Use Of Solid-state Fermentation Research Articles

A systematic study on the characteristics and applications of laccases produced by fungi: insights on their potential for biotechnologies

Laccases are polyphenol oxidase enzymes and form the enzyme complex known for their role in wood decomposition and lignin degradation. The present study aimed to systematically review the state-of-the-art trends in scientific publications on laccase enzymes of the last 10 years. The main aspects checked included the laccase-producing fungal genera, the conditions of fungal growth and laccase production, the methods of immobilization, and potential applications of laccase. After applying the systematic search method 177 articles were selected to compound the final database. Although various fungi produce laccase, most studies were Trametes and Pleurotus genera. The submerged fermentation (SmF) has been the most used, however, the use of solid-state fermentation (SSF) appeared as a promising technique to produce laccase when using agro-industrial residues as substrates. Studies on laccase immobilization showed the covalent bonding and entrapment methods were the most used, showing greater efficiency of immobilization and a high number of enzyme reuses. The main use of the laccase was in bioremediation, especially in the discoloration of dyes from the textile industry and the degradation of pharmaceutical waste. Implications and consequences of all these findings in biotechnology and environment, as well as the trends and gaps of laccase research were discussed.

Effect of Purpureocillium lilacinum on inter-root soil microbial community and metabolism of tobacco

BackgroundNumerous chemical pesticides have been used in agricultural production to combat crop diseases and pests. Despite ensuring certain economic advantages, they have also resulted in issues like environmental contamination, declining soil quality, and pesticide residues. Because biological control is environmentally friendly and difficult to acquire resistance to, it has been used in practice recently.MethodsIn this study, we isolated the endophytic fungus Purpureocillium lilacinum from Portulaca oleracea L., which was previously found to have inhibitory effects on soil pathogens in tobacco fields. To find out how the biocontrol agent P. lilacinum affects soil microorganisms and plant metabolism in tobacco cultivation, we used amplicon sequencing technology and gas chromatography-mass spectrometry to look at the structure of soil microbial communities and the networks of interactions between microorganisms and metabolites in the inter-rhizosphere soil of tobacco fields treated with different amounts of P. lilacinum.ResultsThe findings showed that there was a trend toward less microbial diversity among inter-root microorganisms as solid-state fermentation (SSF) products of P. lilacinum increased; however, submerged fermentation (SmF) had no discernible impact on microbial diversity when compared to the direct use of SSF. Additionally, the relationship between inter-root fungi and volatile compounds in tobacco leaves was dominated by a negative correlation.ConclusionsThe result demonstrated that P. lilacinum’s antagonistic interaction in the inter-rhizosphere microbial community was dominant and valuable for biopesticide application. P. lilacinum can work more effectively on tobacco roots by using SSF products. P. lilacinum’s opposition to fungal colonies may enhance the volatile chemicals in tobacco leaves. These provide some implications for the biocontrol application of P. lilacinum.

Bioengineered Enzymes and Precision Fermentation in the Food Industry.

Enzymes have been used in the food processing industry for many years. However, the use of native enzymes is not conducive to high activity, efficiency, range of substrates, and adaptability to harsh food processing conditions. The advent of enzyme engineering approaches such as rational design, directed evolution, and semi-rational design provided much-needed impetus for tailor-made enzymes with improved or novel catalytic properties. Production of designer enzymes became further refined with the emergence of synthetic biology and gene editing techniques and a plethora of other tools such as artificial intelligence, and computational and bioinformatics analyses which have paved the way for what is referred to as precision fermentation for the production of these designer enzymes more efficiently. With all the technologies available, the bottleneck is now in the scale-up production of these enzymes. There is generally a lack of accessibility thereof of large-scale capabilities and know-how. This review is aimed at highlighting these various enzyme-engineering strategies and the associated scale-up challenges, including safety concerns surrounding genetically modified microorganisms and the use of cell-free systems to circumvent this issue. The use of solid-state fermentation (SSF) is also addressed as a potentially low-cost production system, amenable to customization and employing inexpensive feedstocks as substrate.

Growth performance and nutrient composition of black soldier fly larvae reared on solid-state fermentation substrates with various white rot fungi

Abstract. Fitriana EL, Jayanegara A, Astuti DA, Laconi EB. 2022. Growth performance and nutrient composition of black soldier fly larvae reared on solid-state fermentation substrates with various white rot fungi. Biodiversitas 23: 4894-4905. This study examines agricultural byproducts fermented with various rot fungi as a substrate of black soldier fly larvae. The experiment used a three-way randomized design with four replications. The first factor was substrate (cacao pod husk and oil palm frond), the second factor was fungi (Phanerochaete chrysosporium, Trametes versicolor, and Pleurotus sajor-caju), and the third factor was environment condition (light and without light). Approximately 1200 of 7-day-old larvae were added to 600 g substrate treatment. The results showed that the solid-state fermentation substrates had the highest waste reduction parameters and level of consumption. Larvae in the experiment had a crude protein level of 38.84-58.88% of dry matter. Glutamic acid is the most abundant non-essential amino acid in larvae, while leucine is the most abundant essential amino acid. Solid-state fermentation substrate could improve the fatty acid profile quality of larvae in relation to high linoleic acid, polyunsaturated fatty acid, and unsaturated fatty acid percentages, especially CPH-pc substrate. The use of solid-state fermentation in larvae rearing decreased lignin in the cacao pod husk substrate. In conclusion, cacao pod husk substrate solid-state fermented with white rot fungi is not supported the optimum growth performance of BSFL but increases fatty acid profile quality.

Novel green approaches towards utilization of flower waste: A review

Floriculture has been associated with culture and heritage since very ancient time in our country. According to Tata Energy Research Institute (TERI), the waste generation rate in India is increasing at the rate of 1.0 -1.33 per cent annually. In most of the developing countries like India, the floral waste generation occurs largely during worships, festivals, ceremonies etc. from temples, mosque, flower market, flower exhibition and wedding halls find their way into the garbage as a waste causing various environmental problems. This paper discusses the use of solid state fermentation to convert floral wastes into a variety of value-added products like as compost, biogas, bioethanol, dyes, food products, biosurfactants production, incense sticks and handmade paper production etc. and floral waste can thus be converted into wealth.

Filamentous fungi isolated from Tunisian olive mill wastes: use of solid-state fermentation for enzyme production

Olive mill wastewaters and olive cake are effluents generated during olive oil production process. They represent a major disposal and potentially severe pollution problem for the industry, also promising source of substances of high value. The aim of this study is the valorization of olive mill wastes (OMWW, olive cake, olive twigs and leaves) to produce enzymes with high industrial and biotechnological potential, by the solid-state fermentation technique (SSF), from isolated fungi present in olive mill wastewater and olive cake. A total of 47 strains were isolated and purified from these two residues. The metabolic potential of isolated strains was study by testing the hydrolytic enzymes activities of lipase, protease, amylase, cellulase, invertase, phytase and tannase on agar plate media containing different substrate. The monitoring of SSF has shown that the metabolic activity of these strains is extremely rapid using this technique. Our fungi collection contains a diversity of strains capable to producing a variety of enzymes of biotechnological interest.

Advanced assessments on innovative methods to improve the bioaccessibility of polyphenols in wheat

Abstract Polyphenolic compounds (PCs) are a group of natural antioxidants found in plants. Because most of the PCs exist in bound form in wheat, the efficiency of the solvent extraction method is very low. In general, there are three types of methods used to extract the bound PCs or convert them to free ones and as a result, increase the bioaccessibility of these non-extractable compounds. Although chemical extraction is the most common method used in this way, the enzymatic treatment also accomplishes with less efficiency to break down the linkages between cell wall components and PCs. In recent years, the use of solid state fermentation have been growing up and several microorganisms have been used to produce hydrolyzing enzymes and convert bound PCs to extractable ones. By using this method, the biological effects of the sample originated from the antioxidant property of PCs can be increased in fermented wheat cultivars. This paper provides an overview of different modifications and treatments emphasizing the effect of solid state fermentation as an alternative method for increasing the content of free PCs in wheat.

Conditions Affecting Lingzhi or Reishi Medicinal Mushroom Ganoderma lucidum (Agaricomycetes) Basidiome Quality, Morphogenesis, and Biodegradation of Wood By-products in Argentina.

Solid-state fermentation (SSF) with the medicinal higher Basidiomycete Ganoderma lucidum was studied as a strategy to use pine (Pinus radiata D. Don) and poplar (Populus nigra L.) wood chips and sawdust. Fruiting bodies were produced and the value of the biotransformed substrate was assessed. The highest mushroom yield (63 g dry weight per kilogram of dry substrate) was obtained with poplar sawdust and wood chips. Immersion of the bioreactors was a simple watering method that obtained suitable yields. Two morphological types were induced using 2 different incandescent light intensities. High light irradiation induced the highest valued mushroom morphology (as a whole product). Time course study of substrate biodegradation and mycelial growth dynamics indicated that the trophophase lasted 20 days and presented laccase activity of 0.01-0.03 units · g-1. The activity at idiophase was 10 times higher. Aqueous and alkali extracts, as well as carbohydrase enzyme profile activity, revealed differences in the properties of the residual substrate; some related to the substrate source are considered to be of concern for further use of this pretreated biomass. In view of the results obtained, we propose use of SSF of pine and poplar with G. lucidum to profitably recycle softwood by-products from the timber industry.

COMPARATIVE STUDY OF ANTIOXIDANT ACTIVITIES, COSMECEUTICAL PROPERTIES AND PHENOLIC ACIDS COMPOSITION OF FERMENTED RICE BRAN AND COCONUT TESTA

Malaysia produces large number of agro-industrial products annually including rice and coconut. Along with the production, rice bran and coconut testa are also abundantly generated as by-products. Solid state fermentation (SSF) is a simple biotechnological technique that has been employed for production of bioactive compounds. The use of SSF on agro-industrial by-products is economically viable and helps to reduce the environmental impact of the by-products disposal. In this present study, rice bran (RB), coconut testa (CT) and a combination of rice bran and coconut testa (RBCT) were fermented with single culture of Aspergillus oryzae and Rhizopus oligosporus. The effect of fungal SSF on the cosmeceutical-related activities and phenolic acid content of the fermented RB, CT and RBCT extracts was determined. The results showed that total phenolic content (TPC) as well as antioxidant activity measured using the ferric reducing ability of plasma (FRAP) of all substrates increased upon fermentation. Contrary to the TPC and FRAP assays results, the radical-scavenging activity of the fermented samples as measured by using the DPPH (2,2-diphenyl-1-picrylhydrazyl) method, were varied. Tyrosinase inhibition activity was found to be the highest in the extract of RB fermented with A. oryzae (56.18%) compared to other extracts. The same extract showed the highest elastase inhibition activity with a value of 60.27%. High performance liquid chromatography (HPLC) analyses showed ferulic, sinapic, caffeic and protocatechuic acids were significantly increased in RB fermented with A. oryzae. The results of this study suggest that fungal solid-state fermentation of rice bran may have the potential to be exploited in producing different types of active metabolites that are useful for cosmeceutical industries

From Wastes to High Value Added Products: Novel Aspects of SSF in the Production of Enzymes

Solid-state fermentation (SSF), a process that occurs in the absence or near absence of water, has been used for the production of various high value added products such as enzymes and other organic components. This paper reviews the recent studies reported on the use of SSF for the production of enzymes: lipases, proteases, cellulases, hemicellulases, ligninases, glucoamylases, pectinases, and inulinases. The microorganisms used for fermentation are mostly fungi, and substrates are waste materials from the agriculture and food industry. This shows the advantages of SSF from an economical and environmental viewpoint. The paper provides an update on several issues, viz. wastes, microorganisms, and issues of scaling up and controlling the process of fermentation in solid state.

Enzymatic Hydrolysis of Sugarcane Bagasse Using Enzyme Extract and Whole Solid-state Fermentation Medium of Two Newly Isolated Strains of Aspergillus Oryzae

Enzymatic conversion of lignocellulosic biomass into fuels and chemicals will be a key technology in the future. However, in order to make this process economically feasible, it is necessary to improve the efficiency of enzyme production, since the cost of the enzymatic cocktails significantly influences the viability of the overall process. In this sense, the use of solid-state fermentation (SSF) is particularly advantageous for enzyme production. Here, a comparative study on the enzymatic hydrolysis of steam- exploded sugarcane bagasse (SESB) using enzymatic extract (EE) and whole solid-state fermentation medium (WM) of two newly isolated strains of Aspergillus oryzae (P6B2 and P27C3A) from the Amazon Rainforest was carried out. The biomass conversion using WM from A. oryzae P6B2 was more efficient when compared with the EE, while for A. oryzae P27C3A the conversion yields were similar. The WM from P27C3A supplemented with a low dosage of commercial enzyme resulted in a final conversion of 45% of the theoretical value. Furthermore, the combination of the enzymes from both strains, allowed up to 5 folds improvement in SESB conversion. These results showed that the in-house enzyme production by wild-typeoryzae strains cultivated under SSF can be very advantageous for the enzymatic hydrolysis of biomass and, thus, it can be considered as a potential biotechnological configuration for the production of biofuels.

Growth of fungal strains on coffee industry residues with removal of polyphenolic compounds

Coffee silverskin (CS) and spent coffee grounds (SCG) are abundant residues generated by the coffee industry, but up till now they are practically unutilized being discharged to the environment or burned for elimination. In the present study, the ability of seven different fungal strains from the genera Aspergillus, Mucor, Penicillium, and Neurospora, to grow and release phenolic compounds from these residues under solid-state cultivation conditions was evaluated. All the strains were able to growth on CS and SCG, but Penicillium purpurogenum, Neurospora crassa and Mucor were able to release the highest amounts of phenolic compounds from both residues. Phenolic compounds have wide-range applications in the food and pharmaceutical industries, and thus, the extraction of these compounds could be a valuable alternative for the exploitation of CS and SCG. Additionally, the use of solid-state fermentation to obtain these compounds has a positive environmental impact because the use of chemical solvents is avoided.

Effect of substrate and culture conditions on the production of amylase and pullulanase by thermophilic Clostridium thermosulforegenes SVM17 in solid state fermentation

The endo acting enzyme with dual specificity towards α-1,4- and α-1,6-glycosidic linkages are named as amylopullulanase. The production of extracellular thermostable amylopullulanase by Clostridium thermosulfurogenes SVM17 was investigated in solid state fermentation (SSF). Coarse type wheat bran was found to be the best substrate among ten easily available complex organic substrates evaluated. The production of enzyme reached a peak in 72 h. A high level of enzyme was produced in wheat bran moistened with PYE medium with a moisture content of 73 %. The optimum temperature and pH for amylopullulanase production was 60 °C and 7.5, respectively. An inoculum size of 20 % resulted in maximum production of amylopullulanase. Under the optimum conditions the strain showed a maximum of 17,227 and 21,526 U of amylase and pullulanase activity, respectively per kilogram of bacterial bran (BB). The enzyme production was high in SSF than that in SmF. The use of SSF for the production of thermostable amylopullulanase by C. thermosulfurogenes SVM17 could, therefore led to reduction in the overall cost of enzyme production.

Comparative production of cellulases by mutants of Penicillium janthinellum NCIM 1171 and its application in hydrolysis of Avicel and cellulose

Mutants of Penicillium janthinellum NCIM 1171 were evaluated for cellulase production using both submerged fermentation (SmF) and solid state fermentation (SSF). Mutant EU2D-21 gave highest yields of cellulases in both SmF and SSF. Hydrolysis of Avicel and cellulose were compared using SmF and SSF derived enzyme preparations obtained from EU2D-21. Surprisingly, the use of SSF derived preparation gave less hydrolysis compared to SmF derived enzymes. This may be due to inactivation of β-glucosidase at 50°C in SSF derived enzyme preparations. SmF derived enzyme preparations contained both thermostable and thermosensitive β-glucosidases where as SSF derived enzyme preparations contained predominantly thermosensitive β-glucosidase. This is the first report on less thermostability of SSF derived β-glucosidase which is the main reason for getting less hydrolysis.

Bioethanol production from grape and sugar beet pomaces by solid-state fermentation

A suitable alternative to replace fossil fuels is the production of bioethanol from agroindustrial waste. Grape pomace is the most abundant residue in San Juan and sugar beet pomace could be important in the region. Solid-State Fermentation (SSF) is a technology that allows transforming agroindustrial waste into many valuable bioproducts, like ethanol. This work reports a laboratory scale SSF to obtain alcohol from grape and sugar beet pomace by means of Saccharomyces cerevisiae yeasts. The initial conditions of the culture medium were: sugars 16.5% (p/p); pH 4.5; humidity 68% (p/p). Cultures were inoculated with 10 8 cells/g of pomace, and incubated in anaerobic environment, at 28 °C, during 96 h. SSF showed ethanol maximum concentrations at 48 h and ethanol yield on sugars consumed was more than 82%. Yield attained creates expectation about the use of SSF to obtain fuel alcohol.

Production of polyhydroxyalkanoates (PHAs) from waste materials and by-products by submerged and solid-state fermentation

Polyhydroxyalkanoates are biodegradable polymers produced by prokaryotic organisms from renewable resources. The production of PHAs by submerged fermentation processes has been intensively studied over the last 30 years. In recent years, alternative strategies have been proposed, such as the use of solid-state fermentation or the production of PHAs in transgenic plants. This paper gives an overview of submerged and solid-state fermentation processes used to produce PHAs from waste materials and by-products. The use of these low-cost raw materials has the potential to reduce PHA production costs, because the raw material costs contribute a significant part of production costs in traditional PHA production processes.

Improving Bioethanol Yield: The Use of Solid-State Fermentation Products Grown on DDGS

In a climate of uncertain prices for grain and fuel, conservation and efficiency are more important than ever. Many agro-industrial residues are excellent substrates for solid-state fermentation (SSF). Dried distillers grains with solubles (DDGS) are an important co-product of the ethanol industry and have potential as a substrate for solid-state fermentation. SSF products were produced using DDGS from a distillery and DDGS from a fuel ethanol plant as the fungal growth substrate. The spent grains were inoculated with GRAS (Generally Regarded As Safe) organisms Aspergillus orzyae and Rhizopus oligosporus. After growth, the resultant SSF product was dried and used as an enzyme complex supplement that was added to laboratory scale standard fuel ethanol corn mash fermentations. Improved ethanol yields were consistently observed. The use of solid-state fermentation to produce unique enzyme complexes on DDGS offers a novel way to increase the value of the DDGS and to enhance bioethanol fermentation.

Fungal Generation of Organic Acids for Removal of Lead from Contaminated Soil

The use of solid-state fermentation (SFF) of low cost substrates by fungal species to generate organic acid solutions for washing of lead from a contaminated soil was evaluated in this study. SFF filtrates were generated by fermentation of four substrates (corn cobs, apple pomace, rice and hay) with three fungal species (Aspergillus niger NRRL 2001 (A. niger 1), Aspergillus niger ATCC 64065 (A. niger 2), Aspergillus foetidus NRRL 337) at three fermentation times. The concentration and speciation of organic acids of the filtrates was found to be a function of the substrate type, the fungal species and the fermentation time. Fermentation of rice resulted in the highest concentrations of citric acid while fermentation of corn cobs, apple pomace and hay tended to generate oxalic acid with an increasing fraction of this acid as the fermentation progressed. Batch extraction tests that employed the SSF filtrates revealed that soluble lead concentrations as high as 35 mg/l could be achieved. Filtrates containing elevated concentrations of citric acid resulted in the greatest lead extraction while oxalic acid inhibited solubilization. Due to the buffering of pH that was provided by the soil in the batch tests this factor did not appear to influence lead extraction. Lead extraction was observed over an extended period of time in a column test. Lead extraction was strongly influenced by the pH of the soil column and less strongly influenced by the organic acid content of the SSF filtrate. The speciation of organic acids was substantially modified from primarily citric acid in the SSF filtrate to gluconic acid in the soil column discharge.

Production of poly(3-hydroxybutyrate) by solid-state fermentation with Ralstonia eutropha

The use of solid-state fermentation is examined as a low-cost technology for the production of poly(hydroxyalkanoates) (PHAs) by Ralstonia eutropha. Two agroindustrial residues (babassu and soy cake) were evaluated as culture media. The maximum poly(hydroxybutyrate) (PHB) yield was 1.2 mg g(-1) medium on soy cake in 36 h, and 0.7 mg g(-1) medium on babassu cake in 84 h. Addition of 2.5% (w/w) sugar cane molasses to soy cake increased PHB production to 4.9 mg g(-1) medium in 60 h. Under these conditions, the PHB content of the dry biomass was 39% (w/w). The present results indicate that solid-state fermentation could be a promising alternative for producing biodegradable polymers at low cost.

Development of a bionematicide with Paecilomyces lilacinus to control Meloidogyne incognita.

Root-knot disease caused by Meloidogyne incognita is a matter of grave concern because it affects several economically important crop plants. The use of solid-state fermentation (SSF) may help to elaborate efficient formulations with fungi to be employed in the biologic control of nematodes. Attempts were made to select low-cost substrates for spore production of a strain of Paecilomyces lilacinus with known nematicide capacity. Coffee husks, cassava bagasse, and defatted soybean cake were utilized as substrates, and sugarcane bagasse was used as support. Fermentations were carried out in flasks covered with filter paper at 28 degrees C for 10 d. The products obtained by SSF were evaluated for their nematicide activity in pot experiments containing one seedling of the plant Coleus inoculated with the nematode M. incognita. The plants were evaluated 2 mo after inoculation. Fermented products showed a reduction in the number of nematodes. The best results were obtained with defatted soybean cake, which showed almost 100% reduction in the number of nematodes; the reduction with coffee husk was 80% and with cassava bagasse was about 60%.