultative psychrophilic and mesophilic yeasts. Production of ω-3 and ω-6 polyunsaturated fatty acids by fermentative way. Obligate psychrophilic, facultative psychrophilic and mesophilic yeasts were cultured in a carbon rich medium at different temperatures to investigate if growth parameters, lipid accumulation and fatty acid composition were adaptive and/or acclimatory responses. Acclimation of facultative psychrophiles and mesophiles to lower temperature negatively affected their specific growth rate. Obligate psychrophiles exhibited the highest biomass yield (YX/S), followed by facultative psychrophiles, then by mesophiles. The growth temperature did not influence the YX/S of facultative psychrophiles and mesophiles. Acclimation to lower temperature caused the increase in lipid yield (YL/X) in mesophilic yeasts, but did not affect YL/X in facultative psychrophiles. Similar YL/X were found in both facultative and obligated psychrophiles, suggesting that lipid accumulation is not a distinctive character of adaptation to permanently cold environments. The extent of unsaturation of fatty acids was one major adaptive feature of the yeasts which colonize permanently cold ecosystems. Remarkable amounts of α-linolenic acid were found in obligate psychrophiles at the expenses of linoleic acid, whereas it was generally scarce or absent in all the others strains. Increased unsaturation of fatty acids was also an acclimatory response of mesophiles and facultative psychrophiles to lower temperature. It’s well known that omega-3 polyunsaturated fatty acids (PUFAs) display a variety of beneficial effects on various organ systems and diseases, therefore a process for the microbial production of omega-3 PUFAs would be of great interest. This work sought also to investigate if one of the better psychrophilic yeast, Rhodotorula glacialis DBVPG 4785, stimulated by acclamatory responses, produced omega-3 PUFAs. In fact, the adaptation of psychrophilic yeasts to cold niches is related to the production of higher amounts of lipids and to increased unsaturation degree of fatty acids, presumably to maintain membrane fluidity and functionality at low temperatures. Bioreactor fermentations of Rhodotorula glacialis DBVPG 4785 were carried out at 25, 20, 15, 10, 5, 0, and -3°C in a complex medium with high C:N ratio for 15 days. High biomass production was attained at all the temperatures with a similar biomass/glucose yield (YXS), between 0.40 and 0.45, but the specific growth rate of the strain decreased as the temperature diminished. The coefficients YL/X have been measured between a minimum of 0.50 to a maximum of 0.67, but it was not possible to show a clear effect of temperature. Similarly, the coefficient YL/S ranges from a minimum of 0.22 to a maximum of 0.28: again, it does not appear to be any significant changes due to temperature. Among omega-3 PUFAs, only α-linolenic acid (ALA, 18:3n-3) was found at temperatures below to 0°C, while, it’s remarkable, that the worthy arachidonic acid (C20:4,n-6), stearidonic acid (C20:4,n-3) C22:0 and docosahexaenoic acid (C22:6n-3) were produced only at the late exponential phase and the stationary phase of batch fermentations at 0 and -3°C. The docosahexaenoic acid (DHA) is a beneficial omega-3 PUFA that is usually found in fatty fish and fish oils. The results herein reported improve the knowledge about the responses which enable psychrophilic yeasts to cope with cold and may support exploitation of these strains as a new resource for biotechnological applications.