In-package cold atmospheric plasma processing for shelf-life extension of gilthead seabream (Sparus aurata) fillets.

In-package cold atmospheric plasma (CAP) processing, which refers to the generation of CAP inside a sealed package, enables a local disinfecting reaction, allowing no post-process contamination and extending the shelf-life (SL) of perishable food products, such as fresh fish. In the present study, four in-package CAP treatments, differing in frequency and processing time, were applied on fresh gilthead seabream (Sparus aurata) fillets, prepacked in low-permeability pouches. Fish SL was evaluated during isothermal storage at 2°C, whereas untreated packaged fillets were used as control samples. The SL assessment of the fish fillets was based on microbial enumeration of total aerobic mesophilic count (TMC), total aerobic psychrotrophic count (TPC), Pseudomonas spp., Enterobacteriaceae, and lactic acid bacteria (LAB), pH measurement, determination of color and texture parameters, lipid oxidation, total volatile basic nitrogen (TVB-N) measurement, and sensory evaluation. All CAP treatments were effective against microbial inhibition in fish fillets, especially regarding TMC, TPC, and Pseudomonas spp., resulting in maximum reduction of 1.49, 1.24, and 1.43 log CFU/g, respectively, compared to the control samples after 16 days of storage. However, minor effect was observed against Enterobacteriaceae and no effect against LAB. CAP processing did not affect the color and texture parameters of fish fillets, and TVB-N production was slightly reduced in CAP-treated samples; however, lipid oxidation was accelerated, especially at the more intense processing conditions, by a maximum of 75.5%. The results of the study indicated that in-package CAP processing could be effectively applied for inhibiting spoilage during refrigerated storage and extending SL of fresh fish fillets. PRACTICAL APPLICATION: In-package cold atmospheric plasma (CAP) processing was tested on gilthead seabream fillets, a highly perishable product with high commercial potential if its shelf-life can be extended through minimal processing. The food industry could benefit from in-package CAP technology as it is a cost effective nonthermal processing method while preventing post-processing contamination of the products. Although in-package CAP processing has not been extensively tested on fish, this study examined the quality and shelf-life of a highly perishable fish species, and the results could be further used as a reference for processing optimization of the CAP treatments.

Scale-up and fed-batch cultivation strategy for the enhanced co-production of microbial lipids and carotenoids using renewable waste feedstock.

Agro-industrial by-product valorization as a feedstock for the bioproduction of high-value products has demonstrated a feasible alternative to handle the environmental impact of waste. Oleaginous yeasts are promising cell factories for the industrial production of lipids and carotenoids. Since oleaginous yeasts are aerobic microorganisms, studying the volumetric mass transfer (kLa) could facilitate the scale-up and operation of bioreactors to grant the industrial availability of biocompounds. Scale-up experiments were performed to assess the simultaneous production of lipids and carotenoids using the yeast Sporobolomyces roseus CFGU-S005 and comparing the yields in batch and fed-batch mode cultivation using agro-waste hydrolysate in a 7 L bench-top bioreactor. The results indicate that oxygen availability in the fermentation affected the simultaneous production of metabolites. The highest production of lipids (3.4 g/L) was attained using the kLa value of 22.44 h-1, while higher carotenoid accumulation of 2.58 mg/L resulted when agitation speed was increased to 350 rpm (kLa 32.16 h-1). The adapted fed-batch mode in the fermentation increased the production yields two times. The fatty acid profile was affected according to supplied aeration and after the fed-batch cultivation mode. This study showed the scale-up potential of the bioprocess using the strain S. roseus in the obtention of microbial oil and carotenoids by the valorization of agro-industrial byproducts as a carbon source.

Fermentation development using fruit waste derived mixed sugars for poly(3-hydroxybutyrate) production and property evaluation.

Free sugars from fruit wastes were evaluated for the production of poly(3-hydroxybutyrate) (PHB) in Paraburkholderia sacchari fed-batch bioreactor fermentations. Different initial sugar concentration, carbon to inorganic phosphorus (C/IP) ratio, IP addition during feeding and volumetric oxygen transfer coefficient (kLa) were evaluated to promote PHB production. The highest intracellular PHB accumulation (66.6%), PHB concentration (108.3 g/L), productivity (3.28 g/(L·h)) and yield (0.33 g/g) were achieved at 40 g/L initial sugars, C/IP 26.5, 202.6 h-1kLa value and 20% IP supplementation in the feeding solution. The effect of different microbial mass harvesting time on PHB properties showed no influence in weight average molecular weight and thermal properties. The harvest time influenced the tensile strength that was reduced from 28.7 MPa at 22 h to 13.3 MPa at 36 h. The elongation at break and Young's modulus were in the range 3.6-14.8% and 830-2000 MPa, respectively.

Transcriptional regulation in key metabolic pathways of Actinobacillus succinogenes in the presence of electricity.

The potential of renewable energy application via direct electrode interaction for the production of bio-based chemicals is a promising technology. The utilization of extracellular energy in pure culture fermentations aims in intracellular redox balance regulation in order to improve fermentation efficiency. This work evaluates the impact of a bioelectrochemical system in succinic acid fermentation and the metabolic response of Actinobacillus succinogenes. The metabolic pathway regulation of A. succinogenes was evaluated via RNA expression of the key enzymes that participate in TCA cycle, pyruvate metabolism and oxidative phosphorylation. The genes that were significantly overexpressed in BES compared to non-BES were phosphoenolpyruvate carboxykinase (0.4-fold change), inorganic pyrophosphatase (2.3-fold change) and hydrogenase (2.2-fold change) and the genes that were significantly underexpressed were fumarase (-0.94-fold change), pyruvate kinase (-6.9-fold change), all subunits of fumarate reductase (-2.1 to -1.17-fold change), cytochromes I and II (-1.25 and -1.02-fold change, respectively) and two C4-carboxylic acid transporters.

Fermentative bioconversion of food waste into biopolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) using Cupriavidus necator.

Dependency on plastic commodities has led to a recurrent increase in their global production every year. Conventionally, plastic products are derived from fossil fuels, leading to severe environmental concerns. The recent coronavirus disease 2019 pandemic has triggered an increase in medical waste. Conversely, it has disrupted the supply chain of personal protective equipment (PPE). Valorisation of food waste was performed to cultivate C. necator for fermentative production of biopolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). The increase in biomass, PHBV yield and molar 3-hydroxy valerate (3HV) content was estimated after feeding volatile fatty acids. The fed-batch fermentation strategy reported in this study produced 15.65 ± 0.14 g/L of biomass with 5.32 g/L of PHBV with 50% molar 3HV content. This is a crucial finding, as molar concentration of 3HV can be modulated to suit the specification of biopolymer (film or fabric). The strategy applied in this study addresses the issue of global food waste burden and subsequently generates biopolymer PHBV, turning waste to wealth.

Techno-economic optimization of a process superstructure for lignin valorization.

Lignin, the most abundant aromatic biopolymer on Earth, is often considered a biorefinery by-product, despite its potential to be valorized into high-added-value chemicals and fuels. In this work, an integrated superstructure-based optimization model was set up and optimized using mixed-integer non-linear programming for the conversion of technical lignin to three main biobased products: aromatic monomers, phenol-formaldehyde resins, and aromatic aldehydes/acids. Several alternative conversion pathways were simultaneously compared to assess the profitability of lignins-based processes by predicting the performance of technologies with different TRL. Upon employing key technologies such as hydrothermal liquefaction, dissolution in solvent, or high-temperature electrolysis, the technical lignins could have a market value of 200 €/t when the market price for aromatic monomers, resins, and vanillin is at least 2.0, 0.8, and 15.0 €/kg, respectively. When lower product selling prices were considered, the aromatic monomers and the resins were not profitable as target products.

Sustainable Production of Novel Oleogels Valorizing Microbial Oil Rich in Carotenoids Derived from Spent Coffee Grounds.

Sustainable food systems that employ renewable resources without competition with the food chain are drivers for the bioeconomy era. This study reports the valorization of microwave-pretreated spent coffee grounds (SCGs) to produce oleogels rich in bioactive compounds. Microbial oil rich in carotenoids (MOC) was produced under batch fermentation of Rhodosporidium toruloides using SCG enzymatic hydrolysates. Candelilla wax (CLW) could structure MOC and sunflower oil at a 3.3-fold lower concentration than that of carnauba wax (CBW). MOC-based oleogels with 10% CBW and 3% CLW showed an elastic-dominant and gel-like structure (tan δ ≪ 1), providing gelation and oil binding capacity (>95%). Dendritic structures of CBW-based oleogels and evenly distributed rod-like crystals of CLW-based ones were observed via polarized light microscopy. MOC-based oleogels exhibited similar Fourier-transform infrared spectroscopy spectra. X-ray diffractograms of oleogels were distinguished by the oil type that presented ß'-type polymorphism. MOC-based oleogels could be applied in confectionary products and spreads as substitutes for trans fatty acids, reformulating fat-containing food products.

Development of biodegradable films using sunflower protein isolates and bacterial nanocellulose as innovative food packaging materials for fresh fruit preservation.

This study presents the valorization of side streams from the sunflower-based biodiesel industry for the production of bio-based and biodegradable food packaging following circular economy principles. Bacterial cellulose (BC) was produced via fermentation in 6 L static tray bioreactors using nutrient-rich supplements derived from the enzymatic hydrolysis of sunflower meal (SFM) combined with crude glycerol as carbon source. Novel biofilms were produced using either matrices of protein isolates extracted from sunflower meal (SFMPI) alone or SFMPI matrices reinforced with nanocellulose biofillers of commercial or bacterial origin. Acid hydrolysis was employed for ex-situ modification of BC to nanostructures (56 nm). The biofilms reinforced with bacterial nanocellulose structures (SFMPI-BNC) showed 64.5% higher tensile strength, 75.5% higher Young's modulus, 131.5% higher elongation at break, 32.5% lower water solubility and 14.1% lower water vapor permeability than the biofilms produced only with SFMPI. The biofilms were evaluated on fresh strawberries packaging showing that the SFMPI-BNC-based films lead to effective preservation at 10 °C considering microbial growth and physicochemical profile (weight loss, chemical characterization, color, firmness and respiration activity). The SFMPI-BNC-based films could be applied in fresh fruit packaging applications.

Biorefinery development, techno-economic evaluation and environmental impact analysis for the conversion of the organic fraction of municipal solid waste into succinic acid and value-added fractions.

The organic fraction of municipal solid waste (OFMSW) was used for biorefinery development within a circular bioeconomy context towards extraction of lipids/fats and proteins with 100% and 68% recovery yields, respectively, as well as succinic acid (SA) production. A nutrient-rich hydrolysate (89.1 g/L sugars) produced using crude enzymes derived via solid state fermentation of Aspergillus awamori, was employed in Actinobacillus succinogenes fermentation leading to 31.7 gSA/L with 0.68 g/g yield and 0.67 g/L/h productivity. The SA minimum selling price ($1.13-2.39/kgSA) considering 60,000 tSA/year production varied depending on co-product market prices and OFMSW management fees. The biorefinery using 1000 kg OFMSW contributes 35% lower CO2 emissions than conventional processes for the production of 105 kg vegetable oil, 87 kg vegetable protein and 206.4 kg fossil-SA considering also the CO2 emissions due to OFMSW landfilling. The proposed OFMSW biorefinery leads to cost-competitive SA production with lower CO2 emissions for OFMSW treatment.

Effect of a Carotenoid Extract from Citrus reticulata By-Products on the Immune-Oxidative Status of Broilers.

Although carotenoids generally possess antimicrobial and antioxidant properties, the in vivo synergistic action of carotenoid blends derived from plant-based by-products has not been thoroughly studied. Therefore, the carotenoid characterization and antimicrobial potential of Citrus reticulata extract as well as the impact of this carotenoid-rich extract (CCE) dietary supplementation on the performance, meat quality, and immune-oxidative status of broiler chickens were determined. One hundred and twenty one-day-old hatched chicks (Ross 308) were allocated to two dietary groups, with four replicate pens of 15 birds each. Birds were fed either a basal diet (CON) or the basal diet supplemented with 0.1% CCE (25 mg carotenoid extract included in 1 g of soluble starch) for 42 d. ß-Cryptoxanthin, ß-Carotene, Zeaxanthin, and Lutein were the prevailing carotenoid compounds in the Citrus reticulata extract. The CCE feed additive exerted inhibitory properties against both Gram-positive (Staphylococcus aureus) and negative (Klebsiella oxytoca, Escherichia coli, and Salmonella typhimurium) bacteria. Both the broiler performance and meat quality did not substantially differ, while the breast muscle malondialdehyde (MDA) concentration tended to decrease (p = 0.070) in the CCE-fed broilers. The inclusion of CCE decreased the alanine aminotransferase and MDA concentration, and the activity of glutathione peroxidase, while the activity of superoxide dismutase was increased in the blood. Catalase and NADPH oxidase 2 relative transcript levels were significantly downregulated in the livers of the CCE-fed broilers. Additionally, Interleukin 1ß and tumor necrosis factor (TNF) relative transcript levels were downregulated in the livers of the CCE- fed broilers, while TNF and interferon γ (IFNG) tended to decrease in the spleens and bursa of Fabricius, respectively. The present study provided new insights regarding the beneficial properties of carotenoids contained in Citrus reticulata in broilers' immune-oxidative status. These promising outcomes could be the basis for further research under field conditions.

Prospects on bio-based 2,3-butanediol and acetoin production: Recent progress and advances.

The bio-based platform chemicals 2,3-butanediol (BDO) and acetoin have various applications in chemical, cosmetics, food, agriculture, and pharmaceutical industries, whereas the derivatives of BDO could be used as fuel additives, polymer and synthetic rubber production. This review summarizes the novel technological developments in adapting genetic and metabolic engineering strategies for selection and construction of chassis strains for BDO and acetoin production. The valorization of renewable feedstocks and bioprocess development for the upstream and downstream stages of bio-based BDO and acetoin production are discussed. The techno-economic aspects evaluating the viability and industrial potential of bio-based BDO production are presented. The commercialization of bio-based BDO and acetoin production requires the utilization of crude renewable resources, the chassis strains with high fermentation production efficiencies and development of sustainable purification or conversion technologies.

Techno-economic risk assessment, life cycle analysis and life cycle costing for poly(butylene succinate) and poly(lactic acid) production using renewable resources.

The sustainable production of poly(lactic acid) (PLA) or poly(butylene succinate) (PBS) from corn glucose syrup, corn stover and sugar beet pulp (SBP) have been assessed via process design, preliminary techno-economic evaluation, life cycle assessment and life cycle costing (LCC). Cost-competitive PLA and PBS production can be achieved in a SBP-based biorefinery, including separation of crude pectin-rich extract as co-product, leading to minimum selling prices of $1.14/kgPLA and $1.37/kgPBS. Acidification Potential, Eutrophication Potential and Human Toxicity Potential are lower when SBP is used. The LCC of PLA ($1.42/kgPLA) and PBS ($1.72/kgPBS) production from SBP are lower than biaxial oriented polypropylene (BOPP, $1.66/kg) and general purpose polystyrene (GPPS, $2.04/kg) at pectin-rich extract market prices of $3/kg and $4/kg, respectively. Techno-economic risk assessment via Monte-Carlo simulations showed that PLA and PBS could be produced from SBP at the market prices of BOPP ($1.4/kg) and GPPS ($1.72/kg) with 100% probability to achieve a positive Net Present Value at pectin-rich extract market prices of $3/kg and $4/kg, respectively. This study demonstrated that SBP-based biorefinery development ensures sustainable production of PLA and PBS as compared to fossil-derived counterparts and single product bioprocesses using glucose syrup and corn stover.

Integrated biorefinery development using winery waste streams for the production of bacterial cellulose, succinic acid and value-added fractions.

An integrated biorefinery has been developed using winery wastes (grape pomace-GP, stalks-GS, wine lees-WL). Bacterial cellulose was produced from GP extracted free sugars. Grape-seed oil and polyphenols were extracted from GP. Experimental design was employed to optimize lignin removal (50.8%) from mixtures of remaining GP solids and GS via NaOH (1.19% w/v) treatment at 70°C for 30 min. Delignification liquid contained condensed tannins with 76% Stiasny number. Enzymatic hydrolysis produced a sugar-rich hydrolysate (40.2 g/L sugars). Ethanol, antioxidants, tartaric acid and nutrient-rich hydrolysate were produced from WL. The crude hydrolysates were used in fed-batch Actinobacillus succinogenes cultures for 37.2 g/L succinic acid production. The biorefinery produces 42.65 g bacterial cellulose, 24.3 g oil, 40.3 g phenolic-rich extract with 1.41 Antioxidant Activity Index, 80.2 g ethanol, 624.8 g crude tannin extract, 20.03 g tartaric acid and 157.8 g succinic acid from 1 kg of each waste stream.

Techno-economic evaluation and life cycle assessment of a biorefinery using winery waste streams for the production of succinic acid and value-added co-products.

This study presents techno-economic evaluation and life cycle assessment of a novel biorefinery using the three main waste streams generated by wineries for the production of bio-based succinic acid (SA), crude phenolic-rich extract, grape-seed oil, calcium tartrate and crude tannin-rich extract. Process design has been employed for the estimation of material and energy balances and the sizing of unit operations. The Minimum Selling Price of succinic acid production within a winery waste biorefinery ranges from $1.23-2.76/kgSA depending on the market price and the potential end-uses of the extracted fractions. The Global Warming Potential and the Abiotic Depletion Potential of winery waste valorisation through the proposed biorefinery are 1.47 kg CO2-eq per kg dry waste and 25.2 MJ per kg dry waste, respectively. Biorefining of winery waste could lead to the development of a sustainable and novel bioeconomy business model with new market opportunities and efficient waste management.

Immune-Related Gene Expression Profiling of Broiler Chickens Fed Diets Supplemented with Vinification Byproducts: A Valorization Approach II.

The valorization of vinification byproducts portrays a promising bioprocess for the enrichment of animals' diet with bioactive compounds, such as polyphenols, which could regulate the immune response. Therefore, the impact of dietary grounded grape pomace (GGP), wine lees extract (WYC), and grape stem extract (PE) on the relative transcript level of immune related genes of broiler chickens were examined. Two hundred forty, one-day-old as hatched (male/female) chicks (Ross 308) were allocated to four dietary groups, with four replicate pens each with 15 birds. Birds were fed either a basal diet (CON) or the basal diet supplemented with 2.5% GGP, or 0.2% WYC, or 0.1% PE for 42 d. The relative expression of immune-related genes was investigated using a real-time PCR platform. The mRNA levels of Toll-like Receptor 4 (TLR4) were downregulated (p = 0.039) in the liver of broilers fed the GGP-containing diet compared to the CON, while in the spleen of PE-fed broilers, TLR4 was significantly upregulated (p = 0.043). The mRNA levels of interleukin 8 (IL8) tended to upregulate (p = 0.099) in the bursa of Fabricius and were significantly increased (p = 0.036) in the spleen of broilers fed the PE diet. Vinification byproducts depict a promising sustainable source of polyphenols for the poultry feed industry, but more research is needed under field conditions.

Antioxidant Status of Broiler Chickens Fed Diets Supplemented with Vinification By-Products: A Valorization Approach.

Vinification by-products display great potential for utilization as feed additives rich in antioxidant compounds. Thus, the effect of dietary ground grape pomace (GGP), wine lees extract rich in yeast cell walls (WYC), and grape stem extracts (PE) on the relative expression of several genes involved in liver oxidative mechanisms and the oxidative status of the blood and breast muscle of broiler chickens was investigated. In total, 240 one-day-old as hatched chicks (Ross 308) were assigned to four treatments, with four replicate pens and 15 birds in each pen. Birds were fed either a basal diet (CON) or a basal diet supplemented with 25 g/kg GGP, or 2 g/kg WYC, or 1 g starch including 100 mg pure stem extract/kg (PE) for 42 days. The polyphenolic content of vinification by-products was determined using an LC-MS/MS library indicating as prevailing compounds procyanidin B1 and B2, gallic acid, caftaric acid, (+)-catechin, quercetin, and trans-resveratrol. Body weight and feed consumption were not significantly affected. The relative transcript level of GPX1 and SOD1 tended to increase in the liver of WYC-fed broilers, while NOX2 tended to decrease in the PE group. SOD activity in blood plasma was significantly increased in WYC and PE compared to the CON group. The total antioxidant capacity measured with FRAP assay showed significantly higher values in the breast muscle of PE-fed broilers, while the malondialdehyde concentration was significantly decreased in both WYC- and PE-fed broilers compared to the CON group. The exploitation of vinification by-products as feed additives appears to be a promising strategy to improve waste valorization and supply animals with bioactive molecules capable of improving animals' oxidative status and products' oxidative stability.

Volumetric oxygen transfer coefficient as fermentation control parameter to manipulate the production of either acetoin or D-2,3-butanediol using bakery waste.

The formation of either acetoin or D-2,3-butanediol (D-BDO) by Bacillus amyloliquefaciens cultivated on bakery waste hydrolysates has been evaluated in bioreactor cultures by varying the volumetric oxygen transfer coefficient (kLa). The highest D-BDO production (55.2 g L-1) was attained in batch fermentations with kLa value of 64 h-1. Batch fermentations performed at 203 h-1 led to the highest productivity (2.16 g L-1h-1) and acetoin production (47.4 g L-1). The utilization of bakery waste hydrolysate in fed-batch cultures conducted at kLa of 110 h-1 led to combined production of acetoin, meso-BDO and D-BDO (103.9 g L-1). Higher kLa value (200 h-1) resulted to 65.9 g L-1 acetoin with 1.57 g L-1h-1 productivity. It has been demonstrated that the kLa value may divert the bacterial metabolism towards high acetoin or D-BDO production during fermentation carried out in crude bakery waste hydrolysates.

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.

Biodiesel production using microbial lipids derived from food waste discarded by catering services.

Biodiesel production using microbial oil derived from food waste discarded by the hospilatity sector could provide a sustainable replacement for diesel fuel. Discarded potato peels were used in solid-state fermentations of Aspergillus awamori for the production of glucoamylase (30 U/g) and protease (50 U/g). Hospitality food waste hydrolysis led to 98% (w/w) starch to glucose conversion yields. Crude hydrolyzates were used in shake flask fermentations with the oleaginous yeast Rhodosporidium toruloides Y-27012 leading to 32.9 g/L total dry weight (TDW) with 36.4% (w/w) intracellular lipid content. Fed-bath bioreactor cultures resulted in TDW of 53.9 g/L and lipid concentration of 26.7 g/L. Principal component analysis showed a fatty acid profile similar to soybean oil and solid food waste oil. Microbial oil was transesterified into biodiesel with satisfactory performance considering the European standard EN 14214. This work demonstrated that valorization of food waste for biodiesel production is feasible.

Chemical Profiling, Bioactivity Evaluation and the Discovery of a Novel Biopigment Produced by Penicillium purpurogenum CBS 113139.

Biobased pigments are environmentally friendly alternatives to synthetic variants with an increased market demand. Production of pigments via fermentation is a promising process, yet optimization of the production yield and rate is crucial. Herein, we evaluated the potential of Penicillium purpurogenum to produce biobased pigments. Optimum sugar concentration was 30 g/L and optimum C:N ratio was 36:1 resulting in the production of 4.1-4.5 AU (namely Pigment Complex A). Supplementation with ammonium nitrate resulted in the production of 4.1-4.9 AU (namely Pigment Complex B). Pigments showed excellent pH stability. The major biopigments in Pigment Complex A were N-threonyl-rubropunctamin or the acid form of PP-R (red pigment), N-GABA-PP-V (violet pigment), PP-O (orange pigment) and monascorubrin. In Pigment Complex B, a novel biopigment annotated as N-GLA-PP-V was identified. Its basic structure contains a polyketide azaphilone with the same carboxyl-monascorubramine base structure as PP-V (violet pigment) and γ-carboxyglutamic acid (GLA). The pigments were not cytotoxic up to 250 µg/mL.