Studies on the formation of long-chain dicarboxylic acids from puren-alkanes by a mutant ofCandida tropicalis

Summary Individualn-alkanes, from C₁₁–C₁₆, were metabolized by a mutant ofCandida tropicalis. This strain was selected for its inability to grow in the presence of dodecanedioic acid and dodecane as the sole carbon source. Transformations were studied in fed-batch cultures. Undecane was only poorly transformed, but from dodecane to hexadecane high transformation yields were achieved. Maximum yield of acid-precipitable long-chain dioic acids was obtained with tridecane as substrate. All the products were mixtures of different acids. Besides the ,-alkanedioic acids, the 3-hydroxy derivatives of long-chain ,-alkanedioic acids and dioic acids with a shortened carbon chain were found.     

Hydroxylase regulation in Candida tropicalis grown on alkanes.

Candida tropicalis synthesizes a hydroxylase (3 to 5 nmol of product formed per minute per milligram of protein) and a cytochrome P-450 (0.10 to 0.13 nmol per milligram of protein) during growth on n-tetradecane. A three- to four-fold increase in the level of NADPH cytochrome c reductase is also observed in those cells as compared to the level of cells grown on glycerol. The most efficient inducers of the hydroxylase and of cytochrome P-450 are straight-chain alkanes having at least 10 carbon atoms. Alkenes and higher alcohols are also good inducers. There is little or no growth on ramified hydrocarbons such as pristane and on long-chain aldehydes and fatty acids. The partial inhibition of growth on decane is probably due to the denaturation of the microsomal electron carrier systems by the fatty acid formed by hydroxylation of the decane in the yeast.     

Studies on mixing in horizontal rotating tubular bioreactor

In previous investigations on mixing in a horizontal rotating tubular bioreactor (HRTB) the structured “spiral flow” model was developed which contained four adjustable parameters [1, 2]. In order to incorporate the mixing model in a semifundamental scale-up procedure it was necessary to make a relation between the adjustable model parameters and process parameters of the bioreactor expressed as dimensionless numbers. Mathematical equations which relate adjustable model parameters with dimensionless numbers were developed by non-linear and surface regression methods. These equations were applied to develop the prediction systems for adjustable model parameters. In total, nine systems of equations for the prediction of the adjustable model parameters were established and examined by simulation. Three of them (SC-2, SC-6 and SC-9) were selected as adequate to describe the mixing performance of HRTB in a wide range of process conditions.     

Studies on mixing in horizontal rotating tubular bioreactor

A horizontal rotating tubular bioreactor (HRTB) is a combination of a “thin-layer bioreactor” and a “biodisc” reactor. Its interior is divided by O-ring shaped partition walls. Mixing properties of this new type of the bioreactor were investigated by using a temperature step method. The mixing simulations were done by Runge-Kutta-Fehlberg numerical integration. Adjustable parameters of the “spiral flow” model were optimised by Monte-Carlo method. In this investigation, the structured “spiral flow” model (containing four adjustable parameters) was tested in a wide range of experimental conditions. The results show that the structured “spiral flow” model is capable to describe the mixing in HRTB in the whole range of both bioreactor operational parameters (n and D).     

Studies on mixing in horizontal rotating tubular bioreactor

Mixing studies in the horizontal rotating tubular bioreactor (HRTB) were done to explore the influences of the liquid level (H _M =0.050.08 m) and the distance between the partition walls (D _S =0.020.07 m) on the mixing performance in the bioreactor described by the “spiral flow” model. The optimised adjustable parameters of the model were correlated with the process parameters of the bioreactor expressed as dimensionless numbers: Reynolds rotation number (Re _N ) and Reynolds axial flow number (Re _D ). The polynomial coefficients of the correlations were correlated further with the liquid level in the bioreactor (H _M ) and the distance between the partition walls (D _S ). In that way, three modified prediction systems (SC-2A, SC-6A and SC-9A) were established. The analysis based on different criteria selected the prediction system SC-9A as the most suitable to describe the mixing performance of HRTB.     

Electron microscopic study of Candida tropicalis yeasts grown on a medium containing selenium]

The yeast Candida tropicalis 303 was cultivated on a medium containing selenium, and studied by electron microscopy. The vacuoles of these cells contained electron-dense granules. The correlation between the increase in the number of the electron-dense granules in the cells of C. tropicalis 303 and the biomass suggests the presence of Se0 in the Granules.     

Discrete particle simulations predicting mixing behavior of solid substrate particles in a rotating drum fermenter.

A soft-sphere discrete particle model was used to simulate mixing behavior of solid substrate particles in a slow rotating drum for solid-state fermentation. In this approach, forces acting on and subsequent motion of individual particles can be predicted. The (2D) simulations were qualitatively and quantitatively validated by mixing experiments using video and image analysis techniques. It was found that the simulations successfully predicted the mixing progress as a function of the degree of filling and size of the drum. It is shown that only relatively large, straight baffles perpendicular to the drum wall (67% of the drum radius) increase the mixing performance of the rotating drum. Considering the different aspects of mixing dealt with in this work, it is concluded that the soft sphere discrete particle model can serve as a valuable tool for investigating mixing of solid substrate particles. Finally, it is expected that this model may evolve into a potential tool for design and scale-up of mixed solid-state fermenters.     

Oxygen transfer and mycelial growth in a tubular loop fermentor

A 22 m long. 20 liter tubular loop fermentor (TLF) has been tested for oxygen transfer characteristics and as a reactor for mycelial growth. Model calculations show that the flow pressure drop has an important influence on the axial oxygen profiles. A design model that accounts for this influence is presented. Using the model, K_L a values are calculated from the results of sulfite oxidation experiments. These are correlated with power consumption and aeration rates. The K_L a dependence on aeration rate was found to be less than found with tank reactors. The growth kinetics of three metabolite-producing mycelial organisms in the TLF are presented: a Streptomyces, a Fusarium, and a Acrophialophora. In order to determine the influence of reactor type on the growth and product formation, these cultures have been grown in tanks and shake flasks. The antibiotic, product spectrum of Streptomyces is compared on the basis of inhibition tests and it is shown that the distribution of products is reactor dependent. The Fusarium culture produced a previously unknown metabolite, whose concentration in the loop fermentor was four times higher than in a shake flask. The Acrophialophora culture grew twice as fast in the loop fermentor, but produced essentially none of the specific product. Power Consumptions of up to 8 kW/m³ in the tubular fermentor did not appear to harm the mycelia.     

Citric acid production and morphology of Aspergillus niger as functions of the mixing intensity in a stirred tank and a tubular loop bioreactor

The relationship between Aspergillus niger morphology and citric acid production was investigated in two reactor systems with different configurations, a tubular loop and a stirred tank bioreactor, with operating volumes of 6 and 8 dm³, respectively. Morphology was quantified by image analysis. In each system, morphology, characterized by the parameter P (mean convex perimeter of clumps), and citric acid production, were agitation-dependent and closely linked. Increased agitation caused a reduction of clump sizes and results when both reactors demonstrate that the parameter P should not exceed a threshold value in order to achieve increased productivities. The results obtained from the two reactors were in agreement, both qualitatively and quantitatively. Reducing the fundamentally different mixing conditions of the two bioreactors to the order of the dimensionless mixing parameter relative mixing time (τ_m), results showed that the loop simulated the stirred tank. Also, relationships valid for one system accurately described the results obtained from the other system, demonstrating the validity of the relationship between morphology and productivity for the particular fermentation, regardless of the reactor type. Previous attempts to evaluate the use of loop configurations as scale-up tools and their performance as bioreactors, neglected the morphology of the producer micro-organisms. This study demonstrated the close link between morphology and productivity for citric acid production by A. niger, and identified a morphology parameter that was used successfully to characterize the process performance.     

The influence of fluid mixing upon respiratory patterns for extended growth of a methylotroph in an air-lift fermenter

The influence of methanol dispersion and fluid mixing upon respiratory patterns observed during unlimited fedbatch growth of the methylotrophic bacterium Methylophilus methylotrophus has been investigated. A concentric tube air-lift fermenter was employed for which the mixing and fluid circulation patterns have been well characterized. Respiratory quotients showed a marked dependence upon the position in the vessel at which methanol was injected, the volumetric rate of such methanol addition, the fluid circulation time, and the local mixing behavior; the latter two factors of which are both determined by the air throughput. Such variations are discussed on the basis of simple mixing concepts and observations of fluid dispersion.     

Pressure change and gas mixing induced by oscillations in a closed system

In an attempt to delineate some mechanical behaviors found in branching airways, pressure transmission, gas motion, and mixing were studied during high-frequency oscillation (HFO) in an idealized system consisting of a large straight tube and a rigid sphere linked together by a small straight tube. Depending on the frequency f, and on the unsteadiness dimensionless parameter alpha, pressure amplitude in the large tube is either strongly attenuated or amplified in the sphere. This finding may provide a theoretical basis for the pressure resonance phenomenon observed in the lung by previous investigators. Gas compression in the closed volume causes convective mixing throughout the system. The measured dispersion was found to be proportional to f(VT/A)2, in agreement with a recent report. However, bulk convective mixing was sufficient to explain the dispersion for oscillatory volumes (VT) as small as 80 percent of the small tube volume, as has been previously suggested.     

Physiological response to feeding in little penguins

Specific dynamic action (SDA), the increase in metabolic rate above resting levels that accompanies the processes of digestion and assimilation of food, can form a substantial part of the daily energy budget of free-ranging animals. We measured heart rate (fH) and rate of oxygen consumption (VO2) in 12 little penguins while they digested a meal of sardines in order to determine whether they show specific dynamic action. In contrast to some studies of other penguin species, little penguins showed a substantial SDA, the magnitude of which was proportional to the size of the meal. The energy utilized in SDA was equivalent to 13.4% of the available energy content of the fish. Furthermore, animals such as penguins that forage in a cold environment will probably expend further energy in heating their food to body temperature to facilitate efficient digestion. It is estimated that this additional energy expenditure was equivalent to 1.6%-2.3% of the available energy content of the fish, depending on the time of year and therefore the temperature of the water. Changes in fH during digestion were qualitatively similar to those in VO2, implying that there were no substantial circulatory adjustments during digestion and that the relationship between fH and VO2 in penguins is unaffected by digestive state.     

辣椒对镉胁迫的生理响应

以福湘5号、兴蔬215和川腾1号这3个辣椒品种为材料,用含有0mg/L、100mg/L、200mg/L、300mg/L重金属镉的1/2霍格兰培养液对具10片真叶的辣椒苗进行处理,7d后测定各项生理指标。结果表明,镉对辣椒的生长有很强的毒害作用,随着镉胁迫浓度的升高,3个辣椒品种的生物量、叶片叶绿素含量、电导率、丙二醛含量和活性氧清除系统酶活性都受到了不同程度的影响。在镉胁迫下,辣椒植株的株高、根长、鲜重受到明显抑制,根系发育畸形,叶片黄化。镉胁迫的毒害作用在3个辣椒品种间有差异。其中川腾1号耐镉胁迫能力较强,福湘5号次之,兴蔬215最差。     

Physiological response to a breed evaluation field test in Icelandic horses

This study examined the response in terms of heart rate (HR), respiratory rate (RR), haematocrit (Htc), rectal temperature (RT), and some plasma variables in Icelandic horses of different sexes and ages performing the riding assessment in a breed evaluation field test (BEFT). The study was conducted in Iceland on 266 horses (180 mares and 86 stallions, divided into four age groups; 4, 5, 6 and ⩾7 years old). RT and RR were recorded and blood samples were taken before the warm-up and after the riding assessment. Horse HR, velocity and distance were recorded during the warm-up, the riding assessment and a 5-min recovery period. The distance covered in the BEFT was 2.9±0.4 km (range: 1.8 to 3.8 km, n=248), the duration was 9:37±1:22 min:s (range: 5:07 to 15:32 min:s, n=260) and the average speed was 17.8±1.4 km/h (range: 13.2 to 21.3 km/h, n=248). Average HR was 184±13 b.p.m. (range: 138 to 210 b.p.m., n=102) and peak HR 224±9 b.p.m. (range: 195 to 238 b.p.m., n=102), and 36% of the BEFT was performed at HR ⩾200 b.p.m. Post-exercise plasma lactate concentration (Lac) was 18.0±6.5 mmol/l (range: 2.1 to 34.4 mmol/l, n=266), and there was an increase in total plasma protein, plasma creatine kinase and aspartate amino transferase concentration, as well as RR, RT and Htc. Stallions covered a longer total distance (in the warm-up and BEFT) (P<0.05), at a faster speed during BEFT (P<0.001) than mares and had higher Htc and lower HR and post-exercise Lac values. There were few effects of age, but the 4- and 5-year-old horses had lower Htc than older horses and 4-year-old horses had higher post-exercise RR than older horses, although they were ridden for a shorter distance, shorter duration and at lower peak velocity (P<0.1). The results showed that the riding assessment in the BEFT is a high-intensity exercise. The results also showed that aerobic fitness was higher in stallions and that age had a limited effect on the physiological response. It is suggested that these results should be used as a guide for the development of training programmes and fitness tests in Icelandic horses that would improve both performance and welfare of the horse.     

YATP value in Candida tropicalis grown on n-alkanes, fatty acids, and acetate.

The amount of ATP produced during n-alkane, fatty acid, or acetate metabolism in Candida tropicalis has been established from the P/O ratios measured on isolated mitochondria, yield on substrate and carbon balance. For these three kinds of substrates YATP value has been found to be close to 4, although Ysub on acetate is very different from those found with n-alkanes or fatty acids.     

Mycosis Due to Tropicoporus tropicalis (= Inonotus tropicalis) in a Domestic Dog

Mycopathologia - Mycelial basidiomycetes rarely produce mycoses in animals including humans. We report a case of a 9-year-old female mongrel dog with lesions in the prescapular lymph nodes. The...     

Hydrodynamics and mixing in down flow jet loop bioreactor with a non-Newtonian fluid

Gas holdup and liquid circulation time were measured in a down flow jet loop bioreactor with a non-Newtonian fluid. It was observed that the circulation time decreases with increase in nozzle diameter, draft tube to column diameter ratio and shear thinning of the media. The gas holdup increases with increase in gas and liquid velocities. The optimum draft tube to column diameter ratio was found to be 0.438. Correlations for gas holdup and circulation time involving operational and geometrical variables were presented.     

On the existence of a hysteresis loop in open and closed end pores

We have studied the adsorption of argon at 87 K in slit pores of finite length with a smooth graphitic potential, open at both ends or closed at one end. Simulations were carried out using conventional GCMC (grand canonical Monte Carlo) or kMC (kinetic Monte Carlo) in the canonical ensemble with extremely long Markov chain, of at least 2 × 10⁸ configurations; selected simulations with much longer Markov chains do not show any change in the results. When the pore width is in the micropore range (0.65 nm), type I isotherms are obtained for both pore models and for both simulation methods. However, wider pores (1, 2 and 3 nm in width) all exhibit hysteresis loops in the GCMC simulations, while in the canonical ensemble simulations, the isotherms pass through a sigmoid van der Waals type loop in the transition region. This loop locates the true equilibrium transition. For the pores with one closed end, this transition is close to, or coincides with, the adsorption branch of the GCMC hysteresis loop, but for the open-ended pores, it is more closely associated with the desorption branch. In a separate study of adsorption hysteresis in an infinitely long slit pore, using both simulation techniques, the van der Waals loop follows the adsorption branch of the GCMC isotherm to the transition, then reverts to a long vertical section that falls midway between the two hysteresis branches and finally moves to the desorption transition close to the evaporation pressure. An examination of molecular distributions inside the pores reveals two coexisting phases in the canonical simulations, whereas in the grand canonical simulations, the molecules are uniformly distributed along the length of the pores.     

Isotropic sequence order learning using a novel linear algorithm in a closed loop behavioural system

In this article, we present an isotropic algorithm for sequence order learning. Its central goal is to learn the causal relation between two (or more) inputs in order to react to the earliest incoming signal after successful learning (like in typical classical conditioning situations). We implement this algorithm in a behaving system (a robot) thereby creating a closed loop situation where the learner's actions influence its own sensor inputs to the end of creating an autonomous agent. Autonomous behaviour implies that learning goals are internally defined within the organism's capabilities. Standard learning models for sequence learning (e.g. temporal difference (TD)-learning) need an externally defined reward. This, however, is in conflict with the requirement of an implicitly defined internal goal in autonomous behaviour. Therefore, in this study we present a system in which the external reward is replaced by a reflex loop. This loop explicitly includes the environment. Every reflex loop has the inherent disadvantage, which is that its re-actions occur each time just after a reflex-eliciting sensor event and thus 'too late'. However, a reflex can serve as the internal reference for sequence order learning, which has the task of eliminating this disadvantage by creating earlier anticipatory actions. In our system learning is achieved by modifying synaptic weights of a linear neuron with a correlation based learning rule which involves the derivative of the neuron's output. All input lines are entirely isotropic. The synaptic weight change curve of this rule is strongly related to the temporal Hebb learning rule, which was found in spike timing experiments. We find that after learning the reflex loop is replaced in functional terms with an earlier anticipatory action (and pathway). In addition, we observed that the synaptic weights stabilise as soon as the reflex remains silent.