Production of Beta-Carotene from Synthetic Medium by Blakeslea trispora in Fed-batch Culture

Abstract

The production of β-carotene from a synthetic medium by Blakeslea trispora in fed-batch culture was investigated. A maximum β-carotene concentration of 85.0?mg?L^?1 with productivity of 0.16?mg?L^?1?h^?1 and specific β-carotene production rate of 0.01?mg?g^?1?h^?1 was obtained by feeding the cells constantly with olive oil, cottonseed oil, soybean oil, antioxidant, and low concentration of growth factors at feeding rate of 4.2?mL?h^?1 from the beginning of the fermentation. In this case, the fed-batch culture supported high values of biomass dry weight (11.0?g?L^?1) and sugar utilization (0.976?g?g^?1). The morphology of the fungus was studied during growth in fed-batch fermentation system using an image analysis system. Zygospores are the morphological forms, which are responsible for the production of the pigment. The highest percentage of zygospores (11.44%) was correlated with the highest percentage of intracellular β-carotene (0.72%) in the total biomass dry weight. Moreover, high percentages of vacuolated hyphae, evacuated cells, and degenerated hyphae of the microorganism were observed. This was due to the formation of high amount of H₂O₂ by exposure of the microorganism to high dissolved oxygen concentration.     

Evaluation of the conditions of carotenoids production in a synthetic medium by Sporidiobolus salmonicolor (CBS 2636) in a bioreactor

This work studied the cultivation conditions for the production of carotenoids by Sporidiobolus salmonicolor (CBS 2636) in a bioreactor. A Plackett–Burman design was used for the screening of the most important factors, followed by a complete second order design, to maximise the concentration of total carotenoids. The maximum concentration of 3425.9 μg L^?1 of total carotenoids was obtained in a medium containing 80 g L^?1 glucose, 15 g L^?1 peptone and 5 g L^?1 malt extract, with an aeration rate 1.5 vvm, 180 r.p.m., 25 °C and an initial pH of 4.0. Fermentation kinetics showed that the maximum concentration of total carotenoids was reached after 90 h of fermentation. Carotenoid bio‐production was partially associated with cell growth. The specific carotenoid production (Y_P/X) was 238 μg carotenoids/g cells, whereas Y_P/S (substrate to product yield) was 41.3 μg g^?1. The specific growth rate (μ_x) was 0.045 h^?1. The highest cell and total carotenoid productivity were 0.19 g L^?1 h^?1 and 56.9 μg L^?1 h^?1, respectively.     

Xylitol bioproduction from wheat straw: hemicellulose hydrolysis and hydrolyzate fermentation

A 2² central composite design with five center points was performed to estimate the effects of temperature (120, 130 and 140 °C) and acid loading (100, 150 and 200 mg g^?1) on the yield of monomeric xylose recovery from wheat straw hemicellulose (Y_S/RM). Under the best hydrolysis condition (140 °C and 200 mg g^?1), a Y_S/RM of 0.26 g g^?1 was achieved. After vacuum concentration and detoxification by pH alteration and active charcoal adsorption, the hydrolyzate was used as source of xylose for xylitol bioproduction in a stirred tank reactor. A xylitol production of 30.8 g L^?1 was achieved after 54 h^?1 of fermentation, resulting in a productivity (Q_P) of 0.57 g L^?1 h^?1 and bioconversion yield (Y_P/S) of 0.88 g g^?1. The maximum specific rates of xylose consumption and xylitol production were 0.19 and 0.15 g g^?1 h^?1, respectively. Copyright © 2006 Society of Chemical Industry     

Relative Oxidative Stability of Diacylglycerol and Triacylglycerol Oils

To compare the oxidative stability between diacylglycerol (DAG) oil and conventional triacylglycerol (TAG) oil (that is, soybean oil), the prepared stripped diacylglycerol oil (SDO) and soybean oil (SSBO) were stored at 60 °C in the dark for 144 h. During storage peroxide values (POVs), contents of aldehydes, unsaturated fatty acids were measured to evaluate the oxidative stabilities of the 2 oils. The results showed the content of C18:2, C18:3, and total unsaturated fatty acid decreased faster in DAG oil than in soybean oil, whereas the decreased rate of C18:1 was similar in 2 oils. Also, both rate constants (K₁ and K₂) obtained from POV (K₁) and total aldehydes (K₂) indicated that DAG oil (K₁ = 3.22 mmol/mol FA h^?1, K₂ = 0.023 h^?1) was oxidized more rapidly than soybean oil (K₁ = 2.56 mmol/mol FA h^?1, K₂ = 0.021 h^?1), which was mainly due to the difference of acylglycerol composition of the 2 oils along with higher C18:3 (9.6%) in SDO than SSBO (5.7%). It is concluded that DAG was more easily oxidized than soybean oil at 60 °C in the dark for 144 h.     

UPLC-ESI-MS/MS for determining trans- and cis-vitamin K1 in infant formulas: method and applications

A simple and sensitive method of determining vitamin K₁ isomers (cis-and trans-forms) was developed for routine monitoring of supplemented infant formulas. The proposed method involves the applications of the lipase hydrolysis and the liquid-to-liquid extraction. After the hydrolysis and extraction procedures, the vitamin K₁-enriched extract was directly injected into the ultra-performance liquid chromatography–tandem mass spectrometry system (UPLC-MS/MS). The components were detected using electrospray ionization (ESI) in positive-ion and quantified by multiple-reaction monitoring (MRM) mode. trans-vitamin K₁ was separated from the biologically inactive cis-isomer through a C₃₀ column (4.6?×?150?mm, 3?μm), and vitamin K₁-d₇ was used as an internal standard. Calibration curves were linear over the range of 9.3–464.75?ng?ml^?1 for trans-vitamin K₁ (r ²?>?0.999) and 1.71–85.25?ng?ml^?1 for cis-vitamin K₁ (r ²?>?0.999). The limit of detection for trans- and cis-vitamin K₁ was 0.011?μg 100?g^?1 and 0.01?μg 100?g^?1. The limit of quantification was 0.037?μg 100?g^?1 and 0.031?μg 100?g^?1, respectively. The intra- and inter-batch variations (RSD%) were less than 5?%. The proposed method was applied to determine vitamin K₁ isomers in milk-based, soy-based and rice-based infant formulas, and the cis-vitamin K₁ isomer contributes to 7.05–17.21?% of the total vitamin K₁ in certain infant formulas.     

Integral utilisation of barley husk for the production of food additives

A new and effective chemical–biotechnological process for the global utilisation of barley husk (obtained from the spent grains in the brewing process) is reported. With the proposed process the three main components of the lignocellulosic residue (cellulose, hemicellulose and lignin) are utilised. A first treatment with sulfuric acid (pre‐hydrolysis) allowed the solubilisation of hemicelluloses to give xylose and glucose‐containing liquors (suitable to make fermentation media for the continuous lactic acid (LA) production with L. pentosus) and a solid phase containing cellulose and lignin. In this set of experiments, a maximum volumetric productivity (Q_P) = 2.077 g L^?1 h^?1 and product yield (Y_P/S) = 0.62 g g^?1 were obtained for a dilution rate of 0.01 h^?1. The solid residues from pre‐hydrolysis were treated with NaOH in order to increase their cellulase digestibility, and dissolve the lignin content. The cellulose residue was used as substrates for lactic acid production by simultaneous saccharification and fermentation (SSF) in media containing Trichoderma reesei cellulases and Lactobacillus rhamnosus cells using the complete MRS broth or a cheaper medium. In both cases similar LA concentrations and volumetric productivities were achieved (P = 73.4–71.0 g L^?1 and Q_P = 1.28–1.25 g L^?1 h^?1, respectively), where P is LA concentration. The lignin solution obtained after the alkaline treatment was extracted with ethyl acetate in order to obtain the phenolic components. The extract obtained at pH 3 showed three times more antioxidant activity than the one extracted at pH 12.8, with an EC₅₀ of 1.396 g L^?1 for pH 3 and 4.604 g L^?1 for pH 12.8. The best extracts showed twice antioxidant activity than BHT. Copyright © 2007 Society of Chemical Industry     

EFFECT OF PHYSICAL FACTORS ON ACETIC ACID PRODUCTION IN BRETTANOMYCES STRAINS

Four species of Brettanomyces (intermedius, bruxellensis, custersianus, clausenii) were examined to ascertain their acetic acid production capacity. The results showed that B. bruxellensis was the strain with the best volumetric productivity and specific production rate (P = 0.065 gL^?1 h^?1; V_p = 0.43 gg^?1h^?1). The best kinetic parameters were reached (P = 0.133 gL^?1 h^?1; Y_p/s = 0.23; P_max = 11.64 gL^?1) at an airflow of 288 Lh^?1 (0.6 vvm, OTR = 124 mgO₂L^?1 h^?1), and substrate inhibition was not observed. The influence of temperature and agitation on acetic acid production by B. bruxellensis in a glucose medium was investigated at different levels, 26, 30, 34C and 250, 350, 450 rpm, respectively. Temperature and agitation were shown to be deci‐sive factors (P < 0.05) in acetic acid production at 288 Lh^?1(0.6 vvm, OTR = 124 mgO₂L^?1 h). The optimal conditions for a high volumetric productivity were 30C and 250 rpm, respectively.     

Physiological approach to heterologous human serum albumin production by Kluyveromyces lactis in chemostat culture

Production of recombinant human serum albumin (rHSA) controlled by the constitutive promoter phosphoglycerate kinase was studied in Kluyveromyces lactis. It was governed by both cell concentration and glycolytic flow. The triggering of the fermentation metabolism by unfavourable culture conditions (pH, pO₂, D) caused a decrease in the synthesis of the heterologous protein. The highest productivity (75 mg 1^?1 per h) and rHSA concentration (62 mg 1^?1) were obtained in chemostat culture with a dilution rate of 0·12 h^?1 and with 38 g 1^?1 dry weight.     

Ultrasound‐assisted extraction of ginsenosides in supercritical CO2 reverse microemulsions

Ultrasound‐assisted extraction of ginsenosides from ginseng in supercritical CO₂ reverse microemulsions formed by bis(2‐ethylhexyl) sodium sulfosuccinate (AOT) was studied. Prior to extraction the ginseng was soaked in water for 12 h. It was found that ultrasound significantly enhanced supercritical CO₂ reverse microemulsion extraction. The ginsenoside extraction yield from supercritical CO₂ reverse microemulsion with ultrasound of 20 kHz, 15.2 W cm^?2 and 3/6 s was 2.63 times that without ultrasound at 24 MPa extraction pressure, 45 °C extraction temperature, 4 h extraction time, 5 MPa separation pressure, 55 °C separation temperature and 2 L h^?1 CO₂ flow rate with 140 mL of 0.07 mol L^?1 AOT/ethanol. The maximum extraction yields of ginsenosides from different concentrations of reverse microemulsions were obtained at different ultrasonic intensities. The extraction yield with 20 kHz ultrasound was higher than that with 38 kHz ultrasound at suitably low intensity; however, it was lower at high intensity. The yield improvement may be basically attributed to the mechanical and thermal effects of ultrasound. Copyright © 2006 Society of Chemical Industry     

Assessment of the Influence of the Decanter Set-up During Continuous Processing of Olives at Different Pigmentation Index

New generation decanters allow operators to make real-time adjustments during the virgin olive oil extraction process in order to gain the best performance possible. However, the opportunity to act in line requires a deep understanding of the consequences of changing machine parameters. To this purpose, an experiment was carried out at industrial scale. The decanter feed rate (Fr), ranging from 4075 to 5820 kg h^?1, the bowl/screw conveyor differential speed (?n), set at 18 and 22, and two ripening degrees of the olives were considered as process variables. Two combinations Fr-?n, namely 4620 kg h^?1 at ?n-18 and 5210 kg h^?1 at ?n-22, were found to similarly maximize the process efficiency, regardless the raw material features. After pointing out the best working settings, the corresponding virgin olive oils were compared. The analysis of variance showed that peroxide value, K ₂₃₂, K ₂₇₀, phenols, chlorophylls, β-tocopherol, fruity and bitter notes, and C6 volatile compounds were significantly affected by the machine parameters. An inverse proportionality was observed between the combination Fr-?n and the phenolic compounds. On the whole, the sampling factor exerted a larger influence on the product quality than the decanter set-up.     

Production of (2E)-hexenal by a hydroperoxide lyase from Amaranthus tricolor and salt-adding steam distillation for the separation

(2E)-Hexenal is widely used in flavors and perfumes; however, it is mainly derived from chemical synthesis. In this study, hydroperoxide lyase (HPL) naturally originated from Amaranthus tricolor was used to catalyze the reaction to generate (2E)-hexenal from 13-hydroperoxy-9Z, 11E, 15 Z-octadecatrienoic acid (13-HPOT), and salt-adding steam distillation was used for the separation of (2E)-hexenal. A maximum yield of (2E)-hexenal that reached 1,156.4?mg?L^?1 was obtained with a high substrate concentration (40?mM 13-HPOT) under the following conditions: 10?min, pH 7.5, 20?°C, HPL 16 U?mL^?1, butylated hydroxytoluene (BHT) 1?mM, and dithiothreitol (DTT) 15?mM. Then the separation of (2E)-hexenal was conducted by salt-adding steam distillation. It was found that AlCl₃ had the greatest effect on the separation of (2E)-hexenal, followed by CaCl₂ and NaCl. The distillate yield with the addition of AlCl₃ was 93.2?%, while the distillate yield without the addition of salt was 81.2?%. The distillate concentration with AlCl₃ was 6,464.2?mg?L^?1, while the distillate concentration without the addition of salt was 4,490.3?mg?L^?1. The addition of salt improved the efficiency of the steam distillation. This study was greatly meaningful for providing a green method to the large-scale production of (2E)-hexenal.     

Microchannel emulsification: A promising technique towards encapsulation of functional compounds

ABSTRACT

This review provides an overview of microchannel emulsification (MCE) for production of functional monodispersed emulsion droplets. The main emphasis has been put on functional bioactives encapsulation using grooved-type and straight-through microchannel array plates. MCE successfully encapsulates the bioactives like β-carotene, oleuropein, γ-oryzanol, β-sitosterol, L-ascorbic acid and ascorbic acid derivatives, vitamin D and quercetin. These bioactives were encapsulated in a variety of delivery systems like simple and multiple emulsions, polymeric particles, microgels, solid lipid particles and functional vesicles. The droplet generation process in MCE is based upon spontaneous transformation of interfaces rather than high energy shear stress systems. The scale-up of MCE can increase the productivity of monodispersed droplets >100 L h^?1 and makes it a promising tool at industrial level.     

Continuous beer fermentation – diacetyl as a villain

This work tested the viability of producing beer of good quality after maturation of green beer obtained by primary continuous fermentation of high‐gravity wort using an airlift bioreactor with flocculated biomass. Fermentation performance of the tested setup was unique as it reached a maximum saccharide consumption rate of 9.43 g L^?1 h^?1 and an ethanol productivity of 3.75 g L^?1 h^?1. Despite the high levels of diacetyl present in the green beer, a regular maturation was able to reduce it to below threshold values in up to 15 days. It was observed that diacetyl production was strongly correlated with wort composition injected into the system, rather than with the large amount of biomass immobilized in the bioreactor (up to 727 × 10⁶ cell mL^?1). Organoleptic tests showed that the maturated beer had no major defects. Copyright © 2015 The Institute of Brewing & Distilling     

Selection of lactic acid bacteria for acidification of brown juice (grass juice), with the aim of making a durable substrate for L‐lysine fermentation

BACKGROUND: Brown juice is a waste product from the green crop‐drying industry. Heating and pressing of green crops prior to drying produces a juice rich in nutrients, which can be converted to a stable universal fermentation medium by lowering its pH to 4.0–4.5 via lactic acid fermentation. The aim of this study was to select a strain of lactic acid bacteria for industrial acidification of brown juice. RESULTS: Several strains were tested in fermentation experiments and Lactobacillus salivarius ssp. salivarius DSM 20555 was found to be the best choice. It showed high growth rates at temperatures ranging from 40 to 48 °C, with an optimum temperature of 46 °C (µ_max = 2.2 h^?1), and maintained a high productivity at pH 4.0 in continuous fermentation. The highest productivity of 7.23 g L^?1 h^?1 was found at a dilution rate of 1.0 h^?1. CONCLUSION: With today's increased focus on utilisation of residues from food and agro‐industry, coupled with restrictions on their use as fertiliser for farmlands, lactic acid fermentation could play a significant role in conservation of these nutrient‐rich liquids. This study shows that Lb. salivarius ssp. salivarius DSM 20555 is a very promising micro‐organism for use in such a process. Copyright © 2008 Society of Chemical Industry     

The Effect of the Electric Field on Lag Phase, β-Galactosidase Production and Plasmid Stability of a Recombinant Saccharomyces cerevisiae Strain Growing on Lactose

Ethanol and ??-galactosidase production from cheese whey may significantly contribute to minimise environmental problems while producing value from low-cost raw materials. In this work, the recombinant Saccharomyces cerevisiae NCYC869-A3/pVK1.1 flocculent strain expressing the lacA gene (coding for ??-galactosidase) of Aspergillus niger under ADHI promoter and terminator was used. This strain shows high ethanol and ??-galactosidase productivities when grown on lactose. Batch cultures were performed using SSlactose medium with 50?g?L^?1 lactose in a 2-L bioreactor under aerobic and microaerophilic conditions. Temperature was maintained at 30?°C and pH?4.0. In order to determine the effect of an electric field in the fermentation profile, titanium electrodes were placed inside the bioreactor and different electric field values (from 0.5 to 2?V?cm^?1) were applied. For all experiments, ??-galactosidase activity, biomass, protein, lactose, glucose, galactose and ethanol concentrations were measured. Finally, lag phase duration and specific growth rate were calculated. Significant changes in lag phase duration and biomass yield were found when using 2?V?cm^?1. Results show that the electric field enhances the early stages of fermentation kinetics, thus indicating that its application may improve industrial fermentations?? productivity. The increase in electric field intensity led to plasmid instability thus decreasing ??-galactosidase production.     

Effects of pH and dissolved oxygen on the synthesis of γ‐glutamyltranspeptidase from Bacillus subtilis SK 11.004

BACKGROUND: γ‐Glutamyltranspeptidase (GGT; EC 2.3.2.2) is a widely distributed enzyme that is of interest in the food industry. In this study the effects of pH and dissolved oxygen (DO) on GGT synthesis from Bacillus subtilis SK 11.004 were investigated. RESULTS: GGT production increased to 0.5 U mL^?1 when the pH value was controlled at 6.5. The control of a single DO level revealed that the highest specific growth rate (3.42 h^?1) and GGT production rate (0.40 U g^?1 mL^?1) were obtained at DO levels of 40 and 10% respectively. To satisfy the different oxygen demands at different stages of cell growth and GGT synthesis, a stage DO level control strategy was designed as follows: 40% from 0 to 4 h, 30% from 4 to 6 h and 10% from 6 to 18 h. Furthermore, the maximum biomass (2.27 g L^?1) and GGT production (3.05 U mL^?1) could be obtained using a fermentation strategy combining a constant pH value with stage DO level control. CONCLUSION: The proposed fermentation strategy resulted in a 13.7‐fold increase in GGT production. This finding should be of great importance for the industrial production of GGT. Copyright © 2011 Society of Chemical Industry     

Micellar Electrokinetic Chromatography Method for Determination of the Ten Water-Soluble Vitamins in Food Supplements

The separation and determination of the ten water-soluble vitamins by using capillary electrophoresis in the micellar electrokinetic chromatography in a single run are proposed. The method uses low toxicity and cost solvent (ethanol) as modifier of background electrolyte (BGE) attending to the Green Chemistry principles. The electrophoretic method uses 10.0 % (v/v) ethanol, 2.0 % (w/v) SDS, 0.02 mol?L^?1 borate at pH 8.70 as BGE. The standard and real sample solutions were injected in the eletrophoretic system by hydrodynamic injection under pressure of 0.80 psi for 8 s, and the separation was carried out in a fused silica capillary under a potential of 28 kV at 25 °C; the analytical signals were monitored at 214 nm. The analytical method is precise (r.s.d.?<?6 %), accurate (better than 9 %), selective, sensitive, robust, simple, and presents high analytical frequency as ten water-soluble vitamins were separated in only 18 min, with migration times of 5.75?±?0.02, 6.81?±?0.02, 8.13?±?0.04, 8.80?±?0.07, 8.98?±?0.06, 11.10?±?0.08, 11.34?±?0.05, 13.85?±?0.15, 14.82?±?0.04, and 17.85?±?0.30 min. Detection and quantification limits of 0.34, 0.32, 0.27, 0.20, 2.50, 4.98, 4.92, 0.30, 0.86 and 0.28 mg?L^?1 and 1.02, 0.97, 0.83, 0.62, 7.56, 15.09, 14.91, 0.90, 2.59 and 0.83 mg?L^?1, for vitamins PP (nicotinamide), B₁₂ (cyanocobalamin), B₂ (riboflavin), B₆ (pyridoxine), B₈ (biotin), C (ascorbic acid), B₅ (pantothenic acid), B₃ (nicotinic acid), B₁ (thiamine), and B₉ (folic acid), respectively. Excellent recoveries (intra and inter-day) were obtained and, when the method was applied to food supplement analyses the results were in agreement with the conventional HPLC methods.     

Substrate specificity of a recombinant d-lyxose isomerase from Providencia stuartii for monosaccharides

The specific activity and catalytic efficiency (k_cat/K_m) of the recombinant putative protein from Providencia stuartii was the highest for d-lyxose among the aldose substrates, indicating that it is a d-lyxose isomerase. Gel filtration analysis suggested that the native enzyme is a dimer with a molecular mass of 44 kDa. The maximal activity for d-lyxose isomerization was observed at pH 7.5 and 45 °C in the presence of 1 mM Mn²⁺. The enzyme exhibited high isomerization activity for aldose substrates with the C2 and C3 hydroxyl groups in the left-hand configuration, such as d-lyxose, d-mannose, l-ribose, d-talose, and l-allose (listed in decreasing order of activity). The enzyme exhibited the highest activity for d-xylulose among all pentoses and hexoses. Thus, d-lyxose was produced at 288 g/l from 500 g/l d-xylulose by d-lyxose isomerase at pH 7.5 and 45 °C for 2 h, with a conversion yield of 58 % and a volumetric productivity of 144 g l^− 1 h^− 1. The observed k_cat/K_m (920 mM^− 1 s^− 1) of P. stuartiid-lyxose isomerase for d-xylulose is higher than any of the k_cat/K_m values previously reported for sugar and sugar phosphate isomerases with monosaccharide substrates. These results suggest that the enzyme will be useful as an industrial producer of d-lyxose.     

The relationship between muscle fiber characteristics and meat quality traits of highly marbled Hanwoo (Korean native cattle) steers

To investigate the relationships between muscle fiber characteristics and meat quality traits of Korean native cattle, Hanwoo, Longissimus dorsi (LD), Psoas major (PM) and Semimembranosus (SM) muscles obtained from 18 Hanwoo steers and the muscle fiber characteristics were measured by histochemical analysis. Fiber number, area percentages and density of type IIA and IIB were lower in SM muscle, but higher in PM muscle than other muscles. LD muscle had higher pH_24h, L^? value and fat content whereas SM muscle had lower L^? value and fat content. The lowest WBSF with longer sarcomere length was observed in PM muscle, while SM muscle showed the highest WBSF with shorter sarcomere length. Consequently, the percentage of type I and IIB were highly correlated with meat quality traits and inversely correlated with fat content, L^? value and WBSF. Fiber number and area percentage of type I had a positive correlation with fat content and L^? value and a negative correlation with WBSF. These results suggest that Hanwoo steers had high marbling, more lightness and tenderness when the percentage of type I was high and the percentage of type IIB was low in muscle.     

Enzymic production of feruloyl xylo-oligosaccharides from corn cobs by a family 10 xylanase from Thermoascus aurantiacus

Feruloyl xylo-oligosaccharides (FeXOSs) were obtained from corn cobs (CCs) by treatment with a Thermoascus aurantiacus family 10-β-d-endoxylanase. CCs subjected to a sonication (ST-CC) or an autoclave treatment in order to enhance enzymic access and FeXOS production. Enzymic FeXOS production was increased by CCs thermal treatment up to 3.5 times. The kinetic parameters K_E and V_max, describing enzyme-dependent production rates of FeXOSs from CCs, were estimated at 130±20 nM and 290±10 nM h⁻¹, respectively. The reaction parameters affecting the FeXOS production, such as substrate concentration, enzyme loading and reaction time, have been investigated. The maximum FeXOS production was observed after 24 h incubation of 100 g L⁻¹ AT-CC with 570 nM xylanase. The obtained yield was 12 μmol of FeXOSs per gram of CC.