Postprandial metabolic increment of southern bluefin tuna Thunnus maccoyii ingesting high or low‐lipid sardines Sardinops sagax

This study examined the postprandial metabolism and swimming speed of southern bluefin tuna Thunnus maccoyii when fed sardines Sardinops sagax of either high‐lipid and high‐energy content or low‐lipid and low‐energy content. Five groups of two or three T. maccoyii (mean ±s.e. mass = 19·8 ± 0·5 kg, n = 14) were fed either low [2·2% lipid, 5·5 MJ kg^?1 gross energy (GE)] or high‐lipid (12·9%, 9·2 MJ kg^?1 GE) S. sagax. Before feeding, T. maccoyii swam at 0·74 ± 0·03 body lengths s^?1 (n = 5) and their routine metabolic rate was 305 ± 15 mg kg^?1 h^?1. Swimming speed and metabolic rate of T. maccoyii increased following feeding. Thunnus maccoyii swam 1·3 and 1·8 times faster during digestion of low and high‐lipid S. sagax, respectively. Postprandial peak metabolic rate, duration of elevated metabolism and total postprandial metabolic increment were all greater for T. maccoyii that ingested high‐lipid S. sagax. When total postprandial increment is represented as a proportion of ingested energy, there was no difference between high and low‐lipid meals, equating to between 30 and 35% of ingested energy. It was estimated that increased postprandial swimming costs account for 25 and 46% of the total postprandial metabolic response for low and high‐lipid S. sagax meals, respectively. Specific dynamic action (SDA) accounts for c. 20% of ingested energy regardless of S. sagax lipid level. These results confirm that the postprandial metabolic increment of T. maccoyii is greater than most other fish species. Much of the high cost of postprandial metabolic increment can be attributed to increased postprandial swimming costs. For T. maccoyii, it appears that activity and SDA are not independent, which complicates bioenergetic evaluation. High postprandial metabolic costs accentuate the great energetic requirements of T. maccoyii.     

Effect of dilution rate and methanol‐glycerol mixed feeding on heterologous Rhizopus oryzae lipase production with Pichia pastoris Mut+ phenotype in continuous culture

The induction using substrate mixtures is an operational strategy for improving the productivity of heterologous protein production with Pichia pastoris. Glycerol as a cosubstrate allows for growth at a higher specific growth rate, but also has been reported to be repressor of the expression from the AOX1 promoter. Thus, further insights about the effects of glycerol are required for designing the induction stage with mixed substrates. The production of Rhizopus oryzae lipase (ROL) was used as a model system to investigate the application of methanol‐glycerol feeding mixtures in fast metabolizing methanol phenotype. Cultures were performed in a simple chemostat system and the response surface methodology was used for the evaluation of both dilution rate and methanol‐glycerol feeding composition as experimental factors. Our results indicate that productivity and yield of ROL are strongly affected by dilution rate, with no interaction effect between the involved factors. Productivity showed the highest value around 0.04–0.06 h^?1, while ROL yield decreased along the whole dilution rate range evaluated (0.03–0.1 h^?1). Compared to production level achieved with methanol‐only feeding, the highest specific productivity was similar in mixed feeding (0.9 UA g‐biomass^?1 h^?1), but volumetric productivity was 70% higher. Kinetic analysis showed that these results are explained by the effects of dilution rate on specific methanol uptake rate, instead of a repressor effect caused by glycerol feeding. It is concluded that despite the effect of dilution rate on ROL yield, mixed feeding strategy is a proper process option to be applied to P. pastoris Mut⁺ phenotype for heterologous protein production. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:707–714, 2015     

Bioenergetic model of planktivorous fish feeding, growth and metabolism: theoretical optimum swimming speed of fish larvae

The feeding activity of an individual fish larva is described by an equation which includes parameters for the area successfully searched, probability of food capture multiplied by the cross-sectional perceptive visual field, larval swimming speed and the time required to consume a unit of food energy. The proportion of ingested food energy used for metabolism increases exponentially with increasing swimming speed. The model predicts that food consumption rate increases asymptotically whereas metabolic rate increases exponentially. This results in a predicted growth rate curve that reaches a maximum at a certain swimming speed and decreases at both higher and lower speeds. The model can be used to predict the influence of type of prey, prey density, water temperature etc. on larval growth. An expression describing how many hours per day fish larvae must forage in order to grow at a certain daily body weight gain allows the limits of environmental conditions for positive, zero and negative growth rate to be set. Results of simulations demonstrated that the optimum swimming speed for maximum growth of coregonid larvae increased with an increase in food density, decrease in water temperature or decrease of prey vulnerability. At optimum ‘theoretical’ swimming speed an increase in water temperature from 5 to 17° C required the food density to be increased from 20 to 80 copepods l^?1 in order to maintain a daily growth increment of 2%. The minimum Artemia density required for maintenance metabolism increased from 10 to 30 items 1¹ over the same temperature increase from 5 to 17° C, and food densities required for 8% growth rates were 26 and 56 Artemia nauplii l^?1 at 5 and 17° C, respectively. Contrary to previous findings, results of the present study suggest that metabolic rates of actively feeding fish larvae may be from 5 to 50 times the standard metabolic rate: earlier studies suggested that a factor of 2–3 may be generally applicable.     

Food consumption and growth of larvae and juveniles of three cyprinid species at different food levels

Synopsis The relationships between food availability, consumption and growth were analyzed from the onset of feeding to an age of 90 days in three cyprinid species. Fish were held at 20 ± 0.5° C and given two (three) constant rations of approximately 30, (40) or 100% dry body weight (dbw) ind^-1 day^-1. Food consisted of living zooplankton, the size of which correlated with fish size. At high food densities consumption rates decreased rapidly with fish size in all three species. At reduced rations, fish consumed most of the food offered until they were larger than 10 mg dbw. In all species and at each feeding level daily rations consumed increased allometrically with body size. Respiration rate, expressed as routine metabolic rate differed insignificantly between the three species. At high ration levels, growth rates of small bleak, Alburnus alburnus, were distinctly lower than those of roach, Rutilus rutilus, and blue bream, Abramis ballerus. At low food supply all three species grew at similar rates. Assimilation efficiency at low food conditions was approximately twice that of the well-fed groups. If the caloric equivalents of oxygen consumption as measured in well-fed fish are applied to fish fed at low rations their energy budgets do not balance. This indicates the limitations of fish larvae in the partitioning of energy for growth or activity at such conditions.     

Comparing a bioenergetics model with feeding rates of caged European starlings

We tested a bioenergetics model integrated within a mortality model that estimates numbers of European starlings (Sturnus vulgaris) poisoned with the avicide, Compound DRC-1339 Concentrate. The bioenergetics model predicted daily metabolic rate. Accuracy and reliability of this variable is critical because other algorithms (e.g., toxicity regressions, feeding behavior) in the mortality model depend on metabolic rate to calculate the amount of DRC-1339 ingested per bird. We tested the bioenergetics model by comparing its estimates of metabolic rate with those generated from measuring feeding rates of caged starlings during a feeding trial conducted outdoors during January 2008. Over the 12-day feeding trial, daily feeding rates of caged starlings indicated that metabolic rates ranged from 157 kJ/bird per day to 305 kJ/bird per day. The bioenergetics model predicted metabolic rates ranging from 208 kJ/bird per day to 274 kJ/bird per day. There was no difference between these 2 independently derived estimates of daily metabolic rate (paired t-test: t₍₁₁₎ = 1.4, P = 0.18). Using 95% confidence intervals calculated from variation of feeding rates among cages (n = 4, 6 birds/cage), the bioenergetics model's estimates were within 95% confidence intervals on 9 of 12 days and greater than the upper 95% confidence interval on 3 days. Daily estimates of metabolic rate were directly correlated between the bioenergetics model and the feeding-rate model (r₁₂ = 0.57, P = 0.05). A broad range of temperatures (−17°C to 14°C), wind speeds (0–40 km/hr), and percent cloud cover (0–100%) were encountered during the feeding trial. The bioenergetics model's predictions appeared robust to varying meteorological conditions typical of winters in middle latitudes of the interior United States. Compound DRC-1339 Concentrate is used by USDA Wildlife Services to manage chronic infestations of starlings at livestock facilities, which occur mainly during fall and winter. Compared to other methods used for estimating DRC-1339 mortality (e.g., counting birds pre- and posttreatment), bioenergetics modeling should improve the mortality model's overall accuracy and precision. © The Wildlife Society, 2011     

Production of xanthan by in-flow cultures of Xanthomonas campestris

The kinetics of xanthan formation in Xanthomonas campestris continuous and fed-batch fermentations was studied along with metabolic changes due to growth rate variation. A maximum growth rate within the range 0.11–0.12 h^–1 was obtained from the continuous culture data in defined medium, producing xanthan at rates up to 0.36 g l^–1 h^–1 corresponding to a maximum 67% glucose conversion at a dilution rate (D) of 0.05 h^–1. Comparatively, fed-batch cultivation was more efficient, producing maximum xanthan at 0.75 g l^–1 h^–1 and 63% glucose conversion at 0.1 h^–1. When reaching D=0.062 h^–1 in continuous cultures, a change was observed and the values of the specific rate of substrate consumption shifted, initiating an uncoupled growth region expressing a lack of balance of the catabolic and anabolic reactions. The deviation was not accompanied by a change in specific xanthan production indicating that xanthan metabolism was not affected by D. For fed-batch-grown X. campestris cells within the range D=0.03–0.1 h^–1, both metabolic parameters changed linearly with the growth rate showing a wide region coupled to growth. Outside that range, glucose accumulated and the specific xanthan production dropped, suggesting substrate inhibition. Correspondence to: J. C. Roseiro     

Feasibility study of exponential feeding strategy in fed-batch cultures for phenol degradation using Cupriavidus taiwanensis

An indigenous phenol-degrading bacterial isolate Cupriavidus taiwanensis R186 was used to degrade phenol from an aqueous solution under fed-batch operation. An exponential feeding strategy combined with dissolved oxygen control was applied based on kinetic characteristics of cell growth and phenol degradation to meet sufficient metabolic needs for cellular growth and achieve the best phenol removal efficiency. Without the stress of phenol inhibition, the optimal set point of specific growth rate of exponential feeding for fed-batch phenol degradation was found to be 0.50–0.55μ_max (μ_max denotes the maximum specific growth rate from Monod model). Meanwhile, the sufficient set point of dissolved oxygen for maximal phenol degradation efficiency was approximately at 10–55% air saturation. With the optimal operation conditions, the best phenol degradation rate was 0.213 g phenol h⁻¹, while a shortest treatment time of 15 h was achieved for complete degradation of 11.35 mM (ca. 3.20 g) of phenol.     

Estimation of daily energetic requirements in young scalloped hammerhead sharks, Sphyrna lewini

Juvenile scalloped hammerhead sharks, Sphyrna lewini, are apex predators within their nursery ground in Kāne‘ohe Bay, Ō‘ahu, Hawai‘i. Understanding daily maintenance requirements of a top-level predator is an important step toward understanding its ecological impact within a nursery ecosystem. Juvenile S. lewini were fed a range of daily ration levels to examine the effect of feeding rate on growth and gross conversion efficiency. The von Bertalanffy growth model yielded the best fit to the data, predicting a maintenance ration of 115 kJ kg⁻¹ day⁻¹ (3.4% body weight (BW) day⁻¹) and a maximum growth rate of 38 kJ kg⁻¹ day⁻¹. This finding is in agreement with the previous prediction of high energetic requirements for S. lewini. In combination with the hypothesized food limitation within Kāne‘ohe Bay, this result may explain the observed high mortality rates of S. lewini. Gross conversion efficiency, K ₁, ranged from −36% to 34%, with maximum efficiency at feeding levels of 5.1% BW day⁻¹. The growth conversion efficiency of S.␣lewini is similar to that of lemon sharks and teleost fishes. Growth rates of juvenile S. lewini are possibly restricted by their high metabolic rate, limited food availability and foraging inexperience. By directly examining the effect of ration size on growth and food conversion, it was possible to resolve discrepancies between earlier studies, which used respiratory metabolism and gut content analyses.     

Improvement of specific growth rate of <Emphasis Type="BoldItalic">Pichia pastoris</Emphasis> for effective porcine interferon-α production with an on-line model-based glycerol feeding strategy

Effective expression of porcine interferon-α (pIFN-α) with recombinant Pichia pastoris was conducted in a bench-scale fermentor. The influence of the glycerol feeding strategy on the specific growth rate and protein production was investigated. The traditional DO-stat feeding strategy led to very low cell growth rate resulting in low dry cell weight (DCW) of about 90 g/L during the subsequent induction phase. The previously reported Artificial Neural Network Pattern Recognition (ANNPR) model-based glycerol feeding strategy improved the cell density to 120 g DCW/L, while the specific growth rate decreased from 0.15 to 0.18 to 0.03–0.08 h⁻¹ during the last 10 h of the glycerol feeding stage leading to a variation of the porcine interferon-α production, as the glycerol feeding scheme had a significant effect on the induction phase. This problem was resolved by an improved ANNPR model-based feeding strategy to maintain the specific growth rate above 0.11 h⁻¹. With this feeding strategy, the pIFN-α concentration reached a level of 1.43 g/L, more than 1.5-fold higher than that obtained with the previously adopted feeding strategy. Our results showed that increasing the specific growth rate favored the target protein production and the glycerol feeding methods directly influenced the induction stage. Consequently, higher cell density and specific growth rate as well as effective porcine interferon-α production have been achieved by our novel glycerol feeding strategy.     

The effect of feeding on the metabolic rate in harbour seals (Phoca vitulina)

The heat increment of feeding was estimated in harbour seals (Phoca vitulina). Seals were given different amounts of herring, ranging from 0.8 to 2.65 kg. The caloric content of the herring ranged from 6575 to 12560 kJ·kg^-1 depending on time of year. Metabolic rate increased within 30 min after feeding, and the magnitude and duration of heat increment of feeding depended on the size of the meal and the caloric content of the herring. Measured heat increment of feeding was up to 14.9% of gross energy intake and metabolic rate increased as much as 46% above resting, postabsorptive metabolic rate for 15 h duration in a harbour seal with a body weight of approximately 40 kg.     

Improving intracellular production of recombinant protein in <Emphasis Type="Italic">Pichia pastoris</Emphasis> using an optimized preinduction glycerol-feeding scheme

High-cell-density production of recombinant growth hormone of Lateolabrax japonicus (rljGH) expressed intracellularly in Pichia pastoris was investigated. In the regular strategy of induction at a cell density of 160 g l⁻¹, short duration of intracellular rljGH accumulation (17 h) resulted in a low final cell density of 226 g l⁻¹. Thus, a novel strategy of induction at a cell density of 320 g l⁻¹ was investigated. In this strategy, the preinduction glycerol-feeding scheme had a significant effect on the post-induction production. Constant glycerol feeding led to a decrease of the specific rljGH production and specific production rate because of low preinduction specific growth rate. This decrease was avoided by exponential glycerol feeding to maintain a preinduction specific growth rate of 0.16 h⁻¹. The results from exponential glycerol feeding indicated that the rljGH production depended on the preinduction specific growth rate. Moreover, mixed feeding of methanol and glycerol during induction improved the specific production rate to 0.07 mg g⁻¹ h⁻¹ from 0.043 mg g⁻¹ h⁻¹. Consequently, both high cell density (428 g l⁻¹) and high rljGH production could be achieved by the novel strategy: growing the cells at the specific growth rate of 0.16 h⁻¹ to the cell density of 320 g l⁻¹ and inducing the expression by mixed feeding.     

Mixed-feed exponential feeding for fed-batch culture of recombinant methylotrophic yeast

A simple, accurate model capable of predicting cell growth and methanol utilization during the mixed substrate fed-batch fermentation of Mut^S recombinant Pichia pastoris was developed and was used to design an exponential feeding strategy for mixed substrate fed-batch fermentation at a constant specific growth rate. Mixed substrate feeding has been shown to boost productivity in recombinant fed-batch culture of P. pastoris, while fixed growth rate exponential feeding during fed-batch culture is a useful tool in process optimization and control.     

Effect of prey concentration on growth of piscivorous Japanese Spanish mackerel Scomberomorus niphonius larvae in the Seto Inland Sea, Japan

Japanese Spanish mackerel, Scomberomorus niphonius, larvae feed almost exclusively on fish larvae from the first‐feeding stage. The relationship between the growth of S. niphonius larvae and concentration of major prey organisms of the larvae, clupeid larvae, was investigated in the Sea of Hiuchi, the central Seto Inland Sea, Japan, from 28 to 29 May 1997. Water temperature, salinity, and the concentration of clupeid larvae had no significant effect on the S. niphonius larval concentration. Mean growth rate of S. niphonius larvae varied between 0.38 and 0.64 mm day^?1. The temperature and salinity had no significant effect on the mean larval growth rate while there existed prey concentration‐dependent growth at lower prey concentration. The relationship between the mean larval growth rate (G_m) and concentration of clupeid larvae (C_C) was expressed by a logarithmic equation: G_m = 0.037 log C_C + 0.441 (n = 16, r² = 0.519, P < 0.01).     

Two-stage glycerol feeding for enhancement of recombinant hG-CSF production in a fed-batch culture of Pichia pastoris

Recombinant hG-CSF was expressed in Pichia pastoris under the control of the AOX1 promoter. In this study, the glycerol feeding rate was adjusted to achieve the maximum attainable specific growth rate before induction. Using a two-stage glycerol feeding method, the specific growth rate was changed from a maximum value of 0.21 h⁻¹ (at the beginning of feeding) to 0.15 h⁻¹ prior to induction. With this approach, the final dry cell wt and rhG-CSF yield achieved was close to 120 g l⁻¹ and 320 mg l⁻¹, respectively. Our study found that the two-stage feeding method allowed the overall productivity of rhG-CSF to increase 2.9 times that of the conventional fed-batch method.     

<Emphasis Type="Italic">Pichia pastoris</Emphasis> fermentation with mixed-feeds of glycerol and methanol: growth kinetics and production improvement

Fed-batch fermentation of a methanol utilization plus (Mut⁺) Pichia pastoris strain typically has a growth phase followed by a production phase (induction phase). In the growth phase glycerol is usually used as carbon for cell growth while in the production phase methanol serves as both inducer and carbon source for recombinant protein expression. Some researchers employed a mixed glycerol-methanol feeding strategy during the induction phase to improve production, but growth kinetics on glycerol and methanol and the interaction between them were not reported. The objective of this paper is to optimize the mixed feeding strategy based on growth kinetic studies using a Mut⁺ Pichia strain, which expresses the heavy-chain fragment C of botulinum neurotoxin serotype C [BoNT/C(Hc)] intracellularly, as a model system. Growth models on glycerol and methanol that describe the relationship between specific growth rate (μ) and specific glycerol/methanol consumption rate (ν _gly, ν _MeOH) were established. A mixed feeding strategy with desired μ _gly/μ _MeOH =1, 2, 3, 4 (desired μ _MeOH set at 0.015 h⁻¹) was employed to study growth interactions and their effect on production. The results show that the optimal desired μ _gly/μ _MeOH is around 2 for obtaining the highest BoNT/C(Hc) protein content in cells: about 3 mg/g wet cells. Electronic Publication     

Growth energetics of juvenile herring,Clupea harengus L.: food conversion efficiency and temperature dependency of metabolic rate

The relationship between rates of food consumption (C) and somatic growth (G) and the effect of temperature (T) on rates of mass lost during food deprivation were examined in 9–10 cm total length (TL) [1.0–1.5 g dry mass (DM)] juvenile Atlantic herring (Clupea harengus L.) in the laboratory. One feeding‐growth trial was conducted at 16°C using groups of herring feeding on known rations of brine shrimp (Artemia spp.) nauplii to quantify gross and net growth efficiency. Rates of mass lost by groups of herring (a proxy for metabolic rate, M) were measured in trials conducted at 9.7, 14.2 and 17.9°C. Gross growth efficiency (GGE = 100*G*C^?1) at 16°C was 25% at the highest rations (5.8–6.6% DM). The maintenance ration (C_main = C at zero G) was equal to 432 J*fish^?1*d^?1 or 2.0% DM*d^?1. At 16°C, net growth efficiency (100*G*(C?C_main)^?1) was 42%. The nucleic acid content (RNA‐DNA ratio, RD) in herring muscle tissue was strongly related to somatic growth (G, % DM*d^?1 = ?0.36*RD² + 3.21*RD ?3.92, r² = 0.90, P < 0.05, n = 8 groups). The effect of T (9.7–17.9°C) on M was described by a second order polynomial equation M = ?1.24*T + 38.2*T ? 218 (J*g DM^?1*d^?1) and M = ?10*T + 310*T ? 1815 (J*fish^?1*d^?1). This was the first study to investigate the influence of temperature on the metabolic rate of juvenile Atlantic herring under stress‐free conditions in the laboratory and provides the first estimates of gross and net growth efficiency for this species feeding on live prey.     

Metabolic state of Zymomonas mobilis in glucose-, fructose-, and xylose-fed continuous cultures as analysed by 13C- and 31P-NMR spectroscopy

The reasons for the well-known significantly different behaviour of the anaerobic, gram-negative, ethanologenic bacterium Zymomonas mobilis during growth on fructose (i.e. decreased growth and ethanol yields, increased by-product formation) as compared to that on its second natural substrate, glucose, have remained unexplained. A xylose-fermenting recombinant strain of Z. mobilis that was recently constructed in our laboratory also unexpectedly displayed an increased formation of by-products and a strongly reduced growth rate as compared to the parent strain. Therefore, a comprehensive study employing recently developed NMR-based methods for the in vivo analysis of intracellular phosphorylated pool sizes and metabolic fluxes was undertaken to enable a global characterization of the intracellular metabolic state of Z. mobilis during growth on ¹³C-labelled glucose, fructose and xylose in defined continuous cultures. The ¹³C-NMR flux analysis indicated that ribose 5-phosphate is synthesized via the nonoxidative pentose phosphate pathway in Z. mobilis, and it identified a metabolic bottleneck in the recombinant xylose-fermenting Z. mobilis strain at the level of heterologous xylulokinase. The ³¹P-NMR analyses revealed a global alteration of the levels of intracellular phosphorylated metabolites during growth on fructose as compared to that on glucose. The results suggest that this is primarily caused by an elevated concentration of intracellular fructose 6-phosphate. Received: 7 January 1999 / Accepted: 22 March 1999     

Growth-associated synthesis of recombinant human glucagon and human growth hormone in high-cell-density cultures of Escherichia coli

Synthesis of two recombinant proteins (human glucagon and human growth hormone) was investigated in fed-batch cultures at high cell concentrations of recombinant Escherichia coli. The glucose-limited growth was achieved without accumulation of metabolic by-products and hence the cellular environment is presumed invariable during growth and recombinant protein synthesis. Via exponential feeding in the two-phase fed-batch operation, the specific cell growth rate was successfully controlled at the desired rates and the fed-batch mode employed is considered appropriate for examining the correlation between the specific growth rate and the efficiency of recombinant product formation in the recombinant E. coli strains. The two recombinant proteins were expressed as fusion proteins and the concentration in the culture broth was increased to 15 g fusion growth hormone l⁻¹ and 7 g fusion glucagon l⁻¹. The fusion growth hormone was initially expressed as soluble protein but seemed to be gradually aggregated into inclusion bodies as the expression level increased, whereas the synthesized fusion glucagon existed as a cytoplasmic soluble protein during the whole induction period. The stressful conditions of cultivation employed (i.e. high-cell-density cultivation at low growth rate) may induce the increased production of various host-derived chaperones and thereby enhance the folding efficiency of synthesized heterologous proteins. The synthesis of the recombinant fusion proteins was strongly growth-dependent and more efficient at a higher specific growth rate. The mechanism linking specific growth rate with recombinant protein productivity is likely to be related to the change in cellular ribosomal content. Received: 27 May 1997 / Received last revision: 31 October 1997 / Accepted: 21 November 1997     

Effects of Chironomus tentans larval growth retardation on adult emergence and ovipositing success: implications for interpreting freshwater sediment bioassays

A laboratory study was conducted with Chironomus tentans to assess the significance of growth retardation of third-to fourth-instar larvae over a 10-d test period on long-term survival, adult emergence, and ovipositing success. Data were intended to provide interpretive guidance for the commonly used growth endpoint in 10-d sediment bioassays with C. tentans. Larval growth was controlled by using six feeding levels ranging from 0.2 to 5.9 mg dry weight Tetrafin® fish food per day. Mean 10-d survival was 88% at all feeding levels, but larval growth decreased significantly (p<0.05) with each decrease in feeding level. Cumulative successful emergence of adult C. tentans decreased significantly with decreasing larval growth. Mean times to emergence always increased with decreasing growth rates and effects were generally more pronounced for females than males. At the lowest 10-d mean growth where successful emergence occurred (0.27 mg), the times to emergence doubled relative to the times observed at the highest 10-d mean growth (1.03 mg). Ten-day larval growth retardation was strongly correlated with reduction in adult emergence success (r ² = 0.96). Growth retardations 64% resulted in 86–100% reductions in adult emergence. Growth retardation in the range of 35 to 50% equated with comparable percent reductions in adult emergence success. Although fewer females successfully emerged at 10-d growths of 0.37 and 0.74 mg relative to the highest 10-d growth (1.03 mg), ovipositing success of these females did not appear to be adversely affected by either their slower growth rates or their lower mean dry weights (0.62 and 0.99 mg, respectively). Growth of second generation larvae did not appear to be affected by maternal growth rate, but rather appeared to be solely correlated with their own feeding level.     

Optimization of culture on the overproduction of TRAIL in high-cell-density culture by recombinant Escherichia coli

Different nutrient-feeding cultures were carried out in producing recombinant protein of truncated tumor necrosis factor related apoptosis-inducing ligand (TRAIL) (114–281 amino acids of TRAIL) in Escherichia coli strain C600/pBV-TRAIL. The effects of preinduction specific growth rate, postinduction carbon source (glucose and glycerol), and feeding strategies were investigated. The higher preinduction specific growth rate (μ=0.22 h⁻¹) contributed to the increase in the TRAIL production, at which TRAIL was accumulated in bacterial cells as 7.2% of total cellular protein, corresponding to 1.99 g l⁻¹ in contrast with 5.1% (1.29 g l⁻¹) at preinduction specific growth rate (μ=0.1 h⁻¹) during high-cell-density culture. Glycerol was superior to glucose as the postinduction carbon source for TRAIL production. Under similar culture conditions, the final concentration of TRAIL was produced 1.59-fold more when glycerol was used as postinduction carbon source than when glucose was used. At the same time, the results showed that it is efficient to adopt the pH-stat feeding strategy at postinduction for the overproduction of TRAIL. The TRAIL production was increased up to 4.51 g l⁻¹, approximately 16.1% of total cellular protein. The mechanisms behind the preinduction specific growth rate effect on the expression level may be ascribed to the leakage secretion of acetate.