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Open AccessArticle

Characterization of the Red Biochromes Produced by the Endophytic Fungus Monascus purpureus CPEF02 with Antimicrobial and Antioxidant Activities

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Fermentation 2023, 9(4), 328; https://doi.org/10.3390/fermentation9040328 - 25 Mar 2023

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Abstract

Food acceptability and appeal are significantly influenced by colour. Harmful effects associated with synthetic colorants are well established, and research is currently focused on developing natural, synthetic chemical-free substitutes from fungal sources, with broad applications in food, medicine, textiles and agriculture. Additionally, the [...] Read more.

Food acceptability and appeal are significantly influenced by colour. Harmful effects associated with synthetic colorants are well established, and research is currently focused on developing natural, synthetic chemical-free substitutes from fungal sources, with broad applications in food, medicine, textiles and agriculture. Additionally, the market’s dearth of natural red colour substitutes requires the creation of novel red pigment alternatives from secure and scalable sources. The goal of the current research was to establish new endophytic marine fungi that are naturally occurring bio-sources of the red pigment. Based on its profuse extracellular red pigment-producing capacity, the fungus CPEF02 was selected and identified as Monascus purpureus CPEF02 via internal transcribed spacer (ITS) sequences and phylogenetic analysis. The chemical moieties of the pigmented extracts were identified by liquid chromatography-high resolution mass spectrometry (LC-HRMS). The optimal culture conditions for maximum pigment production were investigated by surveying various media compositions. The methanolic fungal colourant extract was shown to have substantial antibacterial and antifungal activities against anthropogenic pathogens, Staphylococcus aureus (MTCC 1430), methicillin-resistant Staphylococcus aureus (ATCCBAA811), Salmonella typhimurium (MTCC 3241) and Vibrio cholerae (N16961) at a 100 µg/mL concentration and at a 1 mg/mL concentration for Alternaria solani (ITCC 4632) and Rhizoctonia solani (AG1-IA). This extract also exhibited antioxidant activity against the 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical with an IC₅₀ of 14.42 µg/mL and a Trolox equivalent antioxidant capacity of 0.571 µM Trolox/µg of the methanolic colourant extract. The findings suggested that M. purpureus’s pigment could be a source of an industrially useful natural red colourant. Full article

(This article belongs to the Special Issue Pigment Production in Submerged Fermentation)

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Open AccessArticle

Pigment Production of Chlamydomonas Strains in Response to Norflurazon and ZnO Nanoparticles

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Thanaporn Intha

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Anchalee Sirikhachornkit

Fermentation 2023, 9(2), 193; https://doi.org/10.3390/fermentation9020193 - 19 Feb 2023

Viewed by 759

Abstract

Numerous species of microalgae have been utilized for pigment production. More and more species are gaining popularity due to their ability to accumulate pigments with varying chemical compositions and the fact that some have distinctive byproducts that can be co-produced. Despite the fact [...] Read more.

Numerous species of microalgae have been utilized for pigment production. More and more species are gaining popularity due to their ability to accumulate pigments with varying chemical compositions and the fact that some have distinctive byproducts that can be co-produced. Despite the fact that many of the species have unique by-products and traits, they are not being used economically due to high production costs. Utilizing agricultural and industrial wastewater for algae cultivation is one way to lower manufacturing costs. Herbicide-contaminated wastewater can result from agricultural contamination. Norflurazon is a popular pesticide frequently used for weed control. The presence of norflurazon in water renders that water unusable and requires proper treatment. Nanoparticles of ZnO (ZnO NPs), on the other hand, are utilized in a variety of industrial productions of numerous household goods. Water contaminated with ZnO NPs can present potential risks to human health and the environment. In this study, two field isolates of the green microalga Chlamydomonas reinhardtii, a widely used model organism, were examined for their reaction to these two compounds in order to assess the responses of different natural strains to environmental stresses. Norflurazon at 10 µM had a higher inhibitory effect on growth and pigment production than ZnO NPs at 200 mg L⁻¹. Although both norflurazon and ZnO NPs inhibit cell growth and pigmentation, they do so through distinct processes. Norflurazon induces oxidative stress in cells, resulting in photosystem damage. ZnO nanoparticles, on the other hand, did not cause photosystem damage but rather mechanical cell damage and disintegration. In addition, the physiological responses of the two Chlamydomonas strains were distinct, supporting the utilization of natural algal strains for specific types of environmental pollutants. Full article

(This article belongs to the Special Issue Pigment Production in Submerged Fermentation)

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Open AccessArticle

Production of Pigments under Submerged Culture through Repeated Batch Fermentation of Immobilized Talaromyces atroroseus GH2

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Juan Pablo Ruiz-Sánchez

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Lourdes Morales-Oyervides

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Daniele Giuffrida

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Laurent Dufossé

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Julio César Montañez

Fermentation 2023, 9(2), 171; https://doi.org/10.3390/fermentation9020171 - 14 Feb 2023

Cited by 2 | Viewed by 983

Abstract

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 [...] Read more.

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. Full article

(This article belongs to the Special Issue Pigment Production in Submerged Fermentation)

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Open AccessArticle

Monascus Yellow Pigment Production by Coupled Immobilized-Cell Fermentation and Extractive Fermentation in Nonionic Surfactant Micelle Aqueous Solution

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Fermentation 2023, 9(2), 168; https://doi.org/10.3390/fermentation9020168 - 13 Feb 2023

Viewed by 675

Abstract

Microbial fermentation with immobilized cells possesses many advantages. However, this fermentation mode is restricted to the production of extracellular products. Our previous study demonstrated that the extractive fermentation of Monascus spp. in nonionic surfactant micelle aqueous solution can export Monascus pigments that are [...] Read more.

Microbial fermentation with immobilized cells possesses many advantages. However, this fermentation mode is restricted to the production of extracellular products. Our previous study demonstrated that the extractive fermentation of Monascus spp. in nonionic surfactant micelle aqueous solution can export Monascus pigments that are supposed to be mainly intracellular products to extracellular culture broth and, in the meantime, extracellularly enhance the production of yellow pigments at a low pH condition; consequently, this makes the continuous production of yellow pigments with immobilized Monascus cells feasible. In this study, immobilized-cell fermentation and extractive fermentation in Triton X-100 micelle aqueous solution were successfully combined to continuously produce Monascus yellow pigments extracellularly. We examined the effects of cell immobilization and Triton X-100 on cell growth, pigment production, and pigment composition. In the repeated-batch extractive fermentation with immobilized cells, the biomass in Ca-alginate gel beads continued to grow and reached 21.2 g/L after seven batches, and dominant yellow pigments were produced extracellularly and stable for each batch. The mean productivity of the extracellular yellow pigments reached up to 22.31 AU_{410 nm}/day within the first four batches (13 days) and 19.7 AU_{410 nm}/day within the first seven batches (25 days). The results also provide a new strategy for producing such intracellular products continuously and extracellularly. Full article

(This article belongs to the Special Issue Pigment Production in Submerged Fermentation)

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Open AccessArticle

Highly Active Astaxanthin Production from Waste Molasses by Mutated Rhodosporidium toruloides G17

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Fermentation 2023, 9(2), 148; https://doi.org/10.3390/fermentation9020148 - 02 Feb 2023

Viewed by 834

Abstract

Astaxanthin is increasingly attracting commercial interest for its application in the nutraceutical and pharmaceutical industries. This study aimed to produce astaxanthin from molasses with our newly mutated strain of Rhodosporidium toruloides G17 and to evaluate biological activities of the produced astaxanthin. To maximize [...] Read more.

Astaxanthin is increasingly attracting commercial interest for its application in the nutraceutical and pharmaceutical industries. This study aimed to produce astaxanthin from molasses with our newly mutated strain of Rhodosporidium toruloides G17 and to evaluate biological activities of the produced astaxanthin. To maximize the astaxanthin yield, the response surface methodology was used so as to optimize the culture conditions. A maximum astaxanthin yield of 1262.08 ± 14.58 µg/L was achieved by growing R. toruloides G17 in a molasses-based medium containing 49.39 g/L reducing sugar, 1.00 g/L urea, 4.15 g/L MgSO₄·7H₂O, and 10.05% inoculum ratio. The produced astaxanthin was then purified and studied for its antioxidant and anticancer activities. This compound exhibited 123-fold higher antioxidant activity than α-tocopherol, with an IC₅₀ value of 0.97 ± 0.01 µg/mL. The astaxanthin also showed a potent inhibitory ability against the following three cancer cell lines: HeLa, A549, and MCF7, with IC₅₀ values of 69.07 ± 2.4 µg/mL, 55.60 ± 2.64 µg/mL, and 56.38 ± 4.1 µg/mL, respectively. This study indicates that astaxanthin derived from our newly mutated R. toruloides G17 is a promising anticancer and antioxidant agent for further pharmaceutical applications. Full article

(This article belongs to the Special Issue Pigment Production in Submerged Fermentation)

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Open AccessArticle

Scale-Up of Pigment Production by the Marine-Derived Filamentous Fungus, Talaromyces albobiverticillius 30548, from Shake Flask to Stirred Bioreactor

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Fermentation 2023, 9(1), 77; https://doi.org/10.3390/fermentation9010077 - 16 Jan 2023

Cited by 1 | Viewed by 904

Abstract

Talaromyces albobiverticillius 30548, a marine-derived fungus, produces Monascus-like azaphilone red/orange pigments which have the potential for various industrial applications. The objective of this study was to scale up pigment production in a 2 L bioreactor with a working volume of 1.3 L [...] Read more.

Talaromyces albobiverticillius 30548, a marine-derived fungus, produces Monascus-like azaphilone red/orange pigments which have the potential for various industrial applications. The objective of this study was to scale up pigment production in a 2 L bioreactor with a working volume of 1.3 L media and to compare its biomass growth and pigment production against small volume (500 mL) shake flasks with 200 mL working volume. Additionally, fungal morphology, pigment intensity, fermentation length and duration of pigment production were also compared. Experiments were carried out at laboratory scale in 200 mL shake flasks without controlling pH and oxygen. In parallel, fermentation was performed in a 2 L bioreactor as an initial scale-up to investigate the influence of dissolved oxygen, agitation speed and controlled pH on pigment production and biomass growth of T. albobiverticillius 30548. The highest orange and red pigment production in bioreactor at 24 °C was noticed after 160 h of fermentation (70% pO₂) with 25.95 AU 470 nm for orange pigments and 22.79 AU 500 nm for red pigments, at pH set point 5.0. Meanwhile, the fermentation using 200 mL shake flasks effectively produced orange pigments with 22.39 AU 470 nm and red pigments with 14.84 AU 500 nm at 192 h under the same experimental conditions (24 °C, pH 5.0, 150 rpm). Regarding fungal morphology, growth of fungus in the bioreactor was in the form of pellets, whereas in the shake flasks it grew in the form of filaments. From the observed differences in shake flasks and closed bioreactor, it is known that the bioprocess was significantly influenced by dissolved oxygen saturation and agitation speed in scale-up. Thus, oxygen transfer appears to be the rate-limiting factor, which highly influences overall growth and production of pigments in Talaromyces albobiverticillius 30548 liquid culture. Full article

(This article belongs to the Special Issue Pigment Production in Submerged Fermentation)

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Open AccessArticle

Pilot-Scale Production of the Natural Colorant Laetiporic Acid, Its Stability and Potential Applications

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Fermentation 2022, 8(12), 684; https://doi.org/10.3390/fermentation8120684 - 28 Nov 2022

Cited by 4 | Viewed by 1273

Abstract

Laetiporus sulphureus, a wood-decaying basidiomycete, produces yellow-orange pigments in fruiting bodies and, as was recently shown, in submerged cultivated mycelia. Out of four strains, the most potent laetiporic acid producer was identified and its yield compared in different media. The complex Moser [...] Read more.

Laetiporus sulphureus, a wood-decaying basidiomycete, produces yellow-orange pigments in fruiting bodies and, as was recently shown, in submerged cultivated mycelia. Out of four strains, the most potent laetiporic acid producer was identified and its yield compared in different media. The complex Moser b medium was replaced by potato dextrose broth, achieving higher yields at a lower cost. Cultivation was then scaled up from shake flask to a 7 L stirred tank bioreactor. Optimization of parameters led to increased product concentrations up to 1 g L⁻¹, the highest yield reported so far. An in situ product recovery strategy with a biphasic system was established, increasing the yield by 19% on the shake flask scale. A crude ethanolic extract of the biomass was examined for color stability and application trials. In contrast to what has been suggested in the past, the pigment showed limited long-term stability to oxygen and light, but was stable under storage in the dark at 4 °C under nitrogen. The orange extract was successfully incorporated into different matrices like foods, cosmetics and textiles. Laetiporic acid can potentially replace petrochemical based synthetic dyes, and can thus support the development of a circular bioeconomy. Full article

(This article belongs to the Special Issue Pigment Production in Submerged Fermentation)

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Open AccessArticle

Cultivation of Inonotus hispidus in Stirred Tank and Wave Bag Bioreactors to Produce the Natural Colorant Hispidin

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Fermentation 2022, 8(10), 541; https://doi.org/10.3390/fermentation8100541 - 14 Oct 2022

Cited by 3 | Viewed by 1156

Abstract

Hispidin (6-(3,4-dihydroxystyrl)-4-hydroxy-2-pyrone) production in submerged cultured mycelia of the basidiomycete Inonotus hispidus was doubled in shake flasks through irradiation with white light. The daily addition of 1 mM hydrogen peroxide as a chemical stressor and a repeated supplementation of the shake flask cultures [...] Read more.

Hispidin (6-(3,4-dihydroxystyrl)-4-hydroxy-2-pyrone) production in submerged cultured mycelia of the basidiomycete Inonotus hispidus was doubled in shake flasks through irradiation with white light. The daily addition of 1 mM hydrogen peroxide as a chemical stressor and a repeated supplementation of the shake flask cultures with 2 mM caffeic acid, a biogenetic precursor, further increased the hispidin synthesis. These cultivation conditions were combined and applied to parallel fermentation trials on the 4 L scale using a classical stirred tank bioreactor and a wave bag bioreactor. No significant differences in biomass yield and colorant production were observed. The hispidin concentration in both bioreactors reached 5.5 g·L⁻¹, the highest ever published. Textile dyeing with hispidin was successful, but impeded by its limited light stability in comparison to industrial dyes. However, following the idea of sustainability and the flawless toxicity profile, applications in natural cosmetics, other daily implements, or even therapeutics appear promising. Full article

(This article belongs to the Special Issue Pigment Production in Submerged Fermentation)

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Open AccessArticle

Pigment Production by Paracoccus spp. Strains through Submerged Fermentation of Valorized Lignocellulosic Wastes

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Fermentation 2022, 8(9), 440; https://doi.org/10.3390/fermentation8090440 - 05 Sep 2022

Cited by 1 | Viewed by 1369

Abstract

Due to the increasing emphasis on the circular economy, research in recent years has focused on the feasibility of using biomass as an alternative energy source. Plant biomass is a potential substitute for countering the dependence on depleting fossil-derived energy sources and chemicals. [...] Read more.

Due to the increasing emphasis on the circular economy, research in recent years has focused on the feasibility of using biomass as an alternative energy source. Plant biomass is a potential substitute for countering the dependence on depleting fossil-derived energy sources and chemicals. However, in particular, lignocellulosic waste materials are complex and recalcitrant structures that require effective pretreatment and enzymatic saccharification to release the desired saccharides, which can be further fermented into a plethora of value-added products. In this context, pigment production from waste hydrolysates is a viable ecological approach to producing safe and natural colorings, which are otherwise produced via chemical synthesis and raise health concerns. The present study aims to evaluate two such abundant lignocellulosic wastes, i.e., wheat straw and pinewood sawdust as low-cost feedstocks for carotenoid production with Paracoccus strains. An alkali pretreatment approach, followed by enzymatic saccharification using an indigenous lab-isolated fungal hydrolase, was found to be effective for the release of fermentable sugars from both substrates. The fermentation of the pretreated sawdust hydrolysate by Paracoccus aminophilus CRT1 and Paracoccus kondratievae CRT2 resulted in the highest carotenoid production, 631.33 and 758.82 μg/g dry mass, respectively. Thus, the preliminary but informative research findings of the present work exhibit the potential for sustainable and economically feasible pigment production from lignocellulosic feedstocks after optimal process development on the pilot scale. Full article

(This article belongs to the Special Issue Pigment Production in Submerged Fermentation)

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Open AccessArticle

Colaconema formosanum, Sarcodia suae, and Nostoc commune as Fermentation Substrates for Bioactive Substance Production

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Fermentation 2022, 8(7), 343; https://doi.org/10.3390/fermentation8070343 - 21 Jul 2022

Viewed by 1137

Abstract

Bioactive compounds extracted from natural renewable sources have attracted an increased interest from both industry and academia. Recently, algae have been highlighted as promising sources of bioactive compounds, such as polyphenols, polysaccharides, fatty acids, proteins, and pigments, which can be used as functional [...] Read more.

Bioactive compounds extracted from natural renewable sources have attracted an increased interest from both industry and academia. Recently, algae have been highlighted as promising sources of bioactive compounds, such as polyphenols, polysaccharides, fatty acids, proteins, and pigments, which can be used as functional ingredients in many industrial applications. Therefore, a simple green extraction and purification methodology capable of recovering biocompounds from algal biomass is of extreme importance in commercial production. In this study, we evaluated the application of three valuable algae (Colaconema formosanum, Sarcodia suae, and Nostoc commune) in combination with Pseudoalteromonas haloplanktis (type strain ATCC 14393) for the production of versatile compounds. The results illustrate that after 6 h of first-stage fermentation, the production of phycobiliproteins in C. formosanum was significantly increased by 156.2%, 188.9%, and 254.17% for PE, PC, and APC, respectively. This indicates that the production of phycobiliproteins from algae can be enhanced by P. haloplanktis. Furthermore, we discovered that after S. suae and N. commune were fermented with P. haloplanktis, mannose was produced. In this study, we describe a feasible biorefinery process for the production of phycobiliproteins and mannose by fermenting marine macroalgae with cyanobacteria. We believe it is worth establishing a scale-up technique by applying this fermentation method to the production of phycobiliproteins and mannose in the future. Full article

(This article belongs to the Special Issue Pigment Production in Submerged Fermentation)

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Open AccessArticle

Biodegradation of Methylene Blue Using a Novel Lignin Peroxidase Enzyme Producing Bacteria, Named Bacillus sp. React3, as a Promising Candidate for Dye-Contaminated Wastewater Treatment

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Fermentation 2022, 8(5), 190; https://doi.org/10.3390/fermentation8050190 - 19 Apr 2022

Cited by 5 | Viewed by 3053

Abstract

The emission of methylene blue (MB) from common industries causes risks to human health by making clean drinking water unavailable and hampering environmental safety. A biological approach offering a more cost-efficient and sustainable alternative solution has been studied and demonstrated to be significantly [...] Read more.

The emission of methylene blue (MB) from common industries causes risks to human health by making clean drinking water unavailable and hampering environmental safety. A biological approach offering a more cost-efficient and sustainable alternative solution has been studied and demonstrated to be significantly effective for the removal of MB using promising microbial isolates. Therefore, this study targeted bacterial candidates, namely Bacillus sp. React3, isolated from soil with the potential to decolorize MB. The phenogenic identification of strain React3 was performed by 16S rRNA sequencing, showing a similarity of 98.86% to Bacillus velezensis CR-502T. The ability of this bacterial strain to decolorize MB was proven through both the lignin peroxidase efficiency and accumulation in the biomass of the living cells. MB removal was determined by the reduction in the maximum absorption at a wavelength of 665 nm, which was observed to be up to 99.5% after 48 h of incubation. The optimal conditions for the MB degradation of strain React3 were pH 7, 35 °C, static, 4% inoculum, and 1000 mg/L of MB, with tryptone as a carbon source and yeast extract as a nitrogen source. Full article

(This article belongs to the Special Issue Pigment Production in Submerged Fermentation)

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Open AccessReview

Natural Substrates and Culture Conditions to Produce Pigments from Potential Microbes in Submerged Fermentation

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Fermentation 2022, 8(9), 460; https://doi.org/10.3390/fermentation8090460 - 14 Sep 2022

Cited by 7 | Viewed by 2261

Abstract

Pigments from bacteria, fungi, yeast, cyanobacteria, and microalgae have been gaining more demand in the food, leather, and textile industries due to their natural origin and effective bioactive functions. Mass production of microbial pigments using inexpensive and ecofriendly agro-industrial residues is gaining more [...] Read more.

Pigments from bacteria, fungi, yeast, cyanobacteria, and microalgae have been gaining more demand in the food, leather, and textile industries due to their natural origin and effective bioactive functions. Mass production of microbial pigments using inexpensive and ecofriendly agro-industrial residues is gaining more demand in the current research due to their low cost, natural origin, waste utilization, and high pigment stimulating characteristics. A wide range of natural substrates has been employed in submerged fermentation as carbon and nitrogen sources to enhance the pigment production from these microorganisms to obtain the required quantity of pigments. Submerged fermentation is proven to yield more pigment when added with agro-waste residues. Hence, in this review, aspects of potential pigmented microbes such as diversity, natural substrates that stimulate more pigment production from bacteria, fungi, yeast, and a few microalgae under submerged culture conditions, pigment identification, and ecological functions are detailed for the benefit of industrial personnel, researchers, and other entrepreneurs to explore pigmented microbes for multifaceted applications. In addition, some important aspects of microbial pigments are covered herein to disseminate the knowledge. Full article

(This article belongs to the Special Issue Pigment Production in Submerged Fermentation)

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