Bioproducts from Solid-State Fermentation: Comparison
Please note this is a comparison between Version 2 by Jason Zhu and Version 1 by Nicolás Oiza.

Solid-state fermentation (SSF) is part of the pathway to consolidate waste as a relevant alternative for the valorization of organic waste. The objective of SSF is to produce one or several bioproducts of added value from solid substrates. Solid-state fermentation can use a wide variety of organic waste as substrates thus, it is an excellent candidate in the framework of the circular bioeconomy to change the status of waste from feedstock.

  • solid-state fermentation
  • organic waste
  • bioproducts

1. Introduction

With the increasing implementation of biological treatments applied to organic waste due to a stringent worldwide legislation against landfill and incineration, anaerobic digestion (AD) [1] and composting [2] have become the main accepted ways to treat organic waste in the framework of the Circular Economy as they permit the recovery of energy and materials [3]. However, a new emerging field of research has been raised in recent years: solid-state fermentation (SSF). Although solid-state fermentation (SSF) has been known for decades, the use of organic waste as a substrate has made SSF a technology in which circularity goes one further step. Therefore, the main goal of SSF would be to produce a valuable/marketable product from renewable materials in a sustainable way, substituting for current highly impacting chemicals. In addition, nutrients in the spent solids could be further recovered through composting or AD [4]. Usually, this bioproduct obtained by SSF is also substituting a non-biodegradable chemical with similar properties and lower cost [3].
Solid-state fermentation is, by definition, the cultivation process in which microorganisms grow on solid materials without the presence of a free liquid phase [5]. In practical terms, a bioreactor, typically aerobic, is filled with the solid substrate and inoculated with the strain of interest to produce the desired bioproduct. Once produced, this bioproduct can be recovered, although in some cases the final fermented solid can be used as the end-product [6]. Several types of reactors have been used in SSF development: packed bed reactors, mechanically stirred reactors, tray reactors, and even plug flow configurations [7,8][7][8]. All these configurations have one common objective in the scale-up process: to overcome the mass and heat transfer limitations of organic matter in solid state, which can result in high temperatures that harm the strain of interest [9]. These problems typically appear when SSF is scaled up, and it is one of the main problems for a full SSF use and commercialization [10,11][10][11]. Recently, some models have been published to monitor the mass behavior in SSF reactors, using traditional techniques such as residence time distributions [12], or more complex approaches, such as computer fluid dynamics [13].
Solid-state fermentation is not a new idea and has been traditionally used for some food processes, especially in Asia. Moreover, composting is a very specific type of SSF. However, it is in the last two decades that the technology has arisen as a promising biotechnological tool. Regarding the bioproducts of interest, researchers first approached the production of hydrolytic enzymes by SSF [14]. A wide variety of enzyme families have been produced by SSF: proteases, lipases, cellulases, among others. Beyond these, SSF is being explored to produce materials to substitute chemicals for biodegradable products that offer the same benefits. This is the case of biopesticides [15], biosurfactants [16], aromas [17], and bioplastics [18], among others.

2. Biosurfactants

Biosurfactants refers to surfactants of microbial origin. Like synthetic surfactants, they are composed of a hydrophilic moiety made up of amino acids, peptides, (poly)saccharides, or sugar alcohols, and a hydrophobic moiety consisting of fatty acids. Correspondingly, the significant classes of biosurfactants include glycolipids, lipopeptides and lipoproteins, and polymeric surfactants. In the case of SSF and in submerged fermentation (SmF), this chemical duality is often achieved by the combination of two substrates, the most typical ones being industrial waste lipids (for instance, waste cooking oil or cakes resulting from oil refining) and sugars in different forms [40,41][19][20]. Low molecular weight biosurfactants stand out among the large number of biosurfactants recently synthetized and characterized. Consequently, glycolipids, which are conformed by mono-, di-, tri-, and tetra-saccharides in combination with one or more chains of aliphatic acids or hydroxyaliphatic acids, are the most common. They are classified into trehalolipids, mannosylerythritol lipids (MEL), rhamnolipids, and sophorolipids (SL) [42][21]. Among them, SL, which consist of a hydrophobic fatty acid tail of 16 or 18 carbon atoms and a hydrophilic carbohydrate head sophorose, have been recently reported as biosurfactants that can be produced using SSF in several conditions and in a viable economic way [29][22], and they are even supplied by several companies, although normally produced by SmF. The use of SL is increasing according to the literature, where the yeast Starmerella bombicola is the main reported producer [43][23]. Sophorolipids, in nature, are produced as extracellular storage and antimicrobial agents. From the industrial point of view, SL can be applied as active components and formulating agents in a wide variety of high-end and bulk applications. Probably, the most reported use is as detergent, although applications in personal care have also been published. Other applications in crop protection, food, biohydrometallurgy, and medical fields are being extensively researched [44][24]. However, SSF presents some limitations as it involves a three-phase heterogeneous system. Drawbacks related to poor homogeneity and energy and mass transfer may affect the process yield and become more complex downstream [45,46][25][26]. Therefore, reactor design and process conditions must be defined taking these limitations into account. In this sense, some models have been suggested for SSF systems, from simplified approaches based on oxygen uptake rate (OUR) to complex 2-D models [47,48][27][28]. However, no specific model for SL production by SSF has been published, as has happened with other bioproducts. Production strategies assayed in submerged fermentation can be adapted to SSF processes using waste as substrate to increase the SL yield [49][29]. In this sense, Jiménez-Peñalver et al. [41][20] performed several experiments to produce SL by SSF of oil cake, molasses, and straw (used as bulking agent) inoculated with S. bombicola. The maximum yield was higher than 0.2 g of SL per g dry matter in 10 days, when intermittent mixing was applied to aerated packed bed reactors. The authors also observed that the SL yield correlated well with oxygen consumption, which is often observed in solid-state fermentation and composting of organic wastes [50][30].

3. Biopesticides

Many microorganisms can produce compounds that are lethal for some typical plagues that are a problem for the cultivation of certain crops. Biopesticides have gained importance in recent years in comparison to traditional chemical products, because of their selective action, biodegradability, and in general, their innocuousness to the environment and food chain. Biopesticides are a group that includes biological control agents for plagues of mainly insects and fungi, and they are produced from a wide number of strains. In the case of SSF using waste as substrate, their development is relatively recent [15]. The reason for this is that not all the biopesticide producing strains are able to thrive in a non-sterile solid-state media. This is the typical case of bacteria, which often grow in synthetic media to produce biopesticides [53][31]. One of the most worldwide commercialized biopesticides are those produced from Bacillus thuringiensis (Bt), which has demonstrated rapid growth and effective sporulation that are key to be predominant in organic waste [21][32]. Bacillus thuringiensis is a gram-positive bacterium consisting of several species that, during sporulation, produce crystal proteins, called delta endotoxins, that have insecticidal action against many plagues [25][33]. The SSF process to make Bt to produce endotoxins is under study. Recently, it has been demonstrated that, as Bt is a facultative microorganism, it is convenient to include a limiting oxygen step to favor the production of these compounds under SSF conditions using a complex substrate as biowaste digestate as substrate [54,55][34][35]. This opens a new line of research to use different SSF conditions to enhance the behavior of bacteria in terms of biopesticide production. Contrarily to bacteria, fungi are well-known producers of biopesticides, due to their ability to grow under solid-state conditions. In this case, SSF is an optimal environment for the growth and sporulation of biopesticide producer fungi strains. It is also worthwhile to mention that agricultural waste has been also referred as suitable substrates to promote fungi development [56][36]. In this case, it is also important to note that some strategies to overcome some batch reactors restrictions, such as mass and heat transfer limitations and the continuous production of biopesticide have been already published [22][37]. As the pure continuous mode of operation is highly complicated in SSF, these strategies consist of sequential batch operation, where other problems such as the need for inoculation and the decrease of porosity are also overcome [22][37]. Although these results are published for Trichoderma harzianum, a well-known fungus for producing biopesticides [57][38], it is evident that they can be extrapolated to other fungi and toxins of interest. This is the case of other strains, such as Beauveria bassiana, Trichoderma koningiopsis, or Metarhizium novozealandicum, which have been reported as biopesticide producers using solid substrates [58,59,60][39][40][41], an open field of research to have commercial products. Another main challenge of biopesticide production from SSF is the downstream process. In this case, a singularity is worthwhile to mention. When the biopesticide has a foliar application, a complex downstream is typically necessary [61][42]. However, sometimes a compost-like product with biopesticide products is required, whose action mainly occurs in soil, which makes the resulting SSF solid an end product ready to use, given that sufficient stability and maturity is achieved [62][43].

4. Antibiotics

Antibiotics are essential for current health care and food production systems. Antibiotic is a very general term that includes different types of molecules with biocide activity, such as some biosurfactants or biopesticides (described above). The production of antibiotics through SSF has been successfully explored allowing for the use of different solid side streams as substrates and targeting diverse applications. This has been recently reviewed in two excellent works [63,64][44][45]. Kumar et al. [63][44] reviewed the production by SSF of antibiotics, among other secondary metabolites, including natamycin, sambacide, neomycin, and the biosurfactant surfactin. They reported successful SSF processes based on agro-industrial or food wastes. Barrios et al. [64][45] reviewed lovastatin biosynthesis. Although the mechanism that regulate secondary metabolism is often the same under either submerged or solid-state fermentation, some specific microorganisms display a different physiology in each environment. This is the case for Aspergillus terreus that produces lovastatin and shows a superior performance under solid-state fermentation [64][45]. Fermented oat straw was used as a lovastatin carrier to inhibit methanogenic archaea in the rumen. This allowed for a 38% reduction of methane emissions from beef cattle in in vitro trials [65][46]. Works on SSF often screen different materials as a substrate as per selection purposes. Al Farraj et al. [66][47] assessed the antibiotic production by Streptomyces sp. AS4 using different wastes, such as wheat and rice bran, apple pomace, or pine and orange peels. Antibiotic production ranged from 43 to 209 U/g influenced by the type of substrate. Wheat bran was the most suitable of the tested materials. In contrast, in a work by Vastrad and Neelagund as reported in [63][44], Streptomyces fradiae preferred apple pomace over cotton seed meal, soybean powder or wheat bran, the last showing the lowest yields for neomycin production. El-Housseiny et al., (2021) [67][48] assessed paromomycin production with Streptomyces rimosus through SSF and compared six different substrates. Corn bran outperformed sugarcane bagasse, sunflower seed meal, soybean meal, barley, and wheat bran. In this case paromomycin production was similar by both SSF and SmF; however, the authors highlighted some advantages of SSF over SmF, such as lower costs, energy consumption, and wastewater discharge. Beyond nutrient composition, substrates often play the role of support for microbial growth. In this sense, physical properties such as water holding capacity, air porosity, particle size, surface area or rugosity play a key role in biosynthesis. An in-depth analysis of the effect of substrate characteristics including nutrients and physical characterization is required to establish robust industrial production systems for antibiotics or any other secondary metabolite.

5. Other Products

The recent boost of SSF research has provoked a proliferation of works published on the use of different typologies of bioproducts. Some of these compounds are still in a development stage and usually the main challenges to solve are scale-up and purification. They are compiled and grouped according to their properties, although each month, the reader can find new products obtained by SSF:
(a)
Aromas and flavors: this field is especially interesting, as typically synthetic products or natural products (after a costly extraction) are being used in food and other industries. Recently, some works have been published on the synthesis of molecules with aroma properties. This is the case of 2-phenylethanol, widely used in industry due to its rose-like odor and antibacterial properties, which has been produced via SSF using several agro-industrial wastes (mainly bagasses and molasses) inoculated by Pichia kudriavzevii and Kluyveromyces marxianus [68][49]. This latter strain and its related SSF process for aroma production has been scaled up to pilot scale using different operation strategies (fed-batch, static-batch, and intermittent mixing) with different productivities, which makes it especially interesting [32][50]. Other similar products have been focused on the production of fruit-like odor compounds constituted by a mixture of volatile esters [17].
(b)
Bioplastics: this field has been traditionally related to wastewater research, especially in the case of PHA (polyhydroxyalkanoates) and PHB (polyhydroxybutyrate), which are synthetized from organic components of wastewater as volatile fatty acids generated in the first stage of anaerobic digestion [69,70][51][52]. In the case of SSF, several recent references report how to produce these novel materials by SSF, although this is again an emerging technology. For instance, Llimós et al. [18] used lignocellulosic-derived residues to produce lignocellulolytic enzymes from fungal strains through SSF to hydrolyze the same residue to be used for obtaining sugar-rich hydrolysates that serve as an alternative carbon source for PHA production. The same authors provide some interesting issues to consider when scaling up the SSF process using not-isolated and near-adiabatic bioreactors [71][53]. Other authors have proposed alternative ways of using agrifood by-products in the SSF process, e.g., using dairy processing waste [72][54] and other materials [73][55].
(c)
Antioxidants: this is a property of certain chemicals that is of high value for the food and cosmetics industry, among others. There are different compounds with antioxidant properties, but the most typical ones biologically produced are the family of phenolic compounds [74][56]. In the case of SSF, there are several publications showing the suitability of certain wastes as a substrate to produce antioxidant phenolic compounds, with waste from olive oil production the most referred [75,76][57][58]. Other agricultural wastes, such as fruits- and cereals-derived waste have also been reported in the production of phenolic compounds by SSF [77,78][59][60]. The main problem of these publications is that they were normally focused on the characterization and properties of the product, whereas SSF is performed with a few grams in lab scale-controlled conditions, which again hampers its commercial development [79][61].
(d)
Recent literature is full of other specific bioproducts obtained from SSF of selected organic waste. This is the case of bio-flocculants [38][62], pigments [34][63], or specific compounds [31,36,37][64][65][66]. Although interesting, most of these studies have again been performed at a very small scale.

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