Stability Enhancement of Anthocyanins from Blackcurrant (Ribes Nigrum L.) Pomace through Intermolecular Copigmentation.

Intermolecular copigmentation denotes the interaction between colored anthocyanins and the colorless copigment, which is not bound covalently to the anthocyanin molecule. This is the first study to investigate the effect of intermolecular copigmentation on the stability of individual anthocyanins from dried blackcurrant pomace (DBP) using four pure phenolic acids as copigments (ferulic, caffeic, chlorogenic, and rosmarinic acid). Studies were performed at pH 3.0 and pH 6.0, with a copigment/anthocyanin extract molar ratio of 5:1, during storage at 20 °C. At both pH 3.0 and 6.0, rosmarinic acid showed the strongest hyperchromic and bathochromic effects (p < 0.05) on day 0. However, rosmarinic acid showed low stability during storage. At pH 3.0, chlorogenic acid and control samples were capable of maintaining very high levels of total anthocyanin stability during storage (p < 0.05). On the other hand, ferulic acid and control samples had the longest estimated half-life during storage at pH 6.0. Intermolecular copigmentation successfully increased the half-life, color retention, and antioxidant activity of the anthocyanin solution, with cyanidin-3-O-glucoside (C3G) exhibiting the highest stability at both pH values. Overall, anthocyanins from DBP, in combination with chlorogenic or ferulic acid, showed potential for use in commercial food applications.

Sulforaphane-enriched extracts from glucoraphanin-rich broccoli exert antimicrobial activity against gut pathogens in vitro and innovative cooking methods increase in vivo intestinal delivery of sulforaphane.

PURPOSE: Studies on broccoli (Brassica oleracea var. italica) indicate beneficial effects against a range of chronic diseases, commonly attributed to their bioactive phytochemicals. Sulforaphane, the bioactive form of glucoraphanin, is formed by the action of the indigenous enzyme myrosinase. This study explored the role that digestion and cooking practices play in bioactivity and bioavailability, especially the rarely considered dose delivered to the colon. METHODS: The antimicrobial activity of sulforaphane extracts from raw, cooked broccoli and cooked broccoli plus mustard seeds (as a source myrosinase) was assessed. The persistence of broccoli phytochemicals in the upper gastrointestinal tract was analysed in the ileal fluid of 11 ileostomates fed, in a cross-over design, broccoli soup prepared with and without mustard seeds. RESULTS: The raw broccoli had no antimicrobial activity, except against Bacillus cereus, but cooked broccoli (with and without mustard seeds) showed considerable antimicrobial activity against various tested pathogens. The recovery of sulforaphane in ileal fluids post soup consumption was < 1% but the addition of mustard seeds increased colon-available sulforaphane sixfold. However, when sulforaphane was extracted from the ileal fluid with the highest sulforaphane content and tested against Escherichia coli K12, no inhibitory effects were observed. Analysis of glucosinolates composition in ileal fluids revealed noticeable inter-individual differences, with six "responding" participants showing increases in glucosinolates after broccoli soup consumption. CONCLUSIONS: Sulforaphane-rich broccoli extracts caused potent antimicrobial effects in vitro, and the consumption of sulforaphane-enriched broccoli soup may inhibit bacterial growth in the stomach and upper small intestine, but not in the terminal ileum or the colon.

Determination of the prebiotic activity of wheat arabinogalactan peptide (AGP) using batch culture fermentation.

PURPOSE: To test the prebiotic activity of wheat arabinogalactan-peptide (AGP), which is a soluble dietary fibre composed of arabinogalactan polysaccharide linked to a 15-residue peptide, which accounts for up to 0.4% of the dry weight of wheat flour. METHODS: The prebiotic activity of AGP prepared from white wheat flour was tested using in vitro fermentation by colonic bacteria in automated pH-controlled anaerobic stirred batch cultures and compared to fructooligosaccharide (FOS) and wheat flour arabinoxylan (AX). Bacterial populations were measured using fluorescence in situ hybridisation (flow-FISH) and short chain fatty acid (SCFA) concentrations were measured using HPLC. RESULTS: Fermentation of AGP resulted in a significant bifidogenic activity and increased concentrations of SCFAs, mainly acetate after 24 h of fermentation. CONCLUSIONS: These results were comparable to those obtained with AX and confirm the prebiotic potential of AGP. Furthermore, fermentation of a mixture of AGP and AX was faster compared to the single substrates and more similar to FOS, indicating that combinations of fermentable carbohydrates with different structures are potentially more effective as prebiotics than single substrates.

Development of chitosan-coated agar-gelatin particles for probiotic delivery and targeted release in the gastrointestinal tract.

This study reports the development of a novel and simple formulation for probiotic delivery using chitosan-coated agar-gelatin gel particles. This methodology involves the production of agar-gelatin particles by thermally treating a mixture of agar and gelatin solutions at high temperatures (121 °C) and subsequently coating with chitosan. The particles were able to protect the probiotic strain Lactobacillus plantarum NCIMB 8826 during incubation for 2 h in simulated gastric fluid (pH 2), as no statistically significant loss (P > 0.05) in cell concentration was observed, and also resist dissolution in simulated intestinal fluid (pH 7.2). Interestingly, this protection is related to the fact that the intense thermal treatment affected the physicochemical properties of agars and resulted in the formation of a strong and tight polymer network, as indicated by the X-ray diffraction (XRD) analysis. Using an in vitro faecal batch fermentation model simulating the conditions of the distal part of the large intestine (pH 6.7-6.9), it was demonstrated by quantitative real-time PCR that the majority of L. plantarum cells were released from the agar-gelatin particles within 30 to 48 h. Overall, this work led to the development of a novel methodology for the production of probiotic-containing particles, which is simpler compared with current encapsulation technologies and has a lot of potential to be used for the controlled release of probiotics and potentially other solid bioactives in the large intestine.Key Points• Chitosan gel particles is a simple and scalable method of probiotic encapsulation.• Autoclaving agar-gelatin particles increases their stability at low pH.• Chitosan gel particles protected L. plantarum during gastrointestinal conditions.• Probiotics could be controlled release in the colon using chitosan gel particles.

Adhesion mechanisms mediated by probiotics and prebiotics and their potential impact on human health.

Adhesion ability to the host is a classical selection criterion for potential probiotic bacteria that could result in a transient colonisation that would help to promote immunomodulatory effects, as well as stimulate gut barrier and metabolic functions. In addition, probiotic bacteria have a potential protective role against enteropathogens through different mechanisms including production of antimicrobial compounds, reduction of pathogenic bacterial adhesion and competition for host cell binding sites. The competitive exclusion by probiotic bacteria has a beneficial effect not only on the gut but also in the urogenital tract and oral cavity. On the other hand, prebiotics may also act as barriers to pathogens and toxins by preventing their adhesion to epithelial receptors. In vitro studies with different intestinal cell lines have been widely used along the last decades to assess the adherence ability of probiotic bacteria and pathogen antagonism. However, extrapolation of these results to in vivo conditions still remains unclear, leading to the need of optimisation of more complex in vitro approaches that include interaction with the resident microbiota to address the current limitations. The aim of this mini review is to provide a comprehensive overview on the potential effect of the adhesive properties of probiotics and prebiotics on the host by focusing on the most recent findings related with adhesion and immunomodulatory and antipathogenic effect on human health.

Supercritical Fluid Extraction of Carotenoids from Vegetable Waste Matrices.

The aim of this work was to evaluate a previously-developed model on supercritical fluid extraction (SFE) for carotenoid recovery from carrot peels on various carotenoid-rich fruit and vegetable wastes. To this end, 15 matrices, including flesh and peels of sweet potato, tomato, apricot, pumpkin and peach, as well as flesh and wastes of green, yellow and red peppers, were submitted to SFE under optimised conditions (59 °C, 350 bar, 15 g/min CO2, 15.5% (v/v) ethanol as co-solvent, 30 min of extraction time). The obtained extracts were characterised for their total carotenoid content, antioxidant activity and total carotenoid recovery (TCR). TCR values were greater than 90% w/w for most samples, with ß-carotene being the most successfully extracted compound (TCRs 88⁻100% w/w). More polar carotenoids, such as lutein and lycopene, exhibited lower TCRs. A comparison with literature data suggested that carotenoid extraction is partially dependent on the composition of vegetable matrices, specifically on polysaccharide and moisture content. The results indicated that the optimised SFE conditions can be used as a general model for carotenoid extraction from various fruit and vegetable matrices and as a viable method for adding value to these waste streams by generating carotenoid-rich extracts.

Evaluation of the prebiotic potential of arabinoxylans extracted from wheat distillers' dried grains with solubles (DDGS) and in-process samples.

Distillers' dried grains with solubles (DDGS) is a low-value agro-industrial by-product, rich in arabinoxylans (AX), which is produced by commercial distillery and bioethanol plants. In a first approach, we investigated the prebiotic potential of four fractions comprising arabinoxylan oligosaccharides (AXOS) and xylooligosaccharides (XOS) obtained by enzymatic hydrolysis of AX fractions derived from DDGS and wet solids (in-process sample of DDGS production process). Anaerobic batch cultures in controlled pH conditions were used to test the prebiotic activity of the samples. Results did not show significant differences between the enzymatic treatments used, and all AXOS/XOS were extensively fermented after 24 h. In addition, significant increases (P < 0.05) in Bifidobacterium and total short-chain fatty acids (SCFAs) were observed after 24 h of fermentation. Finally, DDGS-derived hydrolysates were separated on an anionic semi-preparative column to prepare AXOS/XOS fractions with degree of polymerisation (DP) greater than 3. Bifidogenic activity and an increase of SCFAs were again observed after 24 h of fermentation, although this time, the selectivity was higher and the fermentation slower, suggesting that the fermentation of this substrate could take place (at least partially) in the distal part of the colon with highly desirable beneficial effects.

Advancing Food Preservation: Sustainable Green-AgNPs Bionanocomposites in Paper-Starch Flexible Packaging for Prolonged Shelf Life.

In the pursuit of enhancing food packaging, nanotechnology, particularly green silver nanoparticles (G-AgNPs), have gained prominence for its remarkable antimicrobial properties with high potential for food shelf-life extension. Our study aims to develop corn starch-based coating materials reinforced with G-AgNPs. The mechanical properties were examined using a uniaxial tensile tester, revealing that starch coated with the highest G-AgNPs concentration (12.75 ppm) exhibited UTS of 87.6 MPa compared to 48.48 MPa of control paper, a significant (p < 0.02) 65% increase. The assessment of the WVP showcased a statistical reduction in permeability by up to 8% with the incorporation of the hydrophobic layer. Furthermore, antibacterial properties were assessed following ISO 22196:2011, demonstrating a strong and concentration-dependent activity of G-AgNPs against E. coli. All samples successfully disintegrated in both simulated environments (soil and seawater), including samples presenting G-AgNPs. In the food trial analysis, the presence of starch and G-AgNPs significantly reduced weight loss after 6 days, with cherry tomatoes decreasing by 8.59% and green grapes by 6.77% only. The results of this study contribute to the advancement of environmentally friendly packaging materials, aligning with the UN sustainable development goals of reducing food waste and promoting sustainability.

CLSM method for the dynamic observation of pH change within polymer matrices for oral delivery.

If acid-sensitive drugs or cells are administered orally, there is often a reduction in efficacy associated with gastric passage. Formulation into a polymer matrix is a potential method to improve their stability. The visualization of pH within these materials may help better understand the action of these polymer systems and allow comparison of different formulations. We herein describe the development of a novel confocal laser-scanning microscopy (CLSM) method for visualizing pH changes within polymer matrices and demonstrate its applicability to an enteric formulation based on chitosan-coated alginate gels. The system in question is first shown to protect an acid-sensitive bacterial strain to low pH, before being studied by our technique. Prior to this study, it has been claimed that protection by these materials is a result of buffering, but this has not been demonstrated. The visualization of pH within these matrices during exposure to a pH 2.0 simulated gastric solution showed an encroachment of acid from the periphery of the capsule, and a persistence of pHs above 2.0 within the matrix. This implies that the protective effect of the alginate-chitosan matrices is most likely due to a combination of buffering of acid as it enters the polymer matrix and the slowing of acid penetration.

Ionic liquid-mediated regeneration of cellulose dramatically improves decrystallization, TEMPO-mediated oxidation and alkyl/alkenyl succinylation.

This work demonstrated a successful strategy that simple ionic liquids (ILs) mediated pretreatment could effectively reduce crystallinity of cellulose from 71 % to 46 % (by C2MIM.Cl) and 53 % (by C4MIM.Cl). The IL-mediated regeneration of cellulose greatly promoted its reactivity for TEMPO-catalyzed oxidation, which the resulting COO- density (mmol/g) increased from 2.00 for non-IL-treated cellulose to 3.23 (by C2MIM.Cl) and 3.42 (C4MIM.Cl); and degree of oxidation enhanced from 35 % to 59 % and 62 %, respectively. More significantly, the yield of oxidized cellulose increased from 4 % to 45-46 %, by 11-fold. IL-regenerated cellulose can also be directly subjected to alkyl/alkenyl succinylation without TEMPO-mediated oxidation, producing nanoparticles with properties similar to oxidized celluloses (55-74 nm in size, -70-79 mV zeta-potential and 0.23-0.26 PDI); but in a much higher overall yield (87-95 %) than IL-regeneration-coupling-TEMPO-oxidation (34-45 %). Alkyl/alkenyl succinylated TEMPO-oxidized cellulose showed 2-2.5 times higher ABTS* scavenging ability than non-oxidized cellulose; however, alkyl/alkenyl succinylation also resulted in a significant decline in Fe2+ chelating property.

Bread waste - A potential feedstock for sustainable circular biorefineries.

The management of staggering volume of food waste generated (∼1.3 billion tons) is a serious challenge. The readily available untapped food waste can be promising feedstock for setting up biorefineries and one good example is bread waste (BW). The current review emphasis on capability of BW as feedstock for sustainable production of platform and commercially important chemicals. It describes the availability of BW (>100 million tons) to serve as a feedstock for sustainable biorefineries followed by examples of platform chemicals which have been produced using BW including ethanol, lactic acid, succinic acid and 2,3-butanediol through biological route. The BW-based production of these metabolites is compared against 1G and 2G (lignocellulosic biomass) feedstocks. The review also discusses logistic and supply chain challenges associated with use of BW as feedstock. Towards the end, it is concluded with a discussion on life cycle analysis of BW-based production and comparison with other feedstocks.

Silver Bionanocomposites as Active Food Packaging: Recent Advances & Future Trends Tackling the Food Waste Crisis.

Food waste is a pressing global challenge leading to over $1 trillion lost annually and contributing up to 10% of global greenhouse gas emissions. Extensive study has been directed toward the use of active biodegradable packaging materials to improve food quality, minimize plastic use, and encourage sustainable packaging technology development. However, this has been achieved with limited success, which can mainly be attributed to poor material properties and high production costs. In the recent literature, the integration of silver nanoparticles (AgNPs) has shown to improve the properties of biopolymer, prompting the development of bionanocomposites. Furthermore, the antibacterial properties of AgNPs against foodborne pathogens leads towards food shelf-life improvement and provides a route towards reducing food waste. However, few reviews have analyzed AgNPs holistically throughout a portfolio of biopolymers from an industrial perspective. Hence, this review critically analyses the antibacterial, barrier, mechanical, thermal, and water resistance properties of AgNP-based bionanocomposites. These advanced materials are also discussed in terms of food packaging applications and assessed in terms of their performance in enhancing food shelf-life. Finally, the current barriers towards the commercialization of AgNP bionanocomposites are critically discussed to provide an industrial action plan towards the development of sustainable packaging materials to reduce food waste.

Organosolv Pretreatment of Cocoa Pod Husks: Isolation, Analysis, and Use of Lignin from an Abundant Waste Product.

Cocoa pod husks (CPHs) represent an underutilized component of the chocolate manufacturing process. While industry's current focus is understandably on the cocoa beans, the husks make up around 75 wt % of the fruit. Previous studies have been dominated by the carbohydrate polymers present in CPHs, but this work highlights the presence of the biopolymer lignin in this biomass. An optimized organosolv lignin isolation protocol was developed, delivering significant practical improvements. This new protocol may also prove to be useful for agricultural waste-derived biomasses in general. NMR analysis of the high quality lignin led to an improved structural understanding, with evidence provided to support deacetylation of the lignin occurring during the optimized pretreatment. Chemical transformation, using a tosylation, azidation, copper-catalyzed click protocol, delivered a modified lignin oligomer with an organophosphorus motif attached. Thermogravimetric analysis was used to demonstrate the oligomer's potential as a flame-retardant. Preliminary analysis of the other product streams isolated from the CPHs was also carried out.

Influence of fermentation conditions on the surface properties and adhesion of Lactobacillus rhamnosus GG.

BACKGROUND: The surface properties of probiotic bacteria influence to a large extent their interactions within the gut ecosystem. There is limited amount of information on the effect of the production process on the surface properties of probiotic lactobacilli in relation to the mechanisms of their adhesion to the gastrointestinal mucosa. The aim of this work was to investigate the effect of the fermentation pH and temperature on the surface properties and adhesion ability to Caco-2 cells of the probiotic strain Lactobacillus rhamnosus GG. RESULTS: The cells were grown at pH 5, 5.5, 6 (temperature 37°C) and at pH 6.5 (temperature 25°C, 30°C and 37°C), and their surfaces analysed by X-ray photoelectron spectrometry (XPS), Fourier transform infrared spectroscopy (FT-IR) and gel-based proteomics. The results indicated that for all the fermentation conditions, with the exception of pH 5, a higher nitrogen to carbon ratio and a lower phosphate content was observed at the surface of the bacteria, which resulted in a lower surface hydrophobicity and reduced adhesion levels to Caco-2 cells as compared to the control fermentation (pH 6.5, 37°C). A number of adhesive proteins, which have been suggested in previous published works to take part in the adhesion of bacteria to the human gastrointestinal tract, were identified by proteomic analysis, with no significant differences between samples however. CONCLUSIONS: The temperature and the pH of the fermentation influenced the surface composition, hydrophobicity and the levels of adhesion of L. rhamnosus GG to Caco-2 cells. It was deduced from the data that a protein rich surface reduced the adhesion ability of the cells.

Properties of protein isolates extracted by ultrasonication from soybean residue (okara).

Soybean protein extraction was evaluated using conventional (alkaline phosphate buffer) and ultrasonication assisted methods. The impact of the extraction method on protein yield, chemical composition, and structural properties of the protein isolates was assessed. In conventional alkaline extraction, increasing pH values (from 9 to 12) led to an increase in the protein yield, with pH 12 favouring protein extraction, with yields up to 36 % (w/w). Ultrasonication process at the same pH, led to maximum protein extraction yield of 84 % (w/w). Secondary structural changes in ultrasonicated OPI samples were linked to cavitation effects and the duration of the extraction, rather than the intensity of ultrasonication process. Extracted proteins exhibited increased ß-sheet content, improved zeta-potential and smaller particle size of ultrasonicated proteins, rendering them suitable ingredients for tailored food applications.

Production and evaluation of dry alginate-chitosan microcapsules as an enteric delivery vehicle for probiotic bacteria.

This study investigates the production of alginate microcapsules, which have been coated with the polysaccharide chitosan, and evaluates some of their properties with the intention of improving the gastrointestinal viability of a probiotic ( Bifidobacterium breve ) by encapsulation in this system. The microcapsules were dried by a variety of methods, and the most suitable was chosen. The work described in this Article is the first report detailing the effects of drying on the properties of these microcapsules and the viability of the bacteria within relative to wet microcapsules. The pH range over which chitosan and alginate form polyelectrolyte complexes was explored by spectrophotometry, and this extended into swelling studies on the microcapsules over a range of pHs associated with the gastrointestinal tract. It was shown that chitosan stabilizes the alginate microcapsules at pHs above 3, extending the stability of the capsules under these conditions. The effect of chitosan exposure time on the coating thickness was investigated for the first time by confocal laser scanning microscopy, and its penetration into the alginate matrix was shown to be particularly slow. Coating with chitosan was found to increase the survival of B. breve in simulated gastric fluid as well as prolong its release upon exposure to intestinal pH.

Investigation of the factors influencing the survival of Bifidobacterium longum in model acidic solutions and fruit juices.

The survival of Bifidobacterium longum NCIMB 8809 was studied during refrigerated storage for 6weeks in model solutions, based on which a mathematical model was constructed describing cell survival as a function of pH, citric acid, protein and dietary fibre. A Central Composite Design (CCD) was developed studying the influence of four factors at three levels, i.e., pH (3.2-4), citric acid (2-15g/l), protein (0-10g/l), and dietary fibre (0-8g/l). In total, 31 experimental runs were carried out. Analysis of variance (ANOVA) of the regression model demonstrated that the model fitted well the data. From the regression coefficients it was deduced that all four factors had a statistically significant (P<0.05) negative effect on the log decrease [log10N0 week-log10N6 week], with the pH and citric acid being the most influential ones. Cell survival during storage was also investigated in various types of juices, including orange, grapefruit, blackcurrant, pineapple, pomegranate and strawberry. The highest cell survival (less than 0.4log decrease) after 6weeks of storage was observed in orange and pineapple, both of which had a pH of about 3.8. Although the pH of grapefruit and blackcurrant was similar (pH ∼3.2), the log decrease of the former was ∼0.5log, whereas of the latter was ∼0.7log. One reason for this could be the fact that grapefruit contained a high amount of citric acid (15.3g/l). The log decrease in pomegranate and strawberry juices was extremely high (∼8logs). The mathematical model was able to predict adequately the cell survival in orange, grapefruit, blackcurrant, and pineapple juices. However, the model failed to predict the cell survival in pomegranate and strawberry, most likely due to the very high levels of phenolic compounds in these two juices.

Electrosprayed mucoadhesive alginate-chitosan microcapsules for gastrointestinal delivery of probiotics.

Besides viability protection, a sufficiently prolonged gastrointestinal retention of probiotics has emerged as critically important in improving the functional effectiveness of gastrointestinal delivery of these microorganisms. In this work, we formulated pure, resistant starch-reinforced and chitosan-coated alginate microparticles using an electrospray technique and evaluated their performance as mucoadhesive probiotic formulations for gastrointestinal delivery. In addition, we designed and successfully validated a novel experimental set-up of in vitro wash-off mucoadhesion test, using a portable and low-cost USB microscope for fluorescence imaging. In our test, pure chitosan microparticles (positive control) exhibited the greatest mucoadhesive property, whereas the alginate-resistant starch ones (negative control) were the least retentive on a gastric mucosa. These electrosprayed formulations were spherically shaped, with a size range of 30-600 µm (60-1300 µm with chitosan coating). Moreover, model probiotic Lactobacillus plantarum loaded in alginate-starch formulations was better protected against simulated gastric conditions than in alginate ones, but not better than in the chitosan-coated ones.

Acetic acid buffer as extraction medium for free and bound phenolics from dried blackcurrant (Ribes nigrum L.) skins.

The aim of this study was to investigate the effects of different solvent and extraction temperatures on the free and bound phenolic compounds and antioxidant activity of dried blackcurrant skins (DBS). Apart from acetic acid buffer solution, different solvent systems, including water, methanol, and mixtures of methanol/water, were also employed and the effects of solvent and temperature (30 and 50 °C) on the free and bound forms of anthocyanins, hydroxycinnamic acids, and flavonols yield were assessed. The results showed that among all solvents, acetic acid buffer resulted in the highest free anthocyanin content (1,712.3 ± 56.1 mg/100 g) (P < 0.05) after 2 hr extraction at 50 °C from DBS, while lower amounts of bound anthocyanins and anthocyanidins were detected after acid hydrolysis. Acetic acid buffer extracts exhibited the highest free hydroxycinnamic acid content (268.0 ± 4.5 mg/100 g), total phenolic content (3702.2 ± 259.3 mg GAE/100 g), and DPPH activity (60.7 ± 2.0% of inhibition). However, their free flavonol content was slightly lower (60.2 ± 0.8 mg/100 g) compared to 100% methanol at 30 and 50 °C (71.4 ± 1.5 mg/100 g and 71.5 ± 6.2 mg/100 g, respectively). Two-way ANOVA indicated interactions between solvent and temperature (P < 0.05), which suggested that the relationship between solvent and phenolic compounds depends on the extraction temperature. PRACTICAL APPLICATION: Overall, acetic acid buffer is more environmental friendly, efficient, and cost effective than other solvents, thus, offering an improved extraction method for phytochemicals as valuable ingredients for nutraceutical applications, from underutilized dried blackcurrant skins (DBS).

Optimised Production and Extraction of Astaxanthin from the Yeast Xanthophyllomyces dendrorhous.

Currently, astaxanthin demand is fulfilled by chemical synthesis using petroleum-based feedstocks. As such, alternative pathways of natural astaxanthin production attracts much research interest. This study aimed at optimising bioreactor operation parameters for astaxanthin production and evaluating strategies for its subsequent extraction. The effect of pH and agitation was evident, as a significant reduction in both biomass and astaxanthin production was observed when the culture pH was not controlled and a low agitation speed was applied. At controlled pH conditions and a high agitation speed, a significant increase in biomass (16.4 g/L) and astaxanthin production (3.6 mg/L) was obtained. Enzymatic yeast cell lysis using two commercial enzymes (Accellerase 1500 and Glucanex) was optimised using the central composite design of experiment (DoE). Accellerase 1500 led to mild cell disruption and only 9% (w/w) astaxanthin extraction. However, Glucanex treatment resulted in complete astaxanthin extractability, compared to standard extraction method (DMSO/acetone). When supercritical CO2 was employed as an extraction solvent in Accellerase-pre-treated Xanthophyllomyces dendrorhous cells, astaxanthin extraction increased 2.5-fold. Overall, the study showed that extraction conditions can be tailored towards targeted pigments present in complex mixtures, such as in microbial cells.