Ganoderma lucidum Mycelia Mass and Bioactive Compounds Production through Grape Pomace and Cheese Whey Valorization.

Numerous compounds obtained from the medicinal mushroom Ganoderma lucidum have evidenced renowned bioactive characteristics. Controlled fermentation to generate fungal mycelia confers several advantages, specifically when the valorization of agro-industrial streams as fermentation feedstocks is included. Submerged fermentation of a newly isolated Greek strain of G. lucidum was performed using conventional synthetic media and, also, grape pomace extract (GPE) and cheese whey permeate (CWP) under static and shaking conditions. Under shaking conditions, maximum biomass with GPE and supplementation with organic nitrogen reached 17.8 g/L. The addition of an elicitor in CWP resulted in a significant improvement in biomass production that exceeded synthetic media. Overall, agitation demonstrated a positive impact on biomass productivity and, therefore, on process optimization. Crude intracellular and extracellular polysaccharides were extracted and evaluated regarding antioxidant activity and polysaccharide and protein content. FTIR analysis confirmed the preliminary chemical characterization of the crude extracts. This study introduces the design of a bioprocessing scenario to utilize food industry by-products as onset feedstocks for fungal bioconversions to obtain potential bioactive molecules within the concept of bioeconomy.

Natural Carotenoids: Recent Advances on Separation from Microbial Biomass and Methods of Analysis.

Biotechnologically produced carotenoids occupy an important place in the scientific research. Owing to their role as natural pigments and their high antioxidant properties, microbial carotenoids have been proposed as alternatives to their synthetic counterparts. To this end, many studies are focusing on their efficient and sustainable production from renewable substrates. Besides the development of an efficient upstream process, their separation and purification as well as their analysis from the microbial biomass confers another important aspect. Currently, the use of organic solvents constitutes the main extraction process; however, environmental concerns along with potential toxicity towards human health necessitate the employment of "greener" techniques. Hence, many research groups are focusing on applying emerging technologies such as ultrasounds, microwaves, ionic liquids or eutectic solvents for the separation of carotenoids from microbial cells. This review aims to summarize the progress on both the biotechnological production of carotenoids and the methods for their effective extraction. In the framework of circular economy and sustainability, the focus is given on green recovery methods targeting high-value applications such as novel functional foods and pharmaceuticals. Finally, methods for carotenoids identification and quantification are also discussed in order to create a roadmap for successful carotenoids analysis.

Novel Probiotic/Bacterial Cellulose Biocatalyst for the Development of Functional Dairy Beverage.

The development of innovative functional products with potential health benefits, under the concept of bio-economy, is flourishing. This study undertook an evaluation of non-dairy lactobacilli Lactiplantibacillus pentosus B329 and Lactiplantibacillus plantarum 820 as "ready to use" starter cultures. Lactic acid bacteria (LAB) cultures were evaluated for their fermentation efficiency, before and after freeze-drying, using cheese whey (CW) as a fermentation substrate and subsequent immobilization on bacteria cellulose (BC) to produce a novel biocatalyst. The biocatalyst was applied in functional sour milk production and compared with free cells via the assessment of physicochemical and microbiological properties and sensory evaluation. Evidently, LAB strains exhibited high fermentative activity before and after freeze-drying. Results of a 5-month storage stability test showed that viability was 19% enhanced by immobilization on BC, supporting the concept of "ready to use" cultures for the production of fermented beverages. Likewise, sour milk produced by the BC biocatalyst presented higher organoleptic scores, compared to the free cells case, whereas immobilization on BC enhanced probiotic viability during post-fermentation storage (4 °C, 28 days). The obtained high viability (>107 log cfu/g) demonstrated the efficacy of the proposed bioprocess for the production of functional/probiotic-rich beverages. Ultimately, this work presents a consolidated scheme that includes the advantages and the cooperative effect of probiotic LAB strains combined with a functional biopolymer (BC) towards the formulation of novel functional products that coincide with the pillars of food systems sustainability.

Sustainable Food Systems: The Case of Functional Compounds towards the Development of Clean Label Food Products.

The addition of natural components with functional properties in novel food formulations confers one of the main challenges that the modern food industry is called to face. New EU directives and the global turn to circular economy models are also pressing the agro-industrial sector to adopt cradle-to-cradle approaches for their by-products and waste streams. This review aims to present the concept of "sustainable functional compounds", emphasizing on some main bioactive compounds that could be recovered or biotechnologically produced from renewable resources. Herein, and in view of their efficient and "greener" production and extraction, emerging technologies, together with their possible advantages or drawbacks, are presented and discussed. Μodern examples of novel, clean label food products that are composed of sustainable functional compounds are summarized. Finally, some action plans towards the establishment of sustainable food systems are suggested.

Prebiotic Potential of a New Sweetener Based on Galactooligosaccharides and Modified Mogrosides.

This study was conducted to investigate the sweetness intensity and the potential fecal microbiome modulation of galactooligosaccharides in combination with enzymatically modified mogrosides (mMV-GOS), both generated through a patented single-pot synthesis. Sweetness intensity was performed in vivo by trained sensory panelists. The impact on the human fecal microbiome was evaluated by in vitro pH-controlled batch fermentation, and bacterial populations and organic acid concentrations were measured by qPCR and GC-FID, respectively. Significant growth (p ≤ 0.05) during the fermentation at 10 h of bacterial populations includes Bifidobacterium (8.49 ± 0.44 CFU/mL), Bacteroides (9.73 ± 0.32 CFU/mL), Enterococcus (8.17 ± 0.42 CFU/mL), and Clostridium coccoides (6.15 ± 0.11 CFU/mL) as compared to the negative control counts for each bacterial group (7.94 ± 0.27, 7.84 ± 1.11, 7.52 ± 0.37, and 5.81 ± 0.08 CFU/mL, respectively) at the same time of fermentation. Likewise, the corresponding significant increase in production of SCFA in mMV-GOS at 10 h of fermentation, mainly seen in acetate (20.32 ± 2.56 mM) and propionate (9.49 ± 1.44 mM) production compared to a negative control at the same time (8.15 ± 1.97 and 1.86 ± 0.24 mM), is in line with a positive control (short-chain fructooligosaccharides; 46.74 ± 12.13 and 6.51 ± 1.91 mM, respectively) revealing a selective fermentation. In conclusion, these substrates could be considered as novel candidate prebiotic sweeteners, foreseeing a feasible and innovative approach targeting the sucrose content reduction in food. This new ingredient could provide health benefits when evaluated in human studies by combining sweetness and prebiotic fiber functionality.

Whey protein films reinforced with bacterial cellulose nanowhiskers: Improving edible film properties via a circular economy approach.

Edible films were developed using whey protein concentrate (WPC) and a natural bio-polymer, namely bacterial cellulose (BC). BC was produced via fermentation from orange peels and subsequently acid-hydrolyzed to obtain BC nanowhiskers (BCNW) with high crystallinity (XRD analysis). Morphology of BCNW was analyzed by SEM, TEM, and AFM. WPC/BCNW film composites, containing different amounts of BCNW (0.5-15%, w/w) were developed and characterized. WPC/BCNW film composite was analyzed by Raman spectroscopy, indicating the successful incorporation and the homogenous distribution of BCNW into the WPC film matrix. Mechanical characterization showed that BCNW behaved as a reinforcing filler in the WPC film, increasing tensile strength and Young's modulus by 32% and 80%, respectively. In addition, water vapor permeability was reduced by 33.9% upon the addition of 0.5% BCNW. This study presented a sustainable approach towards the production of WPC films with improved tensile and water barrier properties, suggesting its potential application as a packaging material.

Biosurfactant Production from Lactobacilli: an Insight on the Interpretation of Prevailing Assessment Methods.

Biosurfactants constitute amphiphilic molecules, receiving increased attention as environmentally benign, biodegradable alternatives to substitute for the petroleum derived counterparts in food, pharmaceutical and cosmetics applications. However, their high production cost hinders industrial production. In this study, fifty GRAS lactobacilli strains were screened for their ability to produce biosurfactants, implementing different substrates. Cheese whey permeate (CWP) was also assessed as a low-cost and inherent lactobacilli substrate, aiming to mitigate its polluting impact, expand valorization strategies, alleviate costs deriving from commercial supplements and enhance overall sustainability. Surface tension, emulsification activity (E24) and oil displacement were deployed to identify the most promising candidates. Results reveal surface tension as the most robust method and underline the effect of substrate on biosurfactant synthesis. Likewise, this study indicates the fundamental role of including the final fermentation substrate (CWP) during strain selection to avoid misinterpretation of results and enhance subsequent bioprocess integration.

Techno-economic evaluation and life-cycle assessment of poly(3-hydroxybutyrate) production within a biorefinery concept using sunflower-based biodiesel industry by-products.

This study presents techno-economic evaluation of a biorefinery concept using biodiesel industry by-products (sunflower meal and crude glycerol) to produce poly(3-hydroxybutyrate) (PHB), crude phenolic extracts (CPE) and protein isolate (PI). The PHB production cost at two annual production capacities ($12.5/kg for 2,500 t PHB/year and $7.8/kg for 25,000 t PHB/year) was not cost-competitive to current PHB production processes when the revenues derived from co-products were not considered. Sensitivity analysis projected the economic viability of a biorefinery concept that could achieve a minimum selling price of $1.1/kg PHB similar to polypropylene. The annual PHB production capacity and the identification of marketable end-uses with respective market prices for the co-products CPE and PI were crucial in attaining process profitability. Greenhouse gas emissions (ca. 0.64 kg CO2-eq/kg PHB) and abiotic depletion potential (61.7 MJ/kg PHB) were lower than polypropylene. Biorefining of sunflower meal and crude glycerol could lead to sustainable PHB production.

Advances in Food and Byproducts Processing towards a Sustainable Bioeconomy.

The bioeconomy concept was initially focused on resource substitution, aiming to mitigate the depletion of fossil resources and confer an alternative approach for resource utilization [...].

Development of a Circular Oriented Bioprocess for Microbial Oil Production Using Diversified Mixed Confectionery Side-Streams.

Diversified mixed confectionery waste streams were utilized in a two-stage bioprocess to formulate a nutrient-rich fermentation media for microbial oil production. Solid-state fermentation was conducted for the production of crude enzyme consortia to be subsequently applied in hydrolytic reactions to break down starch, disaccharides, and proteins into monosaccharides, amino acids, and peptides. Crude hydrolysates were evaluated in bioconversion processes using the red yeast Rhodosporidium toruloides DSM 4444 both in batch and fed-batch mode. Under nitrogen-limiting conditions, during fed-batch cultures, the concentration of microbial lipids reached 16.6-17 g·L-1 with the intracellular content being more than 40% (w/w) in both hydrolysates applied. R. toruloides was able to metabolize mixed carbon sources without catabolite repression. The fatty acid profile of the produced lipids was altered based on the substrate employed in the bioconversion process. Microbial lipids were rich in polyunsaturated fatty acids, with oleic acid being the major fatty acid (61.7%, w/w). This study showed that mixed food side-streams could be valorized for the production of microbial oil with high unsaturation degree, pointing towards the potential to produce tailor-made lipids for specific food applications. Likewise, the proposed process conforms unequivocally to the principles of the circular economy, as the entire quantity of confectionery by-products are implemented to generate added-value compounds that will find applications in the same original industry, thus closing the loop.

Cheese Whey Processing: Integrated Biorefinery Concepts and Emerging Food Applications.

Cheese whey constitutes one of the most polluting by-products of the food industry, due to its high organic load. Thus, in order to mitigate the environmental concerns, a large number of valorization approaches have been reported; mainly targeting the recovery of whey proteins and whey lactose from cheese whey for further exploitation as renewable resources. Most studies are predominantly focused on the separate implementation, either of whey protein or lactose, to configure processes that will formulate value-added products. Likewise, approaches for cheese whey valorization, so far, do not exploit the full potential of cheese whey, particularly with respect to food applications. Nonetheless, within the concept of integrated biorefinery design and the transition to circular economy, it is imperative to develop consolidated bioprocesses that will foster a holistic exploitation of cheese whey. Therefore, the aim of this article is to elaborate on the recent advances regarding the conversion of whey to high value-added products, focusing on food applications. Moreover, novel integrated biorefining concepts are proposed, to inaugurate the complete exploitation of cheese whey to formulate novel products with diversified end applications. Within the context of circular economy, it is envisaged that high value-added products will be reintroduced in the food supply chain, thereby enhancing sustainability and creating "zero waste" processes.

Evaluation of 1,3-propanediol production by twoCitrobacter freundiistrains using crude glycerol and soybean cake hydrolysate.

Biodiesel production processes using soybean as feedstock generates soybean cake and crude glycerol as by-products. These by-product streams were used as sole feedstocks for the production of 1,3-propanediol (PDO) using two bacterial strains of Citrobacter freundii. Soybean cake has been converted into a nutrient-rich hydrolysate by crude enzymes produced via solid state fermentation. The effect of initial glycerol and free amino nitrogen concentration on bacterial growth and PDO production has been evaluated in batch bioreactor cultures showing that C. freundii VK-19 is a more efficient PDO producer than C. freundii FMCC-8. The cultivation of C. freundii VK-19 in fed-batch bioreactor cultures using crude glycerol and soybean cake hydrolysates led to PDO concentration of 47.4 g/L with yield and productivity of 0.49 g/g and 1.01 g/L/h, respectively. The effect of PDO, metabolic by-products, and sodium and potassium salts on bacterial growth was evaluated showing that potassium salts initially enhance bacterial growth, whereas sodium salts cause significant inhibition to bacterial growth. Soybean cake hydrolysate and crude glycerol could be utilized for PDO production, but the fermentation efficiency is influenced by the catalyst used during biodiesel production.

Probiotics in Food Systems: Significance and Emerging Strategies Towards Improved Viability and Delivery of Enhanced Beneficial Value.

Preserving the efficacy of probiotic bacteria exhibits paramount challenges that need to be addressed during the development of functional food products. Several factors have been claimed to be responsible for reducing the viability of probiotics including matrix acidity, level of oxygen in products, presence of other lactic acid bacteria, and sensitivity to metabolites produced by other competing bacteria. Several approaches are undertaken to improve and sustain microbial cell viability, like strain selection, immobilization technologies, synbiotics development etc. Among them, cell immobilization in various carriers, including composite carrier matrix systems has recently attracted interest targeting to protect probiotics from different types of environmental stress (e.g., pH and heat treatments). Likewise, to successfully deliver the probiotics in the large intestine, cells must survive food processing and storage, and withstand the stress conditions encountered in the upper gastrointestinal tract. Hence, the appropriate selection of probiotics and their effective delivery remains a technological challenge with special focus on sustaining the viability of the probiotic culture in the formulated product. Development of synbiotic combinations exhibits another approach of functional food to stimulate the growth of probiotics. The aim of the current review is to summarize the strategies and the novel techniques adopted to enhance the viability of probiotics.

Upgrading Grape Pomace through Pleurotus spp. Cultivation for the Production of Enzymes and Fruiting Bodies.

Grape pomace, a by-product derived from winery industries, was used as fermentation media for the production of added-value products through the cultivation of two Pleurotus species. Solid-state (SSF), semiliquid (SLF), and submerged (SmF) fermentations were carried out using grape pomace as substrate. The effect of the different fermentations on the consumption of phenolic compounds, the production of mycelial mass and enzymes was evaluated using P. ostreatus and P. pulmonarius. The production of fungal biomass and enzymes was influenced by the fermentation mode. The maximum biomass values of ~0.5 g/g were obtained for both P. pulmonarius and P. ostreatus in SmF. Laccase production was induced in SSF and a maximum activity of 26.247 U/g was determined for P. ostreatus, whereas the highest endoglucanase activity (0.93 U/g) was obtained in the SmF of the same fungi. Analysis of phenolic compounds showed that both strains were able to degrade up to 79% of total phenolic content, regardless the culture conditions. Grape pomace was also evaluated as substrate for mushroom production. P. pulmonarius recorded the highest yield and biological efficiency of 14.4% and 31.4%, respectively. This study showed that mushroom cultivation could upgrade winery by-products towards the production of valuable food products.

Wood-Derived Dietary Fibers Promote Beneficial Human Gut Microbiota.

Woody biomass is a sustainable and virtually unlimited source of hemicellulosic polysaccharides. The predominant hemicelluloses in softwood and hardwood are galactoglucomannan (GGM) and arabinoglucuronoxylan (AGX), respectively. Based on the structure similarity with common dietary fibers, GGM and AGX may be postulated to have prebiotic properties, conferring a health benefit on the host through specific modulation of the gut microbiota. In this study, we evaluated the prebiotic potential of acetylated GGM (AcGGM) and highly acetylated AGX (AcAGX) obtained from Norwegian lignocellulosic feedstocks in vitro In pure culture, both substrates selectively promoted the growth of Bifidobacterium, Lactobacillus, and Bacteroides species in a manner consistent with the presence of genetic loci for the utilization of ß-manno-oligosaccharides/ß-mannans and xylo-oligosaccharides/xylans. The prebiotic potential of AcGGM and AcAGX was further assessed in a pH-controlled batch culture fermentation system inoculated with healthy adult human feces. Results were compared with those obtained with a commercial fructo-oligosaccharide (FOS) mixture. Similarly to FOS, both substrates significantly increased (P < 0.05) the Bifidobacterium population. Other bacterial groups enumerated were unaffected with the exception of an increase in the growth of members of the Bacteroides-Prevotella group, Faecalibacterium prausnitzii, and clostridial cluster IX (P < 0.05). Compared to the other substrates, AcGGM promoted butyrogenic fermentation whereas AcAGX was more propiogenic. Although further in vivo confirmation is necessary, these results demonstrate that both AcGGM and AcAGX from lignocellulosic feedstocks can be used to direct the promotion of beneficial bacteria, thus exhibiting a promising prebiotic ability to improve or restore gut health.IMPORTANCE The architecture of the gut bacterial ecosystem has a profound effect on the physiology and well-being of the host. Modulation of the gut microbiota and the intestinal microenvironment via administration of prebiotics represents a valuable strategy to promote host health. This work provides insights into the ability of two novel wood-derived preparations, AcGGM and AcAGX, to influence human gut microbiota composition and activity. These compounds were selectively fermented by commensal bacteria such as Bifidobacterium, Bacteroides-Prevotella, F. prausnitzii, and clostridial cluster IX spp. This promoted the microbial synthesis of acetate, propionate, and butyrate, which are beneficial to the microbial ecosystem and host colonic epithelial cells. Thus, our results demonstrate potential prebiotic properties for both AcGGM and AcAGX from lignocellulosic feedstocks. These findings represent pivotal requirements for rationally designing intervention strategies based on the dietary supplementation of AcGGM and AcAGX to improve or restore gut health.

In vitro fermentation properties of pectins and enzymatic-modified pectins obtained from different renewable bioresources.

The suitability of artichoke and sunflower by-products as renewable sources of pectic compounds with prebiotic potential was evaluated by studying their ability to modulate the human faecal microbiota in vitro. Bacterial populations and short-chain fatty acid (SCFA) production were measured. Reduction of the molecular weight of artichoke pectin resulted in greater stimulation of the growth of Bifidobacterium, Lactobacillus and Bacteroides/Prevotella, whilst this effect was observed only in Bacteroides/Prevotella for sunflower samples. In contrast, the degree of methoxylation did not have any impact on fermentability properties or SCFA production, regardless of the origin of pectic compounds. Although further in vivo studies should be conducted, either pectin or enzymatically-modified pectin from sunflower and artichoke by-products might be considered as prebiotic candidates for human consumption showing similar ability to promote the in vitro growth of beneficial gut bacteria as compared to well-recognized prebiotics such as inulin or fructo-oligosaccharides.

Valorization of By-Products from Palm Oil Mills for the Production of Generic Fermentation Media for Microbial Oil Synthesis.

This study demonstrates the production of a generic nutrient-rich feedstock using by-product streams from palm oil production that could be used as a substitute for commercial fermentation supplements. Solid-state fermentations of palm kernel cake (PKC) and palm-pressed fiber (PPF) were conducted in tray bioreactors and a rotating drum bioreactor by the fungal strain Aspergillus oryzae for the production of crude enzymes. The production of protease was optimized (319.3 U/g) at an initial moisture content of 55 %, when PKC was used as the sole substrate. The highest free amino nitrogen (FAN) production (5.6 mg/g) obtained via PKC hydrolysis using the crude enzymes produced via solid-state fermentation was achieved at 50 °C. Three initial PKC concentrations (48.7, 73.7, and 98.7 g/L) were tested in hydrolysis experiments, leading to total Kjeldahl nitrogen to FAN conversion yields up to 27.9 %. Sequential solid-state fermentation followed by hydrolysis was carried out in the same rotating drum bioreactor, leading to the production of 136.7 U/g of protease activity during fermentation and 196.5 mg/L of FAN during hydrolysis. Microbial oil production was successfully achieved with the oleaginous yeast strain Lipomyces starkeyi DSM 70296 cultivated on the produced PKC hydrolysate mixed with commercial carbon sources, including glucose, xylose, mannose, galactose, and arabinose.

Valorization of bakery waste for biocolorant and enzyme production by Monascus purpureus.

A concept of utilizing bakery waste as a nutrient source for the fermentative production of bio-colorant by Monasucus purpureus has been developed. The proposed ideas provide an innovative approach to establish a system or method to reduce the bakery waste problem encountered by Hong Kong or other countries. Bakery waste collected from bakery store was used in submerged and solid-state fermentation of Monascus purpureus to produce bio-colorant, which could potentially be applied in food and textile industries. The feasibility of utilizing bakery waste hydrolysate deriving from hydrolytic reactions by Aspergillus awamori and Aspergillus oryzae for fermentative pigment production was investigated. Preliminary data from bakery waste hydrolysate experiment presented that the highest pigment yield (about 24AU/g glucose) was obtained with bakery waste hydrolysate containing 5g/L initial glucose. Results from the solid state fermentation studies presented that the highest activity of glucoamylase and protease achieved was 8U/g and 117U/g respectively, at an initial moisture content of 55% and 65% respectively at 30°C incubation temperature. The outcome from this study demonstrated that Monascus purpureus constitutes a promising host for bio-colorant and enzyme production using recovered sugars and amino acids from bakery waste.

Sunflower-based biorefinery: poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) production from crude glycerol, sunflower meal and levulinic acid.

Polyhydroxybutyrate (PHB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] production was developed in bioreactor cultures using the strain Cupriavidus necator DSM 7237 cultivated on crude glycerol, sunflower meal (SFM) hydrolysates and levulinic acid as the sole fermentation feedstocks. Bacterial growth and PHB production was influenced significantly by the free amino nitrogen and inorganic phosphorus content of the SFM hydrolysate. Fed-batch bioreactor fermentations led to the production of 27gL(-1) PHB with an intracellular content of 72.9% (w/w). Continuous feeding of levulinic acid led to the production of up to 23.4gL(-1) P(3HB-co-3HV) with an intracellular content of 66.4% (w/w) and a 3HV content of 22.5mol%. A maximum 3HV content of 31mol% was achieved at earlier fermentation time (53h). Thus, levulinic acid could be combined with biodiesel industry by-products for the production of high P(3HB-co-3HV) concentration, intracellular content and industrially useful 3HV content.

Biorefinery development through utilization of biodiesel industry by-products as sole fermentation feedstock for 1,3-propanediol production.

Rapeseed meal (RSM) hydrolysate was evaluated as substitute for commercial nutrient supplements in 1,3-propanediol (PDO) fermentation using the strain Clostridium butyricum VPI 1718. RSM was enzymatically converted into a generic fermentation feedstock, enriched in amino acids, peptides and various micro-nutrients, using crude enzyme consortia produced via solid state fermentation by a fungal strain of Aspergillus oryzae. Initial free amino nitrogen concentration influenced PDO production in batch cultures. RSM hydrolysates were compared with commercial nutrient supplements regarding PDO production in fed-batch cultures carried out in a bench-scale bioreactor. The utilization of RSM hydrolysates in repeated batch cultivation resulted in a PDO concentration of 65.5 g/L with an overall productivity of 1.15 g/L/h that was almost 2 times higher than the productivity achieved when yeast extract was used as nutrient supplement.