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.     

Xanthan gum produced by Xanthomonas campestris from cheese whey: production optimisation and rheological characterisation

BACKGROUND: This aim of this study was the production and rheological characterisation of xanthan gum by Xanthomonas campestris pv. mangiferaeindicae IBSBF 1230 using industrial media and experimental design techniques in a bench bioreactor. RESULTS: The optimised conditions for the production of xanthan starting with 900 mL of cheese whey were 1 g L^?1 magnesium sulphate, 20 g L^?1 potassium phosphate, 28 °C temperature and initial pH 7.2 at 390 rpm agitation and 1.5 vvm aeration, resulting in 36 g L^?1 gum in 72 h. The highest viscosity obtained in the production optimisation study was 1831.34 mPa s at 25 °C with 30 g L^?1 gum. The use of CaCl₂ resulted in the highest solution viscosity under conditions of 25 °C, 1 g L^?1 salt and 46.8 g L^?1 gum, with a value of 1704.53 mPa s. CONCLUSION: In this study, cheese whey, a by‐product of the dairy industry, was used as substrate in the production of xanthan gum, a valuable product in food applications, with optimised high gum production in a bioreactor and a wide range of viscosity values. Copyright © 2009 Society of Chemical Industry     

Evaluation of aeration and substrate concentration on the production of carotenoids by <Emphasis Type="Italic">Sporidiobolus salmonicolor</Emphasis> (CBS 2636) in bioreactor

This study aimed at optimising the cultivation conditions for the production of carotenoids by Sporidiobolus salmonicolor (CBS 2636) in a bioreactor. The maximum content of total carotenoids in the full factorial design 22 was 3131.3 μg/L in synthetic medium with 80 g/L of glucose, 15 g/L of peptone, 5 g/L of malt extract, aeration of 1.5 vvm, agitation of 180 rpm, initial pH of 4.0 at 25 °C. In the kinetic study, we could observe that the bioproduction of carotenoids is associated with cell growth in the exponential phase, and the average specific growth (μ) in bioreactor is 0.046 h⁻¹ with a maximum yield of 0.19 g cells/L h. The maximum yield of carotenoids (60.0 μg/Lh) is observed at 50-h bioproduction. The conversion factor for total organic carbon (TOC) in cells (Y_X/_SCOT) was 2.97 g/g (0–50 h) and 0.254 g/g (50–100 h), the conversion factor glucose into cells (Y_X/Sglicose) was 0.168 g/g (0–100 h). The specific production of carotenoids (Y_P/_X) was 390 μg of carotenoids per gram of cells, the conversion factor of carbon in the product (Y_P/SCOT) was 107.8 μg/g (0–50 h) and 34.4 μg/g (50–100 h), whereas the factor Y_P/_Sglicose was 69.59 μg/g. The agitation and aeration provided better homogeneity in the culture medium, and hence greater availability of nutrients and oxygen, leading to higher production of carotenoids.     

Optimization of the Production of Total Carotenoids by Sporidiobolus salmonicolor (CBS 2636) Using Response Surface Technique

This work aimed at evaluating the conditions of growth and the recovery of total carotenoids produced by Sporidiobolus salmonicolor (CBS 2636). Optimization of carotenoid production was achieved by experimental design technique. A Plackett–Burman design was used, followed by a complete second-order design, to optimize the concentration of total carotenoids in a conventional medium. Maximum concentration of 1,019 μg l⁻¹ of total carotenoids was obtained in a medium containing 40 g l⁻¹ glucose, 10 g l⁻¹ malt extract, and 14 g l⁻¹ peptone, at 180 rpm, 25 °C, and initial pH of 4.0. So far, no previous systematic study using microorganisms of the genus Sporobolomyces (formerly Sporidiobolus) for production of carotenoids has been reported. In this study, very good yields of carotenoids (1 mg l⁻¹) could be obtained after optimization of fermentation medium and operation conditions.     

Chemical composition and sensory analysis of cheese whey‐based beverages using kefir grains as starter culture

The aim of the present work was to evaluate the use of the kefir grains as a starter culture for tradicional milk kefir beverage and for cheese whey‐based beverages production. Fermentation was performed by inoculating kefir grains in milk (ML), cheese whey (CW) and deproteinised cheese whey (DCW). Erlenmeyers containing kefir grains and different substrates were statically incubated for 72 h at 25 °C. Lactose, ethanol, lactic acid, acetic acid, acetaldehyde, ethyl acetate, isoamyl alcohol, isobutanol, 1‐propanol, isopentyl alcohol and 1‐hexanol were identified and quantified by high‐performance liquid chromatography and GC‐FID. The results showed that kefir grains were able to utilise lactose in 60 h from ML and 72 h from CW and DCW and produce similar amounts of ethanol (～12 g L^?1), lactic acid (～6 g L^?1) and acetic acid (～1.5 g L^?1) to those obtained during milk fermentation. Based on the chemical characteristics and acceptance in the sensory analysis, the kefir grains showed potential to be used for developing cheese whey‐based beverages.     

Assessment of Cell Disruption and Carotenoids Extraction from Sporidiobolus salmonicolor (CBS 2636)

The increasing demand for carotenoids by industries has drawn attention to their bio-production. Since pigments are intracellular, extraction steps are then needed after cell cultivation. In this work, different strategies for extraction of carotenoid pigments from Sporidiobolus salmonicolor (CBS 2636) were studied. Different solvents (dimethyl sulfoxide, petroleum ether, hexane, ethyl acetate, chloroform, and acetone), liquid N₂, and diatomaceous earth were used to disrupt the cell and thus release the intracellular carotenoids. The results of this study showed that when multiple solvents were used, a synergistic effect on the extent of carotenoids recovery was obtained. Maximum concentration of total carotenoids (913 μg/L) was obtained in the treatment using liquid N₂ and dimethyl sulfoxide to disrupt the cell, followed by the extraction with a solution of acetone/methanol (7:3, v/v).     

Assessment of Carotenoids Recovery through Cell Rupture of <Emphasis Type="Italic">Sporidiobolus salmonicolor</Emphasis> CBS 2636 Using Compressed Fluids

The increasing demand for carotenoids by industries has drawn attention to their bio-production. Since pigments are intracellular, extraction steps are then needed after cell cultivation. In this work, different strategies for extraction of carotenoid pigments from Sporidiobolus salmonicolor (CBS 2636) were investigated. The cell rupture was carried out using dimethyl sulfoxide (DMSO), two pure compressed fluids, supercritical carbon dioxide and propane, and also a combination of pressurized fluid treatment followed by liquid DMSO. Dichloromethane, ethanol, ethyl acetate, and acetone were tested for the carotenoids extraction. Results obtained show that when multiple solvents were used a synergetic effect on the extent of carotenoids recovery was verified. Maximum concentration of total carotenoids (2,875 μg/L) was obtained in the treatment using supercritical CO₂ (300 bar/120 min) followed by dimethyl sulfoxide to disrupt the cell, and then the extraction with a solution of acetone/methanol (7:3, v/v).     

Biotechnological potential of industrial wastes for economical citric acid bioproduction by Aspergillus niger through submerged fermentation

Submerged citric acid (CA) bioproduction was carried out by Aspergillus niger NRRL‐567 using various industrial wastes, such as brewery spent liquid (BSL), lactoserum and starch industry water sludge. CA bioproduction was carried out by varying the temperature (25–35 °C), pH (3–5), addition of inducers, incubation time and supplementation with different proportions of apple pomace ultrafiltration sludge (APS). The results indicated that under the best conditions with 3% (v/v) methanol, the optimal concentration of 11.34 g L^?1 CA was recorded using BSL at pH 3.5 and temperature 30 °C after 120‐h incubation period. Supplementation of methanol resulted in an increase of 56% CA production. Meanwhile, under similar conditions, higher concentration of 18.34 g L^?1 CA was reported with the supplementation of BSL with 40% (v/v) APS having suspended solids concentration of 30 g L^?1. The present study demonstrated the potential of BSL supplemented with APS as an alternative cheap substrate for CA fermentation.     

Fractionation of angiotensin I converting enzyme inhibitory activity from pea and whey protein in vitro gastrointestinal digests

In vitro gastrointestinal digestion of pea and whey protein produced high angiotensin I converting enzyme (ACE) inhibitory activity with IC₅₀ values of 0.070 and 0.041 mg protein ml^?1 respectively. Ultrafiltration/centrifugation using a membrane with a molecular weight cut‐off of 3000 Da decreased the IC₅₀ value to 0.055 mg protein ml^?1 for pea permeate and 0.014 mg protein ml^?1 for whey permeate. Further fractionation by reverse phase HPLC gave IC₅₀ values as low as 0.016 mg protein ml^?1 for pea and 0.003 mg protein ml^?1 for whey. Consequently, these purification steps enriched the ACE inhibitory activity of the pea digest more than four times and that of the whey digest more than 13 times. HPLC profiles after digestion and ultrafiltration indicate that high ACE inhibitory activity is due to short and more hydrophobic peptides. The results also suggest that potent ACE inhibitory peptides were present alongside low active peptides in whey hydrolysate, while all peptides had more or less the same ACE inhibitory activity in pea hydrolysate. In addition, the hydrolysates and enriched fractions will resist in vivo gastrointestinal digestion after oral administration. Hence these ACE inhibitory peptides, as part of functional foods, can play significant roles in the prevention and treatment of hypertension. Copyright © 2004 Society of Chemical Industry     

Antioxidant and melanogenesis‐inhibitory activities of collagen peptide from jellyfish (Rhopilema esculentum)

BACKGROUND: Jellyfish collagen was hydrolysed with trypsin and properase E, and jellyfish collagen peptide (JCP) was purified from the enzymatic hydrolysate using ion exchange chromatography and gel filtration. The antioxidant activity of JCP in a linoleic acid emulsion system, its superoxide anion‐ and hydroxyl radical‐scavenging activities and its copper‐chelating ability were evaluated in vitro. Initial investigations of JCP's ability to inhibit melanogenesis were carried out using cultured B16 melanoma cells. RESULTS: The molecular weight distribution of JCP was from 400 to 1200 Da. Amino acid analysis showed that JCP was rich in Gly, Pro, Ser, Ala, Glu and Asp and had a total hydrophobic amino acid content of 384.2 g kg^?1. JCP showed high antioxidant activity (IC₅₀147.8 µg mL^?1), superoxide anion‐scavenging activity (IC₅₀21.9 µg mL^?1), hydroxyl radical‐scavenging activity (IC₅₀16.7 µg mL^?1) and copper‐chelating ability (IC₅₀88.7 µg mL^?1) in vitro. It also significantly inhibited intracellular tyrosinase activity, decreased melanin content and enhanced glutathione synthesis (P < 0.05). Furthermore, JCP decreased intracellular cAMP levels and suppressed tyrosinase mRNA expression. CONCLUSION: Based on the results of this study, JCP exerts anti‐melanogenic actions via its antioxidant properties and copper‐chelating ability. JCP could be used as a natural skin‐lightening agent in the medicine and food industries. Copyright © 2009 Society of Chemical Industry     

Production of detoxified sorghum straw hydrolysates for fermentative purposes

Sorghum straw can be hydrolysed to obtain monosaccharide solutions, mainly containing xylose. The usual biotechnological application of xylose is its bioconversion to xylitol. The global process from straw to xylitol can give an added value to the sorghum straw. The process has the following sequential steps: reduction of size, acid hydrolysis, neutralization, detoxification, fermentation, recovery and purification. This work deals with the optimization of the detoxification process of sorghum straw hydrolysates with activated charcoal. The variables evaluated were pH (1–5), contact time (20–60 min) and activated charcoal charge (20–33 g kg^?1). Mathematical models were obtained through a factorial experimental design. The models suggest that optimal conditions for detoxification are pH 1, contact time of 29 min and a charcoal charge of 33 g kg^?1. These conditions allowed hydrolysates with 54.2 g xylose L^?1, 13.5 g glucose L^?1, 12 g arabinose L^?1, 0.2 g furfural L^?1 and 0.0 g acetic acid L^?1 to be obtained. The results suggest that performing the detoxification step before the neutralization step gave the best outcome. Fermentations by Candida parapsilosis NRRL Y‐2315 were performed and it was confirmed that the treated hydrolysate is suitable for xylitol production, yielding up to 17 g L^?1 of this polyol. Copyright © 2006 Society of Chemical Industry     

Carotenoids production from a newly isolated <Emphasis Type="Italic">Sporidiobolus pararoseus</Emphasis> strain by submerged fermentation

This work aimed at evaluating the total carotenoids production by a newly isolated Sporidiobolus pararoseus. Bioproduction was carried out in an orbital shaker, using 10% (w/v) of inoculum (25 °C, 180 rpm for 35 h), incubated for 120 h in a dark room. Liquid N₂ and dimethylsulphoxide (DMSO) were used for cell rupture, and carotenoids were extracted with a solution of acetone/methanol (7:3, v/v). Optimization of carotenoids bioproduction was achieved by experimental design technique. Initially, a Plackett–Burman design was used for the screening of the most important factors, after the statistical analysis, a complete second-order design was carried out to optimize the concentration of total carotenoids in a conventional medium. Maximum concentration of 856 μg/L of total carotenoids was obtained in a medium containing 60 g/L of glucose, 15 g/L of peptone, and 15 g/L of malt extract, 25 °C, initial pH 4.0 and 180 rpm. Fermentation kinetics showed that the maximum concentration of total carotenoids was reached after 102 h of fermentation and that carotenoids bioproduction was associated with cell growth.     

Production of carotenoids by Rhodotorula glutinis MT-5 in submerged fermentation using the extract from waste loquat kernels as substrate

BACKGROUND: The aim of present study was to investigate the feasibility of the hydrolysate extracts from waste loquat kernels as substrate in submerged culture of yeast Rhodotorula glutinis MT‐5 for carotenoid production. RESULTS: Loquat kernel was found to have high protein (22.5%) and total carbohydrate (71.2%) contents. Dried and powdered loquat kernels were subjected to acid hydrolysis with 2 mol L^?1 HCl. The hydrolysate obtained was used for the preparation of loquat kernel extract and detoxified loquat kernel extract. The detoxification of hydrolysate was performed with Ca(OH)₂. Among the 10 R. glutinis isolates, the MT‐5 was found to be best in order to produce carotenoid using the extract as substrate. Production media prepared with detoxified loquat kernel extract or loquat kernel extract gave maximum biomass concentrations of 12.64 and 11.37 g L^?1, and maximum carotenoid concentrations of 72.36 and 62.73 mg L^?1, respectively. CONCLUSION: This study has provided effective processes for the conversion of waste material of plant origin to the extracts which are very rich in term of total fermentable sugar. The practicability of the extracts as fermentation substrate was proven in carotenoid production. To the best of our knowledge, this is the first report on use of this waste material as a substrate in yeast fermentations. Copyright © 2011 Society of Chemical Industry     

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     

Comparison of bioethanol and beta‐galactosidase production by Kluyveromyces and Saccharomyces strains grown in cheese whey

The production of ethanol and beta‐galactosidase by Kluyveromyces marxianus and Saccharomyces fragilis strains grown in cheese whey was evaluated. The conditions for fermentation in 50 g/L (3.3% lactose) and 150 g/L (8.8% lactose) cheese whey were 100 rpm for 24 h at 30, 35 and 40 °C. Saccharomyces fragilis IZ 275 in 8.8% lactose at 40 °C resulted in 3.90% ethanol. Kluyveromyces marxianus CCT 3172 showed higher beta‐galactosidase, 1.10 U/mg, at 30 °C. Therefore, the choice of cultivation conditions and the most suitable species is important for obtaining high yields of the products of interest.     

Fate of aflatoxin M1 in cheese whey processing

Aflatoxin M₁ (AFM₁) is an important mycotoxin frequently found in milk and in dairy products. It is a minor metabolic product of Aspergillus flavus and A parasiticus. However, it occurs in dairy products as a metabolite formed in cows from aflatoxin B₁ contained in animal feeds. In cheese production, AFM₁ distributes between curd and whey, being present in products derived from cheese whey processing. In this study, cheese whey from dairy processing was artificially contaminated with the mycotoxin at about 0.1 µg l⁻¹. Ultra‐filtration experiments of whey were carried out in order to determinate AFM₁ distribution between retentate (protein‐rich fraction) and permeate (lactose‐rich fraction). Recoveries of AFM₁ in retentate were 72.6–86.4% while, in permeate, recoveries were in the range 2.4–14.7%. Partition coefficients of AFM₁, lactose and protein were calculated to determine whether there was an interaction between AFM₁ and protein. In all experiments, AFM₁ partition coefficient was lower than 1, whilst for lactose coefficients close to 1 were determined, showing an affinity of aflatoxin M₁ to the protein‐rich fraction (retentate). Copyright © 2005 Society of Chemical Industry     

Optimisation of hydrolysis of purple sea urchin (Strongylocentrotus nudus) gonad by response surface methodology and evaluation of in vitro antioxidant activity of the hydrolysate

BACKGROUND: Hydrolysates prepared from sea urchin (Strongylocentrotus nudus) gonad by enzymatic treatment showed strong 1,1‐diphenyl‐2‐picrylhydrazyl radical scavenging activity and reducing power. RESULTS: Hydrolysis of S. nudus gonad by the commercial protease papain was optimised for maximum degree of hydrolysis (DH) and trichloroacetic acid‐soluble peptide index (TCA‐SPI) using response surface methodology. Results showed that the optimal conditions were the following: temperature of 48.83 °C, pH of 6.92, enzyme‐to‐substrate ratio of 3143 U g^?1, and substrate concentration of 83.5 g L^?1. Under these conditions, a DH of 27.96 ± 0.54% and a TCA‐SPI of 57.32 ± 0.63% were obtained. The hydrolysate prepared in the optimal conditions was fractionated by an ultra‐filtration system and the resultant fraction below 10 kDa was found to effectively scavenge hydroxyl radical (EC₅₀ = 13.29 ± 0.33 mg mL^?1) and hydrogen peroxide (EC₅₀ = 16.40 ± 0.37 mg mL^?1), inhibit lipid peroxidation (EC₅₀ = 11.05 ± 0.62 mg mL^?1), chelate Fe²⁺ (EC₅₀ = 7.26 ± 0.44 mg mL^?1), and protect mice macrophages against death induced by tert‐butyl hydroperoxide. CONCLUSION: Hydrolysates prepared from S. nudus gonad have the potential to be applied as natural antioxidant agents. Copyright © 2012 Society of Chemical Industry     

Batch kinetics and modelling of ethanolic fermentation of whey

The fermentation of whey by Kluyveromyces marxianus strain MTCC 1288 was studied using varying lactose concentrations at constant temperature and pH. The increase in substrate concentration up to a certain limit was accompanied by an increase in ethanol formation, for example, at a substrate concentration of 10 g L^?1, the production of ethanol was 0.618 g L^?1 whereas at 50 g L^?1 it was 3.98 g L^?1. However, an increase in lactose concentration to 100 g L^?1 led to a drastic decrease in product formation and substrate utilization. The maximum ethanol yield was obtained with an initial lactose concentration of 50 g L^?1. A method of batch kinetics was utilized to formulate a mathematical model using substrate and product inhibition constants. The model successfully simulated the batch kinetics observed at S₀ = 10 and 50 g L^?1 but failed in case of S₀ = 100 g L^?1 because of strong substrate inhibition.     

Production of whey protein isolate hydrolysate fractions with enriched ACE-inhibitory activity

A study on the enrichment of angiotensin-converting enzyme (ACE) inhibitory activity in whey protein isolate (WPI) hydrolysate fractions is presented. A previously identified low molecular mass fraction (1 kDa permeate) of an enzymatically hydrolysed heat-treated WPI with elevated ACE-inhibition (IC₅₀ = 0.23 g L⁻¹) was subjected to cascade membrane ultrafiltration (UF) and diafiltration steps at lab-scale. Assaying for ACE-inhibition revealed that the 1 kDa retentate demonstrated the highest ACE-inhibitory activity (IC₅₀ = 0.17 g L⁻¹). Isoelectric focussing (IEF) of the hydrolysate fraction further increased ACE-inhibition in fractions collected within the pH range 6.1–6.6. Overall, both UF and IEF enriched the ACE inhibitory activity in the original fraction by ∼52%, demonstrating the potential for enrichment of bio-functional activities in enzymatic hydrolysates of whey proteins.     

Low‐field NMR: A tool for studying protein aggregation

Low‐field nuclear magnetic resonance (NMR) spin–spin relaxation (T₂) measurements were used to study the denaturation and aggregation of β‐lactoglobulin (β‐LG) solutions of varying concentrations (1–80 g L^?1) as they were heated at temperatures ranging from ambient up to 90 °C. For concentrations of 1–10 g L^?1, the T₂ of β‐LG solutions did not change, even after heating to 90 °C. A decrease in T₂ was only observed when solutions having higher concentrations (20–80 g L^?1) were heated. Circular dichroism (CD) spectroscopy and fluorescence tests using the dye 1‐anilino‐8‐naphthalene sulfonate (ANS) on 0.2 and 1 g L^?1 solutions, respectively, indicated there were changes in the protein's secondary and tertiary conformations when the β‐LG solutions reached 70 °C and above. In addition, dynamic light scattering (DLS) showed that protein aggregation occurred only at concentrations above 10 g L^?1 and for heating at 70 °C and above. The hydrodynamic radius increased as T₂ decreased. When excess 2‐mercaptoethanol was added, the changes in both T₂ and the hydrodynamic radius followed the same trend for all β‐LG protein concentrations between 1 and 40 g L^?1. These observations led to the conclusion that the changes in T₂ were due to protein aggregation, not protein unfolding. Copyright © 2007 Society of Chemical Industry