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Bioprocessing for Biomolecules Production (2020, Wiley)_部分2

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内容提示: 25512Research and Production of Ingredients Using UnconventionalRaw Materials as Alternative SubstratesSusanaRodríguez-CoutoIKERBASQUE,BasqueFoundationforScience,MariaDiazdeHaro3,48013,Bilbao,Spain12.1 IntroductionHugeamountsofvegetablewastesaregeneratedworldwidebytheagricultural,forestry,and food-processing industries, causing serious environmental pollution, including asignif i cant amount of greenhouse gas emission (Oelofse and Nahman 2013). T h is isparticularly problematic in countries whose economy ...

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25512Research and Production of Ingredients Using UnconventionalRaw Materials as Alternative SubstratesSusanaRodríguez-CoutoIKERBASQUE,BasqueFoundationforScience,MariaDiazdeHaro3,48013,Bilbao,Spain12.1 IntroductionHugeamountsofvegetablewastesaregeneratedworldwidebytheagricultural,forestry,and food-processing industries, causing serious environmental pollution, including asignif i cant amount of greenhouse gas emission (Oelofse and Nahman 2013). T h is isparticularly problematic in countries whose economy is mainly based on agricultureand farming. Most of these wastes are used as animal feed or burned (Mussatto et al.2012). However, they are a good source of bioactive compounds (Ayala-Zavala andGonzález-Aguilar 2011; Gornas and Rudzinska 2016; Ravindran and Jaiswal 2016;Banarjee et al. 2017), which can have potential as food additives and/or nutraceuticals.In addition, the demand of food additives from natural sources is growing considerablydue to the increasing trend in consuming foods and supplements with health benef i tproperties.Table12.1liststheannualworldwidegenerationofseveralbiologicalwastes.T h e European 2020 growth strategy, launched in 2010, has set the goal of shifting themodelsofproductionandconsumptionfromlineartocircular(Imbert2017).T h eEuro-pean Union (EU) generates 1000 million tons of agricultural waste, 500 million tons ofgarden and forestry waste, and 250 million tons of food waste from the food-processingindustry per year (Stabnikova et al. 2010). In this context, the management of industrialbiological wastes presents a great challenge. Current management of biological wastesis costly and has a negative impact on the environment. T h erefore, valorization of suchwastesinanenvironmentallyfriendlyandcost-ef f ectivewayisneeded.Inthissense,therecovery of added-value compounds from biological wastes by solid-state fermentation(SSF)usingfungalstrainsisconsideredasapromisingapproach(Farinas2015).Inaddi-tion, the bioproducts obtained can be labeled as natural, which would make them moreattractive for the consumers.T h e potential of biological wastes as raw materials to produce added-value com-pounds has been reviewed by several authors. Mirabella et al. (2014) published anextensive review on the potential use of wastes from the food industry. T h e authorsconcluded that the sustainability of the whole recovery process must be assessed beforeconsidering its implementation. Also, Gowe (2015) analyzed the potential of fruit andvegetable wastes to obtain bioactive compounds of commercial interest for the foodBioprocessing for Biomolecules Production, First Edition.Edited by Gustavo Molina, Vijai Kumar Gupta, Brahma N. Singh, and Nicholas Gathergood.© 2020 John Wiley & Sons Ltd. Published 2020 by John Wiley & Sons Ltd. 256 12 Research and Production of Ingredients Using Unconventional Raw Materials as Alternative SubstratesTable 12.1 Major biological wastes produced worldwide.Waste Annual production ReferenceRice (Oryza sativa) straw 731 million tons Karimi et al. (2006)Wheat (Triticum aestivum)straw529 million tons Govumoni et al. (2013)Sugarcane (Saccharumof f i cinarum) bagasse200 million tons Lachos-Pérez et al. (2016)Wheat (Triticum aestivum) bran 150 million tons Prückler et al. (2014)Corn (Zea mays) cobs 144 million tons da Silva et al. (2015)Banana (Musa paradisiaca) peel 57.6 million tons Ahmad and Danish (2018)Citrus wastes 15 million tons Marin et al. (2007)Potato (Solanum tuberosum)waste12 million tons El-Boushy and Van der Poel (1994)Apple (Malus domestica)pomace8 million tons FAO (2011)Grape (Vitis vinifera) pomace 6 million tons Wadhwa et al. (2013)Tomato (Solanum lycopersicum)waste320000 tons Encinar et al. (2008)industry. Similarly, Helkar et al. (2016) assessed the potential of the food industrybyproducts as raw materials for the development of functional food ingredients. T h eyindicated that the ef f i cient utilization of these byproducts could make the food industrysustainable. More recently, Kowalska et al. (2017) reviewed the potential of agro-wastesas sources of new natural ingredients for the food, pharmaceutical, and cosmeticindustry. T h ey pointed out that despite the need to reduce agro-wastes and exploittheir potential ef f ectively, at present few agro-wastes are utilized properly in the foodindustry as a source of new natural ingredients. T h erefore, there is a gap for researchand innovation with commercial potential in this area.12.2 Solid-State Fermentation (SSF)SSF is a fermentation process in which the microorganisms grow on a solid support inan environment with none or very little free-f l owing water (Pandey 2003). SSF is knownfromancienttimesandhasbeenmainlyusedforfoodprocessing(e.g.cheese,koji,soya)and is still used to produce important biomolecules and products for many industries,including food, pharmaceutical, textile, biochemical, and bioenergy (Pandey 2003; Soc-col and Vandenberghe 2003).SSF has received more attention by researchers in the last decades due to theincreasing interest in producing added-value products from organic wastes to replacenon-renewable materials and make the industrial chemical processes cleaner. T h einterest in SSF comes from its simplicity and closeness to the natural living habitatsof many microorganisms (Wang and Yang 2007). In addition, several studies haveshown that SSF processes can lead to higher yields, higher productivities, or bet-ter product characteristics than those attained by submerged fermentation (SmF) 12.3 Production of Food Ingredients from Unconventional Raw Materials by SSF 257(a) (b)Figure 12.1 The white-rot fungus Trametespubescens grown on solidstate fermentation (SSF)conditions using sunf l ower seed shells as a support-substrate (a) and on submerged fermentation(SmF) forming pellets (b).methods. Moreover, the low water content of the SSF reduces the fermenter size,downstream processing, stirring, and sterilization, thus, making the process moreeconomical (Pandey 2003; Raghavarao et al. 2003; Hölker and Lenz 2005; Nigam 2009).In Figure 12.1 photographs of a white-rot fungus grown under SSF and SmF conditionsare shown.However, although the scale-up methods for SmF are well developed, this is not thecaseforSSF.T h emaindif f i cultiestothescale-upofSSFprocessesarisefromtheintenseheatgeneration,thewaterloss,andtheheterogeneityofthesystem(Mitchelletal.2000;di Luccio et al. 2004). Hence several solid-state bioreactor designs have been developedto overcome these problems, but few have been used at large-scale (Durand 2003).T h e successful development of SSF processes depends on several aspects such as thetypeofmicroorganism,thetypeofsubstrate,andtheprocessparameters.InFigure12.2,the general processes involved in the valorization of organic waste by SSF are summa-rized (Yazid et al. 2017). Soccol et al. (2017) reviewed recent developments and innova-tionsinSSFandassertedthatthepracticabilityandadvantagesofSSFmustbeevaluatedfor each process, and the economic viability will depend on a thorough comparisonbetween SSF and SmF processes. Table 12.2 presents the advantages and disadvantagesof SSF over SmF (Pérez-Guerra et al. 2003).12.3 Production of Food Ingredients from UnconventionalRaw Materials by SSFIn Table 12.3 the production of dif f erent food ingredients from unconventional rawmaterials by SSF processes in the last f i ve years is presented.12.3.1 Organic AcidsOrganic acids are one of the most versatile additives in the food and beverage indus-try due to their solubility, hygroscopy, buf f ering, and chelation abilities. T h us, they are 258 12 Research and Production of Ingredients Using Unconventional Raw Materials as Alternative SubstratesCheck:MoistureParticle sizepH etc.Pre-treatment?SSFBiological wastes:• Agriculture• Food industry• ForestrySolid-liquid extraction(other down streamstrategy)High added-valuebioproducts@feedstocksFermented medium(to be directly used)Monitor:AerationTemperatureetc.Remaining residues• Composting• Anaerobic digestionBiogas and/or compostInoculum?• Mechanically• Chemically• Biologically• Bacteria• FungiYesYesNoNoFigure 12.2 Flowchart of valorization of biological wastes to produce added-value products bysolidstate fermentation (SSF) (after Yazid et al. 2017).widely used in food and beverage industries as preservatives and acidulants. Usually,organic acids are produced commercially by either chemical synthesis or SmF. How-ever, SSF is a very promising technique where high concentrations of the product canbeattainedwiththeuseoflow-costsubstrates,suchasthosefromtheagricultural,food,andforestryindustries,resultinginprocesseswitheconomicandenvironmentaladvan-tages (Vandenberghe et al. 2018). In fact, SSF has been successfully used for many yearsto produce citric and lactic acid on a large scale (Mussatto et al. 2012).Certik et al. (2013) tested four Mucor strains and dif f erent cereal substrates(wheat bran, rye bran, oat f l akes, barley groats, and spent malt grain) to producegamma-linoleic acid under SSF conditions. Mucor circinelloides was the best producerof gamma-linoleic acid among the tested strains when grown on rye bran/spent malgrains at a ratio of 3:1. Furthermore, they observed that the addition of sunf l ower oilat 30% led to the highest amount of gamma-linoleic acid in the fermented substrate(24.2g/kg).Dhillon et al. (2013) produced citric acid by cultivation of Aspergillus niger under SSFconditions in a 12-L rotating drum bioreactor using fresh apple pomace, provided by anapple juice company, as a substrate. T h ey obtained a production of 220.6±13.9g citricacid/kg dry solids operating under optimized conditions (i.e. 3% (v/v) methanol, inter-mittentagitationof1hourevery12hoursat2rpm,1vvmofaerationrateand120hoursof incubation time). Also, Yadegari et al. (2013) studied the production of citric acid by 12.3 Production of Food Ingredients from Unconventional Raw Materials by SSF 259Table 12.2 Advantages and disadvantages of solid-state fermentation (SSF) over submergedfermentation (SmF).Advantages DisadvantagesSimilar or higher yields are obtained than thoseobtained in the corresponding submergedcultures.Only microorganisms that can grow at lowmoisture levels can be used.T h e low availability of water reduces thepossibilities of contamination by bacteria andyeast. T h is allows working in aseptic conditions insome cases.Usually the substrates require pretreatment(size reduction by grinding, rasping orchopping, homogenization, physical, chemicalor enzymatic hydrolysis, cooking or vaportreatment).Similar environment conditions are present tothose of the natural habitats for fungi thatconstitute the main group of microorganismsused in SSF.Biomass determination is very dif f i cult.Higher levels of aeration, especially adequate inthose processes demanding an intensive oxidativemetabolism.T h e solid nature of the substrate causesproblems in the monitoring of the processparameters (pH, moisture content, andsubstrate, oxygen, and biomass concentration).T h e inoculation with spores (in those processesthat involve fungi) facilitates their uniformdispersion through the mediumAgitation may be very dif f i cult. For this reason,static conditions are preferredCulture media are often quite simple. T h esubstrate usually provides all the nutrientsnecessary for growthFrequent need of high inoculum volumesSimple design reactors with few spatialrequirements can be used due to theconcentrated nature of the substratesMany important basic scientif i c andengineering aspects are yet poor characterized.Information about the design and operation ofreactors on a large scale is scarceLow energetic requirements (in some casesautoclaving or vapor treatment, mechanicalagitation and aeration are not necessary)Possibility of contamination by undesirablefungiSmall volumes of polluting ef f l uents. Fewerrequirements of dissolvents are necessary forproduct extraction due to its high concentrationT h e removal of metabolic heat generatedduring growth may be very dif f i cultT h e low moisture availability may favor theproduction of specif i c compounds that may notbe produced or may be poorly produced in SmFExtracts containing products obtained byleaching of fermented solids are often ofviscous natureIn some cases, the products obtained have slightlydif f erent properties (e.g. more thermotolerance)when produced in SSF in comparison to SmFMass transfer limited to dif f usionDue to the concentrated nature of the substrate,smaller reactors in SSF with respect to SmF canbe used to hold the same amounts of substrateIn some SSF, aeration can be dif f i cult due to thehigh solid concentrationSpores have longer lag times due to the needfor germinationCultivation times are longer than in SmFSource: Extracted from Pérez-Guerra et al. (2003). Table 12.3 Some food ingredients produced by solid-state fermentation (SSF) of biological wastes in the last f i ve years.Waste Microorganism Product ReferenceOrganic acidsWheat bran, rye bran, oat f l akes, barleygroats, spent malt grainMucor petrinsularis, Mucordimorphosporus, Mucor circinelloides,Mucor hiemalisGamma-linolenic acid Certik et al. (2013)Apple pomace Aspergillus niger Citric acid Dhillon et al. (2013)Sugarcane bagasse A. niger Citric acid Yadegari et al. (2013)Oil palm empty fruit bunch Trichoderma reesei Humic acids Motta and Santana (2014)Nut shells, rice bran, rice husk, nut oilcake, sugar cane bagasse, fresh orangefruit wastesUstilago maydis Itaconic acid Raf i et al. (2014)Apple pomace Rhizopus oryzae Fumaric acid Das et al. (2015)Rice straw, faba bean straw Aspergillus oryzae, AzotobacterchroococcumOrganic acids Saber et al. (2015)Apple pomace and peanut shell Consortium of Aspergillus ornatus andAlternaria alternataCitric acid Ali et al. (2016)Pulp and solid paper waste R. oryzae Fumaric acid Das et al. (2016)Corn cob powder A. niger Oxalic acid Mai et al. (2016)Citric pulp Fusarium moniliforme, GibberellafukikuroiGibberellic acid De Oliveira et al. (2017)Wheat bran and sugarcane bagasse Endophytic fungi Organic acids Dezam et al. (2017)Phenolic compoundsCof f ee wastes A. niger, Aspergillus ustus, Mucor sp.,Penicillium purpurogenum, NeurosporacrassaPhenolics Machado et al. (2013) Wheat bran R. oryzae Phenolics Dey and Kuhad (2014)Citrus waste Paecilomyces variotii Phenolics Madeira et al. (2014)Rice bran R. oryzae Phenolics Schmidt et al. (2014)Berry pomaces A. niger Polyphenols, lipids Dulf et al. (2015)Tangerine residues Lentinus polychrous Antioxidants Nitayapat et al. (2015)Rice bran Rhizopus oligosporus and MonascuspurpureusAntioxidants Razak et al. (2015)Soybean mean Bacillus amyloliquefaciens, Lactobacillusspp., Saccharomyces cerevisiaePhenolics Chi and Cho (2016)Plum by-products A. nigerR. oligosporusPolyphenols, lipids Dulf et al. (2016)Caulif l ower outer leaves A. niger, A. oryzae, A. sojae, R. oryzae, R.azygosporus. PhanerochaetechrysoporiumKaemferol metabolites Huynh et al. (2016)Fig by-products R. oryzae,Trichoderma sp., A. nigerPhenolics Buenrostro-Figueroa et al. (2017)Apricot pomace A. nigerR. oligosporusPhenolics Dulf et al. (2017)Buckwheat Agaricus strains Phenolics Kang et al. (2017)Psidium guajava leaves Monascus anka and Bacillus sp. Polyphenols Wang et al. (2017)Chokeberry pomace A. nigerR. oligosporusPhenolics Dulf et al. (2018)Grape, apple and pitahaya residues Rhizomucor miehei Phenolics Zambrano et al. (2018)Aroma compoundsFood waste mixtures S.cerevisiae, Kluyveromyces marxianus,kef i r𝜀-pinene Aggelopoulos et al. (2014)Sugarcane bagasse Trichoderma viride Coconut aroma Fadel et al. (2015)Orange peels S .cerevisiae Aroma esters Mantzouridou et al. (2015)(Continued) Table 12.3 (Continued)Waste Microorganism Product ReferenceApple pomace S. cerevisiae, Hanseniasporavalbyensism, Hanseniaspora uvarumAroma compounds Rodriguez-Madrera et al. (2015)Sugracane bagasse, sugar beet molasses K. marxianus Aroma compounds Martinez et al. (2017)PigmentsWheat bran, rye bran, oat f l akes, barleygroats, spent malt grainM. petrinsularis, M. dimorphosporus, M.circinelloides, M. hiemalis𝛽-Carothene Certik et al. (2013)Whole grain, dehulled grain, bransubstratesM. purpureus Pigments Srianta and Harijono (2015)Wheat wastes Yamadazyma guilliermondii, Yarrowialipolytica, Xanthophylomycesdendrorhous, SporidiobulussalmonicolorAstaxanthin Dursun and Dalgic (2016)Olive pomace X. dendrorhous, S. salmonicolor Astaxanthin Erilymaz et al. (2016)Rice, corn, whole sorghum grain,dehulled sorghum grain, sorghum branM. purpureus Pigments Srianta et al. (2016)Rice, corn, whole sorghum grain,dehulled sorghum grain, sorghum branM. purpureus Pigments Srianta et al. (2017) 12.3 Production of Food Ingredients from Unconventional Raw Materials by SSF 263A. niger under SSF conditions using sugarcane bagasse, provided by a sugar factory, asa substrate. T h ey found that pretreating the sugarcane bagasse with sodium hydroxideincreasedthecitricacidproductionfrom75.34gkg −1 substrateto97.81gkg −1 substrate.Motta and Santana (2014) reported the production of humic acid by SSF ofTrichoderma reesei in f i xed-bed columns using empty palm bunch, an underutilizedby-productofthepalmoilindustry,asasubstrate.T h eyfoundahumicacidproductivityof 0.73mg/100g of substrate after 72hours of incubation.Raf i etal.(2014)testeddif f erentagro-foodwastessuchasgroundnutshells,ricebran,rice husk, orange pulp, ground nut oil cake, and sugarcane bagasse as substrates foritaconic acid production by the fungus Ustilago maydis under SSF conditions. All thesubstrates led to promising yields of itaconic acid, rice bran, orange pulp and sugar-cane bagasse producing the highest ones (around 26–27gkg −1 substrate) operating atoptimal conditions (i.e. pH3, moisture 60%, temperature 32 ∘ C and a cultivation time off i vedays).Dasetal.(2015)reportedaproductionof52±2.1goffumaricacidperkgofdryapplepomace (with 50% moisture content) by the fungus Rhizopus oryzae after 14 cultivationdays in tray bioreactors. Later (Das et al. 2016), they tested pulp and paper solid wastefrom a paper industry as a substrate. T h ey found that the particle size highly inf l uencedthe fumaric acid production, a particle size range of 850μm<x≤300μm leading to thehighest production (41.65gkg −1 dry substrate).Saber et al. (2015) investigated the synergistic ef f ect of the f i lamentous fungusAspergillus oryzae and the non-symbiotic nitrogen f i xing bacterium Azotobacterchroococcum as an innovative technique to convert rice straw, faba bean straw, and rockphosphate into organic acids under SSF conditions. T h ey found that citric (15.3mgg −1fermented biomass) and succinic acid (13.7mgg −1 fermented biomass) were the mainorganic acids contained in the fermented biomass, corresponding to the 98.87% of thetotal organic acids detected.Ali et al. (2016) used dif f erent agro-wastes (apple pomace, peanut shell and a mix-ture of both apple pomace and peanut shell at 50:50 ratio) as substrates for SSF toenhancethecitricacidproductionfromsingleandco-cultureconsortiumofAspergillusornatus and Alternaria alternata. Partial optimisation of the co-culture (arginine addi-tion, 30 ∘ C, 25g apple pomace at 50% moisture, pH5 and a cultivation time of 48hours)showed a maximum citric acid yield of 2.644±0.99mgml −1 .Maietal.(2016)studiedtheproductionofoxalicacidbyamethanol-resistantA.nigerstrainundersemiSSFconditionsusingcorncobasasubstrate.Amaximumproductivityof 123.0gkg −1 dry weight of corncob was attained operating at optimal conditions (i.e.,10 6 sporesg −1 dry weight and 5% (w/v) corncob in 0.1N NaOH solution).De Oliveira et al. (2017) investigated the production of the important phytohormonegibberellic acid by the fungi Fusarium moniliforme and Gibberella f j ikuroi using dif-ferent cultivation techniques (SSF, semi SSF, and SmF) at f l ask and bioreactor scaleoperatingwithcitricpulp,a subproduct obtainedfrom the extractionof orangejuice, asasubstrate.SSFledtothehighestproductionofgibberellicacidatbothf l ask(7.60gkg −1dry substrate) and bioreactor scale (7.34gkg −1 dry substrate).Dezametal.(2017)investigatedthepotentialofendophyticfungi,isolatedfromman-grove areas in Sao Paulo (Brazil), to produce organic acids by SSF using wheat bran andsugarcanebagasseassubstrates.T h ehighestyieldoforganicacids(135.5mgg −1 drysub-strate), corresponding mainly to citric acid (106.8mgg −1 dry substrate), was produced 264 12 Research and Production of Ingredients Using Unconventional Raw Materials as Alternative Substratesby the isolate Aspergillus awamori when grown on a mixture of wheat bran and sug-arcane bagasse at a ratio of 1:3. T h ese results pointed out the potential of endophyticfungi for organic acid production using agro-industrial wastes as feedstock under SSFconditions,thus,providingasustainablealternativetotheorganicacidmanufactureandcontributing to the circular economy.12.3.2 Phenolic CompoundsPhenolic compounds have been extensively studied for their application in the foodindustry as preservatives to improve the shelf life of perishable products. T h e currentconcern about the impact of food on health inf l uences the consumer choice of foodbased on its formulation. Consumers prefer phenolic compounds from natural sources.Antioxidants have become increasing popular during the last decades due to theirpotential properties to prevent chronic diseases such as cardiovascular diseases, can-cer, osteoporosis, diabetes mellitus, and neurodegenerative diseases (Pandey and Rizvi2009).Consequently,varioussourcesofdif f erentantioxidantphenoliccompounds,suchas fruits, vegetables, wine, cof f ee, tea, and cereals, have been investigated to replacehealth hazard antioxidant compounds like butylated hydroxyanisole, butylated hydrox-ytoluene, and tertiary butyl hydroquinone in dif f erent food products.Phenolic compounds are commonly extracted from plant materials by organic sol-ventssuchasmethanol,ethylacetate,acetone,andn-hexaneusingvariousconventionalextraction methods (Ignat et al. 2011). However, these methods do not allow the com-plete release of bound phenolics from plant materials and, in addition, they are notenvironmentally friendly. T h us, microbial fermentation processes are considered verypromising to produce antioxidant phenolic compounds due to their cost-ef f ectivenessandenvironmentaladvantages.Table12.3showstheproductionofphenoliccompoundsfrom dif f erent biological wastes by SSF processes.Machado et al. (2013) found that the strains Penicillium purpurogenum GH2, Neu-rosporacrassaATCC10337andMucor sp.3Ppresentedgreatabilitytogrowandreleasephenolic compounds from cof f ee silverskin and spent cof f ee grounds, abundant cof f eewastes from the cof f ee industry, under SSF conditions. T h us, these f i ndings of f er newpossibilities for the use of underutilized wastes.DeyandKuhad(2014)showedthattheextractionofphenoliccompoundsfromwheatimproved by SSF with R. oryzae. T h erefore, fermented wheat may serve as a power-ful source of natural antioxidants. Madeira et al. (2014) developed a process for thebiotransformationofphenoliccompoundsfromcitruswastes,providedbyalocalindus-trial company, by SSF with Paecilomyces variotii. T h is process increased by 73% theantioxidant capacity of the waste. In addition, the commercially interesting bioactivecompounds hesperetin, naringenin, and ellagic acid were also produced.Schmidt et al. (2014) reported the increased in free phenolics by more than 100% ofricebranafterSSFwithR.oryzae.Inaddition,theprof i leofthephenolicschanged,withgallic and ferulic acid showing the highest increase (170 and 765mg/g, respectively).Dulf et al. (2015) showed the increase in phenolic content and antioxidant activity ofberrypomacesbySSFofA.niger.Later,theyobservedthesameforSSFofplum,apricot,and chokeberry pomaces with A. niger and Rhizopus oligosporus (Dulf et al. 2016, 2017,2018). Also, Nitayapat et al. (2015) increased the phenolic content and the antioxidantactivity of tangerine wastes by SSF with Lentinus polychrous. 12.3 Production of Food Ingredients from Unconventional Raw Materials by SSF 265Razak et al. (2015) proved that the phenolic content and antioxidant activity of ricebran could be increased by SSF with fungi such as R. oligosporus and Monascus pur-pureus. Likewise, Chi and Cho (2016) showed that the SSF of soybean meal with Bacil-lus amyloliquefaciens signif i cantly improved its total phenolic content and antioxidantactivity.Huynh et al. (2016) tested caulif l ower outer leaves as a substrate to produce phenoliccompounds by dif f erent f i lamentous fungi, namely A. niger, A. oryzae, Aspergillus sojae,R. oryzae, Rizhopus. azygosporus and Phanerochaete chrysoporium under SSF condi-tions.A.sojaeledtothehighestleveloftotalphenoliccompounds(321mgrutinequiva-lents/100gfreshweight)after1fermentationday,thisvaluebeingthree-foldhigherthanthat found in the unfermented substrate. Also, Bei et al. (2017) reported that the SSF ofoats with Monascus anka considerably increased their content in phenolic compoundsand, in addition, they showed higher antioxidant activities.Buenrostro-Figueroa et al. (2017) investigated the valorization of f i g byproducts froma jam and wine-making company, by SSF with the following fungal strains: R. oryzae,Trichodermasp.,A.niger HT4andA.niger GH1.T h eyfoundthatA.niger HT4ledtothehighest release of total polyphenols. Further, the optimized process (36h, 40 ∘ C, pH5.0,moisturecontent60%,NaNO 3 0.57gl −1 ,KH 2 PO 4 3.04gl −1 ,MgSO 4 ⋅7H 2 O1.52gl −1 andKCl 5.37gl −1 ) released 10.37mg of gallic acid equivalents/g of dry matter.Kang et al. (2017) assessed the ef f ect of SSF of buckwheat with three Agaricus strains(Agaricus blazei Murrill SH26, Agaricus bisporus AS2796, and Agaricus bisporus G1)on its total phenolic content and antioxidant properties. A. blazei supported the high-est phenolic content in buckwheat (18.07mgg −1 ) after 21 fermentation days. Also, theantioxidant properties of buckwheat increased, save for the A. bisporus G1 cultures.Wang et al. (2017) showed that the co-fermentation of guava leaves with Monascusanka and Bacillus sp. under SSF conditions promoted the release of insoluble-boundpolyphenol components. In addition, these polyphenols presented higher antioxidantactivitiesthanthoseextractedfromunfermentedguavaleaves.Moreover,theyobservedthattheantioxidantcapacitiesofthesolublepolyphenolswereconsiderablyincreasedbythis microbial co-fermentation. Recently, Zambrano et al. (2018) also reported that theSSF of dif f erent agro-wastes (black grape pomace, and apple and yellow pitahaya peel,core, peduncle, and seed mixtures) with Rhizomucor miehei increased their extractablephenolic content and improved their phenolic antioxidant properties.12.3.3 Flavor and Aroma CompoundsFlavors are largely used in the food industry to improve food organoleptic properties.Most f l avoring compounds are produced via chemical synthesis or extraction from nat-ural materials. However, since consumers prefer food free of chemical substances, theproduction of aroma compounds by SSF would likely be well received (Table 12.3).Aggelopoulos et al. (2014) reported the production of high amounts of the aromavolatile compound 𝜀-pinene by kef i r grown on a mixture of food wastes (orange pulp,molasses,potatopulpandwhey)underSSFconditionsatayieldof4kgpertonoftreatedsubstrate. Also, Fadel et al. (2015) assessed the ability of Trichoderma viride to pro-ducecoconutaromainSSFusingsugarcanebagasseasasubstrate.T h eanalysisrevealedthat 6-pentyl-𝛼-pyrone was the main volatile compound produced contributing to the 266 12 Research and Production of Ingredients Using Unconventional Raw Materials as Alternative Substratescoconutaroma,thehighestproductionof3.62mgg −1 ofdrymatterafterf i vecultivationdays being attained.Mantzouridou et al. (2015) reported for the f i rst time the feasibility of SSF of orangepeel waste to produce yeast volatile esters with fruity-like character at a high yield(250mgkg −1 of fermented orange peel waste). Likewise, Rodriguez-Madrera et al.(2015)showedthattheSSFofapplepomacewithautochthonousyeaststrainsgenerateddif f erent volatile compounds with application in the food industry. More recently,Martinez et al. (2017) proved the ef f i ciency of the generally recognized as safe (GRAS)strain Kluyveromyces marxianus to produce aroma compounds by SSF using a mixtureof sugarcane bagasse and sugar beet molasses as the only substrate.12.3.4 PigmentsSynthetic colorants are considered hazardous to human health and, thus, only fewclasses are acceptable to be used in the food industry. T h erefore, the production ofnon-hazardous colorants suitable for use in the food industry is needed. In this context,naturalpigmentssecretedbycertainfungalspeciessuchasAspergillus,Fusarium,Peni-cillium,Paecilomyces,andTrichodermaappearasapromisingapproach(Akilandeswariand Pradeep 2016). Furthermore, several natural pigments have antioxidant propertieswhich is very interesting since there is an increasing trend toward the development ofnutraceutical food. In addition, these natural pigments can be produced in SSF usingagro-industrial wastes as substrates for the microorganism, thus, making the processmore cost-ef f i cient and ecological. In Table 12.3 dif f erent natural pigments producedunder SSF are presented.Certik et al. (2013) tested four Mucor strains and dif f erent cereal substrates (wheatbran, rye bran, oat f l akes, barley groats and spent malt grain) to produce the pigment𝛽-carotene under SSF conditions. Mucor circinelloides was the best producer of𝛽-carotene (9.5mgkg −1 ) among the tested strains when grown on rye bran and spentmalt grains at a ratio of 3:1 with glucose addition.Srianta and Harijono (2015) showed that M. purpureus was able to produce yellow,orange, and red pigments when grown on whole sorghum grain, dehulled sorghumgrain, and sorghum bran substrates with soaking under SSF conditions. T h e authorsalso observed that the whole sorghum grain cultures led to the highest ethanol solublepigments, while the fermented bran with soaking cultures contained the highestwater-soluble pigments. T h ey (Srianta et al. 2016) also studied the production andcomposition of the pigments produced by M. purpureus under SSF grown on dif f erentcereal substrates (i.e. rice, corn, whole sorghum grain, dehulled sorghum grain, andsorghum bran). T h e highest pigment production was achieved on rice, followed bydehulled sorghum grain, whole sorghum grain, corn, and f i nally sorghum bran. Twelvepigments were detected on the Monascus-fermented products at dif f erent levels, thered pigment rubropunctamine being the major one (57–87%) except for sorghum brancultures, which produced Yellow II as the major one. Later (Srianta et al. 2017), theyfound that the pigments produced by M. purpureus when grown on rice, corn, andsorghum under SSF conditions presented considerable antioxidant activities. However,the specif i c pigments responsible for these activities were not identif i ed.Dursun and Dalgic (2016) proved the successful use of wheat wastes, one of the mainagro-industrial wastes worldwide, to produce the carotenoid pigment astaxanthin by References 267Yamadazyma guilliermondii, Yarrowia lipolytica, Xanthophyllomyces dendrorhous,and Sporidiobolus salmonicolor under SSF conditions. T h e maximal astaxanthin yield(109.23mgg −1 substrate) was produced by X. dendrorhous when cultured at optimizedconditions (i.e. 20 ∘ C, pH5.5 and 90% moisture). Also, Ery𝚤lmaz et al. (2016) studied theproduction of astaxanthin by the yeasts X. dendrorhous and S. salmonicolor under SSFconditions usingolive pomaceas a substrate. T h e former led to a maximumastaxanthinyield of 220.24±17.47μgg −1 dry substrate.12.4 OutlookManyindustriesgeneratehugequantitiesofbiologicalwastes,causingaseriousenviron-mental concern. T h us, there is an urgent need to change the global perception towardindustrial biological wastes, as they can be ef f ectively utilized in an eco-friendly man-nertoproduceadded-valueproducts(e.g.,foodadditives).Inthissense,manyindustrialandacademiclaboratoriesarefocusingonthevalorizationofindustrialbiowastes.How-ever, further research and optimization studies on valorization technologies focused onfull rather than laboratory studies are needed. Although SSF is a very promising greentechnology for biowaste valorization, research on improving the scale-up facilities inSSF, by the development of robust designs and conf i gurations of bioreactors, processautomation, and online monitoring of parameters, is required.ReferencesAggelopoulos, T., Katsieris, K., Bekatorou, A. et al. (2014). Solid state fermentation of foodwaste mixtures for single cell protein, aroma volatil...

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