A Comparative Study of New Aspergillus Strains for Proteolytic Enzymes Production by Solid State Fermentation.

Proteinase and glycoside hydrolase production is enhanced in solid-state fermentation by manipulating the carbon and nitrogen fluxes in Aspergillus oryzae

An enzymatic extract from Aspergillus niger 3T5B8 was produced by Solid State Fermentation (SSF) in aerated columns, using wheat bran as substrate. A combination of extracts produced using three different process conditions varying temperature, pH and aeration formed the final extract (Mixture). The Mixture was concentrated by an ultrafiltration process that partially purified and provided an efficient recovery of the enzymatic activities of xylanase (88.89%), polygalacturonase (89.3%), β-glucosidase (93.15%), protease (98.68%) and carboxymethylcellulase (CMCase) (98.93%). SDS-PAGE analysis showed 15 visible protein bands in the crude and concentrated Mixture with molecular weights ranging from 15.1 to 104.6 kDa. Thin layer chromatography confirmed the effective action of β-glucosidase and xylanase hydrolysis activities over cellobiose and xylan, respectively. A central composite design (CCD) with two variables and four replicates at the center points was used to determine the optimal temperature and pH for CMCase and β-glucosidase. The optimal temperature was 78.9 °C and pH 3.8 for CMCase and 52.8 °C and pH 4.8 for β-glucosidase, respectively.

Solid state fermentation of waste bread pieces by Aspergillus awamori: Analysing the effects of airflow rate on enzyme production in packed bed bioreactors

Protease production in solid state fermentation with liquid medium recycling in a stacked plate reactor and in a packed bed reactor by a local strain of Rhizopus oryzae

Production of an Alkaline Protease From Nocardiopsis Alba Om-4, a Haloalkaliphilic Actinobacteria in Solid-State Fermentation Using Agricultural Waste Products

This study reports the optimization of milk-clotting protease production from Aspergillus oryzae DRDFS13 under solid-state fermentation (SSF) in both one-variable-at-a-time and response surface methodology (RSM). The production and optimization of milk-clotting protease obtained from Aspergillus oryzae DRDFS13 under solid-state fermentation (SSF) using different agro-industrial wastes as solid substrates were studied. The agro-industrial wastes used included wheat bran, rice bran, pea bran, and grass pea bran. The chemical composition of the best solid substrate was tested using standard methods. Others cultivation parameters were studied, and the results showed that the optimum fermentation medium composed of wheat bran, casein (1% w/w), and glucose (0.5% w/w) and the conditions for maximum milk-clotting protease production were at the moisture content of 55.0%, inoculum of 0.5*106 spores/mL, incubation temperature of 30°C, pH of 6.0, and fermentation time of 5days. The highest milk-clotting activity was obtained from the crude enzyme extracted using 0.1M NaCl and partial purification of the crude enzyme using chilled acetone, and 80% (NH4)2SO4 increased the ratio of MCA/PA from 0.56 to 1.30 and 0.65, respectively. Moreover, the highest MCA (137.58U/mL) was obtained at a casein concentration of 0.5%, pH 4.0, and 25°C, using RSM. Thus, results from the present study showed that the optimization of milk-clotting protease production from A. oryzae DRDFS 13 under SSF by both one-variable-at-a-time and RSM significantly increased the milk-clotting activity. This is the first report from a fungus in the Ethiopian setting and a modest contribution to highlight the potential of harnessing microbial protease enzymes for industrial applications.

The present study evaluated the performance of the fungus Trichoderma reesei to tolerate and biodegrade the herbicide diuron in its agrochemical presentation in agar plates, liquid culture, and solid-state fermentation. The tolerance of T. reesei to diuron was characterized through a non-competitive inhibition model of the fungal radial growth on the PDA agar plate and growth in liquid culture with glucose and ammonium nitrate, showing a higher tolerance to diuron on the PDA agar plate (inhibition constant 98.63 mg L-1) than in liquid culture (inhibition constant 39.4 mg L-1). Diuron biodegradation by T. reesei was characterized through model inhibition by the substrate on agar plate and liquid culture. In liquid culture, the fungus biotransformed diuron into 3,4-dichloroaniline using the amide group from the diuron structure as a carbon and nitrogen source, yielding 0.154mg of biomass per mg of diuron. A mixture of barley straw and agrolite was used as the support and substrate for solid-state fermentation. The diuron removal percentage in solid-state fermentation was fitted by non-multiple linear regression to a parabolic surface response model and reached the higher removal (97.26%) with a specific aeration rate of 1.0 vkgm and inoculum of 2.6 × 108 spores g-1. The diuron removal in solid-state fermentation by sorption on barley straw and agrolite was discarded compared to the removal magnitude of the biosorption and biodegradation mechanisms of Trichoderma reesei. The findings in this work about the tolerance and capability of Trichoderma reesei to remove diuron in liquid and solid culture media demonstrate the potential of the fungus to be implemented in bioremediation technologies of herbicide-polluted sites.

Changes in rice bran bioactives, their bioactivity, bioaccessibility and bioavailability with solid-state fermentation by Rhizopus oryzae

Cow dung and agroresidues were used as the substrates for the production of alkaline proteases by Pseudomonas putida strain AT in solid-state fermentation. Among the various substrates evaluated, cow dung supported maximum (1351±217 U/g) protease production. The optimum conditions for the production of alkaline proteases were a fermentation period of 48 h, 120% (v/w) moisture, pH 9, and the addition of 6% (v/w) inoculum, 1.5% (w/w) trehalose, and 2.0% (w/w) yeast extract to the cow dung substrate. The enzyme was active over a range of temperatures (50–70°C) and pHs (8–10), with maximum activity at 60°C and pH 9. These enzymes showed stability towards surfactants, detergents, and solvent and digested various natural proteins.

Solid-state fermentation of Jatropha seed cake for optimization of lipase, protease and detoxification of anti-nutrients in Jatropha seed cake using Aspergillus versicolor CJS-98

Proteases are a group of enzymes that catalyse the degradation of proteins resulting in the production of their amino acid constituents. In the present study newly isolated Streptomyces sp. was subjected to produce proteases through solid state fermentation while wheat bran was used as substrate. To produce proteases, a local strain Streptomyces sp. was isolated from a soil sample of Ezzemouls saltpan, located in Ain M'lila (East of Algeria). The phenotypic and phylogenetic studies of this strain showed that it represents probably a new species. The SSF production medium for Streptomyces sp. was optimized using Plackett and Burman statistical methods. The results showed a maximum activity on basal wheat bran medium supplemented with 1% fructose. The best SSF humidifying solution was pH 9.0 phosphate buffer at 50% moisture. Protease has an optimum at pH 7.0, which is a typical characteristic of neutral proteases. The optimum temperature was 60°C and proved stable up to 80°C. The results showed that the novel Streptomyces sp. isolate is a good producer of extracellular neutral protease on wheat bran, which can be beneficial for industries. J. Bio-Sci. 29(1): 33-48, 2021 (June)

This study aimed to examine how lactic acid bacteria (LAB) Lactobacillus plantarum mediated solid-state fermentation (SSF) of whole wheat flour (WWF) affected the release of phenolic fractions and their in vitro antioxidant (DPPH•, ABTS•+, H2O2, and ferric reducing power) activities as well as phenolic acids. Results revealed that fermentation techniques successfully released the bound phenolic fractions into free form of the treated wheat and considerably increased the free/bound ration of phenolic fractions. The free and bound phenolic fractions total phenolic, flavonoid and antioxidant (TPC, TFC and TAC) contents were found increased 284.87 % and 39.60 %; 339.92 % and 105.86 %; 157.13 % and 52.43 %, respectively for SSFWG whereas 399.59 % and 103.24 %; 522.18 % and 102.72 %; 226.35 % and 95.67 %, respectively for SSFWF until 72hr and then found decreased. In HPLC analysis, individual phenolics were found increased significantly (P ≤ 0.05) for free phenolic fractions and decreased for bound phenolic fractions. For free phenolics, 3,4-dihydroxy benzoic acid was found increased enormously (164.54 % and 220.47 %, respectively for SSFWG and SSFWF) among the other individual phenolics. After SSF, both free and bound phenolic extracts DPPH•, ABTS•+, H2O2 and ferric reducing power of wheat became stronger. Principal component analysis and correlation matrix showed TPC are strongly correlated with the in vitro antioxidant activities and individual phenolics. The results of this experiment suggest that SSF is an effective bioprocessing technique to improve the phenolic release and antioxidant activities of wheat, which may further enhance the health benefits and utilization of fermented whole wheat flour. Consequently, the food and nutraceutical industries may get inspiration for the utilization of fermented whole wheat flour for future food production to get benefit from the release of phenolics found in fermented wheat and minimize the losses of raw material by-products.

Enzyme production varies in different fermentation systems. Enzyme expression in different fermentation systems yields important information for improving our understanding of enzymatic production induction. Comparative studies between solid-state fermentation (SSF) using agro-industrial waste wheat bran and submerged fermentation (SmF) using synthetic media were carried out to determinate the best parameters for peptidase production by the fungus Aspergillus fumigatus Fresen. Variables tested include: the concentration of carbon and protein nitrogen sources, the size of the inoculum, the pH of the media, temperature, and the length of the fermentation process. The best peptidase production during SSF was obtained after 96 hours using wheat bran at 30 °C with an inoculum of 1 × 106 spores and yielded 1500 active units (U/mL). The best peptidase production using SmF was obtained after periods of 72 and 96 hours of fermentation in media containing 0.5% and 0.25% of casein, respectively, at a pH of 6.0 and at 30 °C and yielded 40 U/mL. We also found examples of catabolite repression of peptidase production under SmF conditions. Biochemical characterization of the peptidases produced by both fermentative processes showed optimum activity at pH 8.0 and 50 °C, and also showed that their proteolytic activity is modulated by surfactants. The enzymatic inhibition profile using phenylmethylsulfonyl fluoride (PMSF) in SmF and SSF indicated that both fermentative processes produced a serine peptidase. Additionally, the inhibitory effect of the ethylene-diaminetetraacetic acid (EDTA) chelating agent on the peptidase produced by SmF indicated that this fermentative process also produced a metallopeptidase.

This study aims to evaluate the impact of solid-state fermentation (SSF) by Trichoderma reesei on the phenolic content, antioxidant and antimicrobial activities of garden cress seeds (GCS). The factorial statistical design was employed to optimize the SSF conditions, incubation time, pH, temperature and moisture, for maximum production of the phenolic content and microbial carbohydrate-cleaving enzymes from GCS. The total phenolic content significantly increased from unfermented GCS (401mg gallic acid equivalent (GAE) 100g-1 ) to fermented GCS (3600mg GAE 100g-1 ) by ninefold. The total antioxidant activity significantly increased in fermented GCS. Fifteen phenolic compounds were detected in fermented GCS with high concentrations compared to 14 in unfermented GCS using high-performance liquid chromatography. A strong correlation between the production of the carbohydrate-cleaving enzymes and the phenolic content of fermented GCS was observed. The phenolic compounds of fermented GCS showed higher antimicrobial activity. The fermented GCS is a powerful source of phenolic compounds with high antioxidant potentials, which can be used as dietary supplement and antimicrobial agent. Solid-state fermentation is a promising technique used for production of added-value bioactive compounds. SSF increased the total phenolic content and antioxidant activity of GCS several folds compared to germination process, which recently studied.

A new approach for proteases production by Aspergillus niger based on the kinetic and thermodynamic parameters of the enzymes obtained