Simultaneous enrichment of cereals with polyunsaturated fatty acids and pigments by fungal solid state fermentations

Four Mucor strains were tested for their ability to grow on four cereal substrates and enriched them with gamma-linolenic acid (GLA) and β-carotene. M. circinelloides CCF-2617 as the best producer accumulated of both GLA and β-carotene in high amounts during utilization of rye bran/spent malt grains (3:1). The first growth phase was characterized by rapid GLA biosynthesis, while distinct β-carotene formation was found in the stationary fungal growth. Therefore various cultivation conditions were tested in order to optimize the yield of either GLA or β-carotene. The fungus grown on cereal substrate supplemented with glucose produced maximal 8.5 mg β-carotene and 12.1 g GLA in 1 kg fermented substrate, respectively. On the other hand, the highest amount of GLA in the fermented substrate (24.2 g/kg) was achieved when 30% of sunflower oil was employed to the substrate. Interestingly, β-carotene biosynthesis was completely inhibited when either whey or linseed oil were added to the substrate.     

Lipase production and Penicillium simplicissimum morphology in solid-state and submerged fermentations

A comparative study of Penicillium simplicissimum morphology and lipase production was performed using solid-state (SSF) and submerged (SmF) fermentation. SSF was carried out on babassu cake as culture medium and SmF on a semi-synthetic medium and a medium based on suspended babassu cake grains. Yield of product on biomass, specific activity and conidia production were 3.3-, 1.3- and 2-fold higher in SSF. In SmF, the type of fungus growth differed according to the medium. Using the semi-synthetic medium, the fungus formed densely interwoven mycelial masses without conidia production, whereas using the babassu-based medium the fungus formed free mycelia and adhered to the surfaces of the grains, producing conidia. The results show that babassu cake induces conidiation in SmF. In SSF, the fungus not only grew on the surface of the grains, producing conidia abundantly, but also effectively colonized and penetrated the babassu particles. The high conidia production and lipase productivity in SSF may be related to the low availability of nutrients or to other stimuli associated with this type of fermentation. Thus, the high production of the thermostable P. simplicissimum lipase, using a non-supplemented, low-cost agro-industrial residue as the culture medium, demonstrates the biotechnological potential of SSF for the production of industrial enzymes.     

昆虫夏滞育的调控及其遗传基础

几十年来对昆虫夏滞育的研究不断深入 ,阐明了不少昆虫夏滞育的生态机理。目前 ,探讨其内在的机制已成为研究的方向。该文着重论述夏滞育的生理生化特征、夏滞育的调控及其遗传基础     

Assessment of physiological conditions in E. coli fermentations by epifluorescent microscopy and image analysis

The development of monitoring methods for assessing the physiological state of microorganisms during recombinant fermentation processes has been encouraged by the need to evaluate the influence of processing conditions in recombinant protein production. In this work, a technique based on microscopy and image analysis was developed that allows the simultaneous quantification of parameters associated with viability and fluorescent protein production in recombinant Escherichia coli fermentations. Images obtained from light microscopy with phase contrast are used to assess the total number of cells in a given sample and, from epifluorescence microscopy, both protein producing and injured cells are evaluated using two different fluorochromes: propidium iodide and enhanced yellow fluorescent protein. This technique revealed the existence of different cell populations in the recombinant E. coli fermentation broth that were evaluated along four batch fermentations, complementing information obtained with standard techniques to study the effects of the temperature and induction time in recombinant protein production processes. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

On-line rheological measurements and control in fungal fermentations

A system for on-line rheological measurements and control in filamentous fermentations is presented. The output signals from the control unit can be used in terms of process control. Diluting the broth in a growth controlled feed pattern was found to influence the viscosity of the broth and lead to process improvements. Just diluting the fermentation broth to keep the viscosity below a preset value was seen to give only temporary process improvements. The higher the viscosity, the less effective the viscosity controlled dilutions. The failure to get full control over the viscosity by the dilution techniques used is caused by the large number of factors influencing the rheological properties of an Aspergillus niger culture. The factors shown in this work to influence the rheological properties of the fermentation broth were the biomass concentration, the specific growth rate, mixing qualities (impeller speed and working volume), and the dissolved oxygen concentration. (c) 1992 John Wiley & Sons, Inc.     

Short Communication: Characterization and succession of yeast populations associated with spontaneous fermentations during the production of Brazilian sugar-cane aguardente

A succession of yeasts was observed during fermentation of aguardente with Saccharomyces cerevisiae being the predominant species. Candida sake, Kluyveromyces marxianus var. drosophilarum and apiculate yeasts were also frequent. Transient yeast species were found in variable numbers, probably due to the daily addition of sugar-cane juice. Killer yeasts were isolated and may have a role in the exclusion of some transient and contaminant species.     

Genomic characterization and selection of wine yeast to conduct industrial fermentations of a white wine produced in a SW Spain winery

Aims: To analyse the diversity of wild yeast in spontaneous fermentations of a white wine and to select the most suitable autochthonous starter yeasts. The selected yeasts would be used for inoculation of industrial fermentations in several years. Methods and Results: Yeasts were characterized by applying electrophoretic karyotyping. This technique was chosen because it can reveal the large‐scale mutations in the yeast genome induced by gross chromosomal rearrangements. This type of mutation is considered one of the main forces behind the rapid evolution of industrial yeasts. A heterogeneous population of yeast strains was observed in the spontaneous fermentations during two consecutive years. Four of the most abundant strains were isolated and tested for microbiological features of industrial importance. The selected autochthonous strains were used as starter yeasts for the following 7 years. In the majority of these experiences, we obtained homogeneous yeast populations, in which the karyotype of one of the inoculated strains – karyotype V – emerged as clearly dominant. Conclusions: The inoculation of the selected strain with karyotype V and a proper handling of the inoculum scaling‐up process led to the substitution of the spontaneous fermentations by controlled fermentations producing a highly satisfactory final product. Significance and Impact of the Study: We monitored the wine yeast population of an industrial system for a total of 9 years. Our work is one of the first examples made at industrial scale showing how molecular techniques can be successfully applied to improve the efficiency of the winemaking process.     

Repeated acidosis challenges and live yeast supplementation shape rumen microbiota and fermentations and modulate inflammatory status in sheep

This study aimed to investigate the impact of repeated acidosis challenges (ACs) and the effect of live yeast supplementation (Saccharomyces cerevisiae I-1077, SC) on rumen fermentation, microbial ecosystem and inflammatory response. The experimental design involved two groups (SC, n=6; Control, n=6) of rumen fistulated wethers that were successively exposed to three ACs of 5 days each, preceded and followed by resting periods (RPs) of 23 days. AC diets consisted of 60% wheat-based concentrate and 40% hay, whereas RPs diets consisted of 20% concentrate and 80% hay. ACs induced changes in rumen fermentative parameters (pH, lactate and volatile fatty-acid concentrations and proportions) as well as in microbiota composition and diversity. The first challenge drove the fermentation pattern towards propionate. During successive challenges, rumen pH measures worsened in the control group and the fermentation profile was characterised by a higher butyrate proportion and changes in the microbiota. The first AC induced a strong release of rumen histamine and lipopolysaccharide that triggered the increase of acute-phase proteins in the plasma. This inflammatory status was maintained during all AC repetitions. Our study suggests that the response of sheep to an acidosis diet is greatly influenced by the feeding history of individuals. In live yeast-supplemented animals, the first AC was as drastic as in control sheep. However, during subsequent challenges, yeast supplementation contributed to stabilise fermentative parameters, promoted protozoal numbers and decreased lactate producing bacteria. At the systemic level, yeast helped normalising the inflammatory status of the animals.     

Succinate production in dual-phase Escherichia coli fermentations depends on the time of transition from aerobic to anaerobic conditions

We examined succinic acid production in Escherichia coli AFP111 using dual-phase fermentations, which comprise an initial aerobic growth phase followed by an anaerobic production phase. AFP111 has mutations in the pfl, ldhA, and ptsG genes, and we additionally transformed this strain with the pyc gene (AFP111/pTrc99A-pyc) to provide metabolic flexibility at the pyruvate node. Aerobic fermentations with these two strains were completed to catalog physiological states during aerobic growth that might influence succinate generation in the anaerobic phase. Activities of six key enzymes were also determined for these aerobic fermentations. From these results, six transition times based on physiological states were selected for studying dual-phase fermentations. The final succinate yield and productivity depend greatly on the physiological state of the cells at the time of transition. Using the best transition time, fermentations achieved a final succinic acid concentration of 99.2 g/l with an overall yield of 110% and productivity of 1.3 g/l h. Journal of Industrial Microbiology & Biotechnology (2002) 28, 325–332 DOI: 10.1038/sj/jim/7000250 Received 01 October 2001/ Accepted in revised form 12 March 2002     

Influence of yeast population on characteristics of the wine obtained in spontaneous and inoculated fermentations of must from <Emphasis Type="Italic">Vitis vinifera</Emphasis> Lado

This work describes the influence of yeast population on the chemical characteristics of wine obtained by spontaneous and inoculated fermentation of must from Vitis vinifera Lado, a minor white grapevine autochthonous to Galicia (NW Spain). The study was carried out for two consecutive years. The results showed that musts derived from Lado presented a high acidity though the potential alcohol level was acceptable. The genetic diversity of S. cerevisiae strains isolated from spontaneous fermentations was low, probably due to must characteristics, although these did not interfere with the implantation of the commercial strains used. Analyses showed that the wines subsequently produced had high alcoholic levels and very high acidities (pH 3.0) as was expected from must composition. Wines obtained from spontaneous fermentations had a lower alcohol content but higher total acidity than those from inoculated fermentations. Monovarietal wines produced from Lado were poorly evaluated in sensorial tests because of their unbalanced structure and sourness; however, when they were mixed with other autochthonous white varieties with less acidity, the resulting wines were well accepted.     

Cell morphology of extrusion foamed poly(lactic acid) using endothermic chemical foaming agent

Poly(lactic acid) (PLA) was foamed with an endothermic chemical foaming agent (CFA) through an extrusion process. The effects of polymer melt flow index, CFA content, and processing speed on the cellular structures, void fraction, and cell-population density of foamed PLA were investigated. The apparent melt viscosity of PLA was measured to understand the effect of melt index on the cell morphology of foamed PLA samples. The void fraction was strongly dependent on the PLA melt index. It increased with increasing melt index, reaching a maximum value, after which it decreased. Melt index showed no significant effect on the cell-population density of foamed samples within the narrow range studied. A gas containment limit was observed in PLA foamed with CFA. Both the void fraction and cell-population density increased with an initial increase in CFA content, reached a maximum value, and then decreased as CFA content continued to increase. The processing speed also affected the morphology of PLA foams. The void fraction reached a maximum value as the extruder’s screw speed increased to 40 rpm and a further increase in the processing speed tended to reduce the void fraction of foamed samples. By contrast, cell-population density increased one order of magnitude by increasing the screw speed from 20 to 120 rpm. The experimental results indicate that a homogeneous and finer cellular morphology could be successfully achieved in PLA foamed in an extrusion process with a proper combination of polymer melt flow index, CFA content, and processing speed.     

Factors influencing productivity of fermentations employing recombinant microorganisms

Advances in molecular biology and the exploitation of recombinant DNA procedures for the expression of heterologous genes in microbial hosts offer an opportunity to develop fermentation processes more scientifically and less empirically than has been feasible previously. Nevertheless, the highly interdisciplinary nature of the task makes this a complex undertaking. The relationships between microbial physiology and plasmid copy number, plasmid stability, and gene expression are not well documented for host/vector systems and appear to be highly system specific. In an effort to account for these complex biological interactions which can exceed our intuitive capabilities, mathematical models have been designed to aid in process analysis and development activities. Fermentation strategies based on certain physiological assumptions are beginning to be reported for recombinant systems. Representative efforts in these areas indicate the broad scope of considerations involved in developing fermentation processes for recombinant microorganisms and the opportunities for further research in this field.     

The physico-chemical basis of leaf wettability in wheat

Summary Wild type wheat (Triticum aestivum L.) and three mutant lines that have reduced glaucousness on the flag leaf sheath have been examined for variations in glaucousness, contact angles, wax chemistry and wax morphology. On the sheath and culm, organs that are glaucous in the wild type, increasing glaucousness is correlated with increasing contact angles, an increasing proportion of -diketones plus hydroxy--diketones in the was and an increasing proportion of wax tubes. Organs that were non-glaucous in all four lines, namely both surfaces of the vegetative leaves and the adaxial surface of the flag leaf, had high contact angles, a dense covering of wax plates and waxes rich in primary alcohols but devoid of -diketones and hydroxy--diketones. The abaxial surface of the flag leaf was the most complex of the organ surfaces studied. In the wild type the glaucousness of the sheath continued onto this surface for 1–2 cm and this was correlated with the other characters studied as it was on the sheath. In the mutants, however, the tubes were absent. Flat ribbons of varying widths, a new wax structure in wheat, as well as various types of plates were found instead. These structures continued to the flag leaf tip and were also present on the abaxial surface of the wild type flag leaf. Changes in contact angle at the tip could not be correlated with the other measured parameters.     

污水处理厂污泥膨胀和污泥发泡的比较分析

活性污泥法由于操作简单、处理效果好被广泛应用于市政污水和工业废水的处理。污泥膨胀和污泥发泡现象影响二次沉淀池的泥水分离过程和生物反应池的微生物量稳定,严重困扰着污水处理厂的正常运行,被称为污水处理厂的"癌症"。本文从污泥膨胀和污泥发泡的定义及分类出发,全面地比较了表征污泥膨胀和污泥发泡的方法、引起污泥膨胀和污泥发泡的丝状细菌种类及控制污泥膨胀和污泥发泡方法的异同,并探讨了污泥膨胀和污泥发泡问题的未来研究方向和控制策略,期望能够为今后污泥膨胀和污泥发泡问题的研究和调控提供有价值的参考。     

Modeling and measurements of fungal growth and morphology in submerged fermentations

Generalizing results from fungal fermentations is difficult due to their high sensitivity toward slight variation in starting conditions, poor reproducibility, and difference in strains. In this study a mathematical model is presented in which oxygen transfer, agitation intensity, dissolved oxygen tension, pellet size, formation of mycelia, the fraction of mycelia in the total biomass, carbohydrate source consumption, and biomass growth are taken into account. Two parameters were estimated from simulation, whereas all others are based on measurements or were taken from literature. Experimental data are obtained from the fermentations in both 2 L and 100 L fermentors at various conditions. Comparison of the simulation with experiments shows that the model can fairly well describe the time course of fungal growth (such as biomass and carbohydrate source concentrations) and fungal morphology (such as pellet size and the fraction of pellets in the total biomass). The model predicts that a stronger agitation intensity leads to a smaller pellet size and a lower fraction of pellets in the total biomass. At the same agitation intensity, pellet size is hardly affected by the dissolved oxygen tension, whereas the fraction of mycelia decreases slightly with an increase of the dissolved oxygen tension in the bulk. All of these are in line with observations at the corresponding conditions.     

Estimation of hyphal tensile strength in production-scale Aspergillus oryzae fungal fermentations.

Fragmentation of filamentous fungal hyphae depends on two phenomena: hydrodynamic stresses, which lead to hyphal breakage, and hyphal tensile strength, which resists breakage. The goal of this study was to use turbulent hydrodynamic theory to develop a correlation that allows experimental data of morphology and hydrodynamics to be used to estimate relative (pseudo) tensile strength (sigma(pseudo)) of filamentous fungi. Fed-batch fermentations were conducted with a recombinant strain of Aspergillus oryzae in 80 m(3) fermentors, and measurements were made of both morphological (equivalent hyphal length, L) and hydrodynamic variables (specific power input, epsilon; kinematic viscosity, v). We found that v increased over 100-fold during these fermentations and, hence, Kolmogorov microscale (lambda) also changed significantly with time. In the impeller discharge zone, where hyphal fragmentation is thought to actually take place, lambda was calculated to be 700-3500 microm, which is large compared to the size of typical fungal hyphae (100-300 microm). This result implies that eddies in the viscous subrange are responsible for fragmentation. Applying turbulent theory for this subrange, it was possible to calculate sigma(pseudo)from morphological and hydrodynamic measurements. Pseudo tensile strength was not constant but increased to a maximum during the first half and then decreased during the second half of each fermentation, presumably due to differences in physiological state. When a literature correlation for hyphal fragmentation rate (k(frag)) was modified by adding a term to account for viscosity and tensile strength, the result was better qualitative agreement with morphological data. Taken together, these results imply hyphal tensile strength can change significantly over the course of large-scale, fed-batch fungal fermentations and that existing fragmentation and morphology models may be improved if they accounted for variations in hyphal tensile strength with time.     

On the physico-chemical basis of voltage-dependent molecular gating mechanisms in biological membranes

Summary The possible nature and theoretical treatment of electric field-induced molecular processes in a membrane are examined. Special attention is given to fairly fast switching phenomena as reflected by asymmetry currents as well as ionic gating in squid axon and similar systems. The apparent charge displacement associated with the underlying mechanisms is argued to be brought about by conformational transitions of integral macromolecular structures. Under these circumstances, voltage changes can actually control the functional state of membranes by direct interference with conformational equilibria. A basic model is quantitatively discussed and shown to account for certain observed asymmetry currents. Effects due to temperature, pressure, or chemical interactions can be readily described. It is indicated how more complicated voltage-dependent membrane processes may be approached along these lines.     

Extracellular fungal polyol lipids: A new class of potential high value lipids

Extracellular fungal glycolipid biosurfactants have attracted attention because productivities can be high, cheap substrates can be used, the molecules are secreted into the medium and the downstream processing is relatively simple. Three classes of extracellular fungal glycolipid biosurfactants have provided most of the scientific advances in this area, namely sophorolipids, mannosylerythritol lipids and cellobioselipids. Polyol lipids, a fourth class of extracellular fungal glycolipid biosurfactants, comprise two groups of molecules: liamocins produced by the yeast-like fungus Aureobasidium pullulans, and polyol esters of fatty acids, produced by some Rhodotorula yeast species. Both are amphiphilic, surface active molecules with potential for commercial development as surfactants for industrial and household applications. The current knowledge of polyol lipids highlights an emerging group of extracellular fungal glycolipid biosurfactants and provides a perspective of what next steps are needed to harness the benefits and applications of this novel group of molecules.     

城市污水处理厂活性污泥生物泡沫研究进展

城市污水处理厂运行过程中一旦发生活性污泥生物泡沫,就会影响污泥沉降和处理厂运行效能,对出水水质、作业安全和公共健康带来一系列挑战。生物泡沫是自活性污泥法诞生以来长期困扰污水处理厂运行的难题。生物泡沫的形成需要气泡、表面活性物质和疏水物质等3点基本的要素,在其中主要富集了诺卡氏型丝状细菌(Nocardioformfilamentousbacteria)和微丝菌(Candidatus Microthrix parvicella)这两种类型微生物。多种环境和运行因素包括温度、溶解氧、pH、污泥龄、特别是营养物质种类和浓度等均会对这些丝状微生物的生长产生影响。抑制丝状细菌生长的常用控制策略包括选择器、生长动力学控制、投加化学药剂以及噬菌体等方法,通过降低两类丝状细菌在生化池中的浓度以期消除生物泡沫现象。本文总结了生物泡沫的类型、成因、表征生物泡沫程度的指标、影响生物泡沫的环境因素以及常用的调控策略的原理及优缺点等,尽可能全面地介绍活性污泥生物泡沫的研究现状,并探讨未来研究方向和控制策略,期望能够为今后研究活性污泥微生物和污水处理厂运行调控提供有价值的参考。     

An investigation into the relationship between physical damage and fungal infection in Colophospermum mopane

Colophospermum mopane trees in Zambia's North Luangwa National Park and an adjacent Game Management Area, were examined for evidence of physical damage caused by browsers, humans, weather, fire, etc. The same trees were assessed for visual signs of fungal colonization, with the aim of investigating any possible relationship between physical damage and attack by fungi. The results of this study suggest that the severity of physical damage had a relatively small influence on the activity of fungi colonizing and utilizing mopane heartwood. The majority of fungi isolated from infected trees were saprophytes, and the common occurrence of these agents in healthy trees suggests that heartwood degradation is not detrimental to the health of the tree. ‘Internal roots’ produced from the cambium inside the hollow trunk, which apparently utilize the detritus accumulated there, is probably an adaptation through which C. mopane can recycle minerals and nutrients.