Perstraction of intracellular pigments through submerged fermentation of Talaromyces spp. in a surfactant rich media: a novel approach for enhanced pigment recovery

A high percentage of the pigments produced by Talaromyces spp. remains inside the cell, which could lead to a high product concentration inhibition. To overcome this issue an extractive fermentation process, perstraction, was suggested, which involves the extraction of the intracellular products out of the cell by using a two-phase system during the fermentation. The present work studied the effect of various surfactants on secretion of intracellular pigments produced by Talaromyces spp. in submerged fermentation. Surfactants used were: non-ionic surfactants (Tween 80, Span 20 and Triton X-100) and a polyethylene glycerol polymer 8000, at different concentrations (5, 20, 35 g/L). The highest extracellular pigment yield (16 OD500nm) was reached using Triton X-100 (35 g/L), which was 44% higher than the control (no surfactant added). The effect of addition time of the selected surfactant was further studied. The highest extracellular pigment concentration (22 OD500nm) was achieved when the surfactant was added at 120 h of fermentation. Kinetics of extracellular and intracellular pigments were examined. Total pigment at the end of the fermentation using Triton X-100 was 27.7% higher than the control, confirming that the use of surfactants partially alleviated the product inhibition during the pigment production cultur

Enzymatic Processes Triggered by PEF for Astaxanthin Extraction From Xanthophyllomyces dendrorhous

The aim of this study was to evaluate the potential of pulsed electric fields (PEF) to improve the extraction of the lipid-soluble astaxanthin from fresh biomass of a wild-type (CECT 11028) and mutant (ATCC 74219) Xanthophyllomyces dendrorhous strain using ethanol as solvent. Inactivation and propidium uptake studies revealed that inactivation is a good index for estimated the proportion of irreversible permeabilized cells when inactivation is higher than 70% in the two strains. Ethanol was ineffective for extracting carotenoids from the PEF-treated cells (20 kV/cm, 135 µs) of the two strains. However, after aqueous incubation of PEF-treated X. dendrorhous ATCC 74219 cells for 12 h, up to 2.4 ± 0.05 mg/g dried weight (d.w.) of carotenoids were extracted in ethanol. From total carotenoid extracted, around 84% corresponded to all-trans astaxanthin. The detection and quantification of esterase activity in the supernatant and the relationship between the percentage of esterase activity quantified and the amount of carotenoids extracted indicate that the extraction of astaxanthin was mediated by enzymatic esterase activity triggered by PEF during incubation. On the other hand, the formation of a large lipid globule into the cytoplasm of PEF-treated X. dendrorhous CECT 11028 cells during aqueous incubation prevented carotenoid extraction. The process developed in this investigation represents a more sustainable and greener method that those previously used for extracting astaxanthin from yeast

Selection of best conditions of inoculum preparation for optimum performance of the pigment production process by Talaromyces spp. using the Taguchi method

Process optimisation techniques increasingly need to be used early on in research and development of processes for new ingredients. There are different approaches and this article illustrates the main issues at stake with a method that is an industry best practice, the Taguchi method, suggesting a procedure to assess the potential impact of its drawbacks. The Taguchi method has been widely used in various industrial sectors because it minimises the experimental requirements to define an optimum region of operation, which is particularly relevant when minimising variability is a target. However, it also has drawbacks, especially the intricate confoundings generated by the experimental designs used. This work reports a process optimisation of the synthesis of red pigments by a fungal strain, Talaromyces spp. using the Taguchi methodology and proposes an approach to assess from validation trials whether the conclusions can be accepted with confidence. The work focused on optimising the inoculum characteristics, and the studied factors were spore age and concentration, agitation speed and incubation time. It was concluded that spore age was the most important factor for both responses, with optimum results at 5 days old, with the best other conditions being spores concentration, 100,000 (spores/mL); agitation, 200 rpm; and incubation time, 84 h. The interactive effects can be considered negligible and therefore this is an example where a simple experimental design approach was successful in speedily indicating conditions able to increase pigment production by 63% compared to an average choice of settings

Influence of granulation process parameters on food tablet properties formulated using natural powders (Opuntia ficus and Chlorella spp.)

In this research two natural health supplements, cactus flour (Opuntia ficus) and microalgae biomass (Chlorella spp.) have been examined as novel ingredients to formulate health supplement tablets. The physical and mechanical properties of the pure powders were quantified, and their ability to be tableted directly without additional processing was investigated. High-shear wet granulation (HSWG) was explored to improve the flowability and compression characteristics of the powders. Using an L9 Taguchi experimental design, the effect of critical process parameters of HSWG was examined on quality attributes of tablets. Tablets were successfully formulated without the addition of excipients following wet granulation. Consistent and acceptable quality tablets were produced based on the optimal processing conditions determined by L9 Taguchi design. The tablet tensile strength achieved was 0.91 ± 0.05 MPa with a disintegration time < 30 min and a friability value of 0.05 ± 0.02%. The tablets obtained in the present study are comparable with commercial available natural supplement tablets in terms of disintegration time, friability and tensile strength. The research provided a basis for the potential use of Cactus (Opuntia spp.) and microalgae (Chlorella sp.) powders as novel ingredients for the development of dietary supplements tablets, and to be used as excipients for the production of pharmaceutical tablets

Perstraction of Intracellular Pigments through Submerged Fermentation of Talaromyces spp. in a Surfactant Rich Media: A Novel Approach for Enhanced Pigment Recovery

A high percentage of the pigments produced by Talaromyces spp. remains inside the cell, which could lead to a high product concentration inhibition. To overcome this issue an extractive fermentation process, perstraction, was suggested, which involves the extraction of the intracellular products out of the cell by using a two-phase system during the fermentation. The present work studied the effect of various surfactants on secretion of intracellular pigments produced by Talaromyces spp. in submerged fermentation. Surfactants used were: non-ionic surfactants (Tween 80, Span 20 and Triton X-100) and a polyethylene glycerol polymer 8000, at different concentrations (5, 20, 35 g/L). The highest extracellular pigment yield (16 OD500nm) was reached using Triton X-100 (35 g/L), which was 44% higher than the control (no surfactant added). The effect of addition time of the selected surfactant was further studied. The highest extracellular pigment concentration (22 OD500nm) was achieved when the surfactant was added at 120 h of fermentation. Kinetics of extracellular and intracellular pigments were examined. Total pigment at the end of the fermentation using Triton X-100 was 27.7% higher than the control, confirming that the use of surfactants partially alleviated the product inhibition during the pigment production culture

Assessment of the Dyeing Properties of the Pigments Produced by Talaromyces spp.

The high production yields of pigments by Talaromyces spp. and their high thermal stability have implied that industrial application interests may emerge in the food and textile industries, as they both involve subjecting the colourants to high temperatures. The present study aimed to assess the potential application of the pigments produced by Talaromyces spp. in the textile area by studying their dyeing properties. Dyeing studies were performed on wool. The dyeing process consisted of three stages: scouring, mordanting, and dyeing. Two different mordants (alum, A; ferric chloride, F) were tested at different concentrations on fabric weight (A: 5, 10, 15%; F: 10, 20, 30%). The mordanting process had a significant effect on the final colour of the dyed fabrics obtained. The values of dyeing rate constant (k), half-time of dyeing (t1/2), and sorption kinetics behaviour were evaluated and discussed. The obtained results showed that pigments produced by Talaromyces spp. could serve as a source for the natural dyeing of wool textiles

Production of Pigments under Submerged Culture through Repeated Batch Fermentation of Immobilized <i>Talaromyces atroroseus</i> GH2

Pigments of natural origin have become a research trend, and fungi provide a readily available alternative source. Moreover, developing novel processes that increase yields, reduce process time and simplify downstream processing is of increased interest. In this sense, this work proposes an alternative for Talaromyces atroroseus GH2 biomass re-utilization to produce pigments through consecutive batches using immobilized mycelium. Different support materials were evaluated for pigment production and immobilization capacity. Then, Taguchi’s method was applied to determine the effect of four factors related to fungal immobilization and pigment production (inoculum concentration, support density, working volume and support volume). Afterward, process kinetics for pigment production using immobilized cells of T. atroroseus GH2 in consecutive batches were evaluated. All evaluated factors were significant and affected pigment production and microorganism growth differently. At improved conditions, immobilization capacity reached 99.01 ± 0.37% and the pigment production was 30% higher than using free cells. Process kinetics showed that the production could continue for three batches and was limited by excessive microorganism growth. Indeed, more studies are still needed, but the immobilization of Talaromyces atroroseus GH2 represents a promising strategy for allowing downstream-processing intensification since immobilized biomass is easily removed from the fermentation media, thus paving the way for the further development of a continuous process

Coproduction of Microbial Oil and Carotenoids within the Circular Bioeconomy Concept: A Sequential Solid-State and Submerged Fermentation Approach

The main objective of integrative biorefinery platforms is to propose efficient green methodologies addressed to obtain high-value compounds with low emissions through biochemical conversions. This work first screened the capacity of various oleaginous yeast to cosynthesize high-value biomolecules such as lipids and carotenoids. Selected strains were evaluated for their ability to coproduce such biocompounds in the waste-based media of agro-food (brewer’s spent grain, pasta processing waste and bakery waste). Carbon and nitrogen source feedstock was obtained through enzymatic hydrolysis of the agro-food waste, where up to 80% of total sugar/starch conversion was obtained. Then, the profitability of the bioprocess for microbial oil (MO) and carotenoids production by Sporobolomyces roseus CFGU-S005 was estimated via simulation using SuperPro Designer®. Results showed the benefits of establishing optimum equipment scheduling by identifying bottlenecks to increase profitability. Sensitivity analysis demonstrated the impact of MO price and batch throughput on process economics. A profitable process was achieved with a MO batch throughput of 3.7 kg/batch (ROI 31%, payback time 3.13 years). The results revealed areas that require further improvement to achieve a sustainable and competitive process for the microbial production of carotenoids and lipids

Microbial Carotenoid Synthesis Optimization in Goat Cheese Whey Using the Robust Taguchi Method: A Sustainable Approach to Help Tackle Vitamin A Deficiency

The work describes the carotenoid synthesis process by Rhodotorula glutinis P4M422 using an agro-industrial waste as the substrate, seeking a biorefinery platform approach for waste utilization to produce high-value molecules. A culture medium based on goat milk whey (GMW) was optimized via the Taguchi method (L9 array). Four factors (ethanol, carbon and nitrogen source, and pH) were evaluated at three levels. The carbon and nitrogen composition were the factors dominating the process performance. Optimized conditions were validated (Urea, 0.3% w/v; pH, 4.5; ethanol, 10% v/v; glucose, 6.0%), and the carotenoid production (4075 µg/L) was almost 200% higher than when using the un-optimized process (2058 µg/L). Provitamin A carotenoids torulene, β–carotene, and γ–carotene (different proportions) were produced under all conditions. The hydrolyzed goat milk whey showed promising expectations as a low-cost source for carotenoid production by Rhodotorula glutinis P4M422. The results are important for the innovative sustainable production of carotenoid-rich matrices for different purposes (nutrition, health promotion, color) and industries (foods, nutricosmetics, nutraceuticals, feeds), notably to help to combat vitamin A deficiency

Medium design from corncob hydrolyzate for pigment production by Talaromyces atroroseus GH2: Kinetics modeling and pigments characterization

The genusTalaromyceshas gained attention due to its ability to produce pigments with potential industrialapplications in different areas. Prosperous application of fungal pigments has challenges to overcome, like de-veloping a cost-effective bioprocess. Using agroindustrial wastes could provide inexpensive substrates and itcontributes to maximize sustainability. Therefore, this study evaluated the feasibility of using corncob as a low-cost substrate for pigment production byTalaromyces atroroseusGH2. An acid hydrolysis treatment was used torelease sugars from corncob. Corncob liquors with enough xylose concentration (&gt; 20 g/L) were investigated asfermentation media with and without the addition of nutrients. Process kinetic modeling was applied andpigments produced in corncob and control media were characterized. The diluted hydrolyzate without nutrientsupplementation showed a pigment production (16.17 ± 0.37 OD500nm) comparable to the control medium(17.26 ± 0.41 OD500nm).Talaromyces atroroseusGH2 was able to co-utilize xylose and glucose in the corncob-based medium. However, growth kinetics patterns differed in both media. In the hydrolyzate medium, biomassgrowth presented an extended lag phase, which requires reduction for future process optimization. Finally,characterized pigments differed among evaluated media, but the pigments produced byTalaromyces atroroseusGH2 were mostlyMonascus’pigments homologous like monascorubrin and rubropunctamine.TalaromycesatroroseusGH2 ability to produce pigments using corncob hydrolyzate makes it a pigment-producing strain foran economically competitive large fermentation scal