Optimization of probiotic and lactic acid production by Lactobacillus plantarum in submerged bioreactor systems

Biomass and lactic acid production by a Lactobacillus plantarum strain isolated from Serrano cheese, a microorganism traditionally used in foods and recognized as a potent probiotic, was optimized. Optimization procedures were carried out in submerged batch bioreactors using cheese whey as the main carbon source. Sequential experimental Plackett–Burman designs followed by central composite design (CCD) were used to assess the influence of temperature, pH, stirring, aeration rate, and concentrations of lactose, peptone, and yeast extract on biomass and lactic acid production. Results showed that temperature, pH, aeration rate, lactose, and peptone were the most influential variables for biomass formation. Under optimized conditions, the CCD for temperature and aeration rate showed that the model predicted maximal biomass production of 14.30 g l⁻¹ (dw) of L. plantarum. At the central point of the CCD, a biomass of 10.2 g l⁻¹ (dw), with conversion rates of 0.10 g of cell g⁻¹ lactose and 1.08 g lactic acid g⁻¹ lactose (w/w), was obtained. These results provide useful information about the optimal cultivation conditions for growing L. plantarum in batch bioreactors in order to boost biomass to be used as industrial probiotic and to obtain high yields of conversion of lactose to lactic acid.     

Towards the design of an optimal strategy for the production of ergosterol from Saccharomyces cerevisiae yeasts

The total yield of ergosterol produced by the fermentation of the yeast Saccharomyces cerevisiae depends on the final amount of yeast biomass and the ergosterol content in the cells. At the same time ergosterol purity—defined as percentage of ergosterol in the total sterols in the yeast—is equally important for efficient downstream processing. This study investigated the development of both the ergosterol content and ergosterol purity in different physiological (metabolic) states of the microorganism S. cerevisiae with the aim of reaching maximal ergosterol productivity. To expose the yeast culture to different physiological states during fermentation an on‐line inference of the current physiological state of the culture was used. The results achieved made it possible to design a new production strategy, which consists of two preferable metabolic states, oxidative‐fermentative growth on glucose followed by oxidative growth on glucose and ethanol simultaneously. Experimental application of this strategy achieved a value of the total efficiency of ergosterol production (defined as product of ergosterol yield coefficient and volumetric productivity), 103.84 × 10^?6 g L^?1h^?1, more than three times higher than with standard baker's yeast fed‐batch cultivations, which attained in average 32.14 × 10^?6 g L^?1h^?1. At the same time the final content of ergosterol in dry biomass was 2.43%, with a purity 86%. These results make the product obtained by the proposed control strategy suitable for effective down‐stream processing. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:838–848, 2017     

A fuzzy logic controller to control nutrient dosage in a fed-batch baker's yeast process

A fuzzy logic controller designed to control glucose feeding in a fed-batch baker's yeast process is presented. Feeding is carried out in portions and the controller determines the time at which glucose should be added and computes the size of the portion to provide the maximum glucose uptake rate. Moreover, the controller detects and prevents the occurrence of overdosage. The experimental results indicate that yield and specific growth rate obtained with the controller approached 55% and 0.13 h^–1, respectively.     

Effects of aeration rate and feed on growth,productivity and nutrient composition of black soldier fly (Hermetia illucens L.) larvae

Black soldier fly larvae (Hermetia illucens L.) are widely cultivated as bioconversion agents. These larvae are known to be highly sensitive to changes in the external environment, such as temperature and relative humidity. Therefore, a rearing hive equipped with an instrumentation system known as the Modular Fly Hive was designed to cultivate black soldier larvae. This study aimed to determine the effects of aeration rate (0 – 0.84 m³/s.) and feed type (coconut endosperm waste and soybean curd) on growth and productivity of black soldier fly cultivated in Modular Fly Hives as well as well as feed digestibility and nutrient composition of the larval biomass. Increasing aeration rate decreased the average temperature and relative humidity within the hives. The results showed that the larvae demonstrated a sigmoidal growth curve for all types of feed and the type of feed significantly affects the average wet weight and length of the larvae. The greatest dry biomass productivity (52.85 ± 3.85 g/m²/day) was observed when the larvae were cultivated using soybean curd residue with an aeration rate of 0.84 m³/s, while the lowest dry biomass productivity (35.27 ± 9.72 g/m²/day) was observed when the larvae were cultivated using coconut endosperm waste without aeration. Proximate analysis revealed that the larval biomass had total protein, lipid, ash and carbohydrate content in the range of 37.20 – 48.60%, 9.61 – 20.02%, 4.80 – 6.40%, 33.86 – 38.70%, respectively. Amino acid in the biomass were dominated by glutamic acid (11.11–12.30%), aspartic acid (8.25 – 10.35%), leucine (8.09 – 8.57%), and lysine (6.74 – 8.14%). Lipid isolated from the larval biomass were mainly composed of lauric acid (28.35–61.68%), linoleic acid (6.27 – 30.29%), palmitic acid (7.62 – 15.23%), and myristic acid (5.05 – 14.34%).     

Production of polysaccharide–peptide complexes by submerged mycelial culture of an entomopathogenic fungus Cordyceps sphecocephala

The effects of medium components and environmental factors on the production of mycelial biomass and polysaccharide–peptide complexes (exobiopolymers) by Cordyceps sphecocephala J-201 were investigated in submerged cultures. The optimal temperature and initial pH for the production of both mycelial biomass and exobiopolymers in flask cultures were found to be 25 °C and pH 4–5, respectively. The optimal combination of the media constituents was as follows (g l⁻¹): sucrose 40, yeast extract 6, polypepton 2, KH₂PO₄ 0.46, K₂HPO₄ 1, and MgSO₄·7H₂O 0.5. The results of bioreactor culture revealed that the maximum concentration of mycelial biomass (28.2 g l⁻¹) was obtained at an agitation speed of 300 rpm and at an aeration rate of 2 vvm, whereas maximum exobiopolymer production (2.5 g l⁻¹) was achieved at a milder agitation speed (150 rpm). There was a significant variance in mycelial morphology between different aeration conditions. Looser mycelial pellets were developed, and their size and hairiness increased as the aeration rate increased from 0.5 to 2.0 vvm, resulting in enhanced exobiopolymer production. The apparent viscosities of fermentation broth increased rapidly towards the end of fermentations at the conditions of high aeration rate and agitation speed, which were mainly due to high amount of mycelial biomass rather than exobiopolymers at the later stages of fermentation. The three different exobiopolymers (FR-I, -II, and -III) were fractionated by a gel filtration chromatography on Sepharose CL-6B. The carbohydrate and protein contents in each fraction were significantly different and the molecular weights of FR-I, FR-II, and FR-III were determined to be 1831, 27, and 2.2 kDa, respectively. The compositional analysis revealed that the three fractions of crude exobiopolymers consisted of acidic and nonpolar amino acids, such as aspartic acid, glutamic acid, glycine, and valine in protein moiety, and of mainly mannose and galactose in sugar moiety.     

Effect of the aeration rate and agitation speed on β-carotene production and morphology of Blakeslea trispora in a stirred tank reactor: mathematical modeling

The effect of aeration rate and agitation speed on β-carotene production and morphology of Blakeslea trispora in a stirred tank reactor was investigated. B. trispora formed hyphae, zygophores and zygospores during the fermentation. The zygospores were the morphological form responsible for β-carotene production. Both aeration and agitation significantly affected β-carotene concentration, productivity, biomass and the volumetric mass transfer coefficient (K_La). The highest β-carotene concentration (1.5 kg m⁻³) and the highest productivity (0.08 kg m⁻³ per day) were obtained at low impeller speed (150 rpm) and high aeration rate (1.5 vvm). Also, maximum productivity (0.08 kg m⁻³ per day) and biomass dry weight (26.4 kg m⁻³) were achieved at high agitation speed (500 rpm) and moderate aeration rate (1.0 vvm). Conversely, the highest value of K_La (0.33 s⁻¹) was observed at high agitation speed (500 rpm) and high aeration rate (1.5 vvm). The experiments were arranged according to a central composite statistical design. Response surface methodology was used to describe the effect of impeller speed and aeration rate on the most important fermentation parameters. In all cases, the fit of the model was found to be good. All fermentation parameters (except biomass concentration) were strongly affected by the interactions among the operation variables. β-Carotene concentration and productivity were significantly influenced by the aeration, agitation, and by the positive or negative quadratic effect of the aeration rate. Biomass concentration was principally related to the aeration rate, agitation speed, and the positive or negative quadratic effect of the impeller speed and aeration rate, respectively. Finally, the volumetric mass transfer coefficient was characterized by the significant effect of the agitation speed, while the aeration rate had a small effect on K_La.     

Interactive global change factors mitigate soil aggregation and carbon change in a semi‐arid grassland

The ongoing global change is multi‐faceted, but the interactive effects of multiple drivers on the persistence of soil carbon (C) are poorly understood. We examined the effects of warming, reactive nitrogen (N) inputs (12 g N m^?2 year^?1) and altered precipitation (+ or ? 30% ambient) on soil aggregates and mineral‐associated C in a 4 year manipulation experiment with a semi‐arid grassland on China's Loess Plateau. Our results showed that in the absence of N inputs, precipitation additions significantly enhanced soil aggregation and promoted the coupling between aggregation and both soil fungal biomass and exchangeable Mg²⁺. However, N inputs negated the promotional effects of increased precipitation, mainly through suppressing fungal growth and altering soil pH and clay‐Mg²⁺‐OC bridging. Warming increased C content in the mineral‐associated fraction, likely by increasing inputs of root‐derived C, and reducing turnover of existing mineral‐associated C due to suppression of fungal growth and soil respiration. Together, our results provide new insights into the potential mechanisms through which multiple global change factors control soil C persistence in arid and semi‐arid grasslands. These findings suggest that the interactive effects among global change factors should be incorporated to predict the soil C dynamics under future global change scenarios.     

Optimization of a Propionibacterium acidipropionici continuous culture utilizing sucrose

To produce propionic acid and vitamin B₁₂ from sucrose, the strain Propionibacterium acidipropionici NRRL B3569 was selected by screening a number of Propionibacterium strains. The nutrient composition and the fermentation conditions for this strain were optimized in continuous culture. The investigations show that within a concentration range of 30–170 g l^–1 of sucrose in the fermentation medium, no significant substrate inhibition occurred. For the production of propionic acid and vitamin B₁₂, concentrations of 1.5 mg FeSO₄·7H₂O g^–1 dry biomass, 0.75 mg cobalt ions g^–1 dry biomass, 0.3 mg 5,6-dimethylbenzimidazole g^–1 dry biomass, and 12 g yeast extract 1^–1 were necessary additions to the sources of nitrogen, phosphate, and magnesium ions. The extra addition of up to 2.8 g betaine g^–1 dry biomass significantly increases the production of vitamin B₁₂. In the optimization of the pH value, temperature, and aeration, it was established that the conditions for propionic acid production and vitamin B₁₂ production are different. Whereas the optimal production of propionic acid took place under completely anaerobic conditions with a pH value of 6.5 and a temperature of 37°C, optimal vitamin B₁₂ production required a temperature of 40°C and aerobic conditions (0.5 vvm aeration at 100 rpm) with a pH value of 6.5.     

Effect of culturing parameters on the vegetative growth of Haematococcus alpinus (strain lcr‐cc‐261f) and modeling of its growth kinetics

The present study investigated the effect of different culture conditions on the vegetative growth of a new species, Haematococcus alpinus (strain LCR‐CC‐261f) using airlift photobioreactors. The influence of culture medium, aeration rates, CO₂ concentration in air‐gas mixture, temperature, light intensities, and wavelengths were investigated to achieve sustainable high cell density cultures. Growth parameters were determined by fitting the data to a form of the logistic equation that included a lag phase. The shear‐sensitive vegetative cells favored lower aeration rates in the photobioreactors. MLA medium increased to 40 mM nitrate produced high density cultures. Temperatures between 12°C and 18°C, 3% (v/v) CO₂ concentration and a narrow photon flux density ranging between 37 and 48 μmol photons · m^?2 · s^?1 were best suited for growth. The wavelength of the light source also impacted growth and a high cell density of 9.6 × 10⁵ cells · mL^?1 was achieved using a mixture of red and blue compared to warm white, red, or blue LEDs.     

Biomass production and energy balance of Miscanthus over a period of 11 years: A case study in a large‐scale farm in Poland

Giant miscanthus (Miscanthus × giganteus Greef and Deuter) and Amur silver grass (Miscanthus sacchariflorus Maxim./Hack) are rhizomatous grasses with a C4 photosynthetic pathway that are widely cultivated as energy crops. For those species to be successfully used in bioenergy generation, their yields have to be maintained at a high level in the long term. The biomass yield (fresh and dry matter [DM] yield) and energy efficiency (energy inputs, energy output, energy gain, and energy efficiency ratio) of giant miscanthus and Amur silver grass were compared in a field experiment conducted in 2007–2017 in North‐Eastern Poland. Both species were characterized by high above‐ground biomass yields, and the productive performance of M. × giganteus was higher in comparison with M. sacchariflorus (15.5 vs. 9.3 Mg DM ha^?1 year^?1 averaged for 1–11 years of growth). In the first year of the experiment, the energy inputs associated with the production of M. × giganteus and M. sacchariflorus were determined at 70.5 and 71.5 GJ/ha, respectively, and rhizomes accounted for around 78%–79% of total energy inputs. In the remaining years of cultivation, the total energy inputs associated with the production of both perennial rhizomatous grasses reached 13.6–15.7 (M. × giganteus) and 16.9–17.5 GJ ha^?1 year^?1 (M. sacchariflorus). Beginning from the second year of cultivation, mineral fertilizers were the predominant energy inputs in the production of M. × giganteus (78%–86%) and M. sacchariflorus (80%–82%). In years 2–11, the energy gain of M. × giganteus reached 50 (year 2) and 264–350 GJ ha^?1 year^?1 (years 3–11), and its energy efficiency ratio was determined at 4.7 (year 2) and 18.6–23.3 (years 3–11). The energy gain and the energy efficiency ratio of M. sacchariflorus biomass in the corresponding periods were determined at 87–234 GJ ha^?1 year^?1 and 6.1–14.3, respectively. Both grasses are significant and environmentally compatible sources of bioenergy, and they can be regarded as potential energy crops for Central‐Eastern Europe.     

Soft sensor based on 2D‐fluorescence and process data enabling real‐time estimation of biomass in Escherichia coli cultivations

In bioprocesses, specific process responses such as the biomass cannot typically be measured directly on‐line, since analytical sampling is associated with unavoidable time delays. Accessing those responses in real‐time is essential for Quality by Design and process analytical technology concepts. Soft sensors overcome these limitations by indirectly measuring the variables of interest using a previously derived model and actual process data in real time. In this study, a biomass soft sensor based on 2D‐fluorescence data and process data, was developed for a comprehensive study with a 20‐L experimental design, for Escherichia coli fed‐batch cultivations. A multivariate adaptive regression splines algorithm was applied to 2D‐fluorescence spectra and process data, to estimate the biomass concentration at any time during the process. Prediction errors of 4.9% (0.99 g/L) for validation and 3.8% (0.69 g/L) for new data (external validation), were obtained. Using principal component and parallel factor analyses on the 2D‐fluorescence data, two potential chemical compounds were identified and directly linked to cell metabolism. The same wavelength pairs were also important predictors for the regression‐model performance. Overall, the proposed soft sensor is a valuable tool for monitoring the process performance on‐line, enabling Quality by Design.     

A study on using fireclay as a biomass carrier in an activated sludge system

By adding a biomass carrier to an activated sludge system, the biomass concentration will increase, and subsequently the organic removal efficiency will be enhanced. In this study, the possibility of using excess sludge from ceramic and tile manufacturing plants as a biomass carrier was investigated. The aim of this study was to determine the effect of using fireclay as a biomass carrier on biomass concentration, organic removal and nitrification efficiency in an activated sludge system. Experiments were conducted by using a bench scale activated sludge system operating in batch and continuous modes. Artificial simulated wastewater was made by using recirculated water in a ceramic manufactutring plant. In the continuous mode, hydraulic detention time in the aeration reactor was 8 and 22 h. In the batch mode, aeration time was 8 and 16 h. Fireclay doses were 500, 1,400 and 2,250 mg l⁻¹, and were added to the reactors in each experiment separately. The reactor with added fireclay was called a Hybrid Biological Reactor (HBR). A reactor without added fireclay was used as a control. Efficiency parameters such as COD, MLVSS and nitrate were measured in the control and HBR reactors according to standard methods. The average concentration of biomass in the HBR reactor was greater than in the control reactor. The total biomass concentration in the HBR reactor (2.25 g l⁻¹ fireclay) in the continuous mode was 3,000 mg l⁻¹ and in the batch mode was 2,400 mg l⁻¹. The attached biomass concentration in the HBR reactor (2.25 g l⁻¹ fireclay) in the continuous mode was 1,500 mg l⁻¹ and in the batch mode was 980 mg l⁻¹. Efficiency for COD removal in the HBR and control reactor was 95 and 55%, respectively. In the HBR reactor, nitrification was enhanced, and the concentration of nitrate was increased by 80%. By increasing the fireclay dose, total and attached biomass was increased. By adding fireclay as a biomass carrier, the efficiency of an activated sludge system to treat wastewater from ceramic manufacturing plants was increased.     

Miscanthus sacchriflorus exhibits sustainable yields and ameliorates soil properties but potassium stocks without any input over a 12‐year period in China

The perennial C₄ Miscanthus spp. is used in China for bio‐fuel production and its ecological functions. However, questions arise as to its economic and environmental sustainability in abandoned farmland where the costs should be very low. Little is known about its yield performance and effects on soil properties when it was harvested annually without any inputs in China. To address these questions, an experiment was implemented for 12 years on annually harvested Miscanthus sacchariflorus planted in 2006 and managed without fertilization, irrigation, or any other inputs. We determined biomass yields each year, biomass allocation, and soil properties before and after its cultivation. Biomass yields of M. sacchariflorus reached a peak value (29.67 t/ha) 3 years after cultivation and was maintained at a stable level (averaged 22.22 t/ha) during 2012–2017. Its root shoot ratio increased due to more biomass allocated below‐ground with time. Long‐term cultivation of M. sacchariflorus increased organic carbon contents, pH (for the absence of fertilization), microbial carbon, nitrogen and phosphorus contents, and soil carbon nitrogen ratios (0–100 cm). Soil bulk density was decreased significantly (p < .05) independent of soil depths. Annual harvest did not reduce total nitrogen and phosphorus, available nitrogen, and potassium, but total the potassium content of soil (0–100 cm). Cultivation of M. sacchariflorus increased available phosphorus contents in 40–100 cm soil and reduced that value in 20–40 cm soil. Biological nitrogen fixation provided ~218.74 kg ha^?1 year^?1 (1 m depth) nitrogen for the system offsetting nitrogen export by biomass harvest and stabilizing nitrogen levels of soil. In conclusion, M. sacchriflorus exhibited sustainable biomass yields and ameliorated soil properties but the decrease of total potassium contents after 12 years’ cultivation without any input. These conclusions could provide important information timely for the government and encourage farmers to promote large‐scale utilization of M. sacchriflorus on the abandoned farmland in China.     

Fermentation and microencapsulation of the nematophagous fungus <Emphasis Type="Italic">Hirsutella rhossiliensis</Emphasis> in a novel type of hollow beads

In this work, fermentation and formulation aspects of the nematophagous fungus Hirsutella rhossiliensis BBA were investigated. When incubated in 2% (w/w) glucose and 0.5% (w/w) yeast extract medium in a 1-L Erlenmeyer flask without baffles, heavy pellet formation was observed. Only 40% of the mycelium had a size less than 500 μm. When a flask with three baffles was used, the portion of mycelium <500 μm rose to 95%. In the next step, the influence of aeration rate and stirrer speed on production of finely dispersed mycelium in a stirred tank reactor was investigated. The best fermentation results were obtained at 0.4 vvm and 400 rpm stirrer speed with 90% mycelium <500 μm and 5 g/L biomass. Then, mycelium was microencapsulated in hollow beads based on sulfoethylcellulose (SEC). Experiments on the capsule nutrient reservoir showed that 15% (w/w) corn gluten and 0.5% (w/w) yeast extract could be replaced with 3% (w/w) autoclaved baker's yeast which was never used as capsule additive before. Radial growth of mycelium out of dried hollow beads containing 1% (w/w) biomass and 3% (w/w) baker's yeast was faster than for alginate beads containing equivalent amounts of biomass and yeast indicating a higher bio-control potential.     

A C6/C5 co‐fermenting Saccharomyces cerevisiae strain with the alleviation of antagonism between xylose utilization and robustness

During second‐generation bioethanol production from lignocellulosic biomass, the desired traits for fermenting microorganisms, such as Saccharomyces cerevisiae, are high xylose utilization and high robustness to inhibitors in lignocellulosic hydrolysates. However, as observed previously, these two traits easily showed the antagonism, one rising and the other falling, in the C6/C5 co‐fermenting S. cerevisiae strain. In this study, LF1 obtained in our previous study is an engineered budding yeast strain with a superior co‐fermentation capacity of glucose and xylose, and was then mutated by atmospheric and room temperature plasma (ARTP) mutagenesis to improve its robustness. The ARTP‐treated cells were grown in 50% (v/v) leachate from lignocellulose pretreatment with high inhibitors content for adaptive evolution. After 30 days, the generated mutant LF1‐6 showed significantly enhanced tolerance, with a six‐fold increase in cell density in the above leachate. Unfortunately, its xylose utilization dropped markedly, indicating the recurrence of the negative correlation between xylose utilization and robustness. To alleviate this antagonism, LF1‐6 cells were iteratively mutated with ARTP mutagenesis and then anaerobically grown using xylose as the sole carbon source, and xylose utilization was restored in the resulting strain 6M‐15. 6M‐15 also exhibited increased co‐fermentation performance of xylose and glucose with the highest ethanol productivity reported to date (0.525 g g^?1 h^?1) in high‐level mixed sugars (80 g L^?1 glucose and 40 g L^?1 xylose) with no inhibitors. Meanwhile, its fermentation time was shortened by 8 h compared to that of LF1. During the fermentation of non‐detoxified lignocellulosic hydrolysate with high inhibitor concentrations at pH ~3.5, 6M‐15 can efficiently convert glucose and xylose with an ethanol yield of 0.43 g g^?1. 6M‐15 is also regarded as a potential chassis cell for further design of a customized strain suitable for production of second‐generation bioethanol or other high value‐added products from lignocellulosic biomass.     

Industrial scale of optimization for the production of carboxymethylcellulase from rice bran by a marine bacterium, Bacillus subtilis subsp. subtilis A-53

Rice bran and yeast extract were found to be the best combination of carbon and nitrogen sources for the production of carboxymethycellulase (CMCase) by Bacillus subtilis subsp. subtlis A-53. Optimal concentrations of rice bran and yeast extract for the production of CMCase were 5.0% (w/v) and 0.10% (w/v), respectively. Optimal temperature and initial pH of medium for cell growth of B. subtilus subsp. subtilis A-53 were 35 °C and 7.3, whereas those for the production of CMCase by B. subtilus subsp. subtilis A-53 were 30 °C and 6.8. Optimal agitation speed and aeration rate in a 7 L bioreactor were 300 rpm and 1.0 vvm, respectively. The optimal agitation speed and aeration rate for the production of CMCase by B. subtilus subsp. subtilis A-53 were lower than those for cell growth. The highest productions of CMCase by B. subtilus subsp. subtilis A-53 in 7 and 100 L bioreactors were 150.3 and 196.8 U mL⁻¹, respectively.     

Antioxidant property and production of exopolysaccharide from Armillaria mellea in submerged cultures: Effect of culture aeration rate

Effects of culture aeration rate on production and antioxidant property of exopolysaccharide (EPS) by Armillaria mellea were investigated in a 5‐L stirred‐tank bioreactor where an optimal biomass aeration rate of 1.2 vvm with 0.22 g/g cell yield and 0.6 vvm EPS formation rate with 7.66 mg/g product yield were achieved. A two‐stage aeration process to maximize the biomass and EPS productions proceeded with a 1.55‐fold enhancement (from 4.28 to 6.65 g/L) in biomass formation and a 2.68‐fold enhancement (from 86.9 to 233.2 mg/L) in the EPS production, as compared with those from the aeration rate of 0.3 vvm. The molecular weights of EPS in cultures of different aeration rates are closely correlated with their protein/polysaccharide ratios (R²=0.830) and EC₅₀ (EC₅₀, the effective concentration where the antioxidant property is 50%) values in antioxidant activity (R²=0.960), reducing power (R²=0.894) and chelating ability (R²=0.954). EPS from the two‐stage aeration rate culture shows a strong antioxidant property by the conjugated diene method, reducing power and chelating ability on ions. Therefore, we present results to regulate and to optimize A. mellea cultures to efficiently produce biomass and EPS. The fermented EPS has the potential to be used as for antioxidant‐related functional foods and pharmaceutical industries.     

Investigation of the potential of biocalorimetry as a process analytical technology (PAT) tool for monitoring and control of Crabtree-negative yeast cultures

Biological reaction calorimetry, also known as biocalorimetry, has led to extensive applications in monitoring and control of different bioprocesses. A simple real-time estimator for biomass and growth rate was formulated, based on in-line measured metabolic heat flow values. The performance of the estimator was tested in a unique bench-scale calorimeter (BioRC1), improved to a sensitivity range of 8 mW l^− 1 in order to facilitate the monitoring of even weakly exothermic biochemical reactions. A proportional–integral feedback control strategy based on these estimators was designed and implemented to control the growth rate of Candida utilis, Kluyveromyces marxianus and Pichia pastoris by regulating an exponential substrate feed. Maintaining a particular specific growth rate throughout a culture is essential for reproducible product quality in industrial bioprocesses and therefore a key sequence for the step from quality by analysis to quality by design. The potential of biocalorimetry as a reliable biomass monitoring tool and as a key part of a robust control strategy for aerobic fed-batch cultures of Crabtree-negative yeast cells in defined growth medium was investigated. Presenting controller errors of less than 4% in the best cases, the approach paves the way for the development of a generally applicable process analytical technology platform for monitoring and control of microbial fed-batch cultures.     

Optimization of fermentation parameters for the biomass and DHA production of Schizochytrium limacinum OUC88 using response surface methodology

Fermentation parameters for biomass and DHA production of Schizochytrium limacinum OUC88 in a fermenter (working volume 7 L) were optimized using Plackett–Burman and central composite rotatable design. Out of 10 factors studied by Plackett–Burman design, 4 influenced the biomass production significantly. Central composite rotatable design was used to optimize the significant factors and response surface plots were generated. Using these response surface plots and point prediction, optimized values of the factors were determined as follows temperature (°C) 23 °C, aeration rate 1.48 L min⁻¹ L⁻¹, agitation 250 rpm and inoculum cells in mid-exponential phase, the maximum yield of DCW and DHA were 24.1 and 4.7 g L⁻¹, respectively. These predicted values were also verified by validation experiments.