Effect of Growth Rate and Nutrient Limitation on the Composition and Biomass Yield of Acinetobacter calcoaceticus

Acinetobacter calcoaceticus was grown on ethanol in a chemostat as a model system for single-cell protein production. The substrate yield coefficient (Y(s), grams of biomass/gram of ethanol), protein yield coefficient (Y(p), grams of protein/gram of ethanol), and biomass composition were measured as a function of the specific growth rate. Nucleic acid, protein, Y(p), and Y(s) all increased at higher growth rates. Although protein content increased only 14% (from 53 to 67%), Y(p) almost doubled over the same range of growth rates. The increase in Y(p) was due to the higher protein content of the biomass and to higher values of Y(s). The higher values of Y(s) were attributed to maintenance metabolism, and the value of the maintenance coefficient was found to be 0.11 g of ethanol per g of cell per h. When A. calcoaceticus was cultivated under a phosphorus limitation protein content, Y(p) and Y(s) were lower than in carbon-limited cultures. It was concluded that a single-cell protein fermentation using A. calcoaceticus should be operated at a high growth rate under ethanol-limiting conditions in order to maximize both the protein content of the biomass and the amount of biomass and/or protein made from the substrate.     

Kinetics of batch ethanol fermentation of cheese-whey powder (CWP) solution as function of substrate and yeast concentrations

A lactose utilizing yeast strain, Kluyveromyces marxianus DSMZ-7239 was used for ethanol formation from cheese-whey powder (CWP) solution in batch experiments. Effects of initial substrate (CWP) and yeast concentrations on the rate and extent of ethanol formation were investigated. The initial pH and oxidation-reduction potential (ORP) was kept at 5 and -250 mV, respectively. The rate and extent of ethanol formation increased with increasing CWP concentration up to 156 g l(-1) (75 g l(-1) sugar) and then decreased for larger CWP concentrations due to substrate inhibition at high sugar concentrations. The ethanol yield coefficient was also maximum (0.54 g EtOH/g sugar) and equal to the theoretical yield at CWP concentration of 156 g l(-1). The growth yield coefficient was found to be Y(x/s)=1.2+/-0.1g biomass g sugar(-1). The rate of sugar utilization and ethanol formation also increased linearly with increasing initial biomass concentrations. A kinetic model describing the rate of sugar utilization and substrate inhibition as function of the initial substrate and the biomass concentrations was developed. The kinetic constants were determined using the experimental data. Model predictions of sugar utilization rates were in good agreement with the experimental data. The results indicated that the initial sugar concentration should be below 75 g l(-1) (CWP<156 g l(-1)) and the initial biomass should be above 850 mg l(-1) to obtain high rates and yields of ethanol formation and to avoid substrate inhibition.     

Evaluation of growth yield of Spirulina (Arthrospira) sp. in photoautotrophic, heterotrophic and mixotrophic cultures

In microbial cultures, both cellular growth rate and yield (defined as the degree of substrate conversion into the biomass) are important. Although effect of culture conditions on growth kinetics has been well documented for various microbial strains, there is almost no literature concerning the effect of environmental conditions on growth equilibrium, expressed as biomass yield coefficients from substrate. The present paper discusses the effect of culture conditions: irradiance (physical substrate) and glucose concentration (chemical substrate) on biomass yield coefficients from two chemical substrates: glucose and nitrate-nitrogen in photoautotrophic, heterotrophic and mixotrophic culture of blue-green alga Spirulina (Arthrospira) sp. The efficiency of substrates incorporation into the biomass can be precisely determined only if the elemental composition of the biomass is known. The experimental results showed that culture conditions had a substantial influence on biomass yield coefficients (biomass yield from glucose and nitrate-nitrogen). It was found that, the increase of irradiance favoured increase of biomass yield coefficient from both, glucose and nitrate-nitrogen. However, in the case of yield from nitrogen in mixotrophic culture, the effect was opposite. The effect of glucose concentration was different: the higher the initial glucose concentration, the lower the biomass yield coefficients from chemical substrates.     

A model for the effects of primary substrates on the kinetics of reductive dehalogenation

A kinetic model that describes substrate interactions during reductive dehalogenation reactions is developed. This model describes how the concentrations of primary electron-donor and -acceptor substrates affect the rates of reductive dehalogenation reactions. A basic model, which considers only exogenous electron-donor and -acceptor substrates, illustrates the fundamental interactions that affect reductive dehalogenation reaction kinetics. Because this basic model cannot accurately describe important phenomena, such as reductive dehalogenation that occurs in the absence of exogenous electron donors, it is expanded to include an endogenous electron donor and additional electron acceptor reactions. This general model more accurately reflects the behavior that has been observed for reductive dehalogenation reactions. Under most conditions, primary electron-donor substrates stimulate the reductive dehalogenation rate, while primary electron acceptors reduce the reaction rate. The effects of primary substrates are incorporated into the kinetic parameters for a Monod-like rate expression. The apparent maximum rate of reductive dehalogenation (q _{m, ap} ) and the apparent half-saturation concentration (K _ap ) increase as the electron donor concentration increases. The electron-acceptor concentration does not affect q _{m, ap} , but K _ap is directly proportional to its concentration.Definitions for model parameters RX halogenated aliphatic substrate - E-M ⁿ reduced dehalogenase - E-M ⁿ⁺² oxidized dehalogenase - [E-M ⁿ ] steady-state concentration of the reduced dehalogenase (moles of reduced dehalogenase per unit volume) - [E-M ⁿ⁺²] steady-state concentration of the oxidized dehalogenase (moles of reduced dehalogenase per unit volume) - DH₂ primary exogenous electron-donor substrate - A primary exogenous electron-acceptor substrate - A2 second primary exogenous electron-acceptor substrate - X biomass concentration (biomass per unit volume) - f fraction of biomass that is comprised of the dehalogenase (moles of dehalogenase per unit biomass) - stoichiometric coefficient for the reductive dehalogenation reaction (moles of dehalogenase oxidized per mole of halogenated substrate reduced) - stoichiometric coefficient for oxidation of the primary electron donor (moles of dehalogenase reduced per mole of donor oxidized) - stoichiometric coefficient for oxidation of the endogenous electron donor (moles of dehalogenase reduced per unit biomass oxidized) - stoichiometric coefficient for reduction of the primary electron acceptor (moles of dehalogenase oxidized per mole of acceptor reduced) - stoichiometric coefficient for reduction of the second electron acceptor (moles of dehalogenase oxidized per mole of acceptor reduced) - r _RX rate of the reductive dehalogenation reaction (moles of halogenated substrate reduced per unit volume per unit time) - r _d1 rate of oxidation of the primary exogenous electron donor (moles of donor oxidized per unit volume per unit time) - r _d2 rate of oxidation of the endogenous electron donor (biomass oxidized per unit volume per unit time) - r _a1 rate of reduction of the primary exogenous electron acceptor (moles of acceptor reduced per unit volume per unit time) - r _a2 rate of reduction of the second primary electron acceptor (moles of acceptor reduced per unit volume per unit time) - k _RX mixed second-order rate coefficient for the reductive dehalogenation reaction (volume per mole dehalogenase per unit time) - k _d1 mixed-second-order rate coefficient for oxidation of the primary electron donor (volume per mole dehalogenase per unit time) - k _d2 mixed-second-order rate coefficient for oxidation of the endogenous electron donor (volume per mole dehalogenase per unit time) - b first-order biomass decay coefficient (biomass oxidized per unit biomass per unit time) - k _a1 mixed-second-order rate coefficient for reduction of the primary electron acceptor (volume per mole dehalogenase per unit time) - k _a2 mixed-second-order rate coefficient for reduction of the second primary electron acceptor (volume per mole dehalogenase per unit time) - q _m,ap apparent maximum specific rate of reductive dehalogenation (moles of RX per unit biomass per unit time) - K _ap apparent half-saturation concentration for the halogenated aliphatic substrate (moles of RX per unit volume) - k _ap apparent pseudo-first-order rate coefficient for reductive dehalogenation (volume per unit biomass per unit time)     

Kinetic behaviour of waste tyre rubber as microorganism support in an anaerobic digester treating cane molasses distillery slops

The kinetics of anaerobic digestion of cane molasses distillery slops was investigated using a continuous-flow bioreactor which contained waste tyre rubber as support, to which the microorganisms became immobilized. Hydraulic retention times (HRT) ranging from 1 to 10 days were investigated at an average influent chemical oxygen demand (COD) concentration of 47.7?g/l. The maximum substrate utilization rate, k, and half saturation coefficient, K _L, were determined to be 1.82?kg COD_removed/kg VSS day and 0.33?kg COD/kg VSS day. The yield coefficient, Y, and sludge decay rate coefficient, K _d, were also determined to be 0.06?kg VSS/kg COD_removed and 0.05?day^-1, respectively. Methane production was maximum (6.75?l/l day) at a 2 day HRT corresponding to a biomass loading rate of 2.578?kg COD/kg VSS day. Biogas yield ranged between 0.51?l/g COD (HRT=2 days) and 0.25?l/g COD (HRT=1?day). In addition, the methane percentage in the biogas varied between 70.5% (HRT=10?days) and 47.5% (HRT=1?day). The close relationship between biomass loading rate and specific substrate utilization rate supported the use of Monod equations. Finally, the experimental values of effluent substrate concentration were reproduced with deviations equal to or less than 10% in every case.     

In situ pulse respirometric methods for the estimation of kinetic and stoichiometric parameters in aerobic microbial communities

In situ pulse respirometry was applied in an activated sludge bubble column treating synthetic wastewater for the estimation of the (i) maximum specific oxygen consumption rate, (ii) substrate affinity constant, (iii) biomass growth yield, (iv) maintenance coefficient, and (v) specific endogenous respiration rate. Parameters obtained from respirometry were compared to those obtained by the chemostat method, based on substrate and biomass measurements, under several dilution rates. The low sensitivity of substrate measurement methods and the difficulties of sampling heterogeneous biomass suspension are critical issues limiting the applicability of the chemostat method. Additionally, the extensive time consuming nature of this method allows concluding that chemostat method presents several disadvantages in comparison with in situ pulse respirometric techniques. Parameters were obtained from respirograms by fitting ASM1 and ASM3 models, and from experiments performed by injecting pulses of increasing substrate concentration. The injection of pulses of increasing concentration was the most adequate method, with several advantages such as a simpler experimental data interpretation, and results with better confidence.Considering the assessment and comparison of the experimental and calculation methods presented, it is recommended that the estimation of kinetic and stoichiometric parameters in mixed aerobic cultures should preferentially be performed by using in situ respirometric techniques.     

A theoretical reassessment of microbial maintenance and implications for microbial ecology modeling

We attempted to reconcile three microbial maintenance models (Herbert, Pirt, and Compromise) through a theoretical reassessment. We provided a rigorous proof that the true growth yield coefficient (Y(G) ) is the ratio of the specific maintenance rate (a in Herbert) to the maintenance coefficient (m in Pirt). Other findings from this study include: (1) the Compromise model is identical to the Herbert for computing microbial growth and substrate consumption, but it expresses the dependence of maintenance on both microbial biomass and substrate; (2) the maximum specific growth rate in the Herbert (μ(max,H) ) is higher than those in the other two models (μ(max,P) and μ(max,C) ), and the difference is the physiological maintenance factor (m(q) =?a); and (3) the overall maintenance coefficient (m(T) ) is more sensitive to m(q) than to the specific growth rate (μ(G) ) and Y(G) . Our critical reassessment of microbial maintenance provides a new approach for quantifying some important components in soil microbial ecology models.     

Substrate and energy costs of the production of exocellular enzymes by Bacillus licheniformis

Substrate and energy costs of the production of exocellular enzymes from glucose and citrate by B. Iicheniformis S1684 as well as molar growth yields corrected for these costs of product formation were calculated using data from chemostat experiments. The calculations showed that 1.46-1.73 mol glucose and 2.31-2.77 mol citrate are needed for formation and excretion of 1 mol protein. Consequently, the values of the maximal product yield from substrate (Y(psm') g/mol) are 80 < Y(psm) < 95 when product is formed from glucose and 50 < Y(psm) < 60 when product is formed from citrate. The higher substrate costs for product formation from citrate are due to a higher level of CO(2) production during protein formation and a higher substrate requirement for the energy supply of product formation and excretion than when product is formed from glucose. The theoretical ATP requirement for protein synthesis could be determined reasonably well, but the energy costs of protein excretion could not be determined exactly. The energy costs of protein formation are higher than those of biomass formation or protein excretion. Molar growth yields corrected for the substrate costs of product formation were high, indicating a high efficiency of growth.Growth and production parameters were determined as well from experimental data of recycling fermentor experiments using a parameter optimization procedure based on a mathematical model describing biomass growth as a linear function of the substrate consumption rate and the rate of product formation as a linear function of biomass growth rate. The fitting procedure yielded two growth and production domains during glucose limitation. In the first domain the values for the maximal growth yield and maintenance coefficient were in agreement with those found in chemostat experiments at corresponding values of Y(spm). Domain 2 could be described best with linear growth and product formation. In domain 2 the rate of product formation decreased and more substrate became available for biomass formation. As a consequence the specific growth rate increased in the shift from domain 1 to 2. Domain 2 behavior most probably is caused by the rel-status of B. Iicheniformis S1684.     

Issues in the culture of the extremely thermophilic methanogen, Methanothermus fervidus

Basic issues in the culture of the extremely thermophilic archaeon, Methanothermus fervidus, have been investigated, including culture medium formulation, substrate yield and product yield coefficient, growth rate and stoichiometry, and H(2) uptake kinetics. The pH optimum for growth of this organism was estimated at 6.9. Growth medium buffered with PIPES instead of bicarbonate supported both increased growth rate and maximum biomass concentration. Substitution of titanium(III) citrate for the reducing agent sodium sulfide improved culture performance as well. However, independent adjustment of iron and nickel concentrations from 11 to 111 muM, respectively, and carbon dioxide partial pressure from 5 to 20 psia did not impact the culture of M. fervidus significantly. An elemental balance approach was utilized to aid in design of a defined medium to support growth to a target maximum biomass concentration of at least 1.0 g dry wt/L. The growth of this organism was limited by H(2) availability in this reformulated culture medium. The maximum growth rate and biomass concentration achieved in anaerobic vials with the defined medium was 0.16 h(-1) and 0.74 g dry wt/L, respectively. This maximum biomass concentration was a 72% improvement over that obtained with a literature-based defined medium. The Monod parameter, K(s), with H(2) as limiting substrate, was estimated at 1.1 +/- 0.4 psia (55 +/- 20 muM in the broth), based on a H(2) consumption study. Representative values for the substrate yield, Y(X/CO(2) ), and product yield coefficient, Y(CH(4)/) (X), were determined experimentally to be 1.78 +/- 0.04 g dry wt/mol CO(2), and 0.52 +/- 0.01 mol CH(4)/g dry wt, respectively. A bench-scale fermentation system suitable for the culture of extremely thermophilic anaerobes was designed and constructed and proved effective for the culture of M. fervidus. (c) 1993 Wiley & Sons, Inc.     

Modeling alginate encapsulation system for biological hydrogen production

Wastewater treatment using encapsulated biomass is a promising approach for high-rate resource recovery. Encapsulation matrices can be customized to achieve desired biomass retention and mass transport performance. This, in turn, facilitates treatment of different waste streams. In this study, a model was developed to describe calcium-alginate beads encapsulating hydrogen-producing biomass, with the goal of enabling appropriate a priori customization of the system. The model was based on a classic diffusion-reaction model, but also included the growth of encapsulated biomass and product inhibition. Experimental data were used to verify the model, which accurately described the effect of hydraulic retention time, bead size, and feed concentration on resource (hydrogen) recovery from brewery wastewater. Sensitivity analyses revealed that the hydrogen production rate was insensitive to substrate diffusivity and bead size, but sensitive to the substrate partition coefficient, initial encapsulated biomass concentration, and the total volume of beads in the reactor, demonstrating that this system was growth-limited rather than diffusion-limited under the tested conditions. Because the model quantifies the relationship between the hydrogen production rate and various input and operating parameters, it should be possible to extend the model to determine the most cost-effective system for optimal performance with a given waste stream.     

Packed bed column fermenter and kinetic modeling for upgrading the nutritional quality of coffee husk in solid-state fermentation.

Studies were carried out to evaluate solid-state fermentation (SSF) for the upgradation of the nutritional quality of coffee husk by degrading the caffeine and tannins present in it. SSF was carried out by Aspergillus niger LPBx in a glass column fermenter using factorial design experiments and surface response methodology to optimize bioprocess parameters such as the substrate pH and moisture content and aeration rate. The first factorial design showed that the moisture content of the substrate and aeration rate were significant factors for the degradation of toxic compounds, which was confirmed by the second factorial design too. The kinetic study showed that the degradation of toxic compounds was related to the development of the mold and its respiration and also to the consumption of the reducing sugars present in coffee husk. From the values obtained experimentally for the oxygen uptake rate and CO(2) evolved, the system determined a biomass yield (Y(x/o)) of 3.811 (g of biomass).(g of consumed O(2))(-1) and a maintenance coefficient (m) of 0.0031 (g of consumed O(2)).(g biomass of biomass)(-1).h(-1). The best results on the degradation of caffeine (90%) and tannins (57%) were achieved when SSF was carried out with a 30 mL.min(-1) aeration rate using coffee husk having a 55% initial moisture content. The inoculation rate did not affect the metabolization of the toxic compounds by the fungal culture. After SSF, the protein content of the husk was increased to 10.6%, which was more than double that of the unfermented husk (5.2%).     

Effect of operating conditions on solid substrate fermentation

In this work the effects of environmental parameters on the performance of solid substrate fermentation (SSF) for protein production are studied. These parameters are (i) air flow rate, (ii) inlet air relative humidity, (iii) inlet air temperature, and (iv) the heat transfer coefficient between the outer wall of the fermentor and the air in the incubator. The air flow is supplied to effect cooling of the fermented mass by evaporation of water. A dynamic model is developed, which permits estimation of biomass content, total dry matter, moisture content, and temperature of the fermented matter. The model includes the effects of temperature and moisture content on both the maximum specific growth rate and the maximum attainable biomass content. The results of the simulation are compared with actual experimental data and show good agreement with them. The most important conclusions are that (i) the evaporative cooling of the biomass is very effective for temperature control and (ii) the air flow rate and the heat transfer coefficient have strong effects but they affect the biomass morphology and are not controllable easily. Also, a simple technique for the determination of the optimum temperature and moisture content profile for cell protein production is applied. The simulated biomass production increases considerably employing the optimum temperature and moisture content profiles. The ultimate goal is to implement the determined effects of the environmental parameters on the SSF biomass production and the temperature and moisture variation profiles to effectively control the SSF and optimize the biomass production. (c) 1993 John Wiley & Sons, Inc.     

Secretion-dependent proteolysis of heterologous protein by recombinant Escherichia coli is connected to an increased activity of the energy-generating dissimilatory pathway

The synthesis of a proteolytically unstable protein, originally designed for periplasmic export in recombinant Escherichia coli BL21(DE3), a strain naturally deficient for the ATP-dependent protease Lon (or La) and the outer membrane protease OmpT, is associated with a severe growth inhibition. This inhibition is not observed in BL21(DE3) synthesizing a closely related but proteolytically stable protein that is sequestered into inclusion bodies. It is shown that the growth inhibition is mainly caused by a slower cell division rate and a reduced growth yield and not by a general loss of cell division competence. Cells proceed with their normal growth characteristics when exposed again to conditions that do not sustain the expression of the heterologous gene. The performance of cells synthesizing either the stable or the degraded protein was also studied in high cell density cultures by employing a new method to calculate the actual specific growth rate, the biomass yield coefficient, and the dissimilated fraction of the carbon substrate in real-time. It is shown that the growth inhibition of cells synthesizing the proteolytically degraded protein is connected to an increased dissimilation of the carbon substrate resulting in a concomitant reduction of the growth rate and the biomass yield coefficient with respect to the carbon source. It is postulated that the increased dissimilation of the carbon substrate by lon-deficient Bl21(DE3) cells synthesizing the proteolytically unstable protein may result from a higher energy demand required for the in vivo degradation of this protein by ATP-dependent proteases different from the protease Lon.     

Carbon conversion efficiency and limits of productive bacterial degradation of methyl tert-butyl ether and related compounds

The utilization of the fuel oxygenate methyl tert-butyl ether (MTBE) and related compounds by microorganisms was investigated in a mainly theoretical study based on the Y(ATP) concept. Experiments were conducted to derive realistic maintenance coefficients and K(s) values needed to calculate substrate fluxes available for biomass production. Aerobic substrate conversion and biomass synthesis were calculated for different putative pathways. The results suggest that MTBE is an effective heterotrophic substrate that can sustain growth yields of up to 0.87 g g(-1), which contradicts previous calculation results (N. Fortin et al., Environ. Microbiol. 3:407-416, 2001). Sufficient energy equivalents were generated in several of the potential assimilatory routes to incorporate carbon into biomass without the necessity to dissimilate additional substrate, efficient energy transduction provided. However, when a growth-related kinetic model was included, the limits of productive degradation became obvious. Depending on the maintenance coefficient m(s) and its associated biomass decay term b, growth-associated carbon conversion became strongly dependent on substrate fluxes. Due to slow degradation kinetics, the calculations predicted relatively high threshold concentrations, S(min), below which growth would not further be supported. S(min) strongly depended on the maximum growth rate mu(ma)(x), and b and was directly correlated with the half maximum rate-associated substrate concentration K(s), meaning that any effect impacting this parameter would also change S(min). The primary metabolic step, catalyzing the cleavage of the ether bond in MTBE, is likely to control the substrate flux in various strains. In addition, deficits in oxygen as an external factor and in reduction equivalents as a cellular variable in this reaction should further increase K(s) and S(min) for MTBE.     

Comparison of maximum cell yield and maintenance coefficients in axenic cultures and activated sludge communities

Comparison of the equations that describe the relationship between the maximum cell yield coefficient, the maintenance coefficient, and the specific growth rate at steady-state conditions revealed that the equations used for axenic cultures are congruent with those commonly used for mixed-culture system such as activated sludge. A unified basis was proposed. The expression of the yield and maintenance coefficients in carbon units according to the unified basis permitted one to evaluate literature data on both axenic and mixed-culture systems. From this it appears that the maximum cell yield ranges from 0.50–0.80 (mg biomass carbon formed/mg substrate carbon used) for both axenic and mixed systems. However, the maintenance coefficient (mg substrate C/mg biomass C·hr) for the axenic cultures was between 0.010 and 0.100, but for activated sludge communities it was between 0.001 and 0.010. Microorganisms were isolated from sludge communities with these apparently low maintenance requirements and grown axenilly. Their maintenance coefficients but not their maximum yield coefficients decreased with decreasing specific growth rates. The consequences of this finding with regard to species selection in mixed-culture systems and the concept of cellular maintenance requirement are discussed.     

Kinetic study of anaerobic digestion of brewery wastewater

A study of the kinetics of the anaerobic digestion of brewery wastewater was carried out using a 1-litre, continuous-flow, completely-mixed, bioreactor operating at 35°C and containing a saponite-immobilized biomass at a concentration of 6·2 g volatile suspended solids (VSS)/litre. The bioreactor worked satisfactorily in a range of hydraulic retention times from 1·2 to 10 days and eliminated more than 95% of the initial chemical oxygen demand (COD) in all instances.

Guiot's kinetic model was used to determine the macroenergetic parameters of the system, and showed it to have a yield coefficient for the biomass (Y) of 0·080 g VSS/g COD and a specific rate of substrate uptake for cell maintenance (m) of 0·045 g COD/g VSS day.

The experimental results showed the rate of substrate uptake (R_s; g COD/g VSS day), correlated with the concentration of biodegradable substrate (S_b; g COD/litre), through an equation of the Michaelis-Menten type.     

Influence of substrate concentration on the macroenergetic parameters of anaerobic digestion of black-olive wastewater

Summary The macroenergetic parameters of the anaerobic digestion of black-olive wastewater, i.e. the yield coefficient for the biomass (Y. g VSS/g COD) and the specific rate of substrate uptake for cell maintenance (m, g COD/g VSS-day) decreased 6 limes and increased 5 times. respectively, when the influent substrate concentration increased from 1.1 to 4.4 g COD/l. This was significant at 95% confidence level. The use of the Guiot kinetic model allows a more accurate prediction of growth yield to be made as it relates substrate utilization to product formation.     

Influence of specific growth rate on biomass yield, productivity, and compostion of Candida utilis in batch and continuous culture.

Candida utilis was grown in batch and continuous culture on prickly pear juice as sole carbon and energy source. In batch culture the maximum specific growth rate (mum) and the substrate yield coefficient (Yps) varied according to sugar concentration. When the fermentation was carried out with 1% sugar, mum and Ys were 0.47/h and 42.6%, respectively. The best yields occurred in a chemostat at the pH range of 3.5 to 4.5 and temperature of 30 C. A beneficial effect on Ys was observed when the dilution rate (D) was increased. At a D of 0.55/h, the productivity was 2.38 g/liter per h. The maintenance coefficient attained a value of 0.09 g of sugar/g of biomass per h. Increases of D produced higher protein contents of the biomass. The information obtained indicates that protein production with Candida utilis, using prickly pear juice, should be carried out a high dilution rates where the Ys and protein content of the cell mass are also higher.     

The effect of pyrogenically modified substrates on mineralizing activity and growth strategies of microorganisms of grey forest soil

The rates of the mineralization processes initiated by the input of plant residues and pyrogenically modified plant material into gray forest soil under forests and meadows were assayed. While meadow plant residues was mineralized more rapidly than the forest floor, decomposition of the pyrogenic material resulted in disproportional changes in CO₂ emission from soils. Statistical treatment showed that the respiratory activity of CO₂ emission by heterotrophic microorganisms, which is a physiological characteristic of microbial communities, is 89% determined by the substrate quality. The maximal specific growth rate, which reflects the functional changes in microbial communities, was affected by the cenosis (36%) and the substrate (30%). Most of the carbon of the original plant material (up to 90%) was removed during the burning of plant substrates. The remaining compounds in the pyrogenically transformed material changed the process of mineralization in soil compared both to the control variant and to soil enriched with plant residues. Input of plant residues and ash into the soil resulted in increased total and active biomass, while the maximal specific growth rate decreased and the generation time for the active biomass increased. In the case of soils with plant residues, these changes in the state of microbial communities were brief and occurred during the period of intense mineralization (0–5 days), while, in soils with plant ash, stable changes were revealed after more prolonged incubation. Experimental determination of the microbial biomass turnover time (MTT) by means of two methods (from the ratio between the microbial biomass and respiration and from microbial specific growth rates) made it possible to determine the economical coefficient Y for microbial communities metabolizing the substrates of different availability. Depending on the experimental variant, the Y values varied from 0.22 to 0.51. Decreased maximal specific growth rate and increased values of Y (the coefficient of efficiency of substrate utilization) showed the predominant contribution of K-strategists in the mineralization of low available substrates in soil. The balance calculations and physiological characteristics of the microbial community suggested that the priming effect was most probable in soils enriched with plant ash.     

Influence of specific growth rate on biomass yield, productivity, and compostion of Candida utilis in batch and continuous culture.

Candida utilis was grown in batch and continuous culture on prickly pear juice as sole carbon and energy source. In batch culture the maximum specific growth rate (mum) and the substrate yield coefficient (Yps) varied according to sugar concentration. When the fermentation was carried out with 1% sugar, mum and Ys were 0.47/h and 42.6%, respectively. The best yields occurred in a chemostat at the pH range of 3.5 to 4.5 and temperature of 30 C. A beneficial effect on Ys was observed when the dilution rate (D) was increased. At a D of 0.55/h, the productivity was 2.38 g/liter per h. The maintenance coefficient attained a value of 0.09 g of sugar/g of biomass per h. Increases of D produced higher protein contents of the biomass. The information obtained indicates that protein production with Candida utilis, using prickly pear juice, should be carried out a high dilution rates where the Ys and protein content of the cell mass are also higher.